Smart Nanomaterials Targeting Pathological Hypoxia 9819917174, 9789819917174

Citation preview

Smart Nanomaterials Technology

Sonam Chawla  Sachidanand Singh  Azamal Husen Editors

Smart Nanomaterials Targeting Pathological Hypoxia

Smart Nanomaterials Technology Series Editors Azamal Husen , Wolaita Sodo University, Wolaita, Ethiopia Mohammad Jawaid, Laboratory of Biocomposite Technology, Universiti Putra Malaysia, INTROP, Serdang, Selangor, Malaysia

Nanotechnology is a rapidly growing scientific field and has attracted a great interest over the last few years because of its abundant applications in different fields like biology, physics and chemistry. This science deals with the production of minute particles called nanomaterials having dimensions between 1 and 100 nm which may serve as building blocks for various physical and biological systems. On the other hand, there is the class of smart materials where the material that can stimuli by external factors and results a new kind of functional properties. The combination of these two classes forms a new class of smart nanomaterials, which produces unique functional material properties and a great opportunity to larger span of application. Smart nanomaterials have been employed by researchers to use it effectively in agricultural production, soil improvement, disease management, energy and environment, medical science, pharmaceuticals, engineering, food, animal husbandry and forestry sectors. This book series in Smart Nanomaterials Technology aims to comprehensively cover topics in the fabrication, synthesis and application of these materials for applications in the following fields: • Energy Systems—Renewable energy, energy storage (supercapacitors and electrochemical cells), hydrogen storage, photocatalytic water splitting for hydrogen production • Biomedical—controlled release of drugs, treatment of various diseases, biosensors, • Agricultural—agricultural production, soil improvement, disease management, animal feed, egg, milk and meat production/processing, • Forestry—wood preservation, protection, disease management • Environment—wastewater treatment, separation of hazardous contaminants from wastewater, indoor air filters

Sonam Chawla · Sachidanand Singh · Azamal Husen Editors

Smart Nanomaterials Targeting Pathological Hypoxia

Editors Sonam Chawla Department of Biotechnology Jaypee Institute of Information Technology Noida, Uttar Pradesh, India

Sachidanand Singh Department of Biotechnology Sankalchand Patel University Visnagar, Gujarat, India

Azamal Husen Wolaita Sodo University Wolaita, Ethiopia

Smart Nanomaterials Technology ISBN 978-981-99-1717-4 ISBN 978-981-99-1718-1 (eBook) https://doi.org/10.1007/978-981-99-1718-1 © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 This work is subject to copyright. All rights are solely and exclusively licensed by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors, and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, expressed or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. This Springer imprint is published by the registered company Springer Nature Singapore Pte Ltd. The registered company address is: 152 Beach Road, #21-01/04 Gateway East, Singapore 189721, Singapore

Preface

Arrival of smart nanomaterials in the ever-expanding field of nanoscience is an important milestone in this arena. The smart nanotechnology has touched all areas of health and disease prevention, and offers unique advancements in the previous issues of concern associated with conventional diagnostics or therapeutics—precision, improved drug delivery and efficacy, safety and toxicity, biocompatibility, and so on. Hypoxia, or suboptimal oxygen availability in tissues, became a commonplace word during the COVID-19 pandemic which hit the world in 2020, wherein “Silent Hypoxia” wreaked havoc with human life. However, clinicians continually dealt with hypoxia as an omnipresent pathological feature of many modern-day diseases such as cancer, cerebral ischemia and stroke, vascular diseases, high-altitude hypoxia, infections, and obstructive pulmonary disorders. This book provides a comprehensive summary of the newest research on smart nanoplatforms responding to hypoxia. We have covered the fundamentals of smart nanomaterials and their applications in recognizing/diagnosing as well as a therapeutic intervention (drug delivery systems) for hypoxia in various clinical settings. Future possibilities to develop bio-nanobots with integration of smart nanotechnology and biotechnology as well as advanced approaches such as use of NMR-based pharmacometabonomics for evaluating the efficacy of smart nanomaterials are also elaborated and discussed. A body of literature also covers the advantages conferred by incorporating smart nanoplatforms in synergy with conventional therapies, especially in the backdrop of anticancer therapy regimes such as photodynamic therapy. The chapters will benefit our readership developing an understanding of the utility of smart/stimuli-responsive/intelligent nanomaterials for clinical applications. We extend our sincere thanks to all contributors for their timely response and excellent contributions. We also express our sincere thanks to our families and colleagues

v

vi

Preface

for their support during compilation of this academic endeavor. We shall be happy to receive comments and criticism, if any, from subject experts and general readers of this book. Gautam Buddha Nagar, India Visnagar, India Wolaita, Ethiopia

Sonam Chawla Sachidanand Singh Azamal Husen

Contents

Smart Nanotechnology in Pathological Hypoxia: An Innovative Avenue for a Clinical Hurdle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Sonam Chawla, Aaysha Gupta, Mahima Bhardwaj, Sachidanand Singh, and Azamal Husen Bioinspired Nanosystems Interacting with the Host Environment: Smart Nanosystems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Shatabdi Basu, Koena Mukherjee, Koel Mukherjee, and Dipak Maity Nanotechnology-Based Approaches to Relieve Tumour Microenvironment Hypoxia via Enhanced Oxygen Delivery . . . . . . . . . . . Manisha Singh, Rashi Rajput, Vinayak Agarwal, Divya Jindal, Pranav Pancham, and Sudha Srivastava Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pooja Kumari, Preeti Sharma, Yogesh Srivastava, and Narendra Kumar Sharma Recent Advancements in the Field of Stimuli-Responsive Polymeric Nanomaterials for Cancer Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . N. Sisubalan, S. Nisha Nandhini, M. Gnanaraj, A. Vijayan, Joe Rithish, C. Karthikeyan, and K. Varaprasad

1

11

35

59

81

Nanopoxia: Antimonene-Based Nanoplatform Targeting Cancer Hypoxia for Precision Cancer Therapy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103 Shikha Srivastava, Sarita Singh, Sanchalika Mishra, Manju Pandey, and Md. Yaqub Khan Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115 Aashna Srivastava, Dharmendra Prajapati, Sachidanand Singh, and Tanvi Jain

vii

viii

Contents

Nanoproteomics: An Approach for the Identification of Molecular Targets Associated with Hypoxia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 J. Deepa Arul Priya, Sumira Malik, Mohammad Khalid, and Akash Gautam Nanomaterial-Mediated Theranostics for Vascular Diseases . . . . . . . . . . . 163 Tejaswini Divanji, Krisha Desai, Bhupendra Prajapathi, and Saritha Shetty Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors in Tumor Diagnosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187 Sudha Srivastava, Namita Sharma, and Manisha Singh Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis . . . 207 Saroj Kumar Das, Nishant Ranjan Chauhan, and Subhash Mehto Multifunctional Hypoxia Imaging Nanoparticles . . . . . . . . . . . . . . . . . . . . . 243 Preeti Sharma, Pooja Kumari, Tikam Chand Dakal, Jyotsana Singh, and Narendra Kumar Sharma Anaerobic Bacteria Mediated Hypoxia Specific Delivery of Nanoparticles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 257 Nisha Sharma and Smriti Gaur Phytoactive Ingredient-Loaded Theranostics . . . . . . . . . . . . . . . . . . . . . . . . . 279 Gurpreet Kaur Smart Nanomaterials for Alleviating the Limitation of Photodynamic Therapy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 295 V. T. Anju, Siddhardha Busi, Madangchanok Imchen, Mahima S. Mohan, and Madhu Dyavaiah Redox Responsive Smart Nanomaterials to Tackle Hypoxia Associated Oxidative Damage and Inflammatory Mediators Using Phytocompounds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313 Noopur Khare, Mahima Bhardwaj, Sonam Chawla, Rahat Praveen, and Sachidanand Singh ROS Responsive Silica Nanoparticles for Controlled and Targeted Drug Delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 327 Dharmendra Prajapati, Anil Patani, Tanvi Jain, Ashish Patel, and Sachidanand Singh New Developments in Nano-theranostics Combined with Intelligent Bio-responsive Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 347 Kopal Jain, Nikita Basant, and Amit Panwar

Contents

ix

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat Tumour Hypoxia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367 Bhupendra G. Prajapati, Amruta Desai, Pooja Desai, Aarohi Deshpande, Aarohi Gherkar, Manas Joshi, and Shama Mujawar NMR-Based Pharmacometabonomics of Nanoparticles for Treating Hypoxia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 383 Isha Gupta, Sonia Gandhi, and Sameer Sapra Nanoceria and Hypoxia: Promises and Challenges . . . . . . . . . . . . . . . . . . . . 399 Aditya Arya, Sneha Singh, and Amit Kumar

About the Editors

Sonam Chawla is an Assistant Professor in Jaypee Institute of Information Technology, NOIDA since 2019. She has previously taught in Abdul Kalam Technical University, Uttar Pradesh, India; and also worked as Guest Lecturer in Lady Irwin College (DU), Delhi, India. She has completed her Ph.D. from Defence Institute of Physiology and Allied Sciences (DIPAS), DRDO, Delhi as CSIR research fellow (CSIR JRF NET 2009). She is the recipient of the Vice-Chancellor’s Medal in her graduation as well as post-graduation. Her area of work is hypoxia biology and ageing. She has guided 7 master’s students and 1 Ph.D. student is working in the domain of anti-ageing phytomolecules. She has delivered several invited talks in national and international events and has >10 peer reviewed publications. Sachidanand Singh is a Professor in the Department of Biotechnology, Sankalchand Patel University, Visnagar, Gujarat, India. His academic background boasts of B.Tech Biotechnology, M.Tech Bioinformatics and Ph.D. in Bioinformatics. He has postdoctoral research experience from the College of Pharmacy, Ohio State University, USA. His specialization area includes Plant Biotechnology, metagenomics, Systems Biology and Drug design. He has more than 13 years of research and academic experience. He has served for a decade in the School of Biotechnology and Health Sciences, Karunya University, Coimbatore, Tamil Nadu, India. He also worked as consultant for college of pharmacy, Ohio State University as a Statistical Genetics during xi

xii

About the Editors

the year 2018. He has published more than 60 research articles in peer reviewed journals and completed one SERB, Govt. of India funded project, two University research grant projects. He has received several awards in his area of research and for his teaching strategies. He has organized different conferences and workshops and received Govt. funds for the same. He has shared his area of expertise with different colleges and universities as invited speakers in different conferences and workshops. He has guided 36 B.Tech and 10 M.Tech projects. Presently 6 Ph.D. students are enrolled under him and 2 Ph.D. graduates are working in the field of herbal products, drug design and integrating with network biology. He has already edited around half-dozen of books for international publishing houses. Azamal Husen served as Professor & Head, Department of Biology, University of Gondar, Ethiopia and is a Foreign Delegate at Wolaita Sodo University, Wolaita, Ethiopia. Earlier, he was a Visiting Faculty of the Forest Research Institute, and the Doon College of Agriculture and Forest at Dehra Dun, India. His research and teaching experience of 20 years involves studies of biogenic nanomaterial fabrication and application, plant responses to environmental stresses and nanomaterials at the physiological, biochemical and molecular levels, herbal medicine, and clonal propagation for improvement of tree species. He has conducted several research projects sponsored by various funding agencies, including the World Bank (FREEP), the National Agricultural Technology Project (NATP), the Indian Council of Agriculture Research (ICAR), the Indian Council of Forest Research Education (ICFRE), and the Japan Bank for International Cooperation (JBIC). He received four fellowships from India and a recognition award from the University of Gondar, Ethiopia, for excellent teaching, research, and community service. Husen has been on the Editorial board and the panel of reviewers of several reputed journals published by Elsevier, Frontiers Media, Taylor & Francis, Springer Nature, RSC, Oxford University Press, Sciendo, The Royal Society, CSIRO, PLOS, MDPI, John Wiley & Sons and UPM Journals. He is on the advisory board of Cambridge Scholars Publishing, UK. He is a Fellow of the Plantae group of the American Society of Plant

About the Editors

xiii

Biologists, and a Member of the International Society of Root Research, Asian Council of Science Editors, and INPST. To his credit are over 200 publications; and he is Editor-in-Chief of the American Journal of Plant Physiology. He is also working as Series Editor of Exploring Medicinal Plants, published by Taylor & Francis Group, USA; Plant Biology, Sustainability, and Climate Change, published by Elsevier, USA; and Smart Nanomaterials Technology, published by Springer Nature Singapore Pte Ltd. Singapore.

Smart Nanotechnology in Pathological Hypoxia: An Innovative Avenue for a Clinical Hurdle Sonam Chawla, Aaysha Gupta, Mahima Bhardwaj, Sachidanand Singh, and Azamal Husen

Abstract Smart nanotechnology has revolutionized the arena of therapeutics and their delivery as well as diagnostics. It is an advancing field, wherein the “smart” nanoparticles respond to a variety of external stimuli—temperature, pH, oxygen concentration, electrical/magnetic field, presence or absence of a metabolite or enzyme, and acquire functionality. Smart nanoplatforms are increasingly being developed to sense and respond molecular and biochemical milieu in hypoxia, and stand to have applications in drug delivery, drug action, overcoming the hypoxiainduced chemo/photoresistance, sensing or imaging hypoxic regions in vivo, etc. The biomedical relevance of having robust hypoxia-responsive smart nanomaterials is immense as hypoxia is a coincident feature of the majority of human infectious and non-communicable diseases. Keywords Hypoxia · Smart nanoparticles · Diagnostics · Therapeutics · Cancer

1 Introduction It was in the 1600 s that the scientific community recognized the “part of air” that was necessary for life—oxygen (Salyer 1989). Oxygen is indispensable to the majority of life forms on earth today, and for the same reason the mechanisms and pathways S. Chawla (B) · A. Gupta Department of Biotechnology, Jaypee Institute of Information Technology, NOIDA, Sector 62, Uttar Pradesh 201309, India e-mail: [email protected]; [email protected] S. Singh Department of Biotechnology, Sankalchand Patel University, Visnagar, Gujarat 384315, India A. Husen Wolaita Sodo University, Wolaita, Ethiopia M. Bhardwaj Department of Biotechnology, Vignan’s Foundation for Science, Technology & Research (Deemed to be University), Vadlamudi Guntur, Andhra Pradesh, India © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_1

1

2

S. Chawla et al.

to sense its paucity, and adapt to its sub-optimal conditions are quick and conserved across life forms (Tirpe et al. 2019). Sub-optimal availability of oxygen, termed as “hypoxia”, can manifest at various levels and is coincident with majority of human diseases, either as a cause or an outcome. Pathological hypoxia can manifest with non-communicable diseases— cardiovascular disease, diabetes, respiratory diseases, cancer, ischemia, rheumatoid arthritis, etc. as well as communicable diseases (Chen et al. 2020; Chen and Gaber 2021; Mora et al. 2022). A case in point—silent hypoxia post-CoVID-19 infection—was prevalent in 20–40% patients and was a major risk factor for patient mortality (Rahman et al. 2021). Bhutta and co-workers classified pathological hypoxia into following categories (Bhutta et al. 2022): • • • •

Hypoxemic hypoxia. Circulatory hypoxia. Anemic hypoxia. Histotoxic hypoxia.

Hypoxemic hypoxia is an outcome of pulmonary failure to adequately oxygenate the blood resulting in low arterial blood oxygen tension. Hypoventilation, impeded alveolar diffusion are some of the causes. Post-COVID silent hypoxia falls in the category of hypoxemic hypoxia, along with others diseases such as pulmonary fibrotic diseases and emphysema (Serrano et al. 2021). Another well-investigated pathological hypoxia situation is that of tumor hypoxia. It is caused by increased diffusion distances between nutritive blood vessels and the tumor cells, abnormally developed tumor microvasculature, and decreased O2 transport capacity of the blood due to disease and/or treatment-related reduced oxygen carrying capacity. The categorization by Butta and coworkers’ blurs in the scenario of tumor hypoxia—it is an overlap of histotoxic hypoxia and anemic hypoxia. A large contribution of hypoxic tumor microenvironment is to enhance aggressiveness, metastatic potential, and leading to poor prognosis (Li et al. 2021a, b). Stroke, the second leading cause of deaths in the world (Feigin et al. 2022) has connotations of circulatory hypoxia—the etiology pointing at vascular disturbances, embolism, as well as hypoxemic hypoxia wherein the respiratory muscles are paralyzed (Ferdinand and Roffe 2016). Further, on the basis of duration and severity of the oxygen deprivation pathological hypoxia can be acute, for example, ischemia incidents in the brain are acute episodes or chronic as in the case of the hypoxic core of a tumor (Rowat et al. 2006; Saxena and Jolly 2019). Another hypoxia-associated manifestation of human diseases is that suffered by high-altitude sojourners on fast ascent, where they encounter hypobaric hypoxia leading to acute mountain sickness, high-altitude pulmonary, and/or cerebral edema (Chawla et al. 2014; Rosales et al. 2022). Thus, it is evident that understanding pathological hypoxia is of immense clinical relevance and for the same reason an in-depth knowledge of hypoxia-responsive and adaptive pathways is important.

Smart Nanotechnology in Pathological Hypoxia: An Innovative Avenue …

3

1.1 Molecular Responses to Hypoxia—Hypoxia-Inducible Factors and Allied Pathways A molecular signature common to all forms of hypoxia described above is activation of hypoxia-inducible factor (HIF). HIF is the primary molecular “oxygen level” sensor and triggers adaptive gene expression changes having implication on tissue, organ, and physiological levels adaptations (Luo et al. 2022). Besides HIF, the cell also activates series of other adaptive pathways—autophagy, metabolism, and energy pathways such as mTOR complex 1 and endoplasmic stress, proteasomal degradations, angiogenesis, and erythropoiesis pathways. All these pathways intersect with the central HIF pathway (Fig. 1) at different junctions and facilitate the cell’s response to hypoxic stress, mainly by reducing oxygen consumption and enhancing oxygen delivery (Lee et al. 2019; Lee et al. 2020; Luo et al. 2022). The readership will rightly question here that when the HIF-triggered pathways are adaptive, then how is it that the same molecular framework takes pathological connotations? Studies point to either a sub-optimal/delayed activation or overt molecular responses—activation of inflammatory pathways, vascular disturbances and fluid imbalance, and pro-oxidant intracellular environment which lead to pathological consequences in various human diseases due to hypoxia (Chawla et al. 2020). For example, a classical example is that of tumor hypoxia, the oxygen-deprived core

Fig. 1 Hypoxia-inducible factor pathway and its intersection with other hypoxia-responsive pathways. The adaptive cellular and metabolic changes brought about by HIF activation can be categorized into those increasing oxygen delivery or reducing cellular oxygen consumption (Kaneisha et al. 2000)

4

S. Chawla et al.

of a tumor has increased activation of HIF 1α and 2α, which in turn enhances the pro-survival and invasive nature of cancerous cells along with conferring a chemoresistant phenotype. Tumor hypoxia is in fact a major challenge to the success of chemotherapy (Wo´zniak et al. 2020) (Table 1).

2 Smart Nanomaterials Of the recent technological advancements made by man, nanotechnology has maximally impacted the human healthcare sector, making immense contribution in therapeutics, diagnostics, drug delivery, imaging, regenerative medicine, wearable medical devices, and many more (Sakamoto et al. 2010; Anjum et al. 2021). A case in point, small molecules have been at the heart of new drug development, however their success is delimited by a number of factors such as biocompatibility and bioavailability, stability in biotic and abiotic conditions (light exposure, temperature, enzymatic breakdown, etc.), undesirable off-target effects, and poor pharmacokinetics (Sao et al. 2021). Nanotechnological interventions such as nanoformulations addressed several challenges in classical drug discovery and delivery, especially the pharmacokinetics and targeted delivery (Saeed et al. 2023; de Almeida et al. 2023). However, the field is still open and warrants further technological advances. “Smart” nanotechnology is a leap in the same direction. It stands on use of smart nanomaterials—nano-sized particles that acquire functionality in response to an external stimulus (light, temperature, pH, magnetic/electric field, chemical, pressure) (Aflori 2021). Figure 2 summarizes the various stimuli that have been tapped till date for generating responsive/smart nanomaterials. In response to the external stimulus, these nanomaterials may change their size (increase/decrease), optical properties, surface area, permeability, surface area, shape, solubility, etc. Five major characteristics of smart nanomaterials are immediacy, transiency, self-actuation, directness, and selectivity. These characteristics confer a superiority to smart nanomaterials and unique advantages (Shehata et al. 2022). Various stimuli-responsive nanomaterials being developed/pursued have been reviewed elsewhere in detail (Thangudu 2020).

2.1 Smart Nanomaterials Targeting Hypoxic Tissues Stimuli-responsive nanomaterial directed against hypoxic tissues is a recent but increasingly recognized strategy to deal with pathological hypoxia in varied clinical settings. For example, as highlighted by Li and co-workers in their seminal review on efficacy of oxygen nanocarriers as anti-tumor therapy, the literature focusing on “oxygen”, “tumor”, and “nano” sciences is increasing steadily since 2015 in PubMed (Li et al. 2021a, b; Arulmozhi et al. 2019). In the following book chapters, we elaborate on the utility, efficacy, and potential applications of smart nanoplatforms in addressing pathological hypoxia in varied

Smart Nanotechnology in Pathological Hypoxia: An Innovative Avenue …

5

Table 1 Summarizes principle human diseases with hypoxia as a coincident pathological feature and the impact on HIF expression Human diseases

Hypoxic response

References

Diabetes

High glucose level suppresses activity of HIF-1α

Zhao et al. (2022)

Non-alcoholic fatty liver disease Aggravates the inhibition of (NAFLD) functioning of hepatocytes

Osteoporosis

Ji et al. (2022)

HIF-1α is upregulated in hepatocytes in non-alcoholic fatty liver disease

Wang et al. (2022)

HIF-1α deficiency in hypoxic conditions promotes inhibition of osteoclast formation

Liu et al. (2020)

HIF-1α promotes viral replication by stimulation of Ca2+ release

Serebrovska et al. (2020)

Infectious diseases Pneumonia-associated COVID-19

Activated HIF-1α decreases expression of angiotensin and results in decreased invasiveness of SARS-CoV-2 Viral hepatitis

Hepatitis B virus X protein Lu et al. (2015) promotes the HIF-1α transcription by activating the mitogen-activated protein kinase pathway

Neoplastic diseases Colon cancer

Hypoxia-induced Orai1 that induced colon cancer

Wang et al. (2018)

Lung cancer

Hypoxia elevates HIF-1α level in lung cancer

Popper (2016)

Breast cancer

Increased HIF-1α

Heer et al. (2020)

Pancreatic cancer

HIF-1α increases malignancy of Tao et al. (2021) tumor by inducing epithelial to mesenchymal transition

Prostate cancer

Enhanced invasiveness of prostate cancer PC3 cells by upregulating HIF-1α expression

Ashton and Bristow (2020)

Atherosclerosis

Increased expression of HIF-1α and HIF-2α

Knutson et al. (2021)

Pulmonary hypertension

HIF-1α accelerates vascular remodeling and promotes PH formation

Ghofrani et al. (2006)

Cardiovascular Diseases

(continued)

6

S. Chawla et al.

Table 1 (continued) Human diseases

Hypoxic response

References

Cardiomyopathy

Increased hypoxic response of critical congenital heart disease

Zhou et al. (2017)

Alzheimer’s disease (AD)

Increased HIF-1α results in increased ROS

Burtscher et al. (2021)

Amyotrophic lateral sclerosis (ALS)

HIF-1α activation promotes the motor neuron decline in amyotrophic lateral sclerosis

Anand et al. (2013)

Neurodegenerative diseases

Fig. 2 Schematic summarizing stimuli that have been tapped for developing hypoxia-targeted nanoplatforms

clinical settings—cancer (Chaps. 3, 5, 6, 7), vascular diseases (Chap. 9), stroke and cerebral ischemia (Chap. 11), and high-altitude hypoxia (Chap. 21). Hypoxia-responsive nanomaterials can be classified into following categories based on the biochemical, molecular, and cellular characteristics of hypoxic tissues (Xia et al. 2022; Li et al. 2021a, b). The list is not exhaustive keeping in mind the evolving interest of the clinicians and nanotechnologists in this field: • • • • •

Oxygen-carrying nanomaterials. Oxygen generators. Redox-responsive nanomaterials. Low pH-responsive nanomaterials. Hypoxia-triggered prodrugs in nanoformulations.

Smart Nanotechnology in Pathological Hypoxia: An Innovative Avenue …

7

• Therapeutic peptides in bioresponsive nanoformulations. • Fe/Mn based or fluorescent nanoplatforms largely used in imaging. • Nanoplatforms targeting cellular population in hypoxic regions (e.g., tumorassociated macrophages). • Nanozymes. • X-ray/Near-Infrared-responsive nanoparticles. • High atomic number elements-based nanoparticles. • Phyto-nanotheranostics. The list also includes smart nanoplatforms that work synergistically with conventional therapies and enhance their efficacy in hypoxic regions. Especially in scenario of tackling the hypoxic tumor microenvironment, hypoxia-targeted smart nanoparticles have assisted in enhancing radiosensitization, photosensitization, and chemosensitivity (Chen et al. 2018; Li et al. 2018). Besides the obvious therapeutic applications, there is an evident application in diagnosing and imaging of hypoxic regions in vivo (Chaps. 10, 12). For example, hypoxia-sensing nanoprobe developed using mesoporous silica-coated lanthanide-doped nanoparticles containing O2 -sensitive iridium (III) complex on the exterior shell by Lv and co-workers. The nanoprobe can facilitate intracellular imaging, addressing the disadvantages of autofluorescence, as well as allow imaging in oxygen gradients (Lv et al. 2015). Further, innovative and pathbreaking applications of smart nanotechnology in understanding and tackling hypoxic microenvironment—such as nanobots and nano metabolomics—have also been elaborated in Chaps. 2 and 20. In conclusion, smart nanotechnology is an innovative new paradigm with immense medical potential, namely, with application in developing therapeutics, diagnostics, and imaging solutions. Hypoxia, a classical pathological feature of a wide range of diseases stands to benefit immensely from stimuli-responsive nanoparticles and the same are discussed in the following texts.

References Aflori M (2021) Smart nanomaterials for biomedical applications—a review. Nanomaterials 11(2):396 Anand A, Thakur K, Gupta PK (2013) ALS and oxidative stress: the neurovascular scenario. Oxid Med Cell Long 635831 Anjum S, Ishaque S, Fatima H, Farooq W, Hano C, Abbasi BH, Anjum I (2021) Emerging applications of nanotechnology in healthcare systems: grand challenges and perspectives. Pharmaceutic Basel 14(8):707 Arulmozhi S, Matchado MS, Snijesh VP, Kumar A, Singh S (2019) An insight into anti-arthritic property of C25H34O7 for rheumatoid arthritis using molecular modelling and molecular dynamics approach. Inform Med Unlock 16:100145 Ashton J, Bristow R (2020) Bad neighbours: hypoxia and genomic instability in prostate cancer. Br J Radiol 93(1115):20200087 Bhutta BS, Alghoula F, Berim I (2022) Hypoxia–statPearls–NCBI bookshelf. Hypoxia–StatPearls– NCBI Bookshelf

8

S. Chawla et al.

Burtscher J, Mallet RT, Burtscher M, Millet GP (2021) Hypoxia and brain aging: Neurodegeneration or neuroprotection? Ageing Res Rev 68:101343 Chawla S, Rahar B, Singh M, Bansal A, Saraswat D, Saxena S (2014) Exogenous sphingosine1-phosphate boosts acclimatization in rats exposed to acute hypobaric hypoxia: assessment of haematological and metabolic effects. PLoS ONE 9(6):e98025 Chawla S, Rahar B, Tulswani R, Saxena S (2020) Preventive preclinical efficacy of intravenously administered sphingosine-1-phosphate (S1P) in strengthening hypoxia adaptive responses to acute and sub-chronic hypobaric hypoxia. Eur J Pharmacol 870:172877 Chen Z, Liu L, Liang R, Luo Z, He H, Wu Z, Tian H, Zheng M, Ma Y, Cai L (2018) Bioinspired hybrid protein oxygen nanocarrier amplified photodynamic therapy for eliciting anti-tumor immunity and abscopal effect. ACS Nano 12(8):8633–8645 Chen Y, Gaber T (2021) Hypoxia/HIF modulates immune responses. Biomedicines 9(3):260 Chen PS, Chiu WT, Hsu PL, Lin SC, Peng IC, Wang CY, Tsai SJ (2020) Pathophysiological implications of hypoxia in human diseases. J Biomed Sci 27(1):63 de Almeida Campos L, Fin MT, Santos KS, de Lima Gualque MW, Freire Cabral AKL, Khalil NM, Fusco-Almeida AM, Mainardes RM, Mendes-Giannini MJS (2023) Nanotechnologybased approaches for voriconazole delivery applied to invasive fungal infections. Pharmaceutics 15(1):266 Feigin VL, Brainin M, Norrving B, Martins S, Sacco RL, Hacke W, Fisher M, Pandian J, Lindsay P (2022) World stroke organization (WSO): global stroke fact sheet 2022. Int J Stroke 17(1):18–29 Ferdinand P, Roffe C (2016) Hypoxia after stroke: a review of experimental and clinical evidence. Exp Transl Stroke Med 7(8):9 Ghofrani HA, Voswinckel R, Reichenberger F, Weissmann N, Schermuly RT, Seeger W, Grimminger F (2006) Hypoxia- and non-hypoxia-related pulmonary hypertension–established and new therapies. Cardiovasc Res 72(1):30–40 de Heer EC, Jalving M, Harris AL (2020) HIFs angiogenesis and metabolism: elusive enemies in breast cancer. J Clin Invest 130(10):5074–5087. https://doi.org/10.1172/JCI137552 Ji Y, Liang Y, Chu PH, Ge M, Yeung SC, Man Ip MS, Wo Mak JC (2022) The effects of intermittent hypoxia on hepatic expression of fatty acid translocase CD36 in lean and diet-induced obese mice. Biomed J S2319–4170(22)00141-X Kanehisa M, Goto S (2000) KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res 28(1):27–30 Knutson AK, Williams AL, Boisvert WA, Shohet RV (2021) HIF in the heart: development, metabolism, ischemia, and atherosclerosis. J Clin Investig 131(17):e137557 Lee JW, Ko J, Ju C, Eltzschig HK (2019) Hypoxia signaling in human diseases and therapeutic targets. Exp Mol Med 51(6):1–13 Lee P, Chandel NS, Simon MC (2020) Cellular adaptation to hypoxia through hypoxia inducible factors and beyond. Nat Rev Mol Cell Biol 21(5):268–283 Li J, Shang W, Li Y, Fu S, Tian J, Lu L (2018) Advanced nanomaterials targeting hypoxia to enhance radiotherapy. Int J Nanomed 13:5925–5936 Li X, Wu Y, Zhang R, Bai W, Ye T, Wang S (2021a) Oxygen-based nanocarriers to modulate tumor hypoxia for ameliorated anti-tumor therapy: fabrications, properties, and future directions. Front Mol Biosci 8:683519 Li Y, Zhao L, Li XF (2021b) Hypoxia and the tumor microenvironment. Technol Cancer Res Treat 20:15330338211036304 Liu X, Ma C, Wang S, Liang Z, Yang J, Zhou J, Shu Y, He Z, Zong J, Wu L, Peng P, Su Y, Gao M, Shen K, Zhao H, Ruan J, Ji S, Yang Y, Tang T, Yang Z, Luo G, Zeng M, Zhang W, He B, Cheng X, Wang G, Wang L, Lyu L (2020) Screening of osteoporosis and sarcopenia in individuals aged 50 years and older at different altitudes in Yunnan province: protocol of a longitudinal cohort study. Front Endocrinol (lausanne) 13:1010102 Lu Y, Sui J, Liu Y, Li R, Ma L, Zhang X, Qin X, Li S (2015) Association between hypoxia-inducible factor-1α gene polymorphisms and risk of chronic hepatitis B and hepatitis B virus-related liver cirrhosis in a Chinese population: a retrospective case-control study. Gene 564(1):96–100

Smart Nanotechnology in Pathological Hypoxia: An Innovative Avenue …

9

Luo Z, Tian M, Yang G, Tan Q, Chen Y, Li G, Zhang Q, Li Y, Wan P, Wu J (2022) Hypoxia signaling in human health and diseases: implications and prospects for therapeutics. Signal Transduct Target Ther 7(1):218. https://doi.org/10.1038/s41392-022-01080-1. PMID: 35798726; PMCID: PMC9261907 Lv W, Yang T, Yu Q, Zhao Q, Zhang KY, Liang H, Liu S, Li F, Huang W (2015) A phosphorescent Iridium(III) complex-modified nanoprobe for hypoxia bioimaging via time-resolved luminescence microscopy. Adv Sci (weinh) 2(10):1500107 Mora C, McKenzie T, Gaw IM, Dean JM, von Hammerstein H, Knudson TA, Setter RO, Smith CZ, Webster KM, Patz JA, Franklin EC (2022) Over half of known human pathogenic diseases can be aggravated by climate change. Nat Clim Chang 12(9):869–875 Popper HH (2016) Progression and metastasis of lung cancer. Cancer Metastasis Rev 35(1):75–91 Rahman A, Tabassum T, Araf Y, Al Nahid A, Ullah MA, Hosen MJ (2021) Silent hypoxia in COVID-19: pathomechanism and possible management strategy. Mol Biol Rep 48(4):3863– 3869 Rosales AM, Shute RJ, Hailes WS, Collins CW, Ruby BC, Slivka DR (2022) Independent effects of acute normobaric hypoxia and hypobaric hypoxia on human physiology. Sci Rep 12(1):19570 Rowat AM, Dennis MS, Wardlaw JM (2006) Hypoxaemia in acute stroke is frequent and worsens outcome. Cerebrovasc Dis 21(3):166–172. https://doi.org/10.1159/000090528. Epub 2005 Dec 23 PMID: 16388191 Saeed S, Khan SU, Gul R (2023) Nanoparticle: a promising player in nanomedicine and its theranostic applications for the treatment of cardiovascular diseases. Curr Probl Cardiol 101599 Sakamoto JH, van de Ven AL, Godin B, Blanco E, Serda RE, Grattoni A, Ziemys A, Bouamrani A, Hu T, Ranganathan SI, De Rosa E, Martinez JO, Smid CA, Buchanan RM, Lee SY, Srinivasan S, Landry M, Meyn A, Tasciotti E, Liu X, Decuzzi P, Ferrari M (2010) Enabling individualized therapy through nanotechnology. Pharmacol Res 62(2):57–89011 Salyer J (1989) History of oxygen. Chest 96(4):959 Sao P, Chand Y, Kumar A, Singh S (2021) Potential drug target identification in porphyromonas gingivalis using in-silico subtractive metabolic pathway analysis. Bangladesh J Med Sci 20(4):887–896 Saxena K, Jolly MK (2019) Acute vs. chronic vs. cyclic hypoxia: their differential dynamics, molecular mechanisms, and effects on tumor progression. Biomolecules 9(8):339 Serebrovska ZO, Chong EY, Serebrovska TV, Tumanovska LV, Xi L (2020) Hypoxia, HIF-1α, and COVID-19: from pathogenic factors to potential therapeutic targets. Acta Pharmacol Sin 41(12):1539–1546 Serrano R, Corbella X, Rello J (2021) Management of hypoxemia in SARS-CoV-2 infection: lessons learned from one year of experience, with a special focus on silent hypoxemia. J Intens Med 1(1):26–30 Shehata N, Abdelkareem MA, Sayed ET, Egirani DE, Opukumo AW (2022) Smart materials: the next generation. In: Olabi A-G (ed.) Encyclopedia of smart materials. Elsevier, pp 288–299. ISBN 9780128157336 Tao J, Yang G, Zhou W, Qiu J, Chen G, Luo W, Zhao F, You L, Zheng L, Zhang T, Zhao Y (2021) Targeting hypoxic tumor microenvironment in pancreatic cancer. J Hematol Oncol 14(1):14 Thangudu S (2020) Next generation nanomaterials: smart nanomaterials, significance, and biomedical applications. In: Khan F (ed) Applications of nanomaterials in human health. Springer, Singapore Tirpe AA, Gulei D, Ciortea SM, Crivii C, Berindan-Neagoe I (2019) Hypoxia: overview on hypoxiamediated mechanisms with a focus on the role of HIF genes. Int J Mol Sci 20(24):6140 Wang Z, Ma L, Su M, Zhou Y, Mao K, Li C, Peng G, Zhou C, Shen B, Dou J (2018) Baicalin induces cellular senescence in human colon cancer cells via upregulation of DEPP and the activation of Ras/Raf/MEK/ERK signaling. Cell Death Dis 9(2):217 Wang CH, Liu HM, Chang ZY, Lee MC, Hsu CH, Lee TY (2022) Antioxidants rich herbal formula Ger-Gen-Chyn-Lian-Tang protects lipotoxicity and ameliorates inflammation signaling through

10

S. Chawla et al.

regulation of mitochondrial biogenesis and mitophagy in nonalcoholic fatty liver disease mice. Front Biosci (landmark Edition) 27(8):242 Wo´zniak M, Pastuch-Gawołek G, Makuch S, Wi´sniewski J, Ziółkowski P, Szeja W, Krawczyk M, Agrawal S (2020) Overcoming hypoxia-induced chemoresistance in cancer using a novel glycoconjugate of methotrexate. Pharmaceuticals (basel) 14(1):13 Xia Y, Duan S, Han C, Jing C, Xiao Z, Li C (2022) Hypoxia-responsive nanomaterials for tumor imaging and therapy. Front Oncol 12:1089446 Zhao M, Wang S, Zuo A, Zhang J, Wen W, Jiang W, Chen H, Liang D, Sun J, Wang M (2022) HIF1α/JMJD1A signaling regulates inflammation and oxidative stress following hyperglycemia and hypoxia-induced vascular cell injury. Cell Mol Biol Lett 26(1):40 Zhou S, Yin X, Jin J, Tan Y, Conklin DJ, Xin Y, Zhang Z, Sun W, Cui T, Cai J, Zheng Y, Cai L (2017) Intermittent hypoxia-induced cardiomyopathy and its prevention by Nrf2 and metallothionein. Free Radical Biol Med 112:224–239 Zhou J, Li M, Hou Y (2022) Engineering of a nanosized biocatalyst for combined tumor starvation and low-temperature photothermal therapy. ACS Nano 12:2858–2872

Bioinspired Nanosystems Interacting with the Host Environment: Smart Nanosystems Shatabdi Basu, Koena Mukherjee, Koel Mukherjee, and Dipak Maity

Abstract The potential application of nanotechnology claimed in the medicinalpharmaceutical sector is spreading rapidly nowadays. In recent times, the development of bioinspired nanoparticles or nanocarriers has gained a lot of popularity. In the context of nanomedicine, drug targeting and delivery and its related pharmaceutical properties enhancement is a major issue. Nanometer ranging complex systems (10–1000 nm) applied in biomedical industries are concerned with two important resources: (i) the host environment and (ii) the pharmaceutically active ingredient. The growing demand for more efficient and iatrogenic-free treatments is demanding a full revision of carrier configuration in drug targeting. This chapter will focus mainly on the smart nanosystems and its applications in targeted drug delivery. Keywords Nanoparticles · Nanocarriers · Drug targeting · Cancer · Nanorobots · Smart nanosystems

1 Introduction In nanotechnology division, studies are performed on the construction and application of particles that are present at the level of nanoscale, ranging between the size of 1 and 100 nm. However, the miniaturization of particles offers enhancement S. Basu · K. Mukherjee Department of Bioengineering and Biotechnology, Birla Institute of Technology Mesra, Ranchi, Jharkhand 835215, India K. Mukherjee Department of Electrical and Instrumentation Engineering, National Institute of Technology, Silchar, Assam, India D. Maity (B) Department of School of Health Sciences & Technology, University of Petroleum and Energy Studies, Dehradun, Uttarakhand 248007, India e-mail: [email protected] Department of School of Engineering and Technology, The Assam Kaziranga University , Jorhat, Assam 785006, India © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_2

11

12

S. Basu et al.

of mechanical, chemical, and biological properties. Nanometre-sized particles can also show remarkable character ordering and assembly of behaviors under specific magnitudes of forces which are relatively different from macro-objects (Emerich and Thanos 2006). Due to the relation between the size and properties of these nanoparticles, they can be applied to many other devices and systems (Roco et al. 2011). Further, development in nanotechnology helps in nanomedicine formulation and its target-specific delivery in a living system which can modernize the industrial sector and help in reducing the process cost. Overall nanotechnological application undoubtedly improves the quality of human life in various aspects. A smart nanosystem refers to a particular arrangement of nanosized particles (nanocarrier) that can trap and deliver low molecular weight biomolecules/drugs to its targeted disease site. Since nanosystems have a high loading capacity and are extremely site specific, they can aggregate in tumor cells and focus on diagnosis and treatment (Huang et al. 2018). Smart nanosystems offer opportunities for cancer medication, targeting, and identifying disease-specific targets, with a nanoscale dimension that must get past the multiple layers of barriers to reach the target (Husen et al. 2017). To obtain an effective nanosystem for delivering drugs to tissues, it should meet the following criteria: (a) controlled size and conformation (i.e., nanometric size and shape monodispersed), (b) high stability to avoid aggregation/clustering, (c) ability to deceive the immune system and thus avoid being removed from blood circulation, and (d) drug-loading capacity (Voliani et al. 2012). The benefit of this approach is that it fundamentally alters systemic biodistribution in a way that non-native drug delivery systems (DDS) do not (Madamsetty et al. 2019). The main part of the smart nanosystems is the smart nanocarriers, which can carry the drug molecules to the site and help in controlled release with the help of external stimuli (Singh et al. 2019a). Bioinspired nanocarriers are primarily used in biomedical applications due to their physical and chemical properties. With Nobel metal centers and encapsulating therapeutic and imaging agents, they have multifunctional properties in therapy and diagnostic area (Lôbo et al. 2021). They can be used to treat a variety of diseases such as cardiovascular disease, diabetes, cancer, neurological disease, and bacterial infection. Theranostic nanoplatforms require the selection of an effective therapeutic agent, and a stable carrier to implant a targeted and sustainable drug release (Madamsetty et al. 2019) on the site. Nanocarriers are low in cost and have less toxicity against normal cells, which includes liposomes, lipid-based nanocarriers, protein-based nanocarriers, and metal-based nanocarriers (Fig. 1). The profits of using nanocarriers contain active drug degeneration protection, effective concentrations in the target tissue, and reduction of undesirable poisonous sides (Borm and Muller-Schulte et al. 2006). Similarly, nanocarriers can also be attached to specific ligands to increase their specificity to target tissues (Vasir et al. 2005). Similarly, nanocarriers also have the advantage over other delivery systems as of regulating cellular efflux pumps, which helps to counteract drug-resistant mechanisms seen in some tumors (Deng et al. 2020). With the advancement in technology, bioinspired nanorobots can also be designed and developed artificially for site-specific drug delivery and interaction at a molecular level (Rajesh et al. 2018). Nanorobots will help to deliver small but precise

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

13

Fig. 1 Schematic diagram of various types of smart nanosystems

medication to patients suffering from different types of diseases (thrombocytopenia, identification of cancer cells, and subsequent eradication of the targeted cancer cells and HIV) and finally eradicate from the human body (Li et al. 2017a, b). The advantage of using nanorobots is to eradicate diseased cells precisely without affecting other healthy cells.

2 Smart Nanosystems in Drug Delivery Targeted administration and controlled release of medicinal agents using nanoparticles are known as nanosystems in drug delivery. Based on the makeup of their platforms, the modern configuration of nanosystems (DDS) can be divided into numerous categories, including polymeric nanoparticles, inorganic nanoparticles, viral nanoparticles, lipid-based nanoparticles, and albumin-bound (nab) nanoparticle technologies (Fig. 1). The medicine delivery system should be designed in such a way that to minimize the negative effects dosage and dosing frequency will be lower. The goal of nanoparticle-mediated delivery is to increase therapeutic effectiveness while reducing cytotoxicity. The following aspects must be taken into consideration for finetuning the nanoparticle packets for efficient drug delivery (Wilczewska et al. 2012). Besides, lowering dosage and reducing nanoparticle toxicity can be accomplished by improving ligand-binding efficiency.

14

S. Basu et al.

Reducing the quantity or frequency of medications also lowers the weight of nanoparticles per weight of the drug, resulting in decreased efficacy (Hainfeld et al. 2014). The relationships between inorganic nanoparticulate systems and biosystems may become problematic as they degrade to their beginning names. After therapy, a significant number of patches may still be present in the body, accumulating essence patches that may be toxic (Kreyling et al. 2004). Unspecified environmental hazardous consequences of nanoparticles are not assumed, however, until the nanoparticles undergo transformation to release free metal ions, according to several recent findings. Copper (Cu), silver (Ag), and titanium (Ti) nanoparticles were designed to release very little or no essence ions in aerobic and anaerobic circumstances. This proves that Cu/Ag/Ti nanoparticles slowly release metal ions (Kreyling et al. 2002). Additionally, by minimizing the leakage of essential ions from the core (Loo et al. 2004), the nanoshell coatings effectively protect against a decline in the cellular environment and subsequently reduce toxicity (Grabowska-Jadach et al. 2020).

3 Targeted Drug Delivery Approaches Due to the small size of nanomaterials (in the range of 10–100 nm), they possess larger surface area and quantum effects than the bulk materials and thereby retain distinctive physical, chemical, and magnetic properties (Lovric et al. 2005a). They also possess a prolonged residence time, and the sustained drug release rate can be obtained by controlling the size and architecture of the nanoparticles (Lovric et al. 2005b). Well-known problem of Blood–Brain Barrier (BBB) has been also solved using nanocarriers-based DDS (Mishra et al. 2016). Targeted drug delivery using nanosystems is an upcoming area of medical sector where drug molecules can be delivered with high specificity and accuracy to the target site (Thakur et al. 2015; Kumar et al. 2017). Previously, in conventional method, drug is used to be delivered to site area where huge drug loss was encountered due to non-specificity. It was also causing death to the normal cells as drugs can’t differentiate between normal and diseased cells. But using nanocarriers, drugs can be reached to its specific location without hampering other good/normal cells as well as less wastages of drug molecules can also be noticed (Tewabe et al. 2021). Targeted drug delivery process includes four main stages, retain, evade, target, and release of the drug molecule. For efficient targeted drug delivery using nanocarriers, drug molecules must be first loaded in the delivery vehicle and then transported to the delivery site (Michaelis et al. 2006). As the delivery site can be any part of the living system, sufficient residence time and retention time should be provided. Finally, drug molecules must be released from the carrier and bind to the particular site/cell or receptor to show the efficient drug-targeting function (Kreuter et al. 2003). As nanocarriers have easy biodistribution property and can easily cross the membranes and reach the target-specific site so they are known as “Magic Bullets” (Strebhardt

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

15

Fig. 2 Different types of targeted drug delivery approaches

and Ullrich 2008). Broadly targeted drug delivery can be classified into five types, provided in Fig. 2. One of the most common types of targeted drug delivery approach is based on the interactions between drug molecules and the targeted sites (Fukunishi et al. 2009). This type can be divided into two sub-types: active targeting and passive targeting (Fig. 3). Active targeting is the process of modifying or functionalizing pharmacological nanocarriers with site-specific targeting ligands such as cancer-specific monoclonal antibodies, folate receptor, etc. so that they can recognize the target site and reach there by self-navigation (Lin et al. 2006). Thus, in active targeting, drugs/agents are only delivered to the site-specific location for which the nanocarriers are designed. More focused and concentrated delivery, less toxicity and potential negative effects are anticipated in this active targeting process (Rani et al. 2014). Depending on the interactions, active targeting has various orders. First-order targeting, which deals with the compartmentalization of organs, and second-order targeting, which deals with cellular targeting, are two levels of active targeting (Nobs et al. 2004), that can be impacted. Intercellular organelle targeting is frequently indicated by third-order targeting (Borm and Kreyling et al. 2004). Thus, active targeting is the selective administration of a medication delivery system to the capillary bed of an organ, tissue, or target location (Tewabe et al. 2021). Body reacts when drug molecule acts as a foreign entity and enters into the system. The same concept is taken in passive targeting where drug targeting occurs due to body’s natural response after encountering drug molecules. In passive targeting, drug immobilized nanoparticles can be accumulated via “Enhanced Permeability and Retention” (EPR) effect through “leaky” vasculature of the pathological site (tumors, infarcts, inflammation with less endothelial barriers) which permits passive extravasation of the nanoparticles from the circulation into the tumor interstitium, where they retain due to poor lymphatic drainage (Nemmar et al. 2002). In contrast,

16

S. Basu et al.

Fig. 3 Active and passive drug targeting via nanoparticles-based nanocarriers

endothelial cells of normal tissue vessels are closely packed and present a greater barrier for nanoparticles penetration (Nemmar et al. 2003). This difference in vascular permeability provides a means for tumor-selective accumulation of nanoparticles. However, EPR mechanism depends on long circulation of the nanoparticles in the blood circulation and their retention ability inside the tumor (Lowery et al. 2006). Hence, the nanoparticles are usually attached with the sugars, peptides, or other small molecules that improve their ability to attach with the tumor surface or to penetrate into the cancerous cells (Lockman et al. 2004). In passive targeting carrier, nanoparticles are absorbed by the reticulo endothelial system in their natural course (RES) and therefore make the biodistribution easy (Lockman et al. 2003). They are one of the best means of passively directing medications to the most important compartments. The physiological state or anatomical barrier of the nanoparticles, along with their size, shape, and surface charge, provides the basis for passive targeting. These have an impact on the passive targeted delivery of the drug or imaging agent by increasing permeability and retention effect in tumor, inflammation, or other leaky vasculature and anatomic barrier. The cost is high, also there is a greater risk in toxicity and might contain some side effects (Borm and Kreyling et al. 2004).

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

17

4 Bioinspired Nanocarriers Bioinspired nanocarriers are those which are fabricated on the basis of morphological and functional insight of biological molecules. Nanoparticles-based nanocarriers like dendrimers, and polymeric or lipid-based carriers, such as liposomes, are various examples of bioinspired nanocarriers (Table 1). These nanocarriers-based DDS are helpful for delivering medications to the intended recipient. They permit delivery of medications in target-specific organs or cells but not in another site. Compared to free drugs site-specific delivery is a significant therapeutic advantage since it prevents the delivery of drugs to the incorrect locations. Utilizing the potential of nanobiotechnology, multimodal theranostics has developed into an encouraging tool for therapeutics and diagnostics. By encasing hydrophobic medications inside the core of the nanocarriers, they contribute to improving the effectiveness of medications. They are able to prevent the medicine from being prematurely degraded, prevent the drug from being exposed to the biological environment too early, enhance cellular penetration, manage pharmacokinetics and biodistribution of drug/DDS, and improve drug absorption in a specific tissue (Esfahani et al. 2018). Due to their small size and ability to change physical properties, such as the charge and shape, they are also considered as effective transport agents to transfer healing chemicals to tissues. Following sections describe details about different types of nanocarriers used in theranostic application.

4.1 Lipid Nanostructures When a drug is suspended in lipid excipients, the formulation is referred to as a lipidbased DDS. They can be fabricated by blending of liquid oil and solid lipids, which creates a unique nanostructure in the matrix. Lipid nanostructures are developed by merging two phases, outer layer and inner layer (as shown in Fig. 4). The outside section is made of a four-component ethanol phase containing ionizable lipid, helper phospholipid, cholesterol, and lipid-anchored PEG (Bazile et al. 1995). On the other hand, the inner section is an acidic aqueous phase containing mRNA (Fig. 4). Doxil was the first authorized liposomal medication that is a nanostructured lipid carrier of anticancer drug (i.e., doxorubicin) for the treatment of ovarian cancer.

4.2 Lipid Nanoparticles (LNPs) Lipid nanoparticles (LNPs) act as motivating tools in the pharmaceutical sector to be used in variety of treatments. LNPs were first interpreted as liposomes, which are a flexible platform for delivering nanomedicine. They have the capacity to recapitulate,

18

S. Basu et al.

Table 1 Different types of bioinspired nanocarriers and their applications Definition

Properties

Applications

References

Lipid-based nanoparticles Nanoemulsion

Long-chain and medium-chain glycerides and fatty acids

Medium- and long-chain fatty acids increase permeation. The cell membrane can be shattered by unsaturated carbon chains. Oleic acid makes the cell membrane’s phospholipids more mobile

Enhances the active Jaiswal et al. medicinal (2015) components’ delivery. Stable isotropic thermodynamic system. Here, a single phase is created by combining two immiscible liquids

Solid lipid nanoparticle

Long-chain triglycerides, fatty acids and phospholipids

Triglycerides slow down the release of peptides more than other glycerides do. Resistance to lipolysis that is reliant on structure encourages improved stability. Tripalmitin has a strong affinity for cell membranes, which causes endocytosis

Solid lipid Vimala and nanoparticles Kannan et al. (SLNs) are (2021) employed as nanocarriers frequently because they have developed into an effective, non-toxic, and adaptable pulp carrier system for various medications, avoiding some of the drawbacks of liposomes and polymeric nanoparticles

Nanocapsule

Medium-chain mono-, di-, and triglycerides, long-chain fatty acids

Lipid derivatives with a medium chain (such as C8/C10) function well as penetration enhancers. Medium-chain fatty acids promote emulsification and aid in the solubilization of peptides and proteins

A vesicular system Yurgel et al. is involved. This (2013) system has active molecules inside a core or cavity structure that is encircled by polymeric material. It is used to treat tumors or stop the spread of cancer. They arrive to the desired tumor site and discharge the information there (continued)

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

19

Table 1 (continued) Definition

Properties

Applications

References

Liposome

Phospholipids, phosphatylglycerol derivates, saturated and unsaturated fatty acids

Phospholipids can self-assemble amphiphilic structures

Drugs that are both Gregoriadis hydrophilic and et al. (1976) hydrophobic can be released from lipid vesicles, which contain phospholipid bilayer

Self-emulsifying drug delivery systems (SEDDS)

A self-microemulsifying drug delivery system is a drug delivery system that uses a microemulsion achieved by chemical rather than mechanical means

Greater bioavailability and quicker release rates are both influenced by improved solubility. Faster release rates increase consumer acceptance of many medications given orally. Greater bioavailability reduces the amount of medicine needed

Improved Garti et al. solubilization of (2016) bioactive compounds, ease of production, thermodynamic stability, and spontaneous generation

Inorganic nanoparticle Single layer Nanotube

The nanotubes consist solely of carbon, sp2 -bonded as in graphene strips rolled to form closed cylinders

Tubules up to 6 nm in diameter are created. One-dimensional conductivity and extremely high tensile strength are just two of the revolutionary mechanical and electrical characteristics that single-layer nanotubes are expected to possess

Compatibility of chemicals with biomolecules. It can be utilized in medical devices as biosensors

Kiang et al. (1995)

Graphene Nanotube

Graphene is a two-dimensional material, basically a single layer of graphite, with carbon atoms arranged in a hexagonal, honeycomb lattice

In general, carbon nanotubes are a sheet of graphene that has been wrapped into a cylinder

Used mostly to Yang et al. create biosensors. (2010) They are employed in everything from the diagnosis of fatal illnesses to the search for biological weapons used in conflict or terrorist acts (continued)

20

S. Basu et al.

Table 1 (continued) Metal Nanoparticle

Definition

Properties

Applications

References

Metal nanoparticles are small, submicron-sized objects formed of pure metals or their compounds, such as gold, platinum, silver, titanium, zinc, cerium, iron, and thallium

High levels of biocompatibility, stability, and the capacity to produce in large quantities without using organic solvents all have a positive impact on biological systems

Key biological Rizvi and molecule Saleh (2018) identification with high sensitivity, safer and more accurate imaging of sick tissues, and innovative therapies

Polymeric nanoparticles Micelle

Colloidal nanostructures called micelles have a hydrophobic core and a hydrophilic exterior

Polymeric micelles can access bodily regions that are difficult for liposomes to access

Acquire more Huang et al. medications than (2018) free ones due to increased vascular permeability, tissues. In order to deliver chemotherapeutics in a regulated and targeted manner with high concentration in the tumor cells and minimal side effects, polymeric micelles can be used. They span a size range of 1–100 nm

Dendrimer

Dendrimers are highly branching nanostructures with an inner core that enclose drugs

They are made from macromolecules such as polymer degradation hydrolysis, enzyme-catalyzed polymer degradation (PAMAM), polyamidoamine polypropyleneimine, and polyaryl ether

They are found Vasir et al. useful in (2005) nanomedicine research. They are used for carrier and delivery system for different types of drugs

(continued)

redeem, and release their contents over a short period of time at specific areas inside the body. LNPs, which are currently gaining attention as dynamic components of the COVID-19 mRNA vaccines, are essential for efficiently guarding and conveying mRNA to cells. A liposomal medication formulation is still in use and has received approval from the medical community (Tenchov et al. 2021). Future developments

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

21

Table 1 (continued) Definition

Properties

Applications

References

PLGA

One of the most widely utilized biodegradable polymers is poly (lactic-co-glycolic acid), or PLGA, since its hydrolysis produces the metabolite monomers lactic acid and glycolic acid

Biocompatibility and biodegradability can be obtained. Sustained release is a possibility, and nanoparticles can be directed to particular cells or organs

Well-described production processes and formulations tailored to different medication types, such as hydrophilic or hydrophobic small molecules or macromolecules

Danhier et al. (2012)

mPEG–PGA

Different kinds of natural and synthetic polymers, which typically have strong biocompatibility and biodegradability, are used to create polymeric nanocarriers Evidence of the function of polymers sensitive to reactive oxygen species (ROS), which are being extensively researched for anticancer therapy

Numerous potentially fatal disorders, such as cancer, neurological diseases, cardiovascular diseases, even viral infections and osteoporosis, can benefit from the features of polymerized NPs

These polymeric Sivadasan nanostructures have et al. (2021) benefits over other nanocarriers in that they are stable in a variety of microenvironments, release medications slowly due to polymer degradation, and can encapsulate a wide variety of pharmaceuticals

PEG–PMT

PEI–PLG

in LNPs, such as solid lipid nanoparticles and cationic lipid–nucleic acid complexes, exhibit more complicated structural arrangements and enhanced physical stabilities.

4.3 Lipid Nanocapsules (LNCs) Lipid nanocapsules (LNCs) are nanocarriers similar to surface active composites that are used to encapsulate a wide variety of operational elements. Phase inversion technology was successfully used to create LNCs within the range of 30–100 nm (Valcourt et al. 2016). With various compositions, properties of LNCs can be changed and its size range helps LNCs to suit in different applications. Though it can be applied in many biomedical sectors LNCs are more commonly used as DDS due to its biometric nature (Mouzouvi et al. 2017). LNCs contain both hydrophilic and hydrophobic regions where the nature of the core is hydrophobic (i.e., oily) and the outside part is made of phospholipid. Wilhelmy plate system and drop tensiometry are two techniques which are extensively used to reduce surface tension. LNCs-based nanocarriers hold great promise for delivering anticancer medications, particularly

22

S. Basu et al.

Fig. 4 Schematic representation of lipid nanostructure consisting of an aqueous core (yellow color) and outside phospholipid layer (blue-colored bi-layer)

for chemotherapy medicines that are water insoluble. During the drug loading, drug molecules can be evenly distributed on the surface or can be clustered in specific areas depending on the application area. It has been observed that during drug loading, the spherical shape changes to ellipsoidal shape that also hinders the drug-loading efficiency (Johnston et al. 2008).

4.4 Lipid-Based Micelles They are lipid molecules in spherical form, and they organize themselves as micelles in aqueous solutions (as shown in Table 1). Though there is small difference between liposome and micelles they can be both applicable for nanodrug delivery system (NDDS) (Bahadori et al. 2017). Micelles are single-layer structure of self-assembly of amphiphilic lipid molecules having hydrophilic heads (pointing outward when dispersed in water) and hydrophobic tails pointing toward the center of the structure. Thus, the micelles can be loaded with only hydrophobic antimicrobials or drug molecules to be delivered. The main advantages of micelles structure are less leakage of drugs due to the hydrophobic core nature and easy to handle (Wang et al. 2020).

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

23

4.5 Viral Vectors Virus has the ability to transfer their genetic material into host cells and this property is needed to construct the viral vectors. Viral vectors are such tools that can deliver gene to specific sites, especially applicable to gene therapy (Warnock et al. 2011). Through viral vectors the genetic material can be preserved and released to specific cell at full capabilities. The overall viral vector size is in nanometric size (generally below 100 nm) (Spencer et al. 2020). To make the viral vector nonlethal to host cell, viral components responsible for pathogenicity can be removed through genetic engineering while keeping all the essential components for the payload discharge in place. To deliver genetic materials, there are different types of viral vectors such as adenovirus, retrovirus, lentivirus, and herpes simplex virus. Irrespective of various promising applications of viral vectors, there are still many challenges to be addressed. After many attempts, only four viral vectors-based gene therapy is commercialized, and many are in preclinical stage. One of the biggest drawbacks of viral vectors is high cost and manufacturing difficulties (Fig. 5).

Fig. 5 Advantages and disadvantages of viral vectors

24

S. Basu et al.

4.6 Liposome-Based Nanocarriers Liposome-based nanocarriers consist of lipid bilayers surrounding an aqueous center in these structures (refer Table 1). They are naturally biodegradable and biocompatible. Comparative to other bioinspired nanocarriers, it is easy to synthesize and hardly harmful in nature. These liposome-based nanocarriers have the additional benefit of allowing their surfaces to alter in accordance with the targeted cancer therapy. They are capable of transporting both small and big molecules. It can include chemotherapy substances that are both hydrophilic and hydrophobic. Encapsulated liposomes and their by-products are currently being used in vitro to absorb and protect compounds with unique solubilities and deliver them to the target site inside and outside the body. On demand, certain liposome-derived carriers are licenced for clinical usage in humans via parenteral, transdermal, and oral delivery methods. Similar research is carried out to develop and improve liposomal systems for application in the nose and lungs. Besides, investigations have been continued to avoid using harmful substances like hazardous organic solvents and thereby to create safe and efficient liposome-based micro- and nanocarriers systems on an industrial scale. Liposomes are often identified based on diameter. Alternatively, classifications are often made according to whether a liposome has mono, oligo, or many lamellar bilayers. Liposomes can contain lipids that are produced synthetically or organically (Metselaar and Storm et al. 2005). As a result, liposomes can have a variety of different qualities and can be used to prevent infections (Minko et al. 2006). Therefore, liposomes can have a wide range of different properties and it is important to categorize them as naturally containing lipid, cation, or anion. It may naturally contain zwitter or fusogen ions to manage infection. If necessary, other lipids such as phosphatidylserine, phosphatidylcholine, and lecithin component are added to the naturally occurring phospholipids in the natural liposomes. Natural lipids have numerous hydrophobic lipid chains and a polar hydrophilic head. Engineered or natural lipids with cationic functionalities such as sulfonium, ammonium, or phosphonium ions are often used to create cationic liposomes. Anionic liposomes contain negatively charged functional groups like carboxylic, phosphoric, or sulfonic acids. Although cationic and anionic liposomes often show pH-dependent zeta potentials, they do not totally switch from having a positive charge to a negative charge. Zwitterionic lipids are capable of complete charge reversal below and above their isoelectric point due to the presence of functional groups that are both acidic and alkaline. The fluidity of the lipid bi-layer and the transport of liposomes with biological membranes are two of its most noticeable features (Wang et al. 2020).

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

25

4.7 Polymeric Nanocarrier Various drug delivery approaches in complex body system are a challenging part for pharmaceutical companies due to biodistribution and biocompatibility issues. Therefore, a new aspect of nanobiotechnology is coming forward where polymeric nanocarriers (PNCs) are designed and applied for specific targets. These types of carriers are multi-functional, can be modeled as per requirement, and same time can be used for theranostic purposes (Daglar et al. 2014; Girija 2018). Generally, polymer is known to be such macromolecule that is composed of many numbers of monomers or small chemical units. Polymer can be natural or synthetic substances such as Arabian gum, guar gum, chitosan, alginates, and alginic acid (ALG). (as natural polymer) (Bhatia 2016) and poly-e-caprolactone (PCL), poly (lactide) (PLA), poly (lactide-co-glycolide) (PLGA), poly (alkyl cyanoacrylate) (PACA), poly (glutamic acid) (PGA), polyethyleneimine (PEI), and polyacrylic acid (PAA) (as synthetic polymer) (Table 1). It has been observed from the studies that synthetic polymer shows more purity and biodegradability property but natural one displays more biocompatibility (Karabasz et al. 2020). Depending on the structural characteristics polymeric nanocarriers can be listed as polymeric micelles, polymersomes, polymeric nanogels, polymeric nanocapsules, and dendrimers. Nanocapsules have two parts, outer is known as shell and inner as core (Zhang et al. 2016). Drug molecules are loaded in core part and shell (polymeric) protects the drug from adverse chemical conditions. Sometimes drug molecules are loaded in shell part for controlled delivery in the body but then the polymeric shell part must be as permeable or semi-permeable in nature. The drug release and delivery are very much dependent on the polymeric shell properties like its pore size, thickness, and biodegradation (Nazila et al. 2016). Due to the nanosize, polymeric nanocarriers can pass through the membranes easily and therefore fit to be a delivery vehicle in living body. According to site of application, these polymers can be designed and engineered and integrated with other receptor molecules. In smart polymeric nanocarriers models, various stimuli-responsive polymers are used which are sensitive to chemical (oxidation–reduction, pH, ion), physical (temperature, ultrasound), and biological (enzymes, glucose, inflammation) stimuli (De et al. 2022). For polymeric nanocarrier preparations, three methods are used mainly layer-bylayer method, nanoprecipitation method, and emulsion templated method (Karabasz et al. 2020). Any of these methods follow the rule of monomer polymerization such that they show multifunctionality and specific delivery to the target sites.

5 Nanorobotics A nanorobot is a robotic device fabricated at a miniature scale to perform highly precise and specialized tasks (Hamdi and Ferreira et al. 2007). Unlike any other robotic device, these biorobots also have sensors, actuators, communication modules,

26

S. Basu et al.

source of power supply and manipulators wherever required. The most difficult part is the power source as the present energy sources cannot be miniaturized at a nanoscale (Wang et al. 2013). The choice of material and design of the biosensor depends on how the sensor is going to interact with the measurement site. Measurement sites can be broadly classified as invasive measurements, non-invasive measurements, and ex vivo and in vitro measurements depending on where the sensors are positioned. The biological type of nanosensors/microsensors will consist of a bioreceptor and a transducer (Niidome et al. 2006). An antibody, an enzyme, a protein, or a DNA strain can be used as a receptor (Yao et al. 2020). The design of a bionanorobot is completely multi-disciplinary, and therefore different researchers are focusing on various approaches to resolve this issue. The complete nanorobot concept and its applications have been divided into two parts, targeted motion and drug release mechanism.

5.1 Targeted Motion

Fig. 6 Classification of motion control for a nanorobot

methods of nanomotion

The robots generally can navigate through a known/unknown environment using information from sensors, actuators, and controllers and the flow of information in a robotic system is shown in Fig. 1. However, the lack of proper nanosensors and nanocontrollers has led researchers to alternate methods of motion control for nanorobots. The methods can be broadly categorized into three subcategories as shown in Fig. 6.

Self-propelled 1. Active metals 2. Enzymes

External field 1. Magnetic field based 2. Light energy based 3. Acoustic energy based 4. Electrical energy based

Hybrid approach 1. Bacteria flagellum based Nanoswimmmers

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

5.1.1

27

Self-propelled Nanorobot

Initially, the generation of self-propelled nanorobots was based on electrochemically reduced metallic correspondent salts inside the micro/nanosymmetrical pores (Paxton et al. 2006). Ma et al. reported the development of a microrobot which was used in catalytic reactions for propulsion (Ma et al. 2015). The enzymes used for such catalytic reactions were catalase, gox, and urease. This design was fully biocompatible (Lai et al. 2006). A tubular nanorobot was developed by a group of researchers (Wu et al. 2015) which covered the anticancer drug. However, due to the usage of certain chemicals like H2 O2 , the design was not biocompatible. Li et al. proposed a similar nanorobot which was Mg based but aimed at the stomach area in a human body (Li et al. 2018). Therefore, although various metal oxide-related nanoparticles are present, their applications toward in vivo study is limited and require urgent attention by the research community.

5.1.2

External Field-Based Propulsion

Another method of controlling the nanorobots inside human body is to use external power source such as magnetic field, light, and acoustic signals. Advantages for such type of propulsion are shorter response time and remote control (Gao et al. 2013). The magnetic field-based propulsion is currently under focus of many researchers as magnetic resonance-based imaging is well established for medical community (Gillies et al. 1994). Han et al. reported three different types of magnetic field (rotating, gradient based, and oscillating magnetic field) for generating propulsion inside body (Han et al. 2018). Likewise, permanent magnets and electromagnets were used to generate the magnetic field (Jeong et al. 2010; Ryan and Diller et al. 2016). However, used helical structure to generate a translational movement. The microrobots which were controlled by rotating magnetic field showed very high drugloading and drug-releasing capabilities. Moreover, the disintegration of the microrobots can be triggered in a biocompatible environment in the presence of magnetic fields. Using three iron core coils, the magnetic field was setup. In order to control the motion of nanorobots using magnetic field, nickel was plated by vacuum vapor deposition on the body. Therefore, the nanorobots can then be aligned along the direction of the applied magnetic field. The swimming effectiveness can also be monitored with various geometrical structures such as normal, curly, and helical (Ali et al. 2017). These nanorobots are inserted into the bloodstream using popular method of DNA injection or lipofection method. After they transported to the targeted site and came into contact with cell, they are drawn inside the cell using cell’s endocytosis method (Fig. 7). Chang et al. reported usage of an alternating current to deliver drug (Chang et al. 2007). Further, it is reported that ultrasounds (in the range of MHz) can also be used in nanorobotics (Wang et al. 2012). The results show that different types of autonomous movement can be achieved by using ultrasonic waves aqueous solution.

28

S. Basu et al.

Fig. 7 Schematic representation of nanorobot administration and capture of cancer cells in blood stream

5.1.3

A Hybrid Approach for Nano-locomotion

Several enzymes like glucose oxidase, catalase, urease, and lipase have been used to get nano-locomotion, or more precisely controlled motion in a nanoscale environment. Blood being the only carrier of biological, chemical components inside the human body, an obvious choice for the researchers was to utilize the blood flow as transport medium for nanorobots to identify infected cells as well as for targeted drug delivery (Li et al. 2017a, b; Wang et al. 2014). However, in order to control the motion of artificial nanorobots within the blood flow, a complete model of blood flow is required. To increase the easiness of moving inside the blood stream, nanorobots are designed based on bacterial morphology with flagella motion.

5.2 Drug Release Mechanism The drug release mechanism is another field of research work where actuation is possible either through chemicals present in the body or external source based. In a tumor cell, pH is lower compared to normal living cells. Many researchers are using this fact to achieve triggered actuation for drug release in the presence of a specific

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

29

pH. For example, Li et al. used a polymer coating which was degraded at a higher pH level (Li et al. 2017a, b). However, external light-triggered drug release is the most popular one as light has a certain penetration capability at cellular level without causing harm to the body. For instance, Bozuyuk et al. reported release of drug in the presence of light and no release when off (Bozuyuk et al. 2018).

6 Potential and Limitations of Smart Nanosystem Smart nanosystem revolutionized the whole medical sector through its particularity and specificity approaches. Nanoparticles can increase the intracellular absorption of medications in cancer cells while evading toxicity in normal cells by using both targeting strategies (Borm et al. 2002) . Smart nanosystems are employed for a variety of purposes, including the distribution of medications and therapeutics for their curative purposes like natural markers, diagnostic goals akin to imaging operations, and gene therapy (Singh et al. 2019). In smart nanosystems, nanocarriers are showing much potential in drugs delivery likewise improved payload, bioavailability, biodistribution, specificity, reduced toxicity, tailored biological responses, transported across barrier, and modified pharmacodynamics (Zeb et al. 2020). Although there are many benefits to using nanoparticles as drug delivery devices, there are still several limitations to be resolved, such as poor oral bioavailability, unsteadiness in rotation, inadequate tissue allotment, and toxicity (Cho et al. 2008). Nanotechnology has some drawbacks, including the fact that its development costs are significant. Though it is extremely valuable, its production is challenging. Nanoparticle risks and dangers include increased DNA damage rates, genotoxic exposure, harm to human organs and tissues, poor impacts on crop plant growth and productivity, and adverse effects on helpful microbes in the environment (Borm et al. 2006). Moreover, unpredictable health difficulties of NPs were recognized to scientists due to their distinctive physicochemical uses in far-off natural complexes. The major hindrance to the development of a nanorobot is the power source. A robotic device cannot function at all without a power source but the present power sources, e.g., Lithium-Polymer (LiPo) batteries are harmful for human body. Another major challenge is the size of the power source. The nano-scaled power source will be able to provide a very little amount of energy to all the other components in a robotic system. Therefore, medical community and engineering researchers need to focus mainly on the powerless actuation and externally applied energy resources like magnetic field-based motion.

7 Conclusion Targeted drug delivery and treatments are two areas where smart nanosystem and bioinspired nanosystem applications are quite useful (Zagotto and Bortoli et al.

30

S. Basu et al.

2021). Technologies utilizing nanoparticles for human administration must be built to communicate with a living host environment (Bourges et al. 2003). The creation of biocompatible nanoparticles that can be combined with the drug for accurate, targeted drug delivery applications is part of the concept behind bioinspired smart medicine delivery carriers. Acknowledgements Authors would like to thank Birla Institute of Technology Mesra, NIT Silchar and UPES Dehradun for all support. All authors have reviewed and agreed to the chapter content. Dipak Maity would like to thank the University of Petroleum and Energy Studies (UPES) for in-house financial support (SEED Funding: UPES/R&D-HS/24022022/08) and all other support.

References Albrecht C, Knaapen AM, Becker A (2005) The crucial role of particle surface reactivity in respirable quartz induced reactive oxygen/nitrogen species formation and APE/Ref-1 induction in rat lung. Respir Res 6:129 Ali J, Cheang UK, Martindale JD, Jabbarzadeh, M, Fu HC, Jun Kim M (2017) Bacteria-inspired nanorobots with flagellar polymorphic transformations and bundling. Sci Rep 7 Alyautdin RN, Petrov VE, Langer K (1997) Delivery of loperamide across the blood-brain barrier with polysorbate 80-coated polybutylcyanoacrylate nanoparticles. J Pharm Res 14:325–358 Bahadori F, Kocyigit A, Onyuksel H, Dag A, Topcu G (2017) Cytotoxic, apoptotic and genotoxic effects of lipid-based and polymeric nano micelles, an in vitro evaluation. Toxics 6:7 Ballou B, Lagerholm BC, Ernst LA (2004) Non-invasive imaging of quantum dots in mice. Bioconjug Chem 15:79–86 Baran ET, Özer N, Hasirci V (2002) In vivo half life of nanoencapsulated L-asparaginase. J Mat Sci Mater Med 13:1113–1121 Bazile D, Prud’Homme C, Bassoullet M-T (1995) Stealth PEG-PLA nanoparticles avoid uptake by the mononuclear phagocytes system. J Pharm Sci 84:493–498 Bhatia S (2016) Natural polymer drug delivery systems: nanoparticles, plants and algae. https://doi. org/10.1007/978-3-319-41129-3 Borm PJ (2002) (2007) Particle toxicology: from coal mining to nanotechnology. Inhal Toxicol 14:311–324 Borm PJ, Kreyling W (2004) Toxicological hazards of inhaled nanoparticles—potential implications for drug delivery. J Nanosc Nanotechnol 4:521–531 Borm PJ, Muller-Schulte D (2006) Nanoparticles in drug delivery and environmental exposure: same size, same risks? Nanomedicine 1:235–249 Borm PJA, Robbins D, Haubold S (2006) The potential risks of nanomaterials: a review carried out for ECETOC. Part Fiber Toxicol 3:11 Bourges JL, Gautier SE, Delie F (2003) Ocular drug delivery targeting the retina and retinal pigment epithelium using polylactide nanoparticles. Invest Ophthalmol vis Sci 44:3562–3569 Bozuyuk U, Yasa O, Yasa IC, Ceylan H, Kizilel S, Sitti M (2018) Light-triggered drug release from 3D-printed magnetic chitosan microswimmers. ACS Nano 12:9617–9625 Chang ST, Paunov VN, Petsev DN, Velev OD (2007) Remotely powered self-propelling particles and micropumps based on miniature diodes. Nat Mater 6:235–240 Cho K, Wang X, Nie S, Chen Z, Shin DM (2008) Therapeutic nanoparticles for drug delivery in cancer. Clin Cancer Res 14:1310–1316 Daglar B, Ozgur E, Corman ME, Uzun L, Demirel GB (2014) Polymeric nanocarriers for expected nanomedicine: current challenges and future prospects. RSC Adv 89

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

31

Danhier F, Ansorena E, Silva JM, Coco R, Le Breton A, Préat V (2012) PLGA-based nanoparticles: an overview of biomedical applications. J Control Release 161(2):505–522 De R, Mahata MK, Kim KT (2022) Structure-based varieties of polymeric nanocarriers and influences of their physicochemical properties on drug delivery profiles. Adv Sci. https://doi.org/10. 1002/advs.202105373 Deng Y, Zhang X, Shen H, He Q, Wu Z, Liao W, Yuan M (2020) Application of the nano-drug delivery system in treatment of cardiovascular diseases. Front Bioeng Biotechnol 7:489 Emerich DF, Thanos CG (2006) The pinpoint promise of nanoparticle-based drug delivery and molecular diagnosis. Biomol Eng 23:171–184 Esfahani DR, Tangen KM, Sadeh M, Seksenyan A, Neisewander BL, Mehta AI, Linninger AA (2018) Systems engineers’ role in biomedical research convection-enhanced drug delivery. Comp Aid Chem Eng 42:271–302 Fukunishi Y (2009) Structure-based drug screening and ligand-based drug screening with machine learning. Combin Chem Amp High Throughput Screen 12:397–408 Gao W, D’Agostino M, Garcia-Gradilla V, Orozco J, Wang J (2013) Multi-fuel driven janus micromotors. Small 9(3):467–471 Garti N, Pinthus E, Aserin A, Spernath A (2016) Improved solubilization and bioavailability of nutraceuticals in nanosized self-assembled liquid vehicles. Encapsul Control Release Technol Food syst 173–203 Gillies GT, Ritter RC, Broaddus WC, Grady MS, Howard MA III, McNeil RG (1994) Magnetic manipulation instrumentation for medical physics research. Rev Sci Instrum 65:533–562 Girija R (2018) Medical applications of polymer/functionalized nanoparticle systems. Elsevier Inc., Amsterdam Grabowska-Jadach I, Drozd M, Kulpi´nska D, Komendacka K, Pietrzak M (2020) Modification of fluorescent nanocrystals with 6-thioguanine: monitoring of drug delivery. Appl Nanosci 10:83– 93 Gregoriadis G (1976) The carrier potential of liposomes in biology and medicine: (First of Two Parts). New England. J Med 295(13):704–710 Hainfeld J, Lin L, Slatkin D, Avraham Dilmanian F, Vadas T, Smilowitz H (2014) Gold nanoparticle hyperthermia reduces radiotherapy dose. Nanomedicine: nanotechnology. Biology and Medicine 10:1609–1617 Hamdi M, Ferreira A (2007) Multiscale design and modeling of nanorobots. In: IEEE/RSJ international conference on intelligent robots and systems. pp 3821–3827 Han K, Shields CW IV, Velev OD (2018) Engineering of self-propelling microbots and microdevices powered by magnetic and electric fields. Adv Func Mater 28:1705953 Huang Y, Mei C, Tian Y, Nie T, Liu Z, Chen T (2018) Bioinspired tumor-homing nanosystem for precise cancer therapy via reprogramming of tumor-associated macrophages. NPG Asia Mater 10:1002–1015 Husen A, Iqbal M, Aref IM (2017) Plant growth and foliar characteristics of faba bean (Viciafaba L.) as affected by indole-acetic acid under water-sufficient and water-deficient conditions. J Environ Biol 38:179–186 Jaiswal M, Dudhe R, Sharma PK (2015) Nanoemulsion: an advanced mode of drug delivery system. 3 Biotech 5:123–127 Jeong S, Choi H, Choi J, Yu C, Park JO, Park S (2010) Novel electromagnetic actuation (EMA) method for 3-dimensional locomotion of intravascular microrobot. Sens Actuat A 157:118–125 Johnston MJ, Edwards K, Karlsson G, Cullis PR (2008) Influence of drug-to-lipid ratio on drug release properties and liposome integrity in liposomal doxorubicin formulations. J Liposome Res 18:145–157 Karabasz A, Bzowska M, Szczepanowicz K (2020) Biomedical applications of multifunctional polymeric nanocarriers: a review of current literature. Int J Nanomed 15:8673–8696 Kiang CH, Goddard WA III, Beyers R, Bethune DS (1995) Carbon nanotubes with single-layer walls. Carbon 33:903–914

32

S. Basu et al.

Kreuter J, Ramge P, Petrov V (2003) Direct evidence that polysorbate-80-coated poly (butylcyanoacrylate) nanoparticles deliver drugs of the CNS via specific mechanisms requiring prior binding of drug to the nanoparticles. Pharm Res 20:409–416 Kreyling WG, Semmler M, Erbe F (2002) Translocation of ultrafine insoluble iridium particles from lung epithelium to extrapulmonary organs is size dependent but very low. J Toxicol Environ Health Part A 65:1513–1530 Kreyling WG, Semmler M, Möller W (2004) Dosimetry and toxicology of ultrafine particles. J Aerosol Med 17:140–152 Kumar A, Nautiyal U, Kaur C, Goel V, Piarchand N (2017) Targeted drug delivery system: current and novel approach. Int J Pharm Med Res 5:448–454 Lai M-K, Chang C-Y, Lien Y-W (2006) Application of gold nanoparticles to microencapsulation of thioridazine. J Control Release 111:352–361 Li J, Angsantikul P, Liu W, Esteban-Fernández de Ávila B, Thamphiwatana S, Xu M, Wang J (2017a) Micromotors spontaneously neutralize gastric acid for pH-responsive payload release. Angew Chem Int Ed 56:2156–2161 Li J, de Avila BEF, Gao W, Zhang L, Wang J (2017b) Micro/nanorobots for biomedicine: delivery, surgery, sensing, and detoxification Sci. Robot 2:1–9 Li T, Zhang A, Shao G, Wei M, Guo B, Zhang G, Wang W (2018) Janus microdimer surface walkers propelled by oscillating magnetic fields. Adv Func Mater 28:1706066 Lin W, Huang YW, Zhou XD (2006) In vitro toxicity of silica nanoparticles in human lung cancer cells. Toxicol Appl Pharmacol 217:252–259 Lôbo GC, Paiva KL, Silva ALG, Simões MM, Radicchi MA, Báo SN (2021) Nanocarriers used in drug delivery to enhance immune system in cancer therapy. Pharmaceutics 13:1167 Lockman PR, Koziara JM, Roder KE (2003) In vivo and in vitro assessment of baseline bloodbrain-barrier parameters in the presence of novel nanoparticles. Pharm Res 20:705–713 Lockman PR, Koziara JM, Mumper RJ (2004) Nanoparticle surface charges alter blood-brain barrier integrity and permeability. J Drug Target 12:635–641 Loo C, Lin A, Hirsch L (2004) Nanoshell-enabled photonics-based imaging and therapy of cancer. Technol Cancer Res Treat 3:33–40 Lovric J, Bazzi HS, Cuie Y (2005a) Differences in subcellular distribution and toxicity of green and red emitting CdTe quantum dots. J Mol Med 83:377–385 Lovric J, Cho SJ, Winnik FM (2005b) Unmodified cadmium telluride quantum dots induce reactive oxygen species formation leading to multiple organelle damage and cell death. Chem Biol 12:1159–1161 Lowery AR, Gobin AM, Es D (2006) Immunonanoshells for targeted photothermal ablation of tumor cells. Int J Nanomed 1:149–154 Ma X, Jannasch A, Albrecht UR, Hahn K, Miguel-López A, Schaffer E, Sánchez S (2015) Enzymepowered hollow mesoporous Janus nanomotors. Nano Lett 15:7043–7050 Madamsetty VS, Mukherjee A, Mukherjee S (2019) Recent trends of the bio-Inspired nanoparticles in cancer theranostics. Front Pharmacol 10:1264 Metselaar JM, Storm G (2005) Liposomes in the treatment of inflammatory disorders. Expert Opin Drug Deliv 2:465–476 Michaelis K, Hoffmann MM, Dreis S (2006) Covalent linkage of apolipoprotein E to albumin naniparticles strongly enhances drug transport into the brain. J Pharmacol Exp Ther 317:1246– 1253 Minko T, Pakunlu RI, Wang Y (2006) New generation of liposomal drugs for cancer. Anticancer Agents Med Chem 6:537–552 Mishra N, Pant P, Porwal A, Jaiswal J, Aquib M (2016) Targeted drug delivery: a review. Am J Pharm Tech Res 6(1) Mouzouvi CR, Umerska A, Bigot AK, Saulnier P (2017) Surface active properties of lipid nanocapsules. PLoS ONE 12:e0179211 Nazila K, Yameen B, Wu J, Farokhad OC (2016) Degradable controlled-release polymers and polymeric nanoparticles: mechanisms of controlling drug release. Chem Rev 116:2602

Bioinspired Nanosystems Interacting with the Host Environment: Smart …

33

Nemmar A, Hoet PH, Vanquickenborne B (2002) Passage of inhaled particles into the blood circulation in humans. Circulation 105:411–414 Nemmar A, Hoylaerts MF, Hoet PH (2003) Size effect of intraytracheally instilled particles on pulmonary inflammation and vascular thrombosis. Toxicol Appl Pharmaco 186:38–45 Niidome T, Yamagata M, Okamoto Y (2006) PEG-modified gold nanorods with a stealth character for in vivo applications. J Control Release 114:343–347 Nobs L, Buchegger F, Gurny R (2004) Poly(lactic acid) nanoparticles labeled with biologically active Neutravidin™ for active targeting. Eur J Pharm Biopharm 58:483–490 Paxton WF, Baker PT, Kline TR, Wang Y, Mallouk TE, Sen A (2006) Catalytically induced electrokinetics for motors and micropumps. J Am Chem Soc 128:14881–14888 Rajesh J, Pavithra G, Manjunath T (2018) Design & development of nanobots for cancer cure applications in bio-medical engineering. Int J Eng Res Technol (IJERT) Ncesc 6:1–7 Rani K, Paliwal S, Kirti R (2014) A review on targeted drug delivery: its entire focus on advanced therapeutics and diagnostics. Sch J App Med Sci 2:328–331 Rizvi SA, Saleh AM (2018) Applications of nanoparticle systems in drug delivery technology. Saudi Pharmaceut J 26:64–70 Roco MC, Mirkin CA, Hersam MC (2011) Nanotechnology research directions for societal needs in 2020: summary of international study. J Nanopart Res 13:897–919 Ryan P, Diller E (2016) Five-degree-of-freedom magnetic control of micro-robots using rotating permanent magnets. In: IEEE international conference on robotics and automation (ICRA). pp 1731–1736 Singh S, Sirbaiya A, Mishra A (2019a) Bioinspired smart nanosystems in advanced therapeutic applications. Pharmaceutical Nanotechnology 7:246–256 Sivadasan D, Sultan MH, Madkhali I, Almoshari Y, Thangavel N (2021) Polymeric lipid hybrid nanoparticles (PLNs) as emerging drug delivery platform—a comprehensive review of their properties, preparation methods, and therapeutic applications. Pharmaceutics 13(80):1291 Spencer A, Torrado M, Custódio B, Silva-Reis S, Santos S, Leiro V, Pêgo A (2020) Breaking barriers: bioinspired strategies for targeted neuronal delivery to the central nervous system. Pharmaceutics 12:192 Strebhardt K, Ullrich A (2008) Paul Ehrlich’s magic bullet concept: 100 years of progress. Nat Rev Cancer 8:473–480 Tenchov R, Bird R, Curtze AE, Zhou Q (2021) Lipid nanoparticles from liposomes to mrna vaccine delivery, a landscape of research diversity and advancement. ACS Nano 15:16982–17015 Tewabe A, Abate A, Tamrie M, Seyfu A, Siraj EA (2021) Targeted drug delivery—from magic bullet to nanomedicine: principles, challenges, and future perspectives. J Multidiscip Healthc 14:1711 Thakur A, Roy A, Chatterjee S, Chakraborty P, Bhattacharya K, Mahata PP (2015) Recent trends in targeted drug delivery. Smgroup. https://doi.org/10.13140/RG.2.1.2443.9762 Valcourt C, Saulnier P, Umerska A, Zanelli MP, Montagu A, Rossines E, Joly-Guillou ML (2016) Synergistic interactions between doxycycline and terpenic components of essential oils encapsulated within lipid nanocapsules against gram negative bacteria. Int J Pharm 498:23–31 Vasir JK, Reddy MK, Labhasetwar VD (2005) Nanosystems in drug targeting: opportunities and challenges. Curr Nanosci 1:47–64 Vimala K, Kannan S (2021) Phyto-drug conjugated nanomaterials enhance apoptotic activity in cancer. Adv Protein Chem Struct Biol 125:275–305 Voliani V, Signore G, Nifosi R, Ricci F, Luin S, Beltram F (2012) Smart delivery and controlled drug release with gold nanoparticles: new frontiers in nanomedicine. Recent Patents Nanomed 2:34–44 Wang W, Li S, Mair L, Ahmed S, Huang TJ, Mallouk TE (2014) Acoustic propulsion of nanorod motors inside living cells. Angew Chem Int Ed 53:3201–3204 Wang DY, Van der Mei HC, Ren Y, Busscher HJ, Shi L (2020) Lipid-based antimicrobial deliverysystems for the treatment of bacterial infections. Front Chem 7:872

34

S. Basu et al.

Wang J (2013) Nanomachines: fundamentals and applications. John Wiley & Sons. ISBN: 978–3– 527–65147–4 Warnock JN, Daigre C, Al-Rubeai M (2011) Introduction to viral vectors. Methods Mol Biol 737:1–25 Wilczewska AZ (2012) Nanoparticles as drug delivery systems. Pharmacol Rep 64:1020–1037 Wu Z, Li J, Esteban-Fernández de Ávila B, Li T, Gao W, He Q, Zhang L, Wang J (2015) Waterpowered cell-mimicking janus micromotor. Adv Funct Mater 25:7497–7501 Yang W, Ratinac KR, Ringer SP, Thordarson P, Gooding JJ, Braet F (2010) Carbon nanomaterials in biosensors: should you use nanotubes or graphene? Angew Chem Int Ed 49:2114–2138 Yao S, Ren P, Song R, Liu Y, Huang Q, Dong J, Zhu Y (2020) Nanomaterial-enabled flexible and stretchable sensing systems: processing, integration, and applications. Adv Mater 32:1902343 Yurgel V, Collares T, Seixas F (2013) Developments in the use of nanocapsules in oncology. Braz J Med Biol Res 46:486–501 Zagotto G, Bortoli M (2021) Drug Design: Where We Are and Future Prospects. Molecules 26:7061 Zeb A, Rana I, Choi HI, Lee CH, Baek SW, Lim CW, Khan N, Arif ST, Sahar NU, Alvi AM, Shah FA, Din FU, Bae ON, Park JS, Kim JK (2020) Potential and applications of nanocarriers for efficient delivery of biopharmaceuticals. In Pharmaceutics 12:1–37 Zhang H (2016) Surface characterization techniques for polyurethane biomaterials. Adv Polyureth Biomater Sci Direct 23–73

Nanotechnology-Based Approaches to Relieve Tumour Microenvironment Hypoxia via Enhanced Oxygen Delivery Manisha Singh, Rashi Rajput, Vinayak Agarwal, Divya Jindal, Pranav Pancham, and Sudha Srivastava

Abstract Contemporary times have observed significant expression of the hypoxiamodulated tumorigenic microenvironment (TME) in facilitating the growth of metastatic cancer. Besides stimulating tumour metastasis, this TME has also been associated with elevated resistance towards oxidative stress (ROS)-generated anticancer therapies, thereby culminating the futile therapeutic expressions. Countering these distinct cancerous physiologies several nanotechnology-based applications and approaches have garnered support that exhibits the capabilities of sustainably delivering O2 molecules to the hypoxic site. Such approaches show promising results as they effectively ameliorate the expression of ROS-centred cancer therapeutics. In this chapter, we present an insight into the current nanostructured material strategies for attenuating hypoxia in the TME while also enhancing the oxygen delivery platform at the same time throughout cancerous physiology. These nanotechnologybased approaches focus on enhancing O2 -supply proficiency mechanisms along with exploring conceptual design considerations and their associated challenges and opportunities. This chapter would indeed provide some vital insight for the fabrication of other nanomaterials technology with O2 -supply capabilities for countering the tumour hypoxia-induced resistance of ROS-mediated cancer treatment, and thus promoting ROS-generated cancer therapeutics.

M. Singh (B) Flinders Health and Medical Research Institute, College of Medicine & Public Health, Flinders University, Adelaide, Australia e-mail: [email protected] M. Singh · V. Agarwal · P. Pancham · S. Srivastava Department of Biotechnology, Jaypee Institute of Information Technology, NOIDA, Uttar Pradesh, India R. Rajput Department of Clinical Medicine, Faculty of Medicine, Health and Human Sciences, Macquarie University, Sydney, Australia D. Jindal Department of Physics, IIT Bombay Monash Research Academy, IIT Bombay, Mumbai, Maharashtra, India © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_3

35

36

M. Singh et al.

Keywords Tumour hypoxia · Nanotechnology · Oxygen supply · Tumour oxygenation · Tumour microenvironment

1 Introduction The migration of cancerous cells from the initially infected area to the different organs of the human body is a basic phenomenon of metastatic cancer. The impeded or obstructed prognosis of the metastasis in cancer patients can cause severe irreversible damage to the infected organs further leading to life loss (Irani 2019). Surveyed estimation from the World Health Organization (WHO) reported in 2020 that more than 10 million deaths worldwide were caused by the progressive cancers diversified in various infected organs such as lung cancer—1.8 million, breast cancer—approximately 6.9 lakhs, colon-rectum cancer—9.2 lakhs and liver cancer—8.3 lakhs with other increasing cases of cervical cancer in women that have in-definitive statistics. These numbers are expected to be elevated with the early developing cancers in children around the world with a rate of 400,000 reported cases every year (WHO 2022). The recent yearly survey of American Cancer Society (ACS) stated the possibility of 1.9 million new reported cases in 2022 with approximately 609,360 deaths alone in United States (Siegel et al. 2022). These statistics prompted cancer researchers to conduct an in-depth investigation into the related mechanisms and develop a therapeutic approach for treating it. With the dissection of cellular mechanism-associated metastatic cancerous progression, one of the major factors is angiogenesis in the tumorous growth (Wang et al. 2010). Angiogenesis is the process of new blood vessel formation to help the cell growth and transporting essential signals related to cell proliferation and differentiation. In cancerous condition, instead of endothelial cells forming the blood vessels, antagonistic melanoma cells form the blood vessels like vascular channel called vasculogenic mimicry for tumour cells to provide nutrition and functional signals for cancerous cell progression and metastasis (Maniotis et al. 1999). The stabilization of tumorous blood vascular channels needs support from the extra cellular components such as fibroblasts, neuroendocrine cells, immuneinflammatory cells and adipose cells alongside extracellular matrix and myofibroblast that therefore develop a tumorigenic microenvironment (TME) (Wang et al. 2017a, b). This TME modulates the nearby non-tumorous cells such as pararhyme and mesenchyme cells by altering their physiological and morphological properties to associate with the progressive tumorigenesis (Martin et al. 2016; Naxerova et al. 2014; Naxerova and Jain 2015). The prominent trigger that contributes in TME of tumorigenesis is hypoxia. Decline in the oxygen level in tissues below 10 mmHg causes hypoxia that further increased stress condition with unmatched oxygen supply to the cells and extension of tumorous cell proliferation (Li et al. 2021). The hypoxiainduced factors associated in this progression are the subunits of HIF-dependent signalling cascade. These transcription factors of HIF-subclass include HIF-1, HIF2 and HIF-3 that are associated with the oxygen-sensitive subunits such as HIF-1α, HIF-2α and HIF-3α, respectively, further dimerised to form HIF-1β subunit. The

Nanotechnology-Based Approaches to Relieve Tumour …

37

most studies of HIF-subunit for hypoxia are HIF-1α and HIF-2α with their active proline residues at P402 and P564 for HIF-1α, and P405 and P531 for HIF-2α. These proline residues are highly susceptible to hydroxylation with present oxygen by the protein called hydroxylase domain-containing protein (PHDs) and stimulate their binding to von Hippel–Lindau tumour suppressor (pVHL) causing the HIF-α degradation. Factor inhibiting HIF-1 (FHI1) is another HIF-α regulatory protein depending on oxygen level. HIF-1α gets hydroxylated on the asparagine residue derived by FHI1 can obstruct its interaction with HIF cofactors such as p300 histone acetylase and CBP, thus to suppress the transcription process of HIF-1 (Petrova et al. 2018). Another factor that is highly associated with the tumorigenesis is reactive oxygen species (ROS) production in the tumour cells inside mitochondria and peroxisome. Moreover, the ROS is partially produced by the transmembrane protein family called nicotinamide adenine dinucleotide phosphate oxidases (NOX) that carry the electrons through the biological membrane for oxygen to superoxide catalyzation that further reduce to superoxide dismutase (SOD) and produce hydrogen peroxide (H2 O2 ) which caused the reduced oxygen tension (Weinberg et al. 2019). The regulation of HIF-1α and ROS production can be the focus for TME and tumorigenesis suppression. Therefore, this review explores the efficient nanotechnology-based therapeutic approached towards these targeted factors in tumorigenesis (Fig. 1).

Fig. 1 Schematic representation showing the HIF activation mechanism in cancer pathology. Abbreviations: prolyl-4-hydroxylase domain (PHD); hypoxia response elements (HREs); factor inhibiting HIF-1 (FIH-1); von Hippel–Lindau (VHL) tumour suppressor

38

M. Singh et al.

2 Nanotechnology-Based Approaches for O2 Delivery in Hypoxic Site Recent advancement in the field of nanobiotechnology has garnered a lot of attention from the scientific community; however, its incorporation in establishing a potential oxygen delivery system to the hypoxic regions in a tumorigenic microenvironment is a major step in facilitating cancer treatment. Conventional therapies associated with cancer treatment face several issues in modulating cancer spread via tumorigenic microenvironment. Issues concerning diffusion of oxygen molecules across the primary avascular tumour have been observed as the primary factor contributing to the development of hypoxic conditions. Moreover, several other deviant elements such as distorted tumour microvasculature along with aberrant blood flow restrain the natural immune system to rectify such oxygen-deprived state. Thereby, it becomes even more necessary to explore a potent and effective approach which could cater to such oxygen-deficit states in the tumorigenic microenvironment (Wu et al. 2022).

2.1 Nanochemical Decomposition for O2 Release Methodology for nanochemical decomposition involves the application of various highly reactive compounds. This chemical decomposition tends to be utilized in H2 O2 decomposition reaction for the synthesis of oxygen molecules; moreover, it aids in balancing the oxygen concentration in the hypoxic microenvironment of the tumours. Furthermore, these highly reactive chemical compounds majorly belong to the family of metallic oxides and some of the chief examples of them are MnO2 (Prasad et al. 2014), CeO2 (Wang et al. 2019), MnO (Kim et al. 2019), CaO2 (Liu et al. 2017a, b), perfluorocarbons (PFCs), etc. Perfluorocarbons (PFCs) have been reported to be an effective method for facilitating O2 release which is one of the several applications of nanochemical decomposition. Owing to their incredibly high oxygen solubility, biochemical and biological no reactivity and reliable bio-safety, a category of fluorocarbon-based organic compounds has indeed been widely recognized as effective oxygen carriers for something like an artificial blood substitute. PFCs are racemic mixture of fluorine and carbon atoms that have been thoroughly investigated using MRI and ultrasonography as blood profile studies, with most of these have been cleared by the authorities for application in clinical settings (Zhao et al. 2020). According to reports, PFC nanodroplet-loaded photosensitizers are appropriate for oxygen-boosted PDT (Oxy-PDT). Red blood cells include a protein called haemoglobin, which appears to be an iron-rich protein that carries oxygen to tissues. According to research, haemoglobin has the capacity to reversibly bind to up to four molecules of oxygen which lead to the production of HbO2 . Carriers which are based on haemoglobin have been employed in a number of anticancer therapies to deliver oxygen into hypoxic tumour tissues. PFCs have an astonishingly high solubility compared to Hb, which

Nanotechnology-Based Approaches to Relieve Tumour …

39

has a low oxygen loading efficiency; they typically contain 40–50% oxygen content in 100 ml of PFCs molecule which is roughly equivalent to 200 mL of blood at room temperature at 1 atm. Because of their own larger surface area and uniform apertures, MOF carriers seem to be excellent options for oxygen storage and separation. A research study conducted by Lin et al. demonstrates neoteric multifunctional nanodrug carriers depending upon a metal–organic framework (MOF). For cancer therapy, MOF moieties were conjugated with O2 molecules in association with UCNPs (upconverting nanoparticles) which leads to unveiling novel avenues in enhancing the treatment efficacy of tumours (Cheng et al. 2015). MnO2 is among the most commonly used metal oxides for in situ oxygen generation in hypoxia tumour therapy. The high interest in MnO2 stems from the fact that O2 can be produced for an extended period of time through its reaction with tumour metabolites as building ingredients. Simultaneously, MnO2 is decomposed into non-harmful, hydro-soluble Mn2 + ions. MnO2 metal oxides accumulate in the body at a much lower rate than other metal-based nanoparticle systems. This specific mathematical expression for this phenomenon is expressed as follows (He et al. 2018). High-reactivity MnO2 nanoparticles react with hydrogen peroxide to produce O2 . A few versatile nanoparticles, such as UCNPs, will be anchored to MnO2 nanosheets, a 2D nanomaterial, to realize synergetic therapy. As a gatekeeper, MnO2 nanoshells prevent early drug seepage in healthy cells and subsequently facilitate drug release as they break down. Manganese dioxide-serum albumin composites have been reported to be biocompatible via biomineralization as the nanocomposite encompasses excellent biocompatibility towards bio-macromolecular proteins (such as human serum albumin and bovine serum albumin). The oxygen production process typically uses MnO2 nanoparticles because of their prominent chemical reactivity and selectivity. A study conducted by Wu et al. reported that the preparation of MnO2 nanoparticles using a simple step of redox reaction of KMnO4 and cationic polyelectrolyte (PAH) contributes to improve biocompatibility. Polyelectrolyte-coated MnO2 nanoparticles have been further coated with BSA. Modified MnO2 nanoparticles with a desired size of 50 nm having a charge of −25 mV have reported to exhibited colloidal stability as well as good biocompatibility (Prasad et al. 2014). Besides that Zhao et al. (2020) explored an alternative approach which demonstrates the application of electrodeposition of amorphous nickel–iron composite (mesoporous) by oxygen electrode. This technique involves the deposition of the nanosheet over the nickel foam substrate, due to the expression of 1000 mA cm−2 current density from electrode to the water. The oxidation takes place around an overpotential of 270 mV in an alkaline medium (Yan et al. 2020).

2.2 Breakdown of Catalase to Generate O2 Catalase (CAT) is a vital enzyme in living cells that protect from oxidative damage by catalysing H2 O2 to produce O2 and water. Catalase, a naturally occurring biozyme

40

M. Singh et al.

found in all biotic systems, is proficiently able to degrade H2 O2 to produce oxygen. Although catalase offers a significant approach for increasing oxygen concentration in tumour tissues, its in vivo instability is an issue due to the presence of diverse physiological proteases, ambiguous protein immunogenicity and so its short halflife. Tumour cells have elevated levels of H2 O2 than healthy cells, which can lead to DNA damage, metastasis, atypical proliferation and angiogenesis. However, due to its brief in vivo half-life, catalase’s instability restricts its use in oxygen production (Zhang et al. 2019). One of the most important signalling molecules, such as cellular growth, proliferation of the cells and also tumour metastasis, is the abnormally high level of H2 O2 , a characteristic of cancer cells (López-Lázaro 2007). An endogenous enzyme called catalase protects cells from oxidative damage by catalysing particular processes. The breakdown of H2 O2 to release oxygen and water. A CAT-based nanoplatform has a strong candidate for catalysing intratumorally H2 O2 decomposition, which will relieve tumour hypoxia (Ji et al. 2022). By simultaneously encapsulating the photosensitizers CAT and methylene blue to form a poly(lactic-co-glycolic acid) (PLGA) core and doping black hole quencher 3 (BHQ-3) into the PLGA shell, Chen et al. (2015) reported H2 O2 -activatable and oxygen-evolving core–shell nanoparticles (HAOPNP) (Cheng et al. 2015). The extra intracellular H2O2 and oxygen production catalysed by CAT permeated the core ever since HAOPNP was internalized into tumour cells, resulting in the rupture of the PLGA nanoshell along with release of the photosensitizer methylene blue. BHQ-3 significantly decreased non-selective harm to healthy cells in this system. This work proposed a unique PDT paradigm for stable hypoxic tumours which are associated with a steady oxygen generation. In recent years, Liu’s team and its collaborators have developed variety of catalase (CAT)-based NPs targeting hypoxic tumour PDT in association with (a) paclitaxel (PTX)-fuelled co-assembly of albumin and CAT; (b) smart multifunctional HSA-Ce6-CAT-PTX nanoparticles’ good intra-tumoral penetration, relief in hypoxia along with combined PDT/chemo (Chen et al. 2017a, b); (c) an in situ free-radical polymerization approach employing photosensitizer meso-tetra (p-hydroxyphenyl) porphin as a cross-linker to develop multifunctional nanotheranostics with elevated enzymatic stability and PDT efficacy (Wang et al. 2018); and (d) self-assembled NPs with a bladder intravesical installation-based PDT (CAT-Ce6/F-PEI), developed by combining flu (CAT-Ce6) (Li et al. 2019a, b, c). A reparative CAT-encapsulated nanosystem (HA-CAT@aCe6) used for improved PDT in a BALB/c nude mouse model carrying MDA-MB-231 tumour cells was described by Zhao et al. in 2020. HA appears to have been covalently coupled to catalase using cyclodextrin to develop HA-CAT (CD). HA and catalyse combination have observed to enhance protease’s physiological durability and encouraged active tumour targeting. The coated HA-CAT has about 15% free catalase enzymatic activity, according to a red catalase kit assay. As catalase carriers, MOFs have been reported to help treat tumour hypoxia better. A versatile double-layered MOF hybrid (BQ-MIL@cat-MIL) had been designed to treat tumour hypoxia. By adopting an in situ growing method, catalase was encapsulated in MIL to produce the outer

Nanotechnology-Based Approaches to Relieve Tumour …

41

layer of the MOF hybrid. The homologous MOF hybrid sustained catalase catalytic expression in hypoxic TME, allowing it to be self-reliant O2 (Zhao et al. 2020). Liu et al. (2021a, b) discovered a catalase-like DNAzyme in nanoscale polymeric matrix based on G-quadruplex/hemin. The polymers were able to generate oxygen in the tumours via the decomposition of H2 O2 , which alleviated tumour hypoxia and augmented PDT to resolve hypoxia-associated resistance (Liu et al. 2021a, b).

2.3 Water-Based Splitting to Produce O2 Scientists have developed water-splitting nanostructures that mimic the photosynthesis process, in which chloroplasts is responsible for generating large oxygen with high efficacy by transferring absorbed light energy to the water in cases of green plants. This method differs from previous methods for breaking down intracellular H2 O2 into oxygen. In fact, the best examples of such materials were modified carbon nitride (C3 N4 ) nanometric and calcium peroxide (CaO2 ) nanoparticles (Cardoso et al. 2021). Zhang and colleagues developed a variety of nanomaterials that use photocatalytic water splitting to trigger oxygen generation for improved PDT (Zheng et al. 2016). Recently, Zhang’s team developed the Fe-C3 N4 @Ru@HOP (FCRH) two-photon stimulated nanocomposite to reduce tumour hypoxia along with improved PDT by producing oxygen in situ. In this study, the only modification to the surface of Ru (II) complex-loaded iron-doped C3 N4 (Fe-C3 N4 ) nanoparticles was a hyperbranched covalent copolymer with poly (ethylene glycol) arms (HOP). When subjected to two-photon light, the HOP nanosystem reports a superior two-photon expression cross section which further leads to work as a two-photon light-harvesting device and a donor of Förster resonance energy transfer (FRET) in oxygen generation. When exposed to 808-nm two-photon irradiation, Ru (II) was stimulated to produce and Fe-C3 N4 was activated to break H2 O for oxygen synthesis. The production of oxygen by Fe-C3 N4 was greatly sped up by the addition of photo-induced electrons from energized Ru (II) to Fe-C3 N4 . It is important to note that two-photon laser inside of this system may have raised penetration depth as well high precision, both of which significantly tend to positively influence the PDT efficiency. This oxygen self-supplementation method thus simultaneously overcame the limitations of tissue penetration and hypoxia in PDT and revealed tremendous potential for spatiotemporally controlled tumour treatment in vivo (Li et al., 2019a, b, c). Recently, Yu et al. demonstrated how oxygen was produced during PDT using tiny calcium peroxide (CaO2 ) nanoparticles by reacting with carbon dioxide. This method avoids the covert issue of the cancer site’s insufficient intrinsically endogenous H2 O2 content to make enriched oxygen. The water-splitting method has distinct advantages over other systems for producing oxygen due to the presence of H2 O within the biological environment, which enables in situ oxygen synthesis. It therefore has a huge potential for clinical applications (Yu et al., 2019) (Fig. 2).

42

M. Singh et al.

Fig. 2 Schematic illustration of the strategies based on nanotechnology for overcoming tumour hypoxia microenvironment: (i) PFC-based oxygen carriers; (ii) Nano-chemical decomposition; (iii) metal–organic frameworks (MOFs); (iv) breakdown of catalase to generate O2

3 Various Strategically Used Nanomaterials for TME Hypoxia 3.1 PFC-Based Oxygen Carriers (PFOCs) and Fe2+ Porphyrin Systems These are hydrocarbon-based carriers, formulated by ideally substituting hydrogen with fluorine or halogen atoms, which provide the ability to attract additional electrons from other atoms and form a strong fluorinated carbon backbone with extreme polar nature. Further, this substitution or addition leads to physical changes in the organic compound. Moreover, due to its bulky shape and extreme polar nature, the

Nanotechnology-Based Approaches to Relieve Tumour …

43

solubility in water is decreased when compared to other hydrocarbon-based derivatives. Further, adding hydrophobic and lipophobic capabilities to the carrier-based formulation and microenvironment target site (Jägers et al. 2021). Tumour microenvironment sites are composed of many tumour cells along with other immunoregulatory cells leading to tumour progression. Meanwhile, ferrocene–porphyrin conjugate, an iron-containing porphyrin, is another potential carrier of active substances which are transported to TME sites and activated by photosensitizers where improved ROS along with antitumor activity was observed in low-dose form (Fiorito et al. 2019; Zhang et al. 2020).

3.2 Artificial Red Blood Cells Substitute (RBCSs) Alleviation of tumour microenvironment by improving oxygen intake capacity has proven beneficial. Therefore, targeting of such sites using semi-synthetic Hb containing RBCSs-based carriers provides an additional capacity to reduce hypoxia conditions. These carriers provide the capability to deliver along with oxygen release regulation into the narrow vascular through chemical modification or liposome encapsulation via biodegradable materials (Banerjee et al. 2022; Wu et al. 2022).

3.3 Metal–Organic Framework (MOFs) These carriers have been found to provide excellent ability to control pore geometry, biodegradability, high loading capacity, controlled release and size of MOFs and its ability to carry and store oxygen (DeCoste et al. 2014). However, the ability to avoid immune-mediated clearance is a constant challenge faced with MOFs. A revolutionary method suggests using extrusion and ultrasonic techniques to coat MOFs with brand-new cell membrane biomimetic MOFs (CMMs). Red blood cell, immune cell, cancer cell, platelet and fusion cell membranes are only a few examples of coating membranes that can vary since they can be endowed with good features like lengthy blood circulation, immunological escape and targeting capacity. In order to target tumour cells, Gao et al. developed UiO-66 MOFs based on RBC membrane coat and developed a biomimetic O2 -evolving nanocarrier by co-extrusion treatment that exhibits anticancer capabilities (Liu et al. 2021a, b).

3.4 Oxygen Microcapsules One of the more feasible options is to deliver oxygen directly to the TME site which is possible due to low solubility properties. These oxygen-based microcapsules are polydopamine-nanoparticle-stabilized carrier systems, which have already exhibited

44

M. Singh et al.

anti-tumour and enhanced tumour therapy (Wu et al. 2023). These carrier systems exhibit excellent biocompatibility along with good dispersity in water with quick increase in oxygen concentration near the target sites and further sustaining the oxygen concentration for a long period of time (Dai et al. 2021).

4 Nanotechnology-Based Therapeutic Approaches for Hypoxia 4.1 Nanoparticles Conquering Hypoxia-Inducing Factor 4.1.1

Direct Inhibition of HIF-1α-Based Nanoparticles

Recently, it was shown that the anticancer medication piceatannol (PIC) inhibits the HIF-1 expression in colon carcinoma cells. It is then supported in tumour-bearing animals. Aljabali et al. loaded the drug into BSA NPs to increase the in vitro cytotoxic activity of invasion prevention and free PIC, and later formation of colonies by cancer cells (Aljabali et al. 2020). A putative HIF inhibitor, acriflavine (ACF), was integrated into micelles and then lipid nanocapsules in a different work by Montigaud et al. The in vivo activity of these nanoparticles was detected at a reduced intensity of delivery in comparison to free ACF, and it was discovered that ACF-loaded nanoparticles were capable of killing 4 T1 cells and decreasing the HIF expression (Montigaud et al. 2018). The most popular HIF-1 inhibitor is HIF-1 siRNA. Wan et al. developed HIF-1 siRNA-specific HIF-1 conjugated graphene oxide nanoparticles to deliver to Patu8988 cells. The authors noted that the genesis, proliferation, migration and metastasis of cancer cells were inhibited as a result of the HIF-1 gene being silenced. Additionally, it decreased the glucose transporter-1 expression in pancreatic cancer cells, promoting the death of tumour cells and extending the longevity of mice with tumours (Wan et al. 2019). By using pH-responsive bonding, Zhu et al. integrated HIF-1 siRNA into CdTe quantum dots (QDs). This route of siRNA delivery to the hypoxic tumour for enhanced treatment efficacy was shown by the abundance of siRNA at the tumour site (Zhu et al. 2015).

4.1.2

Inhibition of HIF-1α with Chemotherapy

By creating a DOX and HIF-1 siRNA co-loaded polyamidoamine dendrimer and conjugating it with PEG via an azobenzene linker, Xie et al. developed a method to improve the penetration of DOX and overcome hypoxia-induced chemoresistance in tumour cells. The PEG layer may cleave in a hypoxic TME, which would reduce particle size and change the charge of the surface to positive. The co-loaded medication DOX had improved cytotoxicity in MCF-7 and A549 cells as a result of the

Nanotechnology-Based Approaches to Relieve Tumour …

45

included siRNA’s ability to successfully decrease HIF-1 expression by downregulating P-gp and MDR-1 (Xie et al. 2018). Meng et al. developed MnO2 NPs in a different study that contained the HIF-1 inhibitor ACF. Mn2 +, oxygen and ACF were produced when MnO2 NPs interacted with H2 O2 within the tumour tissues. Inhibition of HIF-1, VEGF and MMP-9 expression in 4 T1 cells and CT26 cells also contributed to antitumor effectiveness. It also helped X-ray beam irradiation, lowered immunological resistance and reduced T-cell depletion (Meng et al. 2018). To combat in SGC-7901 cells of multidrug resistance, Chen et al. developed chitosan nanoparticles that were also given with HIF-1 siRNA and 5-fluorouracil. Co-loaded NPs showed a better transfection efficiency and silencing of gene than in naked siRNA. Additionally, the combination of siRNA and 5-fluorouracil therapy, with chitosan nanoparticles, may promote SGC-7901 cells’ programmed cell death (Chen et al. 2017b).

4.1.3

Inhibition of HIF-1α with Photodynamic Therapy

A powerful alternative for cancer treatment is the HIF-1 inhibitor and PDT combined regimen. Because PDT necessitates greater oxygen consumption to promote vascular injury, this could exacerbate hypoxic TME. Therefore, the increase of HIF-1 brought on by hypoxia may result in PDT resistance (Ji et al. 2006). Therefore, inhibiting HIF-1 expression can increase PDT effectiveness. Lipid–calcium–phosphate (LCP) nanoparticles were employed by Chen et al. to deliver HIF-1 siRNA to head-and-neck cancers. Following HIF-1 intravenous injection, siRNA-loaded LCP NPs, reduced HIF-1 expression in tumour-bearing animals was seen. Furthermore, compared to HIF-1 inhibition or PDT alone, the addition of these NPs to PDT dramatically boosted tumour regression (Chen et al. 2015).

4.2 Targeting Metastatic Hypoxic Condition by Nanodelivery Against Chemoresistance According to Jiang et al., intravenous treatment of Hb@Lipo (Hb@Lipo) substantially suppressed HIF expression in tumour tissue. Additionally, the synergistic effect of cabazitaxel-loaded liposomes (CBZ@Lipo) and Hb@Lipo may prevent the growth and spread of tumours (Jiang et al. 2019). Additionally, Luo and colleagues were successful in creating ODC-HPOC NPs by combining oxygen-containing haemoglobin, doxorubicin (DOX), with chlorin e6 which is a photosensitizer in human serum albumin. The formation of photodynamic-mediated ROS, which has potent anticancer properties, is considerably increased by these NPs. This reduces the chemoresistance that cancer cells develop as a result of hypoxia (Luo et al. 2018). Similar to liposomes that transported both DOX and Hb (Sivasubramanian & Lo, 2022), tumour cell membrane-coated PLGA NPs that contained both DOX and Hb

46

M. Singh et al.

were also shown to have excellent anticancer efficacies (Tian et al. 2017). The exocytosis of DOX by tumour cells as well as the oxygen enrichment of the tumour and the expression of the MDR gene 1 and P-glycoprotein were all reduced in this study. Song et al. recently employed the PFC family compound perfluorooctyl bromide (PFOB) to make nanoemulsions that may act as oxygen carriers. It’s interesting to note that intravenous delivery of the PFOB nanoemulsions significantly increased tumour oxygenation, breaking chemoresistance and increasing the tumour’s sensitivity to low-dose cisplatin therapy (Song et al. 2019). Enhancing oxygen production inside the tumour tissue and concentrating on a number of other factors that cause tumour hypoxia are potential strategies for overcoming chemoresistance. HIF-1, a transcription factor linked to angiogenesis, metastasis and increased P-gp expression, may be a potential target for cancer therapy. To make hypoxia-responsive NPs to co-load DOX and HIF-1 siRNA, Xie et al. recently used polyamidoamine (PAMAM) dendrimer connected with PEG via the linker known as azoreductase-sensitive azobenzene that may be dissolved in an azoreductase-supplemented hypoxia tumour. The particle size reduced and the surface charge was switched from negative to positive at the hypoxic TME, which assisted in the internalization of DOX and siRNA into the tumour cells (Xie et al. 2018). The siRNA could effectively quiet HIF-1, which in turn reduced the P-gp and MDR-1 expression and increased the cytotoxicity of DOX (Zhu et al. 2015).

4.2.1

Nanoparticle Targeting: Enhanced Chemotherapeutics Cellular Uptake in CA

According to several reports, the hypoxic environment impairs the uptake of therapeutic medicines because it prevents caveolin-1-dependent endocytosis (BourseauGuilmain et al. 2016). Notably, reducing carbonic anhydrase (CAH), and the overexpression in the hypoxic regions of many different tumour types, is the sole option to overcome this challenge. In response to these findings, Li et al. developed a CAH inhibitor utilizing the short peptide N-pepABS, which could self-assemble to generate nanofibers in the TME (Chafe et al. 2019; Li et al. 2019a, b, c). In addition to increasing tumour perfusion, photothermal therapy can enhance the way that medications are absorbed by CA cells (Luo et al. 2017; Wu et al. 2014). The slightly higher temperature within the tumour site can improve the permeability of drugs and carriers via CA cell membrane by clathrin- and caveolae-mediated endocytosis (Phung et al. 2019). In a pH-sensitive/switchable NP, Hung et al. co-loaded DOX with indocyanine green (ICG), a photosensitizer. In this nanosystem, two therapeutic drugs are housed in a PLGA core with a pH-responsive surface made of D-tocopheryl polyethylene glycol succinate functionalized with N-acetyl histidine. Because the NPs penetrated the hypoxic tumour and demonstrated high absorption by the cancer cells in a moderately acidic tumour environment when exposed to NIR radiation, they were able to successfully restrict the growth of the tumour (Hung et al. 2017).

Nanotechnology-Based Approaches to Relieve Tumour …

4.2.2

47

Nanoparticle Targeting: Radiotherapy in Metastatic Hypoxic Environment

Cancer cells can be destroyed by radiation therapy (RT) by ionizing DNA, which causes DNA damage in an environment with enriched oxygen. The hypoxic tumour is susceptible to radio resistance because of its oxygen demand. To maximize the effectiveness of radiation, innovative technology is needed to decide rather increase the oxygen supply or normalize the environment. Yi et al. developed gold@manganese dioxide NPs coated with PEG (Au@MnO2-PEG) to combat RT resistance in the treatment of murine breast cancer. In contrast to Au-PEG NPs, the NPs were capable of generating oxygen in the H2 O2 solution when MnO2 was present. Furthermore, 4 T1 cells were effectively eliminated when a 6 Gy X-ray beam disrupted the doublestrand DNA. The in vivo studies successfully revealed that during radiotherapy, oxygen is produced as well as tumour development is suppressed (Yi et al. 2016). To produce a “double” carrier for oxygen delivery, Gao et al. encased the PFC@PLGA NPs in a red blood cell membrane (PFC@PLGA-RBCM). This nanosystem was effective at accelerating oxygenation and blood flow to the tumour. In the four T1tumour-bearing mouse models, the NP did not impact the tumour’s growth on its own, but when the tumour was exposed to X-rays 24 h after intravenous injection of PFC@PLGA-RBCM, it significantly shrank. According to these data, RT and oxygen-supplying NPs are both required for the effectiveness of the treatment (Gao et al. 2017). Song et al. added PFC to a hollow Bi2 Se3 nanosystem (PEGBi2Se3@PFC) that could absorb NIR radiation and produce heat. These NPs were able to significantly release oxygen into the tumour area while also improving blood perfusion and directly killing cancer cells. This RT maintained the elimination of the tumour even in the oxygen-enriched environment, demonstrating the synergistic effects of both via normalization of the oxygen levels (Song et al. 2016a, b).

4.2.3

Nanoparticle Targeting: Photodynamic Therapy (PDT) in Metastatic Hypoxic Environment

PDT failure is thought to be mostly caused by the hypoxic tumour’s low oxygen level. To date, the following methods have been used to address the absence of hypoxia-restrained PDT efficacy (Li et al. 2018).

Intra-Tumoral Oxygen Concentration: Enhanced PDT One of the most popular therapies to control hypoxic TME and improve the therapeutic effectiveness of PDT is direct oxygen administration to the tumour tissues utilizing appropriate carriers such Hb, PFCs and MOFs. This is why Luo et al. encased Hb-indocyanine green (ICG) complexes in lipid-polymer nanoparticles. The tumour tissue was able to enrich itself in oxygen thanks to this nanocarrier, which boosted

48

M. Singh et al.

the production of ROS during PDT and significantly slowed tumour growth (Luo et al. 2016). For the advancement of PDT against malignancies, various organizations have recently tried to develop alternative Hb-based oxygen delivery systems (Wang et al. 2017a, b, 2015). Since each Hb molecule can only carry four oxygen molecules, these Hb-based devices could only load a specific amount of oxygen. PFCs have been considered essential elements for the development of oxygen delivery systems due to their remarkable capacity to transport oxygen (Riess 2005). For instance, Song et al. recommended using nano-PFC in an oxygenation system that uses ultrasonic (US) technology to change the hypoxic TME and enhance PDT. In this work, tumour-bearing animals with hypertoxic respiration received intravenously administered albumin-stabilized PFC nanodroplets while receiving US therapy. The PFC droplets absorb a lot of oxygen after passing through the lung that can be stored and released into the tumour when provoked by ultrasound. As a result, intra-tumoral oxygenation rises significantly, and PDT and RT are more effective against a variety of tumour forms (Song et al. 2016a, b). By converting H2O2 present in the tumour into oxygen using a variety of NPs, including polymeric NPs (Phua et al. 2019; Zhu et al. 2019), albumin NPs (Chen et al. 2017a, b), gold NPs (Liu et al. 2017a, b; Zhang et al. 2019) and mesoporous silica NPs (Xiang et al. 2019), additional studies have demonstrated an improvement in PDT efficacy. Numerous animal models showed these hopeful results. For instance, Chen et al. created self-assembled NPs utilizing a one-step procedure from HSA, chlorine e6 (Ce6), catalase and paclitaxel (PTX) (HSA-Ce6- Cat-PTX NPs). The generated NPs displayed strong tumour homing, which has been effectively activated by H2 O2 breakdown and increased the amount of oxygen reaching the tumour. Notably, the HSA-Ce6-Cat-PTX NPs could have a potent anticancer impact even at a modest treatment dose (Chen et al. 2017a, b).

Strategies to Reduce O2 Utilization in PDT Reducing the oxygen uptake is a potential technique to increase PDT efficacy because the hypoxic condition is made worse by the increased oxygen usage during PDT treatment, which limits the therapeutic outcomes of PDT. As of late, the main tactic to lower oxygen consumption has been the development of PDT which targets subcellular organelles such as plasma membranes, mitochondria and lysosomes. Due to their important element in the production of energy and oxygen uptake (Li and Graham 2012), in addition to their extreme sensitivity to ROS and heat, mitochondria, sometimes known as the “powerhouses” of cells, have come under growing scrutiny for subcellular organelle-targeted PDT. In this context, it has been discovered that inhibiting mitochondrial oxidative respiration with photosensitizers that target mitochondria, such as metal complexes, IR 780, ICG and boron-dipyrromethene (BODIPY), significantly increases antitumor activity in comparison to the agent’s targeting cytoplasm or other organelles, especially in hypoxic conditions (Lin et al. 2018). For instance, Zhao et al. linked the iridium (III) complex having a triplet excited state with YC-1 to enhance NP accumulation and penetration in the tumour

Nanotechnology-Based Approaches to Relieve Tumour …

49

tissue (HIF-1 inhibitor-loaded NPs) (Jeena et al. 2019; Zhao et al. 2020). They showed that due to the prolonged lifetime of the complex and its capacity for light harvesting, iridium (III)-conjugated NPs may effectively create singlet oxygen and detectable photocytotoxicity in response to the cancer, at low oxygen concentrations. Additionally, the inclusion of YC-1 significantly improved the nanoparticles’ PDT efficiency, providing a rational way to overcome the hypoxic susceptibility to PDT therapy. In a different study, Cheng et al. recommended using chimeric peptideconjugated nanoparticles (M-ChiP) to control the location of the mitochondria and plasma membrane for the photosensitizer protoporphyrin IX (PpIX). The PpIX in mitochondrial delivery and membrane permeability was observed to enhance with the dual-targeting M-Chips, maximizing therapeutic efficacy while minimizing side effects (Wang et al. 2020).

Development of Novel Oxygen-Independent PDT The effectiveness of standard PDT, which requires oxygen to produce cytotoxic ROS, is severely limited by severe hypoxia. The best way to overcome this obstacle is to produce photosensitizers that are photoactivated for becoming cytotoxic without the need for an oxygen source. A photoacid generator (PAG) based on sulfonium was used by Fadhel et al. as a hypoxia-independent PDT agent (Fadhel et al. 2016). The PAG may lower intracellular pH and damage cellular components when properly excited by NIR photons. PAG was pegylated and coupled to silica nanoparticles in this work (siNP-PAG9). The researchers found that the resulting siNPPAG9 NPs were significantly more cytotoxic to cancer cells than either of the free and pegylated forms of PAG and could efficiently deliver siNP-PAG9 to HCT-116 cells. Recent years have seen a rise in the usage of metal deposition on semiconductors as an effective photocatalyst in a variety of fields, including biomedical engineering. As an oxygen-independent photosensitizer for PDT, Fan et al. used an RGD peptide-modified metal–semiconductor nanosystem made by depositing Au on CdSe-seeded/CdS nanorods. When exposed to visible light, the hybrid nanocomposite may successfully accelerate the production of ROS from water without the need for oxygen. The nanocomposite’s ability to target tumours and extend its circulation duration may both benefit from the RGD peptide functionalization. The fact that the oxygen-independent nanosystem was able to overcome the hypoxia-induced PDT resistance was a significant discovery, demonstrating the potential of common semiconductors as oxygen-independent PDT agents (Fan et al. 2017).

Combining PDT and PTT for Modulation of Hypoxic TME Photothermal therapy is a viable candidate to be used in conjunction with PDT to combat the hypoxia caused by PDT because of the increased intra-tumoral perfusion caused by hyperthermia. Lately, Wu et al. coupled PDT and PTT in hollow Au/Ag-MnO2 nanospheres coated with PEG and loaded with Ce6 (AAM HNSs).

50

M. Singh et al.

The breakdown of MnO2 by AAM HNSs in the acidic TME with a greater extent of H2 O2 resulted in a significant concentration of oxygen, which together with PTT relieved tumour hypoxia and improved PDT efficacy. Importantly, compared to any solo medication, the combination therapy demonstrated a stronger antitumor effect (Wu et al. 2019). In EphB4 receptor-targeted hollow gold nanospheres functionalized with ICG-conjugated TNYL peptide, Li et al. simultaneously tested a combination of PTT and PDT. The nanoplatform considerably promotes antitumor efficacy by increasing ROS and heat formation while noticeably decreasing HIF-1 expression when exposed to NIR radiation (Li et al. 2017).

4.3 Nanoparticle Relieving: Lowering Hypoxic TME by Immunotherapy Without a doubt, immunotherapy has emerged as the main foundation of cancer treatment; however, there are still challenges (Jackson et al. 2019). Among these difficulties, hypoxia significantly contributes to the emergence of immunotherapy resistance through the following approaches.

4.3.1

Nanoparticle Relieving: Enhanced Effector Immune Cell Infiltration into TME

When cancer cells are exposed to cytotoxic drugs, photothermal therapy or radiation therapy (RT), immunogenic cell death (ICD) is more easily induced in the tumour tissues because of the movement of antitumor immune cells (Nguyen et al. 2019, 2021). High mobility group box 1 protein (HMGB-1), ATP and calreticulin are just a few examples of damage-associated molecular patterns (DAMPs) that are released into the environment as a result of the ICD effect. These molecules then mediate the placement and maturation of APCs at the tumour, which in turn primes the development of CTLs that are specific for an antigen (Kheirolomoom et al. 2019; Mastria et al. 2018; Tran et al. 2018). Therefore, combining drugs that induce ICD and treat hypoxia in roughly the same nanosystems may enhance the capacity of effector immune cells to enter the tissues of the tumour. In HSA NPs, Chen et al. cointegrated Ce6 and Hb for PDT that relied solely on oxygen. When the photosensitizer and oxygen were administered jointly, the PDT’s efficiency was greatly increased and tumour hypoxia was significantly decreased. By releasing DAMPs like HMGB-1 and ATP, the injection of NPs greatly triggered the ICD effect and produced antigencaptured DCs at malignant cell lymph nodes. The proliferation of activated T and NK cells dramatically decreased lung cancer metastasis, primary tumour growth and distant tumour development (Chen et al. 2018). Im et al. combined the photosensitizer Ce6 along with the DC-activating adjuvant CpG, a toll-like receptor (TLR) 9 agonist, in mesoporous silica NPs coated with glycol chitosan and PEG in order to promote

Nanotechnology-Based Approaches to Relieve Tumour …

51

DC maturation for effective antigen presentation. The Azo linker was destroyed in a hypoxic environment and after being exposed to light, which caused the PEG layer to separate and the release of PDT and CpG agents. The addition of CpG significantly increased DC maturation and antigen presentation, along with significantly decreased the in vivo tumour burden. PDT was also observed to significantly increase ICD (Im et al. 2019).

4.3.2

Nanoparticle Relieving: Enhanced Immune Checkpoint Therapy in TME

It is generally known that the programmed death-1 (PD-1) receptor, which is expressed on activated T-cells, modifies T-cell responses. Peripheral tolerance was induced as a result of the interaction of PD-1 and its corresponding ligands, PDL1 or PD-L2, which impaired T-cell cytotoxic activity (Truong et al. 2019). These ligands are overexpressed by cancer cells as a checkpoint to reduce the function and activity of T-lymphocytes. As previously indicated, in the context of TME, hypoxia directly enhances the expression of PD-L1 in cancer cells in addition to intratumorally MDSCs and macrophages via activation of HIF-1 (Zhou et al. 2019). To effectively prevent tumour growth, the immunological checkpoint that could be affected by the harmonization of the hypoxic conditions may be compromised. Recently, Zou et al. demonstrated that the hypoxia-relieving NP developed by embedding catalase (CAT) and DOX into a zeolitic imidazolate framework 8 (ZCD) and then coating it with the melanoma cell membrane (mZCD) increased the therapeutic efficacy of PD-1. Because the cancer cell membrane is capable of targeting and immune escape, the mZCD NPs could effectively concentrate at the tumour site. HIF-1 and PD-L1 expression was lowered as a result of the conversion of H2 O2 to O2 under the influence of CAT, which helped DOX work more effectively. As a result, the effectiveness of the mZCD treatment sparked the ICD effect, which enhances antigen presentation and primes antigen-specific T-cells. It’s interesting to note that the anti-PDL1 medication regimen enhanced the antitumor immune responses that the mZCD induced, effectively inhibiting tumour growth and metastasis (Zou et al. 2018). In a different investigation, Song et al. paired radiation and anti-CTLA-4 therapy with a stealthy liposome (CAT@liposome) laden with H2 O2 and catalase. The combination radio immunotherapy’s effectiveness was further amplified by the intravenous infusion of the CAT@liposome, which improved tumour oxygenation and caused the immune-suppressive TME to revert (Song et al. 2018).

4.3.3

Nanoparticles Relieve: Enhanced Immune Cell-Mediated Cytotoxicity in TME

It has been shown that the HIF-1 upregulation in tumour hypoxia completely promotes the several molecules expression that help cancer cells resist the cytotoxic effects of immune cells, including Bcl-2 (Liang et al. 2017) and NANOG (Jeter et al.

52

M. Singh et al.

2015), or inhibits the expression of molecules essential for immune surveillance, including MHC-I (Sethumadhavan et al. 2017) and MHC-I chain-related protein A/B (MICA/B) (Schilling et al. 2015). Additionally, the hypoxic TME may reduce the ability of NK and T-cells to kill cancer by accelerating the breakdown of granzyme B released by NK cells (Baginska et al. 2013) and preventing IFN-induced anticancer responses (Murthy et al. 2019). Liang et al. suggested a method based on these findings to encourage the immune cell-induced cytotoxicity towards cancer cells by administering an Au@Pt NP to re-oxygenate the hypoxic TME. The intratumoral H2 O2 was successfully converted into oxygen by the Au@Pt NP. The scientists showed that the tumour’s oxygen enrichment produced noticeably improved immune responses against malignancy (Liang et al. 2017). Additionally, Mahiddine et al. reported increased neutrophil tumoricidal activity, which was followed by a reduction in tumour hypoxia. By reducing the hypoxic TME, these findings offer a promising method for cellular immunotherapy, such as CAR-T cells and CAR-NK cells (Mahiddine et al. 2020).

5 Conclusion Here, we provide an overview of existing nanotechnology-based methods for treating cancer while preventing tumour hypoxia. Despite significant advancements and encouraging outcomes, the majority of efforts are still in the early stages of research. These methods have some drawbacks, such as intravenous injection side effects, H2O2 reliance in H2O2-mediated O2 generation, quick inactivation or instability of natural enzymes and nanozymes, and low light transmission in photoactivated O2 production. Additionally, the most popular approach to treat hypoxic tumours is the combination of numerous therapeutic/diagnostic capabilities and O2 -supply capability into one nanosystem in order to obtain better therapeutic efficacy. As a result, laborious and complicated preparatory processes are frequently required. Prior to clinical trials, the toxicity and immunogenicity of all the relevant components should be thoroughly assessed in order to optimize their potential and reduce their adverse effects.

References Aljabali AAA, Bakshi HA, Hakkim FL, Haggag YA, Al-Batanyeh KM, Al Zoubi MS, Tambuwala MM (2020) Albumin nano-encapsulation of piceatannol enhances its anticancer potential in colon cancer via downregulation of nuclear p65 and HIF-1? Cancers (Basel) 12 Baginska J, Viry E, Berchem G, Poli A, Noman MZ, van Moer K, Janji B (2013) Granzyme B degradation by autophagy decreases tumor cell susceptibility to natural killer-mediated lysis under hypoxia. Proc Natl Acad Sci U S A 110(43):17450–17455

Nanotechnology-Based Approaches to Relieve Tumour …

53

Banerjee U, Wolfe S, O’Boyle Q, Cuddington C, Palmer AF (2022) Scalable production and complete biophysical characterization of poly(ethylene glycol) surface conjugated liposome encapsulated hemoglobin (PEG-LEH). PLoS ONE 17(7):e0269939 Bourseau-Guilmain E, Menard JA, Lindqvist E, Indira Chandran V, Christianson H, Cerezo C, Magaña M, Belting M (2016) Hypoxia regulates global membrane protein endocytosis through caveolin-1 in cancer cells. Nat Commun 7:11371 Cardoso AR, Frasco MF, Serrano V, Fortunato E, Sales MGF (2021) Molecular imprinting on nanozymes for sensing applications. Biosensors (Basel) 11(5) Chafe SC, McDonald PC, Saberi S, Nemirovsky O, Venkateswaran G, Burugu S, Dedhar S (2019) Targeting hypoxia-induced carbonic anhydrase IX enhances immune-checkpoint blockade locally and systemically. Cancer Immunol Res 7(7):1064–1078 Chen WH, Lecaros RL, Tseng YC, Huang L, Hsu YC (2015) Nanoparticle delivery of HIF1α siRNA combined with photodynamic therapy as a potential treatment strategy for head-and-neck cancer. Cancer Lett 359(1):65–74 Chen Q, Chen J, Liang C, Feng L, Dong Z, Song X, Liu Z (2017a) Drug-induced co-assembly of albumin/catalase as smart nano-theranostics for deep intra-tumoral penetration, hypoxia relieve, and synergistic combination therapy. J Control Release 263:79–89 Chen Z, Liu L, Liang R, Luo Z, He H, Wu Z, Cai L (2018) Bioinspired hybrid protein oxygen nanocarrier amplified photodynamic therapy for eliciting anti-tumor immunity and abscopal effect. ACS Nano 12(8):8633–8645 Chen Y, Sun L, Guo D, Wu Z, Chen W (2017b). Co-delivery of hypoxia inducible factor-1α small interfering RNA and 5-fluorouracil to overcome drug resistance in gastric cancer SGC-7901 cells. J Gene Med 19(12) Cheng Y, Cheng H, Jiang C, Qiu X, Wang K, Huan W, Hu Y (2015) Perfluorocarbon nanoparticles enhance reactive oxygen levels and tumour growth inhibition in photodynamic therapy. Nat Commun 6:8785 Dai X, Ruan J, Guo Y, Sun Z, Liu J, Bao X, Zhao P (2021) Enhanced radiotherapy efficacy and induced anti-tumor immunity in HCC by improving hypoxia microenvironment using oxygen microcapsules. Chem Eng J 422:130109 DeCoste JB, Weston MH, Fuller PE, Tovar TM, Peterson GW, LeVan MD, Farha OK (2014) Metal–organic frameworks for oxygen storage. Angwandte Chemie Fadhel AA, Yue X, Ghazvini Zadeh EH, Bondar MV, Belfield KD (2016) Pegylated and nanoparticle-conjugated sulfonium salt photo triggers necrotic cell death. Int J Nanomed 11:6161–6168 Fan J-X, Liu M-D, Li C-X, Hong S, Zheng D-W, Liu X-H, Zhang X-Z (2017) A metal–semiconductor nanocomposite as an efficient oxygen-independent photosensitizer for photodynamic tumor therapy. Nanoscale Horizons 2(6):349–355 Fiorito V, Chiabrando D, Petrillo S, Bertino F, Tolosano E (2019) The multifaceted role of heme in cancer. Front Oncol 9:1540 Gao M, Liang C, Song X, Chen Q, Jin Q, Wang C, Liu Z (2017) Erythrocyte-membrane-enveloped perfluorocarbon as nanoscale artificial red blood cells to relieve tumor hypoxia and enhance cancer radiotherapy. Adv Mater 29(35) He Z, Xiao Y, Zhang J-R, Zhang P, Zhu J-J (2018) In situ formation of large pore silica–MnO2 nanocomposites with H+/H2O2 sensitivity for O2-elevated photodynamic therapy and potential MR imaging. Chem Commun Hung CC, Huang WC, Lin YW, Yu TW, Chen HH, Lin SC, Chiu HC (2017) Erratum: active tumor permeation and uptake of surface charge-switchable theranostic nanoparticles for imagingguided photothermal/chemo combinatorial therapy: erratum. Theranostics 7(3):559–560 Im S, Lee J, Park D, Park A, Kim Y-M, Kim WJ (2019) Hypoxia-triggered transforming immunomodulator for cancer immunotherapy via photodynamically enhanced antigen presentation of dendritic cell. ACS Nano 13(1):476–488 Irani S (2019) Emerging insights into the biology of metastasis: a review article. Iran J Basic Med Sci 22(8):833–847

54

M. Singh et al.

Jackson CM, Choi J, Lim M (2019) Mechanisms of immunotherapy resistance: lessons from glioblastoma. Nat Immunol 20(9):1100–1109 Jägers J, Wrobeln A, Ferenz KB (2021) Perfluorocarbon-based oxygen carriers: from physics to physiology. Pflugers Arch 473(2):139–150 Jeena MT, Kim S, Jin S, Ryu JH (2019) Recent progress in mitochondria-targeted drug and drug-free agents for cancer therapy. Cancers (Basel) 12(1) Jeter CR, Yang T, Wang J, Chao HP, Tang DG (2015) Concise review: NANOG in cancer stem cells and tumor development: an update and outstanding questions. Stem Cells 33(8):2381–2390 Ji Z, Yang G, Shahzidi S, Tkacz-Stachowska K, Suo Z, Nesland JM, Peng Q (2006) Induction of hypoxia-inducible factor-1alpha overexpression by cobalt chloride enhances cellular resistance to photodynamic therapy. Cancer Lett 244(2):182–189 Ji B, Wei M, Yang B (2022) Recent advances in nanomedicines for photodynamic therapy (PDT)driven cancer immunotherapy. Theranostics 12(1):434–458 Jiang MS, Yin XY, Qin B, Xuan SY, Yuan XL, Yin H, You J (2019) Inhibiting hypoxia and chemotherapy-induced cancer cell metastasis under a valid therapeutic effect by an assistance of biomimetic oxygen delivery. Mol Pharm 16(11):4530–4541 Jiao L, Zhang X, Cui J, Peng X, Song F (2019) Three-in-one functional silica nanocarrier with singlet oxygen generation, storage/release, and self-monitoring for enhanced fractional photodynamic therapy. ACS Appl Mater Interfaces 11(29):25750–25757 Kheirolomoom A, Silvestrini MT, Ingham ES, Mahakian LM, Tam SM, Tumbale SK, Ferrara KW (2019) Combining activatable nanodelivery with immunotherapy in a murine breast cancer model. J Control Release 303:42–54 Kim J, Heo JN, Do JY, Chava RK, Kang M (2019) Electrochemical synergies of heterostructured Fe2O3-MnO catalyst for oxygen evolution reaction in alkaline water splitting. Nanomaterials 9(10):1486 Li Z, Graham BH (2012) Measurement of mitochondrial oxygen consumption using a Clark electrode. Methods Mol Biol 837:63–72 Li W, Guo X, Kong F, Zhang H, Luo L, Li Q, You J (2017) Overcoming photodynamic resistance and tumor targeting dual-therapy mediated by indocyanine green conjugated gold nanospheres. J Control Release 258:171–181 Li X, Kwon N, Guo T, Liu Z, Yoon J (2018) Innovative strategies for hypoxic-tumor photodynamic therapy. Angew Chem Int Ed Engl 57(36):11522–11531 Li RQ, Zhang C, Xie BR, Yu WY, Qiu WX, Cheng H, Zhang XZ (2019b) A two-photon excited O(2)evolving nanocomposite for efficient photodynamic therapy against hypoxic tumor. Biomaterials 194:84–93 Li Y, Liu X, Nie X, Yang W, Wang Y, Yu R, Shui J (2019c) Multifunctional organic-inorganic hybrid aerogel for self-cleaning, heat-insulating, and highly efficient microwave absorbing material. Adv Func Mater 29(10):1807624 Li Y, Zhao L, Li XF (2021) Hypoxia and the tumor microenvironment. Technol Cancer Res Treat 20:15330338211036304 Li J, Shi K, Sabet ZF, Fu W, Zhou H, Xu S, Chen C (2019a) New power of self-assembling carbonic anhydrase inhibitor: Short peptide-constructed nanofibers inspire hypoxic cancer therapy. Sci Adv 5(9):eaax0937 Liang H, Wu Y, Ou XY, Li JY, Li J (2017) Au@Pt nanoparticles as catalase mimics to attenuate tumor hypoxia and enhance immune cell-mediated cytotoxicity. Nanotechnology 28(46):465702 Lin F, Bao YW, Wu FG (2018) Improving the phototherapeutic efficiencies of molecular and nanoscale materials by targeting mitochondria. Molecules 23(11) Liu C-P, Wu T-H, Liu C-Y, Chen K-C, Chen Y-X, Chen G-S, Lin S-Y (2017a) Self-supplying O2 through the catalase-like activity of gold nanoclusters for photodynamic therapy against hypoxic cancer cells. Small 13(26):1700278 Liu J, Huang J, Zhang L, Lei J (2021a) Multifunctional metal–organic framework heterostructures for enhanced cancer therapy. Chem Soc Rev 50(2):1188–1218

Nanotechnology-Based Approaches to Relieve Tumour …

55

Liu W, Yan Q, Xia C, Wang X, Kumar A, Wang Y, Liu J (2021b) Recent advances in cell membrane coated metal–organic frameworks (MOFs) for tumor therapy. J Mater Chem B 9(22):4459–4474 Liu LH, Zhang YH, Qiu WX, Zhang L, Gao F, Li B, Zhang XZ (2017b). Dual-stage light amplified photodynamic therapy against hypoxic tumor based on an O(2) self-sufficient nanoplatform. Small 13(37) López-Lázaro M (2007) Dual role of hydrogen peroxide in cancer: possible relevance to cancer chemoprevention and therapy. Cancer Lett 252(1):1–8 Luo Z, Zheng M, Zhao P, Chen Z, Siu F, Gong P, Cai L (2016) Self-monitoring artificial red cells with sufficient oxygen supply for enhanced photodynamic therapy. Sci Rep 6(1):23393 Luo D, Carter KA, Miranda D, Lovell JF (2017) Chemophototherapy: an emerging treatment option for solid tumors. Adv Sci (weinh) 4(1):1600106 Luo Z, Tian H, Liu L, Chen Z, Liang R, Chen Z, Cai L (2018) Tumor-targeted hybrid protein oxygen carrier to simultaneously enhance hypoxia-dampened chemotherapy and photodynamic therapy at a single dose. Theranostics 8(13):3584–3596 Mahiddine K, Blaisdell A, Ma S, Créquer-Grandhomme A, Lowell CA, Erlebacher A (2020) Relief of tumor hypoxia unleashes the tumoricidal potential of neutrophils. J Clin Invest 130(1):389– 403 Maniotis AJ, Folberg R, Hess A, Seftor EA, Gardner LM, Pe’er J, Hendrix MJ (1999) Vascular channel formation by human melanoma cells in vivo and in vitro: vasculogenic mimicry. Am J Pathol 155(3):739–752 Martin JD, Fukumura D, Duda DG, Boucher Y, Jain RK (2016) Reengineering the tumor microenvironment to alleviate hypoxia and overcome cancer heterogeneity. Cold Spring Harb Perspect Med 6(12) Mastria EM, Cai LY, Kan MJ, Li X, Schaal JL, Fiering S, Chilkoti A (2018) Nanoparticle formulation improves doxorubicin efficacy by enhancing host antitumor immunity. J Control Release 269:364–373 Meng L, Cheng Y, Tong X, Gan S, Ding Y, Zhang Y, Yuan A (2018) Tumor oxygenation and hypoxia inducible factor-1 functional inhibition via a reactive oxygen species responsive nanoplatform for enhancing radiation therapy and abscopal effects. ACS Nano 12(8):8308–8322 Montigaud Y, Ucakar B, Krishnamachary B, Bhujwalla ZM, Feron O, Préat V, Danhier F, Gallez B, Danhier P (2018) Optimized acriflavine-loaded lipid nanocapsules as a safe and effective delivery system to treat breast cancer. Int J Pharm 551:322–328 Murthy A, Gerber SA, Koch CJ, Lord EM (2019) Intratumoral hypoxia reduces IFN-γ-mediated immunity and MHC class I induction in a preclinical tumor model. Immunohorizons 3(4):149– 160 Naxerova K, Jain RK (2015) Using tumour phylogenetics to identify the roots of metastasis in humans. Nat Rev Clin Oncol 12(5):258–272 Naxerova K, Brachtel E, Salk JJ, Seese AM, Power K, Abbasi B, Jain RK (2014) Hypermutable DNA chronicles the evolution of human colon cancer. Proc Natl Acad Sci U S A 111(18):E1889-1898 Nguyen HT, Byeon JH, Phung CD, Pham LM, Ku SK, Yong CS, Kim JO (2019) Method for the instant in-flight manufacture of black phosphorus to assemble Core@Shell nanocomposites for targeted photoimmunotherapy. ACS Appl Mater Interfaces 11(28):24959–24970 Nguyen HT, Phung CD, Tran TH, Pham TT, Pham LM, Nguyen TT, Kim JO (2021) Manipulating immune system using nanoparticles for an effective cancer treatment: combination of targeted therapy and checkpoint blockage miRNA. J Control Release 329:524–537 Petrova et al., 2018 Petrova V, Annicchiarico-Petruzzelli M, Melino G, Amelio I (2018) The hypoxic tumour microenvironment. Oncogenesis 7:1–10 Phua SZF, Yang G, Lim WQ, Verma A, Chen H, Thanabalu T, Zhao Y (2019) Catalase-integrated hyaluronic acid as nanocarriers for enhanced photodynamic therapy in solid tumor. ACS Nano 13(4):4742–4751 Phung DC, Nguyen HT, Phuong Tran TT, Jin SG, Yong CS, Truong DH, Kim JO (2019) Combined hyperthermia and chemotherapy as a synergistic anticancer treatment. J Pharm Investig 49(5):519–526

56

M. Singh et al.

Prasad P, Gordijo CR, Abbasi AZ, Maeda A, Ip A, Rauth AM, Wu XY (2014) Multifunctional albumin-MnO2 nanoparticles modulate solid tumor microenvironment by attenuating hypoxia, acidosis, vascular endothelial growth factor and enhance radiation response. ACS Nano 8(4):3202–3212 Riess JG (2005) Understanding the fundamentals of perfluorocarbons and perfluorocarbon emulsions relevant to in vivo oxygen delivery. Artif Cells Blood Substit Immobil Biotechnol 33(1):47–63 Schilling D, Tetzlaff F, Konrad S, Li W, Multhoff G (2015) A hypoxia-induced decrease of either MICA/B or Hsp70 on the membrane of tumor cells mediates immune escape from NK cells. Cell Stress Chaperones 20(1):139–147 Sethumadhavan S, Silva M, Philbrook P, Nguyen T, Hatfield SM, Ohta A, Sitkovsky MV (2017) Hypoxia and hypoxia-inducible factor (HIF) downregulate antigen-presenting MHC class I molecules limiting tumor cell recognition by T cells. PLoS ONE 12(11):e0187314 Siegel RL, Miller KD, Fuchs HE, Jemal A (2022) Cancer statistics, 2022. CA Cancer J Clinic 72(1):7–33 Sivasubramanian M, Lo LW (2022) Assessment of nanoparticle-mediated tumor oxygen modulation by photoacoustic imaging. Biosensors (Basel) 12(5) Song G, Liang C, Yi X, Zhao Q, Cheng L, Yang K, Liu Z (2016a) Perfluorocarbon-loaded hollow Bi2Se3 nanoparticles for timely supply of oxygen under near-infrared light to enhance the radiotherapy of cancer. Adv Mater 28(14):2716–2723 Song X, Feng L, Liang C, Yang K, Liu Z (2016b) Ultrasound triggered tumor oxygenation with oxygen-shuttle nanoperfluorocarbon to overcome hypoxia-associated resistance in cancer therapies. Nano Lett 16(10):6145–6153 Song X, Xu J, Liang C, Chao Y, Jin Q, Wang C, Liu Z (2018) Self-supplied tumor oxygenation through separated liposomal delivery of H(2)O(2) and catalase for enhanced radioimmunotherapy of cancer. Nano Lett 18(10):6360–6368 Song D, Beringhs AO, Zhuang Z, Joshi G, Tran TH, Claffey KP, Lu X (2019) Overcoming hypoxiainduced chemoresistance to cisplatin through tumor oxygenation monitored by optical imaging. Nanotheranostics 3(2):223–235 Tian H, Luo Z, Liu L, Zheng M, Chen Z, Ma A, Cai L (2017) Cancer cell membranebiomimetic oxygen nanocarrier for breaking hypoxia-induced chemoresistance. Adv Func Mater 27(38):1703197 Tran TH, Tran TTP, Nguyen HT, Phung CD, Jeong JH, Stenzel MH, Kim JO (2018) Nanoparticles for dendritic cell-based immunotherapy. Int J Pharm 542(1–2):253–265 Truong DH, Tran TTP, Nguyen HT, Phung CD, Pham TT, Yong CS, Tran TH (2019) Modulating T-cell-based cancer immunotherapy via particulate systems. J Drug Target 27(2):145–163 Wan R, Luan Y, Wu Z, Deng S, Peng M, Xu L, Fan G (2019) The target therapeutic effect of functionalized graphene oxide nanoparticles graphene oxide-polyethylene glycol-folic acid1-pyrenemethylamine hydrochloride-mediated RNA interference of HIF-1α gene in human pancreatic cancer cells. J Biomater Appl 34(2):155–177 Wang W, Lin P, Han C, Cai W, Zhao X, Sun B (2010) Vasculogenic mimicry contributes to lymph node metastasis of laryngeal squamous cell carcinoma. J Exp Clin Cancer Res 29(1):60 Wang S, Yuan F, Chen K, Chen G, Tu K, Wang H, Wang LQ (2015) Synthesis of hemoglobin conjugated polymeric micelle: a ZnPc carrier with oxygen self-compensating ability for photodynamic therapy. Biomacromol 16(9):2693–2700 Wang M, Zhao J, Zhang L, Wei F, Lian Y, Wu Y, Guo C (2017a) Role of tumor microenvironment in tumorigenesis. J Cancer 8(5):761–773 Wang P, Li X, Yao C, Wang W, Zhao M, El-Toni AM, Zhang F (2017b) Orthogonal near-infrared upconversion co-regulated site-specific O(2) delivery and photodynamic therapy for hypoxia tumor by using red blood cell microcarriers. Biomaterials 125:90–100 Wang H, Chao Y, Liu J, Zhu W, Wang G, Xu L, Liu Z (2018) Photosensitizer-crosslinked in-situ polymerization on catalase for tumor hypoxia modulation & enhanced photodynamic therapy. Biomaterials 181:310–317

Nanotechnology-Based Approaches to Relieve Tumour …

57

Wang K, Xiang Y, Pan W, Wang H, Li N, Tang B (2020) Dual-targeted photothermal agents for enhanced cancer therapy. Chem Sci 11(31):8055–8072 Wang X, Liu T, Yu J (2019) A limiting current oxygen sensor constituted of(CeO2)0.95(Y2O3)0.05 as solid electrolyte layer and(CeO2)0.75(ZrO2)0.25 as dense diffusion barrier layer. Sensors (Basel) 19(16) Weinberg F, Ramnath N, Nagrath D (2019) Reactive oxygen species in the tumor microenvironment: an overview. Cancers (Basel) 11(8) WHO (2022) Cancer Wu P-C, Shieh D-B, Cheng F-Y (2014) Nanomaterial-mediated photothermal cancer treatment: the pivotal role of cellular uptake on photothermal therapeutic efficacy. RSC Adv 4(95):53297– 53306 Wu K, Zhao H, Sun Z, Wang B, Tang X, Dai Y, Huang W (2019) Endogenous oxygen generating multifunctional theranostic nanoplatform for enhanced photodynamic-photothermal therapy and multimodal imaging. Theranostics 9(25):7697–7713 Wu J, Song J, Yin X, Tang J, Zhang J, Wang X, Liang T (2022) Recent advancements of nanotechnology-based strategies for overcoming tumor microenvironment hypoxia. Front Biosci (landmark Ed) 27(5):145 Wu J, Wang X, Chen L, Wang J, Zhang J, Tang J, Liang T (2023) Oxygen microcapsules improve immune checkpoint blockade by ameliorating hypoxia condition in pancreatic ductal adenocarcinoma. Bioact Mater 20:259–270 Xiang H, Lin H, Yu L, Chen Y (2019) Hypoxia-irrelevant photonic thermodynamic cancer nanomedicine. ACS Nano 13(2):2223–2235 Xie Z, Guo W, Guo N, Huangfu M, Liu H, Lin M, Gao J (2018) Targeting tumor hypoxia with stimulus-responsive nanocarriers in overcoming drug resistance and monitoring anticancer efficacy. Acta Biomater 71:351–362 Yan Z, Liu H, Hao Z, Yu M, Chen X, Chen J (2020) Electrodeposition of (hydro)oxides for an oxygen evolution electrode. Chem Sci 11(39):10614–10625 Yi X, Chen L, Zhong X, Gao R, Qian Y, Wu F, Yang K (2016) Core–shell Au@MnO2 nanoparticles for enhanced radiotherapy via improving the tumor oxygenation. Nano Res 9(11):3267–3278 Yu Q, Huang T, Liu C, Zhao M, Xie M, Li G, Zhao Q (2019) Oxygen self-sufficient NIR-activatable liposomes for tumor hypoxia regulation and photodynamic therapy. Chem Sci 10(39):9091–9098 Zhang L, Yang XQ, Wei JS, Li X, Wang H, Zhao YD (2019) Intelligent gold nanostars for in vivo CT imaging and catalase-enhanced synergistic photodynamic & photothermal tumor therapy. Theranostics 9(19):5424–5442 Zhang Q, He J, Yu W, Li Y, Liu Z, Zhou B, Liu Y (2020) A promising anticancer drug: a photosensitizer based on the porphyrin skeleton. RSC Med Chem 11(4):427–437 Zhao J, Sun S, Li X, Zhang W, Gou S (2020) Enhancing photodynamic therapy efficacy of upconversion-based nanoparticles conjugated with a long-lived triplet excited state Iridium(III)Naphthalimide complex: toward highly enhanced hypoxia-inducible Factor-1. ACS Appl Bio Mater 3(1):252–262 Zhao L, Fu C, Tan L, Li T, Zhong H, Meng X (2020) Advanced nanotechnology for hypoxiaassociated antitumor therapy. Nanoscale Zheng D-W, Li B, Li C-X, Fan J-X, Lei Q, Li C, Zhang X-Z (2016) Carbon-dot-decorated carbon nitride nanoparticles for enhanced photodynamic therapy against hypoxic tumor via water splitting. ACS Nano 10(9):8715–8722 Zhou L, Cha G, Chen L, Yang C, Xu D, Ge M (2019) HIF1α/PD-L1 axis mediates hypoxiainduced cell apoptosis and tumor progression in follicular thyroid carcinoma. Onco Targets Ther 12:6461–6470 Zhu H, Zhang S, Ling Y, Meng G, Yang Y, Zhang W (2015) pH-responsive hybrid quantum dots for targeting hypoxic tumor siRNA delivery. J Control Release 220(Pt A):529–544 Zhu R, He H, Liu Y, Cao D, Yan J, Duan S, Yin L (2019) Cancer-selective bioreductive chemotherapy mediated by dual hypoxia-responsive nanomedicine upon photodynamic therapy-induced hypoxia aggravation. Biomacromol 20(7):2649–2656

58

M. Singh et al.

Zou M-Z, Liu W-L, Li C-X, Zheng D-W, Zeng J-Y, Gao F, Zhang X-Z (2018) A Multifunctional biomimetic nanoplatform for relieving hypoxia to enhance chemotherapy and inhibit the PD1/PD-L1 axis. Small 14(28):1801120

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials Pooja Kumari, Preeti Sharma, Yogesh Srivastava, and Narendra Kumar Sharma

Abstract Hypoxia is a common symptom of many serious disorders, including cancer, ischemic strokes, and rheumatoid arthritis. It is a natural physiologic barrier seen in the microenvironment of solid tumors that has significantly restricted the therapeutic impact of photodynamic treatment (PDT). For oxygen-consumption photodynamic treatment (PDT), local hypoxia is an unfavorable obstacle in tumors. Angiogenesis, invasion, metastasis, and chemotherapy resistance are all affected by a shortage of oxygen. Furthermore, PDT has the possibility of aggravating hypoxia. Meanwhile, the troublesome aggregation-caused quenching effect affects the photosensitizer (PS) compounds utilized in PDT applications and dramatically reduces the efficiency with which deadly reactive oxygen species are produced. Over recent decades, hypoxia’s potential as a therapeutic target has become more widely recognized. Because of their benefits in safety, target specificity, and tumor penetrability, peptides have been intensively explored to treat these disease conditions. Low drug/energy delivery effectiveness, drug resistance brought on by hypoxia, and tumor nonspecificity can all be mitigated using peptides. Three basic methods have recently been applied to use peptide-based nanomaterials to target hypoxia: (i) using hypoxic microenvironment-sensitive peptide linkers that could be split to liberate medicinal payloads; (ii) the merger foregoing, in which earmarks peptides to direct the system to hypoxic surroundings, allowing for selective cleavage; and (iii) target cellular environments using peptide ligands that are particular for a hypoxic environment, such as receptors of the cell surface that are upregulated. In this paper, we go over the advantages and limitations of using peptide-based hypoxia-targeting nanomaterials in a variety of therapeutic situations. Keywords Peptide-based nanoparticles · Photodynamic therapy · Hypoxia · Cancer · Solid tumors P. Kumari · P. Sharma · N. K. Sharma (B) Department of Bioscience and Biotechnology, Banasthali Vidyapith, Tonk, Rajasthan 304022, India e-mail: [email protected]; [email protected] Y. Srivastava Department of Genetics, University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_4

59

60

P. Kumari et al.

Abbreviations PEG H2 O2 DOX CC O2 SR AVT-NP TPC TG2 ELPs PLA PGA PLGA PCL DA ROS PFOB HPAO SPECT NPs DSPC POPE CRT EPR NADs PDT NI AZR NA AZO

Polyethylene Glycol Hydrogen Peroxide Doxorubicin Cytochrome Oxygen Singlet Oxygen Responsive Angiogenesis Vessel Targeting Nanoparticle Tris Phenyl Chlorine Transglutaminase Elastin-Like polypeptides Polylactic Acid Polyglycolic Acid Polylactic Acid Glycolic Acid Polycaprolactone Dimethylmaleic Anhydride Reactive Oxygen Species Perfluorooctyl Bromide Hydroxyphenyl Propionic Acid-OSu Single Photon Emission Computed Tomography Nanoparticles Distearoyl-Sn Glycero-3-Phosphocholine Palmitoyl Oleoyl phosphatidylethanolamine Calreticulin Enhanced Permeability and Retention Nitrobenzyl Alcohol Derivatives Photodynamic Therapy Nitro Imidazole Azoreductase Nitrobenzyl Alcohol Azobenzene

1 Introduction The “Achilles’ heel” of conventional photodynamic therapy (PDT) is hypoxia, a very common characteristic of most solid tumors (Li et al. 2018a, b) that significantly reduces the effectiveness of treatment completely. The unrestrained growth of tumors guzzles both oxygen nutrients, leaving an inadequate need for O2 and less oxygen to relocate to different body parts. The growth of blood vessels is stimulated by the multiplication of tumor cells, but because these blood vessels have aberrant macro- and microstructures, it is difficult to control the delivery of oxygen (Zhao et al.

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

61

2020). To hasten the growth of tumor cells, the tumor cells directly cause healthy cell necrosis and boost-related pathways, such as development stimulation, tissue infiltration, metastasis, and genomic instability (Semenza 2011). Higher levels of hypoxia in cancers reduce therapeutic response and are correlated with tumor aggressiveness, invasion, forecast, and resistance to radiation and chemotherapy (Brown and Wilson 2004; Zhou et al. 2021). Oxygen heightens the cytotoxicity of the cell damage caused by several anticancer medicines. As a result, hypoxia has evolved into both the primary target of cancer treatment to slow tumor growth and a major obstacle to overcome in the creation of novel medicines that do not lead to treatment resistance (Wilson et al. 2011). Peptides are short oligomers made of amidebonded amino acid units (Dehsorkhi et al. 2014). They are crucial proteins’ building blocks and the reason for the structure and operation of living things. Due to their unusual interaction with other organic components, tissues, and cells, peptides are nanostructured and biodegradable, which enhances their advantages for biomedical properties (Wong et al. 2014). The secondary and tertiary structures of peptides can be precisely tuned thanks to the flexibility of amino acid side chains. Larger cell penetration and self-assembling abilities are caused by this modification. Additionally, these minor assemblies can also result in linkages among linkers, and they also contain helices and sheets (Zhang 2017). Peptides can form nanostructures with a wide surface area, such as nanofibers and micelles, which allow for the coupling of the drug and imaging agent. Thus, cell penetration is increased. Additionally, these peptides’ production can be started under some circumstances, offering flexibility and control. Novel and inventive peptide-based materials have been invented for the treatment of the syndrome (Rudra et al. 2012; Sun et al. 2018a; Zhao et al. 2017). For increasing effectiveness in antitumor treatments, they carry out tasks including drug delivery, detection, cell localization, deep tissue invasion, and immune reactions. By integrating several capacities including reactive cleavage terminal, cellular localization, and other features, peptides might also be involved in elements of synthesized complex molecules (Nasrolahi et al. 2013; Zhang et al. 2015, 2019). These peptides have been put to use in the treatment of disorders associated with hypoxia in three primary ways. The first involves using a targeting ligand for therapy, which typically involves focusing on a particular cell, tissue, or microenvironment to achieve the desired impact of the peptide. Most of these tactics use the peptide itself as the aimed ligand (Dissanayake et al. 2017; Gilad et al. 2016). Additionally, these tactics typically involve a cytotoxic tiny molecular peptide that either promotes cytotoxic effects or provides shielding from hazardous environmental stressors, for example, hypoxia, and provides a way of aiming the target over non-target cells. The second strategy uses hypoxic-sensitive linkers that can release therapeutic or imaging payloads when hypoxia is detected. NTR (nitro reductase), AZR (azoreductase), and QR (quinone reductase) are only a few of the bio-reductases that are overexpressed as a result of the extreme situation of hypoxia, which is inferable stress (Xu et al. 2013, 2017). Small compounds found in cellular environments that have lower functional groups can accept electrons such as nitro, azo, and quinone, and undergo a reduction in a hypoxic environment (Yang et al. 2014). The lowering of these hypoxia-responsive components can change the physicochemical and nanosystem properties, like the

62

P. Kumari et al.

size of particles, fluorescence, and hydrophilicity. In conclusion of this information, remarkable progress in nanotechnology has been made by including hypoxiasensitive moieties such as NI (nitroimidazole) (Thambi et al. 2014), NA (nitrobenzyl alcohol) (Thambi et al. 2016; Zheng et al. 2018), and Azo derivatives (azobenzene) (Zhou et al. 2018) to enhance the reaction to bio-reductase in hypoxic cells (Shah et al. 2018). Peptides can indeed be joined to medications or imaging agents using stimuli-responsive components that will tolerate payload produced in a particular environment into the targeted cells. Combining the above tactics results in the third potential way to use peptides (Pearce et al. 2012).

2 Polypeptide-Based Nanoparticles A type of biological polymer having amino acid structural units is called a polypeptide. Polypeptides have several advantages over conventional synthetic polymers, including the capacity to adjust polarity and charge, the ability to withstand hydrolysis, the ease of synthesis, and the ability for enzyme biodegradation in vivo. These protein-like polymers take on many of the traits that proteins possess, including bioactivity, adaptability, biocompatibility, and hierarchical assembly characteristics (Deng et al. 2014; Ou et al. 2018; Yu et al. 2015). Additionally, the solubility, processability, and antifouling properties of biological materials can be improved when polypeptides and synthetic polymers are combined, considerably expanding their range of applications (Canalle et al. 2010). Hypoxia-reactive strategy like (NI, Azo, or NA) inside the polypeptide side chain might enhance miscibility, hydrophobic drug entrapment, and hypoxia sensibility due to its water-hating (hydrophobic) outcome, proceed in choosy drug release at the hypoxic regions. There have been numerous reports of the use of polypeptides with hypoxia-reactive NI motifs in medication delivery and tumor treatment (Ahmad et al. 2016; Deng et al. 2018; Yin et al. 2018). For improved glucoseresponsive insulin administration, Yu et al. created novel hypoxia and hydrogen peroxide double-sensitized polymersome-depended vesicles. The vesicles, which might contain rDNA and GOX /GOD, are self-combined with a diblock copolymer of PEG and NI-altered polyserine interconnected with thioether. A local hypoxic environment was immediately created during the procedure of enzymatic change of glucose into gluconic acid that encouraged the organic drop of NI to hydrophilic 2aminoimidazoles. The excess H2 O2 was removed by the polymer’s thioether moiety, which also encouraged the disintegration of vesicles and the release of the entrapped insulin. The glucose-responsive polymersomes could be paired with a comfortable microneedle-array patch technology for insulin administration (Yu et al. 2017). A hypoxia-responsive nanomaterial was created by combining a cationic lipid-like substance with NI-modified polypeptide methoxy poly (ethylene glycol)-block-poly (l-glutamine-graft-2-nitroimidazole) (mPEG-b-(PLG-g-NI)), which could be used to transport siRNA to breast tumor cells to silence the expression of the cell partition

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

63

cycle 20 gene (CDC20). Long-lasting blood circulation, significant tumor accumulation, effective CDC20 silencing, and highly effective antitumor activity were all displayed by the delivery nanosystem (Li et al. 2020). The literature frequently refers to nitrobenzyl alcohol derivatives (NADs)based hypoxia-responsive polypeptides. One of these, as investigated by Park and colleagues, was the development of hypoxia-perceptive polymeric micelles constructed of the amphiphilic block copolymer poly (ethylene glycol)-poly (-(4nitro) benzyloxycarbonyl-l-lysine) (PEG-b-PLys-g-NBCF) (Thambi et al. 2016). The copolymer may self-combine into micelles in a liquid environment and entrap doxorubicin (DOX). A 1,6-elimination process swiftly breaks down the DOX-loaded micelles under hypoxic conditions, causing a quick production of DOX into the cells. By combining poly(l-lysine)-block-poly (ethylene glycol) (PCL-b-PEG) and poly(l-lysine)-block-poly (l-nitrobenzyl chloroformate), the Shi group produced a complicated micelle featuring a NA motif (PCL-b-PLL-g-NBCF). The micelle strategy has a hypoxia-responsive shell and a PCL core (NBCF-modified PLL and PEG). The arrangement of this core–shell could slow down the immune system’s ability to remove substances quickly during the blood circulation process. When these micelles arrive at the tumor location, the NBCF-modified PLL can begin to break down in the hypoxic microenvironment, increasing the amount of positively stimulating PLL on the micelle area and permitting the micelles to enter cancer (Zhen et al. 2019). In addition to the DOX-loaded micelles have also enhanced tumor tissue hindrance and penetration actions that are demonstrated further in in-vivo and in-vitro studies. Hypoxia-responsive polymer is another instance in the creation of the mPEG-PLG-NC by Zhang et al. by coupling hydrophobic 4nitrobenzyl (3-aminopropyl) carbamate (AP-NC) to the adjacent cables of methoxy poly (ethylene glycol)-b-poly (-propargyl-l-glutamate) (MPEG-PPLG) co-polymers. The self-collected polymer into the nanomaterials in a liquid solution was able to load DOX with a higher entrapment efficiency of about 97.8% because of the intense interaction among the p-nitrobenzyl group and DOX. The DOX-loaded nanoparticles (PPGN@DOX) responded to hypoxia in vitro by releasing drugs. In a hypoxic setting, 4T1 cells can efficiently internalize PPGN@DOX and release DOX into the cell core. Additionally, there were more effective antitumor consequences and fewer adverse effects in vivo (Zhang et al. 2020). In subsequent research, Sun et al. made polymeric micelles those self-assembled utilizing a NA-modified hypoxia-sensitive polypeptide and the (diethylenetriamine-graft-4nitrobenzyl chloroformate)-l-glutamate [mPEG-b-P(Deta-NBCF) LG] amphiphilic polymer methoxy PEG-block-poly to provide cancer cells access to the metalloprotein cytochrome C (CC). The above mentioned metalloprotein cytochrome C-packed micelles effectively absorb tumor cells, and malignant cells under hypoxic environments (Sun et al. 2020). Most nanocarriers contain a steric PEG protection coating to ensure the constancy of the approach for protein adsorption (Suk et al. 2016). However, the PEG layer typically prevents cells from absorbing nanocarriers, which reduces the effectiveness of photodynamic treatment (PDT) (Blanco et al. 2015). An efficient way to strike a balance between method stability and cellular absorption of nanocarriers is to design a sheddable PEG (dePEGylation). Li et al. described using

64

P. Kumari et al.

Fig. 1 a The hydrophilic PEG block and the hydrophobic polypeptide block were joined with the hypoxia-responsive azo linker to create self-assembling amphiphilic copolymer; Ce6 served as a model photosensitizer; and imidazole and Azo moieties are SR and hypoxia-responsive, correspondingly. b As micelles enter tumor, their understanding toward hypoxia might allow for their DePEGylation and greater cellular uptake, while their sensitivity to singlet oxygen might result in their breakdown and rapid release of Ce6

singlet O2 dual-sensitive multifunctional micelles and hypoxia to improve anticancer PDT. The micelles were made using an amphiphilic methoxy poly (ethylene glycol)azobenzene-poly (aspartic acid) copolymer coupled with imidazole (IM) side chains (MPEG-Azo-PAsp-IM). Under hypoxic conditions, Azo and IM are singlet oxygenresponsive (SR), correspondingly. Chlorin e6 (Ce6), a typical photosensitizer, which may be effectively loaded by the micelles. Polypeptide-derived hypoxia-responsive amphiphilic block co-polymers might gather into nanotransporters with diverse anatomy for the exact production of medicines in hypoxic surroundings. These nanocarriers demonstrated substantial tumor accumulation, higher penetrability, and improved inhibitory efficacy on tumor cells as a result of the passive targeting effect (Fig. 1) (Li et al. 2018a, b). As a result, these sophisticated polypeptide-based hypoxia-targeting nanosystems offer a viable method for treating cancer precisely.

3 Self-Assembled Peptide-Based Nanoparticles Several distinct supramolecular nanostructures, including nanotubes, nanofibers, vesicles, and hydrogels, can be spontaneously built by self-assembling peptides (Zhao et al. 2019). Through non-covalent interactions such as hydrogen bonding, van der Waals interactions, electrostatic interactions, and hydrophobic properties,

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

65

the self-gathered construction is retained at a stable low-energy level (Lee et al. 2019). Examples of internal or external stimuli that can successfully regulate how peptides self-assemble include the potential of hydrogen, temperature, ionic force, metal ion complexation, and enzyme actions (Feng et al. 2017; Wang et al. 2017a, b; Yang et al. 2018). Depending on the characteristics of each self-assembled peptide, these nanostructures can be employed in numerous biomedical applications, like drug distribution, material manufacturing, and renewing medicine (Boekhoven and Stupp 2014). By including hypoxia-responsive motifs/peptide ligands that target specific regions of the peptide, the self-assembled peptide nanoparticle could be applied to hypoxia disease detection and treatment. One illustration is Ikeda et al. explorations of pro-apoptotic peptide amphiphiles which might result in self-assembled nanomaterials (Ikeda et al. 2015). The peptide amphiphile AVPI-NP-C12 consisted of pro-apoptotic AVPI tetrapeptide coupled to the hydrophobic dodecyl chain by a nitrophenyl (NP) linker. AVPI-NP-C12 produced self-assembling nanoparticles with nanofiber messes when poly-l-lysine was present. The electrostatic contact among cations and carboxylate anion of AVPI-NP-C12 is thought to be the mechanism causing this. Sodium dithionite, a tumbling representative used to imitate hypoxic atmosphere, can be used to diminish the NP motif because it causes AVPI-NP-C12 to disintegrate and produce a pro-apoptotic peptide. By blocking apoptosis protein inhibitors, apoptotic peptides can increase the efficacy of antitumor suppositories (Arnt et al. 2002). As a result, pro-apoptotic self-assembling peptide nanostructures may be used in nanomedicine to encapsulate and distribute pharmaceuticals as well as to detect hypoxic situations. The performance of the constructed nanostructures in nanomedicine can be significantly impacted by the charges of peptides. The nanostructures can be shielded by a negative charge from being cleaned before they reach the tumor tissue. The nanostructures, however, are challenging for target cells to ingest because of the cytomembrane’s negative potential (Ernsting et al. 2013). Considering this notion, Wang et al. investigated the use of GA-Cy7-NP, a selective-release “mosaic-type” nanomaterial, to target hypoxic cancer cells. These nanoparticles were made using surfactin and a single aqueous solution conjugation of the dye heptamethine carbocyanine (Cy7) and gambogic acid (GA). The system’s carrier platform is surfactin, while its hypoxic goal group and anticancer drug, correspondingly, are represented by Cy7 and GA. The Cy7 assembly is added to the surface of negatively charged electric nanomaterials made by a surfactant for choosy production of a drug complex into hypoxic malignancy without internalizing elements. In addition, GA-Cy7-NP exhibited characteristics of prolonged circulation and considerable tumor cell absorption. In terms of tumor growth, angiogenesis, and cell proliferation, it has a stronger anticancer impact than the prototype drug (Wang et al. 2019a; b). The study by Zhu and colleagues, who created an angiogenic vesselaiming nanomaterial (AVT-NP) comprising photosensitizer 5-(4-carboxyphenyl), trisphenylchlorin (TPC), angiogenic vessel-aiming cyclopeptide (peptide sequence: CGNSNPKSC), and the bio-reductive substance TPZ, provides another illustration of the use of self-assembled peptides. The photosensitizer can produce a significant number of ROS (reactive oxygen species) when bare to radiation, which will

66

P. Kumari et al.

diminish TPZ to create harmful free radicals. Through the targeting of angiogenesis vessels, in vivo tests showed that AVT-NP can preferentially aggregate around tumor cells. With the chemo-photo synergistic effect, the system also demonstrated improved antitumor effectiveness (Guo et al. 2017). For long-term cancer biological imaging, the intracellular self-assemblage of nanoparticles, which has a great retaining rate and negligible mortality, holds significant potential (Zhang et al. 2019). Li and coworkers developed a peptide-based probe for identifying hypoxic neuroblastoma cells using intracellular transglutaminase (TG2)-driven catalyzed replication and in situ self-assemblage of polymerized elastin-like polypeptides (ELPs). The N- and C-terminal amino-acids polymeric binding locations (such as Q and K) and elastin reactive groups make up the polymeric monomer peptides in this arrangement (XGVGP or GYGXP). To fine-tune the scheme for the extreme influence on TG2catalyzed polymerization into ELPs, a variety of variables, including peptide arrangements, temperature delicacy, and the upper critical solution temperature (UCST), are being investigated. In cells that overexpress TG2, intramolecular polymerization and self-assemblage could boost holding productivity and intracellular buildup (such as HeLa). Because hypoxia increases TG2 expression, FITC-labeled peptide probes could precisely image hypoxic neuroblastoma cells for diagnostic reasons (Fig. 2) (Peng et al. 2019). Self-assembling peptides can construct adaptable supramolecular nanostructures when they are activated by biological circumstances (Peng et al. 2019). With the help of these pericellular nanostructures, it is possible to change how effectively enzymes interrelate with cells and how much of them are engaged by cells (Potocky et al. 2005). A self-assembling peptide-based carbonic anhydrase (CA) inhibitor that precisely networked with the upregulation of CA on hypoxic malignancy sheaths was recently developed by Li et al. This substance improved the inhibition and choosiness of CA suppressor by using the hypoxia-triggered self-assembly technique. Self-gathering CA IX inhibitor N-pepABS was formed by uniting the economically existing CA inhibitor 4-(2-aminoethyl) benzenesulfonamide (ABS) with the self-assembling motif [2-naphthaleneacetic acid-(d)-Phe-(d)Phe-(d)-Lys-OH (N-pep)]. Inhibitors of CA IX can focus on the membrane of malignancy cells that are hypoxic and disrupt those cells by using N-pepABS, which can target CA IX and self-combined into nanofibers. Through CA IX-mediated endocytosis, these CA-triggered nanofibers encourage cellular acceptance. Under low pH conditions, nanofibers may expand during the internalization process to produce larger nanofiber bundles. The bundles will subsequently harm the intracellular acid vesicles and prevent the defensive process of autophagy. These findings demonstrate that extremely choosy poisonousness to hypoxic malignancy cells is produced by the cell milieu-triggered adjustable nanostructure. Additionally, in murine 4T1 breast malignancy cells, N-antimetastatic pepABS and antiangiogenic properties were assessed. Both the amount of the tumor and the number of lung metastases were successfully diminished with N-pepABS. After being treated with N-pepABS, the intact tumor vasculature turned dissociated upon the exposure of endothelium marker CD31 and became muted. Additionally, N-pepABS therapy can fruitfully make tumors responsive to the injection of doxorubicin (Dox), and this has been demonstrated to greatly increase antitumor efficacy (Li et al. 2019a; b).

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

67

Fig. 2 Showing hypoxic neuroblastoma cell imaging, specific transglutaminase 2 (TG2)-catalyzed intracellular polymerization and temperature-induced in situ self-assemblage

4 Peptide and Polymer Conjugate Nanoparticles The type of easy compound known as peptide–polymer conjugates is created when peptides and polymers are covalently linked together. In these constructions, polymer conjugation provides the peptides processability under many situations (such as solvent, temperature, and gravity), whereas the peptide gives the peptide– polymer conjugate precision (such as sequence control and low dispersity) (Taylor and Jayaraman 2020). As a result of their biocompatibility, biodegradability, and automatic power, a variety of polymers, including polylactic acid (PLA), polyglycolic acid (PGA), poly (lactic acid glycolic acid) (PLGA), and poly(-caprolactone) (PCL) united with PEG, have been broadly used in peptide–polymer conjugate studies (Milane et al. 2011; Soleymani Abyaneh et al. 2018). When tumor-aiming peptides are combined through these polymers, the attraction for receptor, cell internalization, and tissue penetrating capacity must be improved (Lee et al. 2019; Teesalu et al. 2013; Vasconcelos et at. 2015). These qualities explain why peptide– polymer conjugate materials are used in various medicinal applications, including medication administration, tumor therapy, gene transfer, and antibacterial coatings (Dube et al. 2014; Sun et al. 2018a, 2018b). Recent years have seen an increase in research on peptide–polymer conjugates used in tumor hypoxia therapy. One such pattern focuses on motifs that react to hypoxia. An idea of such a scheme is an effort of Mallik and colleagues, who established tissue-stifling, hypoxiaresponsive polymersomes that may well transport antitumor drug gemcitabine to

68

P. Kumari et al.

compact tumors. Tissue-stifling peptide iRGD (peptide sequence: CRGDKGPDC)functionalized PLA-PEG polymer and the hypoxia-responsive Azo-integrated PLAPEG polymer make up the polymersomes. Drug-entrapped polymersomes effectively condensed the survival of pancreatic tumor cells (Kulkarni et al. 2018) and triple-negative breast tumor cells (Mamnoon et al. 2021) in round cultures while effectively releasing the entrapped substances in vivo and in vitro under hypoxic environments. Ma et al. carried out additional studies in this area and developed a unique AZR-responsive nanoprobe for the precise tomography of mitophagy in existing cells under hypoxia by entrapping mitochondria-targeted rhodamine spirolactam derivatives (Mito-rHP). Azo-N-[maleimide (polyethylene glycol)2000)-AzoN-[maleimide(polyethylene glycol)2000] is composed of 1,2-distearoyl-sn-glycerol3-phosphoethanolamine. The micelle M was self-assembled using (DSPE-Azo-PEGMal) (hypoxia-responsive amphiphilic polymer) (Mito-rHP). The nanoprobe was then functionalized with a cell-stifling peptide (TATp, RKKRRQRRRC), which permitted the nanoprobe to penetrate the cytoplasm without the need for a receptor and avoid being constrained in the endosome. In hypoxic conditions, the highly expressed AZR interfered with the nanoprobe. The entrapped probe Mito-rHP was able to discover mitochondria after being released. Since mito-rHP is also pH-sensitive, it undergoes protonation during mitophagy, resulting in a fluorescent signal to change from “off” to “on” (Ma et al. 2019). Jiang and colleagues created a hypoxia-responsive drug transport scheme in combination with PDT and bio-reductive chemotherapy in another study using the Azo motif. To create the polymer DATAT-PEG-PLGA, the delicate amphiphilic polymer monomethoxy PEGazobenzene-PLGA (PEG-Azo-PLGA) was primarily created. It was then coupled with TAT peptide (peptide sequence: YGRKKRRQRRRC-NH2) and 2,3-dimethyl maleic anhydride (DA). Then, using PEG-Azo-PLGA and DATAT-PEG-PLGA, TAT + AzoNPs nanoparticles were created. TAT + AzoNPs can maintain the payloads C6e and TPZ and aggregate inside tumor cells because the TAT peptide is linked to the surface. TAT + AzoNPs successfully performed PDT on tumor cells close to arteries under laser irradiation and with an adequate supply of oxygen. In addition, oxygen consumption during PDT created a hypoxic milieu that could cause TPZ to be released by breaking the azobenzene link and speeding up TPZ activation, enhancing the efficacy of the combination therapy in tumor cells further from the vasculature (Ihsanullah et al. 2020). Liu et al. recently discovered ROS (reactive oxygen species)-sensitive aryl boronic ester-depended nanomaterials altered with the RBC membrane and iRGD peptide. Ce6 and TPZ can be co-encapsulated in these nanocarriers to obtain tumor-specific release and cooperative PDT (Liu et al. 2019). Zhao et al. established deep burrowing and O2 self-sufficient PDT nanomaterials that depended upon peptide–polymer complexes for stabilization in the circulation of reactive oxygen species in cancers. These nanomaterials (CNP/IP) are created by mixing peptide (sequence: CRGDK)-PEG-PCL and PEG-PCL and encasing them in IR780, a photosensitizer, and perfluorooctyl bromide (PFOB), a synthetic blood replacement with an advanced oxygen storage volume. The alteration of the CRGDK peptide on the encapsulated IR780 and PFOB greatly enhanced their accumulation and penetration in tumor’s core. The release of oxygen by PFOB in hypoxic areas

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

69

significantly reduced hypoxia and improved the effectiveness of PDT (Zhao et al. 2018). A multifunctional platform for battered anticancer and antimetastasis treatment was created by Song et al. in a different work using 131 I-labeled dendrimers altered in the LyP-1 peptide (peptide sequence: CCGNKRTRGC (C2-C10)) (Song et al. 2020). Generation 5 (G5) poly(amidoamine) dendrimers remained used to create the nanosystem. These dendrimers were subsequently coupled with PEG-LyP-1 and 3-(4, -hydroxyphenyl) propionic acid-OSu (HPAO), and enduring dendrimers being radiolabeled in conjunction with 131 I. Single-photon emission computed tomography (SPECT) imaging diagnostic probe for the 131 I-labeled LyP-1-modified dendrimers showed remarkable biocompatibility. A transporter for antitumor and antimetastatic treatment, the LyP-1 peptide also can identify tumor cells in hypoxic parts and alleviate hypoxia. The nanotheranostics device demonstrated precise targeting in vivo and at tumor locations, and it greatly slowed tumor growth and spread in vitro, further demonstrating its potential therapeutic utility (Fig. 3). In solid tumors, a hypoxic microenvironment typically greatly lowers the chemosensitivity of cancer cells. A novel combination therapy tactic was recently given by Zhang et al. to address the drug confrontation of gastric tumors equally in vitro and in vivo (Zhang et al. 2020a; b). For improving the chemotherapeutic result of apatinib on gastric cancer, PLGA nanoparticles (NPs) coloaded salidroside (Sal) and apatite (Apa). The tumor-recognizable peptide iVR1 was supplementarily improved on the NPs-Apa/Sal to increase the efficacy of medication delivery. By particularly targeting the vascular endothelial development factor receptor 1 (VEGFR1) (Cicatiello et al. 2015), the iVR1 peptide can prevent angiogenesis and the spread of some malignancies. The peptide-altered iVR1-NPs-Apa/Sal demonstrated admirable tumor-aiming drug delivery capabilities and had a potent inhibitory effect on tumor progression in vivo as well as cell proliferation, invasion, and migration.

Fig. 3 Representative diagram of peptide and polymer conjugate nanoparticles (Petho et al. 2020)

70

P. Kumari et al.

5 Peptide-Functionalized Liposomes Lipid bilayers make up their structure. Both the hydrophobic molecule in the bilayer and the hydrophilic compound in their core can be contained by these structures (Gonzalez Gomez and Hosseinidoust. 2020). As a result of their biocompatibility, capacity for self-gathering, and capacity in controlling inactive pointing by increasing the penetrability and retention in cancer regions, liposomes are prospective schemes intended for drug and gene transport, ophthalmology, vaccines, imaging, and cosmetics (Ahmed et al. 2019; Akbarzadeh et al. 2013; Zununi Vahed et al. 2017). A targeting peptide added to liposomes improves the biocompatibility of nanocomplexes and increases their aiming specificity and transfection efficacy in cells (Fig. 4) (Yu-Wai-Man et al. 2016). These peptideliposomal systems that target hypoxia have been investigated for medication delivery (Kale and Torchilin et al. 2007), gene delivery (Ko et al. 2009), and tumor therapy (Wang et al. 2019a; b). Liposomes made of 1,2-distearoyl-sn-glycerol-3-phosphocholine (DSPC), palmitoyl oleoyl phosphatidylethanolamine (POPE)-Azo-PEG, a hypoxia-responsive PEGylated lipid, and an iRGD peptide-linked lipid DSPE-PEG-iRGD were created by Kulkarni et al. Liposomes were able to transport the anticancer medication to the hypoxic cores by penetrating deeper thanks to the iRGD peptide on the surface. The integrity of the lipid membrane was disrupted under hypoxic conditions, releasing the medication from liposomes and increasing the cytotoxicity of the prepared pancreatic tumor cell (Kulkarni et al. 2016). This was due to the Azo moieties of hypoxiaresponsive lipids being diminished. For the treatment of hypoxic tumors, liposomebased nanoplatforms with diverse receptive and functional capabilities have been reported. The amphiphilic molecule (mPEG-Ce6-C18), lecithin, and DSPE-PEG-cRGD (peptide sequence: RGD-D-FK) were utilized to self-assemble a liposome; according to a study by Dai et al. ICG/TPZ@Ce6-GdIII, a multiuse theranostic liposome can be created by encapsulating indocyanine green (ICG) and the hypoxia-activated prodrug tirapazamine (TPZ) in a liposome. These multifunctional liposomes can be employed as PTT (photo-thermal therapy)-PDT-stimulated anticancer mediators in addition to being a multimodal imaging contrast agent. Effectively reducing adverse effects on normal cells is possible using cRGD targeting and photothermally activated PDT. Additionally, TPZ could boost the medicinal effect, permitting this special nanosystem to selectively treat tumors using a variety of techniques (Dai et al. 2019). Wang et al. achieved hypoxia-activated chemotherapy in conjunction with PDT in contradiction to metastatic breast tumors by instantaneously transporting of ICG and TPZ to dense tumors by iRGD-altered liposomes (Wang et al. 2017a; b). In a different work, a double endoplasmic reticulum aims strategy to reach PDT-PTT-immunotherapy. The modified ER-targeting padaxin (FAL) peptides, oxygen-transporting HB liposomes (FAL-HB-lipo), and hollow gold nanospheres (FAL-ICG-HAuNs) made up the nanoparticle that was intended to inverse hypoxia.

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

71

Fig. 4 Representative diagram of peptide and polymer conjugate nanoparticles (Sonju et al. 2021)

The ER-directing nanoparticles can cause significant endoplasmic reticulum stress and calreticulin (CRT) revelation proceeding the cell shallow when exposed to NIR light. CRT exposure is an indication that promotes an increased immunological retort, with CD8+ T cell creation and the release of cytotoxic cytokines, and increases the growth of naive dendritic cells. Endoplasmic reticulum-directing PDT-PTT enhances immunotherapy related to immunogenic cell demise by direct reactive oxygen species-depended endoplasmic reticulum stress and improved antitumor effectiveness (Li et al. 2019a; b). Numerous fluorescent reductive probes have been developed and effectively used to track the hypoxic condition of tumor cells. These probes take advantage of numerous reductases being overexpressed in hypoxic conditions (Elmes 2016; Yang et al. 2020; Zhu et al. 2019). Fan et al. recently established a liposome-dependent nanoprobe functionalized with a peptide to aim heart cells and entrap them with nitrobenzene-substituted BODIPY (BDP-NO2) for myocardial hypoxia imaging (GGGGDRVYIHPF). The AT1 angiotensin-II type-1 receptor, which is upregulated in ischemic cardiac cells, can be selectively targeted by the technique. The cardiac cells absorb the nanoprobe and release BDP-NO2. The nitro group of BDP-NO2 is converted through NTR from BDP-NO2 to BDPNH2 , which enhances the nanoprobe’s fluorescence under hypoxic circumstances. The nanoprobe can also be used to image hypoxia levels in real time in an animal (mouse) model of myocardial chlorosis (Fan et al. 2019). Yao et al. investigated a liposome nanosystem for combination therapy using real-time therapeutic monitoring

72

P. Kumari et al.

to target hypoxic tumors with great efficacy and to image apoptosis in real time. In this study, cyclopeptide RA-V (deoxybouvardin), a chemotherapeutic medication, antisense oligonucleotides (RX-0047), and a caspase-8 probe were co-entrapped in a pH-sensitive liposome. Through receptor-mediated endocytosis, the acquired liposomes could specifically infiltrate colon cancer cells’ lysosomes, where the lysosomal acidic milieu induced the liposomes. While antisense oligonucleotides may diminish HIF-1a synthesis to lessen tumor hypoxia, released RA-V may cause cancer cells to apoptosis via the mitochondrial pathway. The liposome moreover is used for medicinal self-observation using the fluorescence of the caspase-8 probe (Yao et al. 2019).

6 Polysaccharides Functionalization Study by Peptides A class of ordinary macro-molecular polymers formed from stretched cables of monosaccharide components connected by glycosidic bonds is known as polysaccharides (Xie et al. 2016). These are widely used in biomedicine because they are natural biomaterials that are water-soluble, risk-free, non-hazardous, biodegradable, and non-immunogenic (Li et al. 2015; Swierczewska et al. 2016). Hydrophilic functional sets, like the hydroxy and carboxyl groups found in polysaccharides, are amenable to chemical and biochemical modification. Through non-covalent interactions, they might also encourage bio-adhesion with organic tissues, mainly epithelium and mucosa. By extending the time that drugs stay in the mucosa, these bioadhesive polysaccharide-based nanocarriers can increase the bioavailability of the drugs. To actively target medicinal molecules, polysaccharide nanoparticle surfaces can also be improved with ligands (Laha et al. 2019). Among the recently developed polysaccharide-based nanosystems for hypoxic tumor therapy, peptidefunctionalized polysaccharide nanoparticles have been characterized; however, exploration on this extent is not as extensive (Park et al. 2016; Shin et al. 2018; Uthaman et al. 2020; Zhang et al. 2021). As in this case, Shu et al. produced RoY (peptide sequence: YPHIDSLGHWRR) peptide-modified chitosan chloride hydrogel (CSCl-RoY). CSCl-RoY hydrogel can influence cell endurance, production, relocation, and duct formation beneath hypoxic situations by connecting to 78 kDa glucose-regulated protein (GRP78) receptor proceeding the film of human umbilical vein endothelial cells. The pathways for protein kinase B (Akt) and extracellular signal-regulated kinase (ERK1/signaling) are activated to fix this. The research also demonstrated that Roy peptide treatment can improve heart repair in addition to triggering angiogenesis at the infarct site (Shu et al. 2015). One of the particles impacted by hypoxia is CD73 (ecto-50-nucleotidase), as well as HIF-1a, which is indispensable for the expansion and development of cancer superparamagnetic iron oxide (SPION) nanoparticles laden with siRNA, which was created by Ghalamfarsa et al. as a transfer mechanism used in quietening of CD73 and HIF-1a genes for the treatment of malignancy (Gahalamfarsa et al. 2019). To enhance the steadiness and functioning of the siRNA-loaded SPIONs in this nanosystem,

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

73

trimethyl chitosan (TMC) and thiolated chitosan (TC) were used as encapsulants. To improve cellular absorption and create the nanoparticle TAT-TMCTCSPIONs, the generated TMC-TC-SPIONs was coupled through the TAT peptide (C (Npys) GRKKRRQRRR). These nanomaterials could knowingly lower the appearance of HIF-1a and CD73 in tumor cells, which would significantly lessen the cancer cell’s ability to migrate and proliferate. Additionally, in vivo testing revealed that these nanoparticles could successfully stop angiogenesis and tumor growth (Hajizadeh et al. 2020). A hyperbranched cationic amylopectin derivative (DMAPA-Amyp), which has low cytotoxicity and good blood compatibility, can be combined with 3(dimethylamino)-1-propylamine to form an operative gene transporter (Zhou et al. 2012). In the latest study, Deng et al. developed a RGD (peptide sequence: KYGRGDC)-altered targeted gene nanotransporter, RGD-DMAPA-Amyp, to cure chlorosis stroke. RGD peptides were selected, assured to the nanotransporters, and transported to the peri-infarct vascular endothelial cells using the targeting technique. The HIF-1a-altered variant (HIF-1a-AA) would have crammed through RGDDMAPA-Amyp to reduce cerebral chlorosis. The results proved that this nanocomplex could greatly speed up the creation of new blood vessels and nerve activity in vivo and had good biocompatibility (Deng et al. 2019; Liu et al. 2021).

7 Conclusion and Outlook for the Future Finally, utilizing capabilities of the peptide arrangement in detection and therapy offers circumstances to prevail over the difficulties that hypoxia illnesses pose. In addition to the previously mentioned targeted specificity, tumor penetrability, and orderliness of high drug delivery, peptides also have other advantages. We summarized the most recent advances in this work using a variety of peptide-depended hypoxia-aiming nanomaterials, which includes polypeptides, prefabricated peptides, coalesce of peptide-polymer, peptide-functionalized liposomes, and polysaccharides. Hypoxia-sensitive linkers and peptide ligands, either in combination or separately, are basic tools used to target hypoxia. With the use of this technique, hypoxic cells can be targeted, peptide-based nanomaterials can aggregate in these areas, and the payloads can successfully pass biological barriers. These peptide-based nanomaterials can also increase permeability into angiogenic arteries and tumor tissues, lengthen blood circulation, encourage tumor cell uptake, and boost therapeutic efficacy. Additionally, peptides and their derivatives are readily prefabricated into a variety of nanofunctional substances with various constructions because of intramolecular and intermolecular forces. These materials can enhance the specificity of cancer cells by utilizing the special properties of the hypoxic ambient of cancer cells. Strong evidence from the literature suggests that using such peptidebased nanomaterials might increase the effectiveness of treating hypoxic disorders overall.

74

P. Kumari et al.

This research is still in the early stages, though. Several obstacles currently hinder the therapeutic application of peptide-based nanomaterials for hypoxic disorders. Enzymes may quickly break down peptides, and mononuclear phagocytes can take them up. Peptide circulation times are also shortened by renal filtration. Therefore, it becomes compulsory to contemplate the system motif and manufacturing alteration of such peptide-depended nanomaterials. To specifically and effectively target the hypoxic areas of various hypoxia illnesses, other peptide ligands with novel sequences must be investigated. The increased permeability and retention (EPR) effect is currently being used in various nanomaterials for medical purposes to passively target tumor tissues. The effectiveness of chemotherapy is constrained by low cellular absorption and internalization, despite the EPR effect’s ability to enrich nanocarriers near the tumor site. Consequently, it is vital to relate peptides with different nanoparticles to generate a positively aimed multifunctional nanocomplex system that will intensify cellular absorption and internalization. In hypoxic environments, this will increase the therapeutic synergy of many therapies like PDT, photo-thermal treatment, thermosdynamic treatment, chemotherapy, radioactivity, etc. Importantly, distinct hypoxic microenvironments are present in various hypoxia-related diseases, including tumors, bone tissue inflammation, cardiac dysfunctions, and hemorrhagic strokes. As a result, materials that are specifically induced by the microenvironment and that are based on the characteristics of peptides must be developed. Additionally, the physicochemical properties of nanomaterials may significantly vary as a result of the alteration in the microenvironment in vivo. Therefore, before conducting clinical trials, a systematic valuation of the bioabsorbable, biodistribution, and in vivo intent of these peptidedependent nanoparticles is required. We anticipate and eagerly await the creation of peptide-based hypoxia-addressing nanoparticles because we think they will significantly advance the early detection and management of disorders associated with hypoxia.

References Ahmad Z, Lv S, Tang Z, Shah A, Chen X (2016) Methoxy poly (ethylene glycol)-block-poly (glutamic acid)-graft-6-(2-nitroimidazole) hexyl amine nanoparticles for potential hypoxiaresponsive delivery of doxorubicin. J Biomater Sci Polym Ed 27(1):40–54. https://doi.org/10. 1080/09205063.2015.1107707 Ahmed KS, Hussein SA, Ali AH, Korma SA, Lipeng Q, Jinghua C (2019) Liposome: composition, characterisation, preparation, and recent innovation in clinical applications. J Drug Target 27(7):742–761. https://doi.org/10.1080/1061186X.2018.1527337 Akbarzadeh A, Rezaei-Sadabady R, Davaran S, Joo SW, Zarghami N, Hanifehpour Y, Samiei M, Kouhi M, Nejati-Koshki K (2013) Liposome: classification, preparation, and applications. Nanoscale Res Lett 8(1):102.https://doi.org/10.1186/1556-276X-8-102 Arnt CR, Chiorean MV, Heldebrant MP, Gores GJ, Kaufmann SH (2002) Synthetic Smac/DIABLO peptides enhance the effects of chemotherapeutic agents by binding XIAP and cIAP1 in situ. J Biol Chem 277(46):44236–44243. https://doi.org/10.1074/jbc.M207578200

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

75

Blanco E, Shen H, Ferrari M (2015) Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat Biotechnol 33(9):941–951. https://doi.org/10.1038/nbt.3330 Boekhoven J, Stupp SI (2014) 25th anniversary article: supramolecular materials for regenerative medicine. Adv Mater (Deerfield Beach, Fla.) 26(11):1642–1659. https://doi.org/10.1002/adma. 201304606 Brown JM, Wilson WR (2004) Exploiting tumour hypoxia in cancer treatment. Nat Rev Cancer 4(6):437–447. https://doi.org/10.1038/nrc1367 Canalle LA, Löwik DW, van Hest JC (2010) Polypeptide-polymer bioconjugates. Chem Soc Rev 39(1):329–353. https://doi.org/10.1039/b807871h Cheng R, Liu L, Xiang Y, Lu Y, Deng L, Zhang H, Santos HA, Cui W (2020) Advanced liposomeloaded scaffolds for therapeutic and tissue engineering applications. Biomaterials 232:119706. https://doi.org/10.1016/j.biomaterials.2019.119706 Cicatiello V, Apicella I, Tudisco L, Tarallo V, Formisano L, Sandomenico A, Kim Y, BastosCarvalho A, Orlandi A, Ambati J, Ruvo M, Bianco R, De Falco S (2015) Powerful anti-tumor and anti-angiogenic activity of a new anti-vascular endothelial growth factor receptor 1 peptide in colorectal cancer models. Oncotarget 6(12):10563–10576. https://doi.org/10.18632/oncota rget.3384 Dai Y, Wang B, Sun Z, Cheng J, Zhao H, Wu K, Sun P, Shen Q, Li M, Fan Q (2019) Multifunctional theranostic liposomes loaded with a hypoxia-activated prodrug for cascade-activated tumor selective combination therapy. ACS Appl Mater Interfaces 11(43):39410–39423. https://doi. org/10.1021/acsami.9b11080 Dehsorkhi A, Castelletto V, Hamley IW (2014) Self-assembling amphiphilic peptides. J Peptide Sci: an off Publication Eur Peptide Soc 20(7):453–467. https://doi.org/10.1002/psc.2633 Deng C, Wu J, Cheng R, Meng F, Klok HA, Zhong Z (2014) Functional polypeptide and hybrid materials: precision synthesis via α-amino acid N-carboxyanhydride polymerization and emerging biomedical applications. Prog Polym Sci 39(2):330–364. https://doi.org/10.1016/j.progpolym sci.2013.10.008 Deng J, Liu F, Wang L, An Y, Gao M, Wang Z, Zhao Y (2018) Hypoxia- and singlet oxygenresponsive chemo-photodynamic Micelles featured with glutathione depletion and aldehyde production. Biomater Sci 7(1):429–441. https://doi.org/10.1039/c8bm01042k Deng L, Zhang F, Wu Y, Luo J, Mao X, Long L, Gou M, Yang L, Deng D (2019) RGD-modified nanocarrier-mediated targeted delivery of HIF-1α-AA plasmid DNA to cerebrovascular endothelial cells for ischemic stroke treatment. ACS Biomater Sci Eng 5(11):6254–6264. https://doi. org/10.1021/acsbiomaterials.9b01362 Dissanayake S, Denny WA, Gamage S, Sarojini V (2017) Recent developments in anticancer drug delivery using cell-penetrating and tumour targeting peptides. J Control Release: off J Control Release Soc 250:62–76. https://doi.org/10.1016/j.jconrel.2017.02.006 Dube N, Seo JW, Dong H, Shu JY, Lund R, Mahakian LM, Ferrara KW, Xu T (2014) Effect of alkyl length of peptide-polymer amphiphile on cargo encapsulation stability and pharmacokinetics of 3-helix micelles. Biomacromol 15(8):2963–2970. https://doi.org/10.1021/bm5005788 Elmes RB (2016) Bioreductive fluorescent imaging agents: applications to tumour hypoxia. Chem Commun (camb) 52(58):8935–8956. https://doi.org/10.1039/c6cc01037g Ernsting MJ, Murakami M, Roy A, Li SD (2013) Factors controlling the pharmacokinetics, biodistribution and intratumoral penetration of nanoparticles. J Control Release: off J Control Release Soc 172(3):782–794. https://doi.org/10.1016/j.jconrel.2013.09.013 Fan Y, Lu M, Yu XA, He M, Zhang Y, Ma XN, Kou J, Yu BY, Tian J (2019) Targeted myocardial hypoxia imaging using a nitroreductase-activatable near-infrared fluorescent nanoprobe. Anal Chem 91(10):6585–6592. https://doi.org/10.1021/acs.analchem.9b00298 Feng Z, Wang H, Chen X, Xu B (2017) Self-assembling ability determines the activity of enzymeinstructed self-assembly for inhibiting cancer cells. J Am Chem Soc 139(43):15377–15384. https://doi.org/10.1021/jacs.7b07147

76

P. Kumari et al.

Ghalamfarsa G, Kazemi MH, Raoofi Mohseni S, Masjedi A, Hojjat-Farsangi M, Azizi G, Yousefi M, Jadidi-Niaragh F (2019) CD73 as a potential opportunity for cancer immunotherapy. Expert Opin Ther Targets 23(2):127–142.https://doi.org/10.1080/14728222.2019.1559829 Gilad Y, Noy E, Senderowitz H, Albeck A, Firer MA, Gellerman G (2016) Synthesis, biological studies and molecular dynamics of new anticancer RGD-based peptide conjugates for targeted drug delivery. Bioorg Med Chem 24(2):294–303. https://doi.org/10.1016/j.bmc.2015.12.020 Gonzalez Gomez A. Hosseinidoust Z (2020) Liposomes for antibiotic encapsulation and delivery. ACS Infect Dis 6(5):896–908.https://doi.org/10.1021/acsinfecdis.9b00357 Guo D, Xu S, Wang N, Jiang H, Huang Y, Jin X, Xue B, Zhang C, Zhu X (2017) Prodrugembedded angiogenic vessel-targeting nanoparticle: a positive feedback amplifier in hypoxiainduced chemo-photo therapy. Biomaterials 144:188–198. https://doi.org/10.1016/j.biomateri als.2017.08.032 Hajizadeh F, Moghadaszadeh Ardebili S, Baghi Moornani M, Masjedi A, Atyabi F, Kiani M, Namdar A, Karpisheh V, Izadi S, Baradaran B, Azizi G, Ghalamfarsa G, Sabz G, Yousefi M, Jadidi-Niaragh F (2020) Silencing of HIF-1α/CD73 axis by siRNA-loaded TAT-chitosan-spion nanoparticles robustly blocks cancer cell progression. Eur J Pharmacol 882:173235.https://doi. org/10.1016/j.ejphar.2020.173235 Ihsanullah KM, Kumar BN, Zhao Y, Muhammad H, Liu Y, Wang L, Liu H, Jiang W (2020) Stepwise-activatable hypoxia triggered nanocarrier-based photodynamic therapy for effective synergistic bioreductive chemotherapy. Biomaterials 245:119982. https://doi.org/10.1016/j.bio materials.2020.119982 Ikeda M, Kawakami M, Kitade Y (2015) Pro-apoptotic peptide amphiphile self-assembled with the assistance of polycations. Chem Lett 44(8):1137–1139. https://doi.org/10.1246/cl.150399 Kale AA, Torchilin VP (2007) “Smart” drug carriers: PEGylated TATp-modified pH-sensitive liposomes. J Liposome Res 17(3–4):197–203. https://doi.org/10.1080/08982100701525035 Ko YT, Hartner WC, Kale A, Torchilin VP (2009) Gene delivery into ischemic myocardium by double-targeted lipoplexes with anti-myosin antibody and TAT peptide. Gene Ther 16(1):52–59. https://doi.org/10.1038/gt.2008.135 Kulkarni P, Haldar MK, Katti P, Dawes C, You S, Choi Y, Mallik S (2016) Hypoxia responsive, tumor penetrating lipid nanoparticles for delivery of chemotherapeutics to pancreatic cancer cell spheroids. Bioconjug Chem 27(8):1830–1838. https://doi.org/10.1021/acs.bioconjchem. 6b00241 Kulkarni P, Haldar MK, Karandish F, Confeld M, Hossain R, Borowicz P, Gange K, Xia L, Sarkar K, Mallik S (2018) Tissue-penetrating, hypoxia-responsive echogenic polymersomes for drug delivery to solid tumors. Chemistry (weinheim an Der Bergstrasse, Germany) 24(48):12490– 12494. https://doi.org/10.1002/chem.201802229 Laha B, Maiti S, Sen KK, Jana S (2019) Nanoscale polysaccharide-based particles for the delivery of therapeutic molecules. In: Green synthesis, characterization and applications of nanoparticles. Elsevier, pp 347–368. https://doi.org/10.1016/B978-0-08-102579-6.00014-9 Lee S, Trinh T, Yoo M, Shin J, Lee H, Kim J, Hwang E, Lim YB, Ryou C (2019) Self-assembling peptides and their application in the treatment of diseases. Int J Mol Sci 20(23):5850. https:// doi.org/10.3390/ijms20235850 Li M, Lin F, Lin Y, Peng W (2015) Extracellular polysaccharide from Bordetella species reduces high glucose-induced macrophage apoptosis via regulating interaction between caveolin-1 and TLR4. Biochem Biophys Res Commun 466(4):748–754. https://doi.org/10.1016/j.bbrc.2015. 09.125 Li J, Meng X, Deng J, Lu D, Zhang X, Chen Y, Zhu J, Fan A, Ding D, Kong D, Wang Z, Zhao Y (2018a) Multifunctional micelles dually responsive to hypoxia and singlet oxygen: enhanced photodynamic therapy via interactively triggered photosensitizer delivery. ACS Appl Mater Interfaces 10(20):17117–17128. https://doi.org/10.1021/acsami.8b06299 Li M, Xia J, Tian R, Wang J, Fan J, Du J, Long S, Song X, Foley JW, Peng X (2018b) Nearinfrared light-initiated molecular superoxide radical generator: rejuvenating photodynamic

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

77

therapy against hypoxic tumors. J Am Chem Soc 140(44):14851–14859. https://doi.org/10. 1021/jacs.8b08658 Li J, Shi K, Sabet ZF, Fu W, Zhou H, Xu S, Liu T, You M, Cao M, Xu M, Cui X, Hu B, Liu Y, Chen C (2019) New power of self-assembling carbonic anhydrase inhibitor: short peptide-constructed nanofibers inspire hypoxic cancer therapy. Sci Adv 5(9):eaax0937. https://doi.org/10.1126/sci adv.aax0937 Li W, Yang J, Luo L, Jiang M, Qin B, Yin H, Zhu C, Yuan X, Zhang J, Luo Z, Du Y, Li Q, Lou Y, Qiu Y, You J (2019b) Targeting photodynamic and photothermal therapy to the endoplasmic reticulum enhances immunogenic cancer cell death. Nat Commun 10(1):3349. https://doi.org/ 10.1038/s41467-019-11269-8 Li Y, Ding J, Xu X, Shi R, Saw PE, Wang J, Chung S, Li W, Aljaeid BM, Lee RJ, Tao W, Teng L, Farokhzad OC, Shi J (2020) Dual hypoxia-targeting RNAi nanomedicine for precision cancer therapy. Nano Lett 20(7):4857–4863. https://doi.org/10.1021/acs.nanolett.0c00757 Liu H, Jiang W, Wang Q, Hang L, Wang Y, Wang Y (2019) ROS-sensitive biomimetic nanocarriers modulate tumor hypoxia for synergistic photodynamic chemotherapy. Biomater Sci 7(9):3706– 3716. https://doi.org/10.1039/c9bm00634f Liu J, Yang Z, Wang J (2021) Recent advances in peptide-based nanomaterials for targeting hypoxia. Nanoscale Adv. https://doi.org/10.1039/D1NA00637A Ma D, Huang C, Zheng J, Zhou W, Tang J, Chen W, Li J, Yang R (2019) Azoreductase-responsive nanoprobe for hypoxia-induced mitophagy imaging. Anal Chem 91(2):1360–1367. https://doi. org/10.1021/acs.analchem.8b03492 Mamnoon B, Loganathan J, Confeld MI, De Fonseka N, Feng L, Froberg J, Choi Y, Tuvin DM, Sathish V, Mallik S (2021) Targeted polymeric nanoparticles for drug delivery to hypoxic, triplenegative breast tumors. ACS Appl Biomater 4(2):1450–1460. https://doi.org/10.1021/acsabm. 0c01336 Milane L, Duan Z, Amiji M (2011) Development of EGFR-targeted polymer blend nanocarriers for combination paclitaxel/lonidamine delivery to treat multi-drug resistance in human breast and ovarian tumor cells. Mol Pharm 8(1):185–203. https://doi.org/10.1021/mp1002653 Nasrolahi Shirazi A, Oh D, Tiwari RK, Sullivan B, Gupta A, Bothun GD, Parang K (2013) Peptide amphiphile containing arginine and fatty acyl chains as molecular transporters. Mol Pharm 10(12):4717–4727.https://doi.org/10.1021/mp400539r Ou Y, Tang ZH, Sun L, Yu HY, Li J, Zhao MH, Xu H (2018) Combretastatin A4/poly (L-glutamic acid)-graft-PEG conjugates self-assembled to nanoparticles. Asian J Pharm Sci 13(2):191–196. https://doi.org/10.1016/j.ajps.2017.10.001 Park W, Bae BC, Na K (2016) A highly tumor-specific light-triggerable drug carrier responds to hypoxic tumor conditions for effective tumor treatment. Biomaterials 77:227–234. https://doi. org/10.1016/j.biomaterials.2015.11.014 Pearce TR, Shroff K, Kokkoli E (2012) Peptide targeted lipid nanoparticles for anticancer drug delivery. Adv Mater (Deerfield Beach, Fla.), 24(28):3803–3710. https://doi.org/10.1002/adma. 201200832 Peng B, Zhao X, Yang MS, Li LL (2019) Intracellular transglutaminase-catalyzed polymerization and assembly for bioimaging of hypoxic neuroblastoma cells. J Mater Chem B 7(37):5626–5632. https://doi.org/10.1039/c9tb01227c Peth˝o L, Kasza G, Lajkó E, Láng O, K˝ohidai L, Iván B, Mez˝o G (2020) Amphiphilic drugpeptide-polymer conjugates based on poly(ethylene glycol) and hyperbranched polyglycerol for epidermal growth factor receptor targeting: the effect of conjugate aggregation on in vitro activity. Soft Matter 16(24):5759–5769. https://doi.org/10.1039/d0sm00428f Potocky TB, Menon AK, Gellman SH (2005) Effects of conformational stability and geometry of guanidinium display on cell entry by beta-peptides. J Am Chem Soc 127(11):3686–3687. https://doi.org/10.1021/ja042566j Rudra JS, Sun T, Bird KC, Daniels MD, Gasiorowski JZ, Chong AS, Collier JH (2012) Modulating adaptive immune responses to peptide self-assemblies. ACS Nano 6(2):1557–1564. https://doi. org/10.1021/nn204530r

78

P. Kumari et al.

Semenza GL (2011) Oxygen sensing, homeostasis, and disease. N Engl J Med 365(6):537–547. https://doi.org/10.1056/NEJMra1011165 Shah A, Malik MS, Khan GS, Nosheen E, Iftikhar FJ, Khan FA, Shukla SS, Akhter MS, Kraatz HB, Aminabhavi TM (2018) Stimuli-responsive peptide-based biomaterials as drug delivery systems. Chem Eng J 353:559–583.https://doi.org/10.1016/j.cej.2018.07.126 Shin SW, Jung W, Choi C, Kim SY, Son A, Kim H, Lee N, Park HC (2018) Fucoidanmanganese dioxide nanoparticles potentiate radiation therapy by co-targeting tumor hypoxia and angiogenesis. Mar Drugs 16(12):510. https://doi.org/10.3390/md16120510 Shu Y, Hao T, Yao F, Qian Y, Wang Y, Yang B, Li J, Wang C (2015) RoY peptide-modified chitosan-based hydrogel to improve angiogenesis and cardiac repair under hypoxia. ACS Appl Mater Interfaces 7(12):6505–6517. https://doi.org/10.1021/acsami.5b01234 Soleymani Abyaneh H, Soleimani AH, Vakili MR, Soudy R, Kaur K, Cuda F, Tavassoli A, Lavasanifar A (2018) Modulation of hypoxia-induced chemoresistance to polymeric micellar cisplatin: the effect of ligand modification of micellar carrier versus inhibition of the mediators of drug resistance. Pharmaceutics 10(4):196.https://doi.org/10.3390/pharmaceutics10040196 Song N, Zhao L, Xu X, Zhu M, Liu C, Sun N, Yang J, Shi X, Zhao J (2020) LyP-1-modified multifunctional dendrimers for targeted antitumor and antimetastasis therapy. ACS Appl Mater Interfaces 12(11):12395–12406. https://doi.org/10.1021/acsami.9b18881 Sonju JJ, Dahal A, Singh SS, Jois SD (2021) Peptide-functionalized liposomes as therapeutic and diagnostic tools for cancer treatment. J Control Release: off J Control Release Soc 329:624–644. https://doi.org/10.1016/j.jconrel.2020.09.055 Suk JS, Xu Q, Kim N, Hanes J, Ensign LM (2016) PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Adv Drug Deliv Rev 99(Pt A):28–51. https://doi. org/10.1016/j.addr.2015.09.012 Sun H, Dong Y, Feijen J, Zhong Z (2018a) Peptide-decorated polymeric nanomedicines for precision cancer therapy. J Control Release: off J Control Release Soc 290:11–27. https://doi.org/10.1016/ j.jconrel.2018.09.029 Sun H, Hong Y, Xi Y, Zou Y, Gao J, Du J (2018b) Synthesis, self-assembly, and biomedical applications of antimicrobial peptide-polymer conjugates. Biomacromol 19(6):1701–1720. https:// doi.org/10.1021/acs.biomac.8b00208 Sun XS, Jang MS, Fu Y, Lee JH, Lee DS, Li Y, Yang HY (2020) Intracellular delivery of cytochrome C using hypoxia-responsive polypeptide micelles for efficient cancer therapy. Mater Sci Eng. C, Mater Biol Appl 114:111069. https://doi.org/10.1016/j.msec.2020.111069 Swierczewska M, Han HS, Kim K, Park JH, Lee S (2016) Polysaccharide-based nanoparticles for theranostic nanomedicine. Adv Drug Deliv Rev 99(Pt A):70–84. https://doi.org/10.1016/j.addr. 2015.11.015 Taylor PA, Jayaraman A (2020) Molecular modeling and simulations of peptide-polymer conjugates. Annu Rev Chem Biomol Eng 11:257–276. https://doi.org/10.1146/annurev-chembioeng-092 319-083243 Teesalu T, Sugahara KN, Ruoslahti E (2013) Tumor-penetrating peptides. Front Oncol 3:216.https:// doi.org/10.3389/fonc.2013.00216 Thambi T, Deepagan VG, Yoon HY, Han HS, Kim SH, Son S, Jo DG, Ahn CH, Suh YD, Kim K, Kwon IC, Lee DS, Park JH (2014) Hypoxia-responsive polymeric nanoparticles for tumortargeted drug delivery. Biomaterials 35(5):1735–1743. https://doi.org/10.1016/j.biomaterials. 2013.11.022 Thambi T, Son S, Lee DS, Park JH (2016) Poly(ethylene glycol)-b-poly(lysine) copolymer bearing nitroaromatics for hypoxia-sensitive drug delivery. Acta Biomater 29:261–270. https://doi.org/ 10.1016/j.actbio.2015.10.011 Uthaman S, Kim Y, Lee JY, Pillarisetti S, Huh KM, Park IK (2020) Self-quenched polysaccharide nanoparticles with a reactive oxygen species-sensitive cascade for enhanced photodynamic therapy. ACS Appl Mater Interfaces 12(25):28004–28013. https://doi.org/10.1021/acsami.0c0 6311

Recent Progress in Hypoxia-Targeting: Peptide-Based Nanomaterials

79

Vasconcelos A, Vega E, Pérez Y, Gómara MJ, García ML, Haro I (2015) Conjugation of cell-penetrating peptides with poly (lactic-co-glycolic acid)-polyethylene glycol nanoparticles improve ocular drug delivery. Int J Nanomed 10:609–631. https://doi.org/10.2147/IJN.S71198 Wang J, Shao F, Li W, Yan J, Liu K, Tao P, Masuda T, Zhang A (2017a) Metal-ion-mediated supramolecular assembly of c3 -peptides. Chem Asian J 12(5):497–502. https://doi.org/10.1002/ asia.201601721 Wang Y, Xie Y, Li J, Peng ZH, Sheinin Y, Zhou J, Oupický D (2017b) Tumor-penetrating nanoparticles for enhanced anticancer activity of combined photodynamic and hypoxia-activated therapy. ACS Nano 11(2):2227–2238. https://doi.org/10.1021/acsnano.6b08731 Wang W, Li X, Wang Z, Zhang J, Dong X, Wu Y, Fang C, Zhou A, Wu Y (2019a) A novel “mosaic-type” nanoparticle for selective drug release targeting hypoxic cancer cells. Nanoscale 11(5):2211–2222. https://doi.org/10.1039/c8nr06452k Wang Y, Fu M, Liu J, Yang Y, Yu Y, Li J, Pan W, Fan L, Li G, Li X, Wang X (2019b) Inhibition of tumor metastasis by targeted daunorubicin and dioscin codelivery liposomes modified with PFV for the treatment of non-small-cell lung cancer. Int J Nanomed 14:4071–4090. https://doi. org/10.2147/IJN.S194304 Wilson WR, Hay MP (2011) Targeting hypoxia in cancer therapy. Nat Rev Cancer 11(6):393–410. https://doi.org/10.1038/nrc3064 Wong S, Shim MS, Kwon YJ (2014) Synthetically designed peptide-based biomaterials with stimuli-responsive and membrane-active properties for biomedical applications. J Mater Chem B 2(6):595–615. https://doi.org/10.1039/c3tb21344g Xie JH, Jin ML, Morris GA, Zha XQ, Chen HQ, Yi Y, Li JE, Wang ZJ, Gao J, Nie SP, Shang P, Xie MY (2016) Advances on bioactive polysaccharides from medicinal plants. Crit Rev Food Sci Nutr 56(Suppl 1):S60–S84.https://doi.org/10.1080/10408398.2015.1069255 Xu K, Wang F, Pan X, Liu R, Ma J, Kong F, Tang B (2013) High selectivity imaging of nitroreductase using a near-infrared fluorescence probe in hypoxic tumor. Chem Commun (camb) 49(25):2554– 2556. https://doi.org/10.1039/c3cc38980d Xu S, Wang Q, Zhang Q, Zhang L, Zuo L, Jiang JD, Hu HY (2017) Real time detection of ESKAPE pathogens by a nitroreductase-triggered fluorescence turn-on probe. Chem Commun (camb) 53(81):11177–11180. https://doi.org/10.1039/c7cc07050k Yang Z, Cao J, He Y, Yang JH, Kim T, Peng X, Kim JS (2014) Macro-/micro-environment-sensitive chemosensing and biological imaging. Chem Soc Rev 43(13):4563–4601. https://doi.org/10. 1039/c4cs00051j Yang S, Xu D, Dong H (2018) Design and fabrication of reduction-sensitive cell penetrating nanofibers for enhanced drug efficacy. J Mater Chem B 6(44):7179–7184. https://doi.org/10. 1039/c8tb00728d Yang Q, Wang S, Li D, Yuan J, Xu J, Shao S (2020) A mitochondria-targeting nitroreductase fluorescent probe with large Stokes shift and long-wavelength emission for imaging hypoxic status in tumor cells. Anal Chim Acta 1103:202–211. https://doi.org/10.1016/j.aca.2019.12.063 Yao Y, Feng L, Wang Z, Chen H, Tan N (2019) Programmed delivery of cyclopeptide RA-V and antisense oligonucleotides for combination therapy on hypoxic tumors and for therapeutic self-monitoring. Biomater Sci 8(1):256–265. https://doi.org/10.1039/c9bm00905a Yin W, Qiang M, Ke W, Han Y, Mukerabigwi JF, Ge Z (2018) Hypoxia-responsive block copolymer radiosensitizers as anticancer drug nanocarriers for enhanced chemoradiotherapy of bulky solid tumors. Biomaterials 181:360–371. https://doi.org/10.1016/j.biomaterials.2018.08.014 Yu H, Tang Z, Zhang D, Song W, Zhang Y, Yang Y, Ahmad Z, Chen X (2015) Pharmacokinetics, biodistribution and in vivo efficacy of cisplatin loaded poly (L-glutamic acid)-g-methoxy poly (ethylene glycol) complex nanoparticles for tumor therapy. J Control Release: off J Control Release Soc 205:89–97. https://doi.org/10.1016/j.jconrel.2014.12.022 Yu J, Qian C, Zhang Y, Cui Z, Zhu Y, Shen Q, Ligler FS, Buse JB, Gu Z (2017) Hypoxia and H2 O2 dual-sensitive vesicles for enhanced glucose-responsive insulin delivery. Nano Lett 17(2):733– 739. https://doi.org/10.1021/acs.nanolett.6b03848

80

P. Kumari et al.

Yu-Wai-Man C, Tagalakis AD, Manunta MD, Hart SL, Khaw PT (2016) Receptor-targeted liposome-peptide-siRNA nanoparticles represent an efficient delivery system for MRTF silencing in conjunctival fibrosis. Sci Rep 6:21881. https://doi.org/10.1038/srep21881 Zhang S (2017) Discovery and design of self-assembling peptides. Interface Focus 7(6):20170028. https://doi.org/10.1098/rsfs.2017.0028 Zhang D, Qi GB, Zhao YX, Qiao SL, Yang C, Wang H (2015) In Situ formation of nanofibers from purpurin18-peptide conjugates and the assembly induced retention effect in tumor sites. Adv Mater (Deerfield Beach, Fla.) 27(40):6125–6130. https://doi.org/10.1002/adma.201502598 Zhang L, Huang Y, Lindstrom AR, Lin TY, Lam KS, Li Y (2019) Peptide-based materials for cancer immunotherapy. Theranostics 9(25):7807–7825.https://doi.org/10.7150/thno.37194 Zhang P, Yang H, Shen W, Liu W, Chen L, Xiao C (2020a) Hypoxia-responsive polypeptide nanoparticles loaded with doxorubicin for breast cancer therapy. ACS Biomater Sci Eng 6(4):2167–2174. https://doi.org/10.1021/acsbiomaterials.0c00125 Zhang Z, Yang W, Ma F, Ma Q, Zhang B, Zhang Y, Liu Y, Liu H, Hua Y (2020b) Enhancing the chemotherapy effect of Apatinib on gastric cancer by co-treating with salidroside to reprogram the tumor hypoxia micro-environment and induce cell apoptosis. Drug Delivery 27(1):691–702. https://doi.org/10.1080/10717544.2020.1754528 Zhang C, Li Q, Wu C, Wang J, Su M, Deng J (2021) Hypoxia-responsive nanogel as IL-12 carrier for anti-cancer therapy. Nanotechnology 32(9):095107. https://doi.org/10.1088/1361-6528/abcdcc Zhao J, Li Q, Hao X, Ren X, Guo J, Feng Y, Shi C (2017) Multi-targeting peptides for gene carriers with high transfection efficiency. J Mater Chem B 5(40):8035–8051. https://doi.org/10.1039/ c7tb02012k Zhao C, Tong Y, Li X, Shao L, Chen L, Lu J, Deng X, Wang X, Wu Y (2018) Photosensitive nanoparticles combining vascular-independent intratumor distribution and on-demand oxygendepot delivery for enhanced cancer photodynamic therapy. Small 14(12):e1703045. https://doi. org/10.1002/smll.201703045 Zhao L, Zou Q, Yan X (2019) Self-assembling peptide-based nanoarchitectonics. Bull Chem Soc Jpn 92(1):70–79. https://doi.org/10.1246/bcsj.20180248 Zhao L, Fu C, Tan L, Li T, Zhong H, Meng X (2020) Advanced nanotechnology for hypoxiaassociated antitumor therapy. Nanoscale 12(5):2855–2874. https://doi.org/10.1039/c9nr09071a Zhen J, Tian S, Liu Q, Zheng C, Zhang, Z., Ding, Y., An, Y., Liu, Y., & Shi, L. (2019). Nanocarriers responsive to a hypoxia gradient facilitate enhanced tumor penetration and improved anti-tumor efficacy. Biomater Sci 7(7):2986–2995.https://doi.org/10.1039/c9bm00461k Zheng J, Shen Y, Xu Z, Yuan Z, He Y, Wei C, Er M, Yin J, Chen H (2018) Near-infrared offon fluorescence probe activated by NTR for in vivo hypoxia imaging. Biosens Bioelectron 119:141–148. https://doi.org/10.1016/j.bios.2018.08.014 Zhou Y, Yang B, Ren X, Liu Z, Deng Z, Chen L, Deng Y, Zhang LM, Yang L (2012) Hyperbranched cationic amylopectin derivatives for gene delivery. Biomaterials 33(18):4731–4740. https://doi. org/10.1016/j.biomaterials.2012.03.014 Zhou Y, Maiti M, Sharma A, Won M, Yu L, Miao LX, Shin J, Podder A, Bobba KN, Han J, Bhuniya S, Kim JS (2018) Azo-based small molecular hypoxia responsive theranostic for tumor-specific imaging and therapy. J Control Release: off J Control Release Soc 288:14–22. https://doi.org/ 10.1016/j.jconrel.2018.08.036 Zhou H, Qin F, Chen C (2021) Designing hypoxia-responsive nanotheranostic agents for tumor imaging and therapy. Adv Healthcare Mater 10(5):e2001277. https://doi.org/10.1002/adhm.202 001277 Zhu C, Zou Z, Huang C, Zheng J, Liu N, Li J, Yang R (2019) Highly selective imaging of lysosomal azoreductase under hypoxia using pH-regulated and target-activated fluorescent nanoprobes. Chem Commun (camb) 55(22):3235–3238. https://doi.org/10.1039/c9cc00462a Zununi Vahed S, Salehi R, Davaran S, Sharifi S (2017) Liposome-based drug co-delivery systems in cancer cells. Mater Sci Eng. C, Mater Biol Appl 71:1327–1341.https://doi.org/10.1016/j.msec. 2016.11.073

Recent Advancements in the Field of Stimuli-Responsive Polymeric Nanomaterials for Cancer Treatment N. Sisubalan, S. Nisha Nandhini, M. Gnanaraj, A. Vijayan, Joe Rithish, C. Karthikeyan, and K. Varaprasad

Abstract Cancer has drawn a serious concern with rapid increase in the cases year by year. In order to control cancer, several treatment methods and surgery are available, such as chemotherapy, radiotherapy, etc. However, they have with them their own disadvantages. This had drawn attention to the development of an efficient treatment strategy. Lately, nanotechnology on the other hand is rapidly growing and started being used for cancer treatment in advanced healthcare applications. Their application in the treatment of cancer was explored in the form of nanomaterials, such as nanogels, nanopolymers, 2D nanosheets, anticancer drug-delivery nanoplatforms, and several nanomedicines. These kinds of nanoplatforms suffer from demerits, such as serious side effects, low efficiency, and reduced target efficiency which demand more research for their practical application. However, stimuli-responsive nanomedicine can overcome these limitations by precisely delivering drugs at specific sites for noninvasive cancer therapy. The prodrug-based smart nanomedicines have the ability to smartly respond to certain types of stimuli and releasing of the desired N. Sisubalan (B) · A. Vijayan · J. Rithish Department of Botany, Bishop Heber College (Autonomous), Affi. To Bharathidasan University, Trichy 620017, Tamil Nadu, India e-mail: [email protected] N. Sisubalan KIRND Institute of Research and Development PVT LTD, Tiruchirappalli, Tamil Nadu 620020, India M. Gnanaraj Department of Biotechnology and Bioinformatics, Bishop Heber College (Autonomous), Affi. To Bharathidasan University, Trichy, Tamil Nadu 620017, India K. Varaprasad Facultad de Ingeniería Arquitectura y Diseño, Universidad San Sebastián, Lientur 1457, 4080871 Concepción, Chile S. N. Nandhini Department of Botany, St. Josephs’s College (Autonomous), Affi. To Bharathidasan University, Trichy, Tamil Nadu, India C. Karthikeyan Department of Chemical and Biochemical Engineering, Dongguk University, Seoul - 04620, Republic of Korea © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_5

81

82

N. Sisubalan et al.

drugs at the target place. The nanopolymers respond to stimuli, such as pH, light, enzyme, and reactive oxygen species and they provide advanced properties to the nanobiomaterials to be used for the next generation of healthcare applications. Stimuli-responsive nanopolymers improve the bioavailability of drugs at the site of cancer. The enzyme-sensitive prodrugs release drugs by the specific proteases that hydrolyzes the amide bonds of proteins at the tumor site. The anticancer strategies formulated must focus on inhibiting both primary tumors and metastasis. This must also be considered while devising anticancer treatments through nanopolymers. The main aim of this chapter is to bring to light the current advances in the treatment of cancer by stimuli-responsive nanopolymers and the associated hindrances in their successful practical clinical application. Keywords Nanomedicines · Nanocarrier · Doxorubicin · Paclitaxel · Nanomicelles · Stimuli-responsive biomaterials

Abbreviations AL HCPT BP-PTX-Gd NPs PLGA DOX PTX RTV HA—PTX + RTV – NMF Vit E TPGS PLGA NIPAM DMAEMA NIPAM-co-DMAEMA BIS TEMED MTX PEG-PCRVP

ATRP AuNRs DOX DOX

Amylated lignin 10-Hydroxycamptothecin Branched polymeric PTX-Gd-based nanoparticles Poly (lactic-co-glycolic acid) hydroxyapatite nanoparticles (HA) Drochloride Paclitaxel (PTX) Ritonavir Paclitaxel and ritonavir nanomicellar formulation Vitamin E TPGS Poly (lactide) co-(glycolide) polymer N-isopropylacrylamide (99%), 2-(dimethylamino) ethyl methacrylate Poly(ethylene glycol) methyl ether methacrylate (methoxy PEG Methacrylate - Poly-g-PEG Bis-acrylamide Tetramethylethylenediamine Methotrexate Poly (ethylene glycol)-b-((2,5-bis[(4carboxylicpiperidylamino) thiophenyl]-croconine)co-4-vinyl pyridine Atom transfer radical-polymerization Gold Nanorod Reduction-responsive doxorubicin Doxorubicin

Recent Advancements in the Field of Stimuli-Responsive Polymeric …

NFX CLM DMF DMSO DCC DMAP DDACT NIPAM CS EDC NHS FITC GSH LCST SEM TEM

83

Nifuroxazide Co-loaded micelles N, N-dimethylformamide Dimethyl sulfoxide Dicyclohexylcarbodiimide 4-(N,N-dimethylamino) pyridine S-1-dodecyl-S’-(α, α’-dimethyl-α”-acetic acid) trithiocarbonate N-isopropylacrylamide Chitosan N-(3-dimethyl-aminopropyl)-N’-ethylcarbodiimide hydrochloride N-hydroxysuccinimide Fluorescein isothiocyanate Glutathione Lower critical solution temperature Scanning electron microscope Transmission electron microscope

1 Introduction Cancer has become a disease of major concern due to the constantly increasing number of cancer patients and the less availability of efficient cures (Varaprasad et al. 2022; Karthikeyan et al. 2021). Though several methods, such as surgery, radiation, monoclonal antibodies, hormonal therapy, and chemotherapy are currently in practice to treat cancer, the associated side effects in the patients still remain a major issue. Chemotherapy faces several challenges including, narrow therapeutic index toxicity, rapid clearance, and organ and poor membrane permeability. It may also cause hair loss, immunosuppression, peripheral neuropathy, infertility, secondary neoplasm, and tumour lysis syndrome. To counteract these problems, structurally, physicochemically, and functionally diversity-rich nanoparticles (NPs), such as dendrimers, carbon nanotubes, polymeric nanoparticles, and polymeric micelles are proposed for drug loading and internalization in target cancer cells. This attempt is to limit drug uptake via normal cells and tissues (Sahoo et al. 2003; Abeylath et al. 2011; Cappuccitti et al. 2022). Nanomedicine, however, has transformed medicine and significantly enhanced the pharmacological and pharmacokinetic profile of unstable anticancer medicines (Buabeid et al. 2020; Bai et al. 2019). Dendrimers are nearly spherical globular macromolecules and nanocarriers with the size of 1–100 nm and their architecture consists of a corona (with peripheral reactive functional groups), a mantle (hyperbranched), and a central core (Tomalia 2005). Dendrimers are a highly attractive class of gene and drug delivery vectors due to their 3D nanostructure (Franiak-Pietryga et al. 2018). Polyamidoamine (PAMAM)

84

N. Sisubalan et al.

and poly (propylene imine) (PPI) are two amine-terminated stimuli-responsive (pHdependent) dendrimers with drug-release behavior. At alkaline pH (high pH), the tertiary amine groups are deprotonated leading to back folding which is the collapse of the dendrimer on itself (Kannan et al. 2014; Kesharwani et al. 2014a, b). Nanoplymers (NPs) are of diameter much smaller than that of animal cells and so can be utilized in NP-mediated cancer imaging, tumor targeting, controlled drug release, multimodality treatment, and gene and photodynamic therapy (Cappuccitti et al. 2022). Due to their properties, such as optimal drug loading, controlled drug release, improved permeability, biocompatibility, and higher patient compliance, polymers are recommended for effective drug administration (Gelperina et al. 2005; Nitta et al. 2013). They play a key role in creating nanocarriers that are more precisely delivered to the tumor location and include a natural anti-cancer drug, such as curcumin. Curcumin-loaded poly (ester amine) nanoparticles increased the drug’s pharmacokinetics while toxicating HeLa and A549 tumor cells. Additionally, it prevented angiogenesis in tumor cells encapsulated in alginate (Ding et al. 2014). However, drug release through polymeric nanocarrier is uncontrollable. The development of stimuliresponsive polymeric nanocarriers has given hope for effective drug delivery applications. The stimuli can be internal (pH, redox) or external (electric and magnetic fields) properties of the biomaterials (Alsehli et al. 2020, Arulmozhi, et al. 2019). Self-assembled nanopolymers have a notable ability to enhance the solubility of hydrophobic drugs and anticancer effects. From recent studies, it had been observed that when specific factors in the tumor microenvironment are activated, physicochemical properties of the bioactive NPs change leading to the spatial release of drugs from tumor cells (Huang et al. 2019; Gong et al. 2021). Chauhan et al. (2017) designed the folic acid-graphene oxide @ gold (FA-GO@Au)-loaded DOX nanocomposites for the release of Au-NPs and DOX at the tumor site through near-infrared mediation. As many biological parameters are simultaneously altered at the tumor location, multibiological response NPs provide greater tumor-specific drug delivery capacities than single-stimulus response NPs (Cheng et al. 2013). The pH of cancer or inflammation site is altered during the pathological conditions and this has been employed to trigger the drug release into a preferred organ or intracellular compartment like a lysosome or endosome (Mura et al. 2013). Nanographene-based drug delivery system had been employed for stimuli-responsive delivery of cancer therapeutic agents, such as pH/light-sensitive graphene oxide-polyethylene glycol (GO-PEG) or ATP-responsive GO formulation as nanocarriers for doxorubicin delivery (Yang et al. 2016). Recent progress in nanomedicine has opened up several nanomaterials to be employed for diagnostics and treatment. The recent studies performed in this area will be covered in this chapter to present the current scenario of stimuli-responsive nanopolymer research for cancer treatment and the mechanism of stimuli-responsive nanopolymers against cytotoxic cells was given in Table 1.

Stimuli pH-responsive

Cathepsin B-responsive

GSH-responsive

GSH-responsive

Name of the nanopolymers

AL-His/HCPT

BP-PTX-Gd NP

HA−ss−PLGA

Polyglutamic acid dendrimers (G2 )

Table 1 Stimuli-responsive nanopolymers with their cytotoxic mechanism

The composite nanoparticles’ ability to target tumors and have stronger cancer therapeutic effects was demonstrated by their cytotoxicity and endocytosis in vitro, which suggested that the Dox@MSNs-(G2 )n had good potential as an anticancer medication (Li et al. 2020)

100 μg/ml

(continued)

According to the results of the cytotoxicity investigation, this stimulus (GSH-induced nanomicellar disintegration) significantly slows down the drug’s release in healthy cells. (Gote et al. 2021)

Exhibited better performance in 4T1 murine breast cancer in mice than Taxol and are nontoxic to adjacent cells/organs (Cai et al. 2020)

0.55 μg

–

The group that received treatment with AL-His/HCPT nanoparticles experienced comparatively sluggish tumor volume growth, indicating a substantial tumor inhibition rate. It suggests that AL-His/HCPT NPs have effective in vivo anti-tumor actions. (Zhao et al. 2020)

Anticancer efficiency

0.97 ± 0.18

IC50 value (μg/mL−1 )

Recent Advancements in the Field of Stimuli-Responsive Polymeric … 85

Stimuli GSH and light

pH and reduction-responsive

pH and enzyme

Name of the nanopolymers

CPAP@DOX

Polymer-Gold Nanorod

Co-prodrug-loaded micelles

Table 1 (continued)

–

3.1 μg mL−1

–

IC50 value (μg/mL−1 )

(continued)

Administration and regulated release of their cargos were made achievable by severing the links between hydrazone and GFLG via stimuli-responsiveness inside the tumor microenvironment (Luo et al. 2020)

The AuNR@PDOX cluster was disassociated into a single AuNR and the pH-responsive PEG-PCRVP shell was broken up, allowing the AuNR to enter deep tumor areas and release the localized administration of DOX chemotherapy, which as a result of the hybrid NPs’ reaction to the shrinking tumor microenvironment (Liu et al. 2019)

It was reported that the supramolecular disintegration was achieved by GSHand light-trigger. The discharge of DOX from the presence of GSH greatly accelerated CPAP@DOX along with light (Li et al. 2019)

Anticancer efficiency

86 N. Sisubalan et al.

Stimuli Thermo- and pH-responsive

Multiple environment-sensitive

Name of the nanopolymers

Poly(N-isopropylacrylamide-co-(2-dimethylamino) ethyl methacrylate)-g-PEG nanoparticle

Polymeric prodrug of gambogic acid

Table 1 (continued)

–

–

IC50 value (μg/mL−1 )

After regulated drug release in the tumor microenvironment, the absorption of micelles by cells was enhanced and was found to be effective in suppressing cancer cell proliferation caused by folate-mediated endocytosis. It was discovered that the drug release from the micelles was temperature-, pH-, and esterase-dependent (Du et al. 2021)

The combined pH and temperature responsiveness of the The drug’s release time and location are controlled by the nanocarrier. A high cumulative release of the MTX was achieved at 45 °C, which is in the hyperthermia temperature range, and a pH of 5.5, which is the tumor acidic pH condition. This demonstrates that temperature and acidic pHs have a significant impact on the drug’s cumulative release (Najafipour et al. 2020)

Anticancer efficiency

Recent Advancements in the Field of Stimuli-Responsive Polymeric … 87

88

N. Sisubalan et al.

2 Smart Polymeric Nanobiomaterials Polymers have been used to deliver the bioactive components at the target site in therapeutic applications. However, to get advanced characteristics, they are modified by several methods and they are used for the development of smart nanobiomaterials.

2.1 The Preparation Process of Stimuli-Responsive Polymers The preparation process of single, dual, and multi-stimuli-responsive nanopolymers were described below.

2.1.1

Single Stimuli-Responsive Polymeric Material

The alkyl amine group was added to an aromatic molecule via the Mannich process. The functional groups of the primary amine group of amylated lignin (AL) and the carboxyl group react to generate an amide bond. The proton peak moved to the lower field when the density of the electron cloud on the amine group decreased. The precipitation process was used to create AL-His/HCPT NPs (Zhao et al. 2020). BPPTX-Gd NPs were created through reversible addition–fragmentation chain transfer (RAFT) polymerization with the use of enzyme-responsive chain transfer agents, and the therapeutic drug paclitaxel was loaded using an enzyme-responsive linker (PTX). Gd(III) chelated in tetraazacyclododecane tetraacetic acids (DOTA) was bound to the polymer chain covalently as a magnetic resonance imaging (MRI) contrast agent. Cyanine 5.5 for fluorescence image was added to the polymer using a thiol–ene click reaction (Zhao et al. 2020) (Scheme 1). Conjugation of hydroxyapatite (HA) nanoparticles with grafted poly(lactic-coglycolic acid) (PLGA) polymers was performed to achieve active targeting. HA has been used as a targeting agent due to its high CD44 receptor expression and efficiently reaches breast cancer cells. Additionally, PLGA is attached to HA via a disulfide (s– s) bond and can be reduced and cleaved at the active site to release the active drug. Therefore, a nanocell formulation of paclitaxel and ritonavir (HA - PTX + RTV NMF) was made with a mixture of polymers such as HA-ss-PLGA and vitamin E TPGS (Vit E-TPGS) (Gote et al. 2021). The cancer drug DOX was modified with FA. DOX-FA was obtained by a 24-h light avoidance reaction under the action of the condensing agent DCC (dicyclohexylcarbodiimide) (Li et al. 2020).

Recent Advancements in the Field of Stimuli-Responsive Polymeric …

89

Scheme 1 Schematic representation of branching pHPMA-based nanomedicine that is cathepsin B-responsive, biodegradable, and theranostic. a The branching pHPMA-PTX-Gd conjugates’ chemical composition. b The conjugate’s hydrophilic and hydrophobic moieties self-assembled to form BP-PTX-Gd NPs. c MR imaging in vivo of BP-PTX-Gd NPs transported to tumor sites through EPR. d Lysosomal endocytosis routes for drug release control, intracellular trafficking, and intracellular biodegradation in the presence of an overexpressed cathepsin B milieu. The released PTX then stopped cells from going through mitosis and triggered the associated apoptotic pathway to cause apoptosis by stabilizing microtubules (Adapted from Cai et al. 2020)

2.1.2

Dual Stimuli-Responsive Polymeric Nanomaterials

Poly(NIPAM-co-DMAEMA)-g-PEG nanoparticles were synthesized by multi-step emulsion polymerization. Bisacrylamide (BIS) was used as a crosslinker and tetramethylethylenediamine (TEMED) as an accelerator. Nanoparticles with an average particle size of 180 nm were assembled by polymerization. Methotrexate (MTX) was encapsulated via a source-diffusion process. They were prepared at 37 °C and pH 7.4, followed by sonication for 10 min and gentle agitation to swell (Najafipour et al. 2020). Ultra-small AuNRs were fabricated and a block polymer, PEG-PCRVP, was synthesized from a single CR and four VP monomers by copolymerization. PEGbromine (Br) was used as a macroinitiator based on the atom transfer radical polymerization (ATRP) method. AuNR bound with PDOX to form AuNR@PDOX through a reduction-responsive disulfide bond (-S–S-). Predetermined ratios of He mixed AuNR@PDOX and PEG-PCRVP in chloroform and dissolved polyvinyl alcohol (PVA) in deionized (DI) water. After subjecting the solution to ultrasonic emulsification, the chloroform phase was evaporated at 25 °C to form core–shell hybrid

90

N. Sisubalan et al.

NPs (Liu et al. 2019). CO-PIM-b-PEG/PEG-FA was synthesized from a mixture of dibromo-functionalized polyimine, K2CO3, PEG/PEG-FA, and acetonitrile/DMSO. PIM-b-PEG/PEG-FA and DOX-loaded micelles were prepared using a nanoprecipitation method. Lyophilization gave the pure product (PIM-b-PEG/PEG-FA). DOXloaded micelles were also synthesized by nanoprecipitation method (Saeedi et al. 2021). DOX/NFX co-loaded micelles (CLM) were synthesized by the thin film hydration method from the block polymers polyOEGMA-block-poly[HPMA-DOX]GFLGKGLFG-poly[HPMA-DOX]-block-polyOEGMA and NFX. The molar ratio dissolved there is 5:1 was dissolved in chloroform and methanol and sonicated. DOX-loaded micelles (DLM) were prepared similarly to CLM. DOX-conjugated macromolecular prodrugs self-assembled into micelles (DLMs) in PBS at pH = 7.4 over a CMC of 0.025 mg/ml (Luo et al. 2020). Functional NP CS-PNIPAM was synthesized from acetylated CS, anhydrous DMF, DDACT, DCC, and DMAP by RAFT polymerization. The CS-g-PNIPAM copolymer was synthesized by grafting NIPAM onto CS, and the K237 peptide was synthesized on CS-g using the EDCNHS technique (via reaction between amino and carboxyl groups). Next, PTX and CS-PNIPAM were dispersed in DMF, stirred, and centrifuged to obtain CS(PTX)-gPNIPAM. CS(PTX)-g-PNPAM was labeled with FITC to facilitate the assessment of cellular uptake (Qian et al. 2019).

2.1.3

Multi-stimuli-Responsive Nanopolymers

HSSG NPs were constructed from highly hydrophilic HA and poorly water-soluble GER. The amphipathic nature of the prodrugs leads to the self-assembly of HSSGprodrug conjugates into NPs in aqueous solution. HSSG with redox responsiveness was synthesized with CYS as a linker for anticancer targeting anticancer drugs. GER bound the chain through GSH-sensitive and -insensitive disulfides and amides, conferring her GSH responsiveness to HSSG NPs (Xia et al. 2022). Chitosan swelled at pH 5 due to protonation of amino groups. Few amino groups were protonated, as most amino groups in chitosan are bound with FA and NH2 -PNIPAM. This results in GFCP micelles with moderately increased particle size. When the temperature was increased to 42 °C, above the lower critical solution temperature (LCST) of the polymer, the micelle shell exhibited a phase (volume) transition from hydrophilic to hydrophobic, and the micelle shell–core structure became progressive. Disrupted to disrupt micelles and increase particle size (Du et al. 2021).

Recent Advancements in the Field of Stimuli-Responsive Polymeric …

91

3 Physicochemical Characterization 3.1 Single Stimuli-Responsive Smart Materials The sizes of AL-His NPs and AL-His/HCPT NPs were 27.37 ± 3.43 and 38.41 ± 6.08 nm, respectively. AL-His/HCPT NPs are larger in size due to drug loading. The drug loading efficiency (DLE) was evaluated using high-performance liquid chromatography and found to be 15.57 ± 1.04 μg for AL-His/HCPT NPs (Zhao et al. 2020). The molecular weight and polydispersity index (PDI) of the BP-PTXGd conjugate were 186 kDa and 2.30, respectively; Molar ratio (2:1:1). Amino acid analysis revealed glycine (Gly), phenylalanine (Phe), and leucine (Leu) in the conjugate, indicating the presence of an enzyme-responsive GFLG tetrapeptide in the polymer backbone. A hydrodynamic diameter of 62.6 ± 1.1 nm was observed for the BP-PTX-Gd NPs by dynamic light scattering (DLS). Nanoparticle self-assembly was primarily driven by a balance of hydrophilic and hydrophilic interactions (Cai et al. 2020, Bhardwaj et al. 2021). The hydrodynamic size range, PDI, and zeta potential were found to be 142.5– 256 nm, 0.2–0.8, and 0.01–0.023 mV, respectively. Formation of HA-ss-PLGA was confirmed by H 1 NMR. The sonication time for the independent variables HA-ssPLGA and Vit E-TPGS with mean values of 22.5 min at 2 wt% and 4 wt%, respectively, reduced the minimum size, PDI, and zeta potential to zero. Close formulations were possible (Gote et al. 2021). MSN had a core–shell structure with a dark iron oxide core and a light silica shell. Its diameter is estimated to be about 20 nm. The size of DOX@MSN did not change after drug loading. The appearance of the dark areas in the mesoporous silica microspheres was Fe3 O4 . The zeta potentials of DOX@MSNs and DOX@MSNs-(G2 )n were 5.03 mV and 1.20 mV, respectively, which are higher than the zeta potential of (G2 )n −13.71 mV, indicating successful loading of DOX@MSN (Li et al. 2020).

3.2 Dual Stimuli-Responsive Smart Materials MTX-loaded P(NIPAM-co-DMAEMA)-g-PEG nanoparticles with a polymer-todrug ratio of 12:1 showed higher encapsulation efficiency and magnitude, with PDI and zeta potential of 285 nm, 0.135, and 5.56 mV, respectively, at room temperature. The swelling rate decreased with increasing temperature. Swelling rates indicated that water absorption in acidic environments was much higher than that in neutral and alkaline solutions (Najafipour et al. 2020). The average CPAP size and PDI were 155.2 nm and 0.305, respectively (Li et al. 2019). The electromagnetic field enhancement of hybrid NPs is stronger than AuNR arrays (fourfold) and PEG-PCRVP NPs (sevenfold) based on finite-difference timedomain method (FDTD) results. Increasing the DOX and NP supply ratio increased the loading level of DOX (Liu et al. 2019). The average size, zeta potential, and

92

N. Sisubalan et al.

CMC of CO-PIM-b-PEG/PEG-FA measured by DLS were 91.67 nm, −8.9, and 2.7 mg/l. SEM and TEM results showed a spherical morphology. DOX release from drug-loaded micelles was highest (50%) in an acidic environment at pH 5.5. This low pH-responsive drug release can be attributed to imine bond cleavage (Liu et al. 2019). The molecular weight and PDI of polyOEGMA-block-poly[HPMA-DOX]GFLGKGLFG-poly[HPMA-DOX]-block-polyOEGMA were 92 kDa and 1.12, respectively. The average sizes of DLM and CLM were 80.60 ± 0.51 nm and 131.33 ± 6.0 nm with PDIs of 0.25 and 0.24, respectively. Both showed a spherical shape under TEM and diameters of 44.30 ± 3.70 nm and 67.30 ± 7.00 nm, respectively. The encapsulation ratio and load capacity of NFX in CLM were 65.4% and 11.5%, respectively. The zeta potentials of DLM and CLM were 4.37 ± 0.99 mV and 5.62 ± 0.92 mV, respectively. Negative charges are said to impede macrophage recognition of the endothelial reticular system and increase blood flow time (Luo et al. 2020). The zeta potentials of CS-g-PNIPAM K237-CS-g-PNIPAM were 14.1 ± 2.1 and 19.1 ± 2.5, respectively. At 37 °C, K237-CS(PTX)-g-PNIPAM showed a strong pH dependence on drug release. After 48 h, at pH 7.4 and pH 5.0, respectively, over 63% and 74% of the PTX that had been taken up were released. At higher temperatures and lower pH levels, drug-loaded NPs showed faster release rates. In the MTT experiment, the drug carriers CS-g-PNIPAM and K237-CS-g-PNIPAM both demonstrated > 90 L viability in L929 cells and MDA-MB-231 (Qian et al. 2019).

3.3 Multi-stimuli-Responsive Smart Materials The HA grafted with GER via CYS’s backbone was visible in the HSSG’s 1 H NMR spectrum. At 5.3 ppm, the GER alkene proton showed its typical peak. HSSG has a polydispersity index of less than 0.200 and a regular spherical form. Incubation for 48 h did not result in a substantial change in HSSG size. The HA shell’s negative surface charge may prevent nonspecific plasma protein adsorption when nanoparticles and blood components interact (Xia et al. 2022). Based on DLS characterization with a submicron particle sizer, the particle size and zeta potential of micelles were determined to be 87.5 1.91 nm and 8.12 1.01 mV, respectively. In the CCK-8 assay, micelles showed anti-proliferative effects on both her MCF-7 and HepG2 cells. The lower the concentration of GA in suspension, the lower its cytotoxicity (Du et al. 2021; Singh et al. 2014).

Recent Advancements in the Field of Stimuli-Responsive Polymeric …

93

4 Anti-tumor Efficiency of Stimuli-Responsive Nanopolymers 4.1 Single Stimuli-Responsive Smart Materials A stimulus-responsive polymer prodrug-based nanotheranostic system was developed by polymerization and conjugation chemistry using cyanine 5.5 and gadolinium chelates (contrast agents) and the branched polymer paclitaxel (PTX) (therapeutic agent). PTX-Gd-based biodegradable branched polymer nanoparticles (BP-PTX-GdNPs) have good biocompatibility and high stability under physiological conditions, but PTX rapidly degrades in the tumor microenvironment, decomposes into and releases in response to stimuli. A cathepsin B-responsive Gly-Phe-Leu-Gly (GFLG) tetrapeptide was used to crosslink poly[N-(2-hydroxypropyl)methacrylamide] (pHPMA) with His PTX-pHPMA polymer chains. Polymer segment with molecular weight and PTX. NP impaired microtubule function, downregulated the antiapoptotic protein Bcl-2, upregulated Bax expression, cleaved caspase-3, cleaved caspase-9, and cleaved PARP and p53 proteins. Similar to free PTX, BP-PTX-Gd-NP was highly cytotoxic to 4T1 cells. BP-PTX-Gd NPs showed prolonged in vivo circulation time and accumulation at the tumor site, as evidenced by MRI, pharmacokinetic analysis, and fluorescence imaging. By imaging chemotherapy, we observed a superior antitumor effect against 4T1 tumors in a mouse model. Thus, BP-PTX-Gd NPs pave the way for theranostic polymer-based therapy of cancers induced by fluorescence or MRI (Cai et al. 2020). The usage of aminated lignin-histidine complexes and His 10hydroxycamptothecin (AL-His/HCPT NPs) to self-assemble novel pH-responsive drug-loaded polymeric nanosystems. Histidine was used to obtain pH responsiveness of evoked drug release due to the acidic tumor microenvironment (endosomal: pH ~ 5–6; Lysosomes: pH ~ 4–5). Drug release increased with decreasing pH and peaked at 72.5% at pH 4.5. The synthesized NPs have a relatively small size of ∼40 nm, biocompatibility, and high drug loading capacity (∼15.57% by weight), making them suitable anti-tumor drug nanocarriers in advanced medical field. The NP-treated 4T1 tumor-bearing mouse model has shown an almost double tumor growth inhibition rate compared with pure HCPT and no severe allergic reactions (Fig. 1) (Zhao et al. 2020). In order to treat triple-negative breast cancer (TNBC) and metastatic breast cancer, paclitaxel (PTX) has been adorned with hyaluronic acid (HA) encapsulated composite nanoparticles and the P-glycoprotein ritonavir (RTV) inhibitor (MBC). HA is a naturally occurring ligand for the overexpressed CD44 receptors seen in breast cancer cells. The poly(lactide)co-(glycolide) polymer and HA can be conjugated together thanks to the disulfide bond (HA-ss-PLGA) (PLGA). When glutathione is present in breast cancer cells, these connections are quickly broken down. RTV supplementation also prevented the metabolism of PTX that is mediated by P-gp and CYP3A4. This leads to MDR reversal and sensitization of the cellular response to PTX. Nanonomicelles can remain stable in circulation for up to

94

N. Sisubalan et al.

Fig. 1 Free HCPT and AL-His/HCPT NPs’ in vivo anticancer efficacy in the mouse model with subcutaneous 4T1 inoculation. a Tumor images taken on day 16 by several groups. b Mice in various groups’ relative tumor volumes. c Throughout the experiment, the body weights of the mice were measured once every 2 days. d The bulk of the tumor has been treated by various groups. When the mice were put to death, the tumor mass was recovered through dissection. e IgE levels. f The WBC changes of various blood groups (Adapted from Zhao et al. 2020)

three days. The in vitro solubility of PTX and drug release from HA-PTX + RTVNMF in pH 6.8 phosphate buffered saline was 76.8. The cumulative release of PTX was higher for HA - PTX + RTV − NMF in response to stimuli of GSH (50 mM) compared with its absence. In in vitro uptake and cytotoxicity study with MBC MCF7 and TNBC cell lines, MDA-MB-231 showed efficient uptake of nanomicelles and PTX compared with MCF-12A cell line (non-neoplastic mammary gland epithelium) (Gote et al. 2021). A Dox@MSNs-(G2 )n dual-targeting nanopolymer prepared with doxorubicin, magnetic suspension silica (MSN), and second-generation polyglutamic acid dendrites. (G2 )n was used as the base material, and the nanostructure was

Recent Advancements in the Field of Stimuli-Responsive Polymeric …

95

achieved by crosslinking with disulfide bonds containing crosslinking agents to efficiently release DOX in cancer cells. enriched with GSH. It is electronegative and has plenty of space to help them absorb DOX (electronegative cancer drug). Folic acid (FA) and magnetic targeting by encapsulation in MSN make Dox@MSNs-(G2 )n very stable. It also improves cell transport and uptake efficiency. Crosslinking by disulfide bonds makes the nanoparticles sensitive to GSH so that they can dissociate and release drugs in cancer cells. The resulting composites exhibited good biocompatibility and low cytotoxicity in CCK-8 assays. Dox@MSNs-(G2 )n’s strong magnetism makes it potential for nuclear magnetic imaging. This allows the simultaneous use of Dox@MSNs-(G2 )n for tumor diagnosis and treatment (Li et al. 2020).

4.2 Dual Stimuli-Responsive Smart Materials Supermolecular drug transporters (CPAPs) based on both GSH-sensitive CD-PEGAzo-PCL and mild CD-PEG-Azo-PCL have been fabricated for intracellular transport of doxorubicin (DOX). The carriers are 70–90 nm in diameter and spherical in shape. CPAP consists of hydrophilic poly(ethylene glycol) (PEG) (CD-PEG) moieties and hydrophobic poly(ε-caprolactone) (PCL) moieties (Azo-PCL). DOX has been encapsulated in the hydrophobic core of CPAP. The GSH reaction is due to a disulfide bond between PEG and b-cyclodextrin (b-CD). The light reaction is obtained by the interaction between b-CD and azobenzene. Drug-loaded CPAP (CPAP@DOX) demonstrated accelerated DOX release upon simultaneous exposure to GSH and light. Studying the cytotoxicity of CPAP and CPAP@DOX with SKOV3 cells and HEK293T cells showed that CPAP@DOX can strongly inhibit cancer cells while reducing toxicity to normal cells. CPAP and CPAP@DOX are biocompatible. CPAP@DOX DDS may therefore offer a promising formulation for regulating medication release (Li et al. 2019). The two-step emulsion polymerization method was used to create the heat- and pH-sensitive poly(N-isopropylacrylamide-co(2-dimethylamino) ethyl methacrylate)-g-PEG (P(NIPAM-co-DMAEMA)-g-PEG). This contains a DMAEMA (pH-sensitive) block, a NIPAM (heat-sensitive) block, and a PEG-based agent. They were loaded with 51% by mass of MTX via the swelling diffusion method. Cytotoxicity testing with the MCF-7 cell line showed that methotrexate-containing nanoparticles (MTX) effectively suppressed tumor growth when combined with hyperthermia (45 °C). The release of MTX at normal physiological pH and temperature (T = 37 °C & pH = 7.4) was less (24%) than drug release (70%) after 48 h at 45 °C and 5,5 pH (tumor acidic pH) (Fig. 2). The constructed nanosystem can be applied for controlled drug delivery and multimodal cancer therapy (Najafipour et al. 2020). The tumor microenvironment provides several biological barriers to prevent drugs from reaching tumor cells. In an attempt to combat this barrier, a pH-sensitive and reduction-sensitive polymer core–shell and gold nanorod (AuNR) (PEG-PCRVP) assembly was fabricated by two-step miniaturization. The absorbance level of hybrid NPs was 14.2% ID g-1, double that of

96

N. Sisubalan et al.

non-reactive hybrid NPs. The reduced sensitive doxorubicin (DOX) precursor was polymerized and coated on AuNR. The high-density PEG molecule and the size of the NPs provide lengthy circulation times, biostability, and a high tumor accumulation efficiency. A local release of DOX occurs at the tumor site as a result of DOXcoated microcleaved AuNRs penetrating the solid tumor after the hybrid NPs have been absorbed by the tumor. On the surface of the AuNR cluster, a linear rise in the photoacoustic (PA) impact of the PA-activated polymer was seen. This shows that the electromagnetic field plays a significant part in the assembly of AuNR. The resulting hybrid NP has the potential to be used as a surface-enhanced Raman scattering imaging agent for real-time study of physiological behaviors in vivo and nanotherapeutic effects of deep tumor penetration. The use of PEG-PCRVP/AuNR@PDOX in the treatment of tumors did not show cancer recurrence (Liu et al. 2019). A folatemodified PEG copolymer has been developed. Castor oil is biodegradable giving a structure such that the copolymer. The redox- and pH-sensitive disulfide and imine bonds, respectively, were incorporated into the backbone of the star copolymer. It selfassembles into stable 91.67 nm micelles and cleaves upon exposure to internal lysosomal pH 5.5 and high concentrations of GSH. The drug loading capacity (DLC) and drug loading efficiency (DLE) of DOX loaded into the hydrophobic core of micelles were 15.93% and 96.13%, respectively. The micelles efficiently release DOX in a redox and pH-activated manner in vitro. The copolymer showed no cytotoxicity against HDF cells and increased cytotoxicity against HeLa cells. These DOX-coated micelles can be used as nanocarriers for hydrophobic drugs (Saeedi et al. 2021). An enzyme/pH-sensitive double-block copolymer (92 kDa MW) was induced to kill primary breast cancer cells and inhibit lung metastasis. It was then conjugated with DOX and subsequently loaded with nifuroxazide (NFX) for self-assembly of co-precursor loaded micelles (CLMs). CLM is biodegradable and has demonstrated a pH/cathepsin B-sensitive drug release pattern. CLM shows reduced in vitro viability, inhibition of cell migration, and invasion of 4T1 breast cancer in mice. In metastatic and lung metastatic 4T1 breast cancer pre-resistant mouse models, CLM has an excellent anti-metastatic effect and good anti-tumor effect. Histological and immunohistochemical immunofluorescence analyze revealed increased apoptosis, Fig. 2 The regulated release of MTX from nanoparticles exhibits dual thermo- and pH-responsive behavior (Adapted from Najafipour et al. (2020)

Recent Advancements in the Field of Stimuli-Responsive Polymeric …

97

inhibited expression of matrix metalloproteinases, and significantly decreased infiltration of MDSCs, leading to inhibition of genetic lung disease. CLM also reduces the cytotoxicity of DOX suggesting its potential use against primary breast cancer and lung metastases (Luo et al. 2020). Chitosan/poly(N-isopropylacrylamide) hybrid NPs functionalized with K237 peptide were fabricated and loaded with PTX. NPs functionalized by synthetic peptides have favorable pH and temperature properties, with enhanced drug release at slightly acidic pH persisting in the tumor microenvironment and enhanced release at higher temperatures. The 3-(4,5-dimethyl-thiazol-yl)-2,5diphenylte-trazolium (MTT) bromide assay (MTT) indicated that peptide-conjugated NPs effectively inhibited breast cancer cell growth compared with peptide-free NPs. Based on confocal microscopy experiments, NPs that can precisely target breast cancer overexpress the KDR/Flk-1 protein MDA-MB-231 are associated with their specific recognition for breast cancer. with KDR/Flk-1 on cancer cells. Functional peptide nanosystems are biocompatible and have the potential to deliver targeted and controlled anticancer drugs in advanced medical applications (Qian et al. 2019).

4.3 Multi-stimuli-Responsive Smart Materials Du et al. (2021) designed several environmentally sensitive polymeric gambogic acid precursors for cancer therapy as shown in Fig. 3. The method is based on chitosan grafting and folic acid–chitosan conjugates are then complexed with thermosensitive amine-terminated poly-N-isopropylacrylamide (NH2-PNIPAM) to develop FACSPN. High drug loading capacity as well as controlled drug release was achieved by conjugation of GA to the graft by esterification. An amphoteric prodrug is formed, the chitosan O-(gambogic acid)-N-(folic acid)-N-(NH2-PNIPAM) (GFCP) graft can self-assemble into micelles. These micelles are biocompatible and stable, and drug release is dependent on esterase, pH, and temperature. Studies on the antitumor effects of GFCP micelles were performed in the H22 tumor-bearing mouse model, testing for tumor cell uptake, cytotoxicity, and mass sphere penetration. When free GAs were injected intravenously, their concentrations in the heart increased rapidly, indicating possible cardiac adverse effects, but when GFCP micelles were used, the concentrations gradually increased and then increased. reduced. Drug release and delivery from micelles were significantly increased and reached a maximum at 0.5 h after introduction into tumor tissue compared with those using free GA. Based on these results, GFCP micelles are likely to enhance the antitumor efficacy of GA both in vivo and in vitro. A redox-sensitive polymeric redox-sensitive hyaluronic acid (HA) geraniol (GER) precursor is produced by combining hydrophobic GER with hydrophilic HA via a disulfide linkage. This effort was made to overcome the low bioavailability and low water solubility of natural RGE. The resulting HSSG self-assembled into a multifunctional nanoparticle approximately 110 nm in diameter with a uniform spherical shape in aqueous solution. Moreover, it maintains its stability in different physiological environments. NPs efficiently release the drug when exposed to buffers that

98

N. Sisubalan et al.

Fig. 3 Antitumor activity in in vivo conditions. a Tumor volumes, b Tumor pictures, c Tumor weights (n = 6; *P 0.05, **P 0.01, ***P 0.001), and e H&E images of tumor tissue of H22-bearing mice following intravenous injection of various treatments (Adapted from Du et al. 2021)

mimic the tumor microenvironment (pH/glutathione/hyaluronidase). Fluorescence microscopy results confirmed that HSSG precursors are selectively absorbed by the human hepatocellular carcinoma cell lines HepG2 and Huh7 through CD44 receptormediated internalization. through targeting. HSSG NPs accumulate at the tumor site in H22 tumor-bearing mice over a longer period of time. They outperformed GER and HCCG NPs in in vitro and in vivo anticancer studies (Xia et al. 2022).

5 Conclusion Recent studies on stimulus-responsive polymer structures in cancer therapy have focused on dual stimulus-responsive polymers. In all studies, the pH-sensitive system was preferred and studied intensively. Several studies have looked at the use of naturally available materials, such as geraniol and lignin in nanodrug-based cancer therapy. Stimuli-responsive nanopolymers for cancer therapy have recently gained

Recent Advancements in the Field of Stimuli-Responsive Polymeric …

99

popularity due to their local drug delivery capabilities, biocompatibility, and tunable physicochemical properties. In addition, their surface can be modified; their composition and size can be controlled. Although the nanopolymers have potential toxicity, limited capacity for hydrophilic drugs, limited storage stability and chemical synthesis, their high loading efficiency and good circulating stability offer Strong hope for cancer diagnosis and treatment. This requires more research to be conducted to eliminate the disadvantages of using nanopolymers in cancer treatment and to use them effectively to use utilize it for practical application in the clinical field. Acknowledgements Dr. Varaprasad Kokkarachedu acknowledges the support from the Fondecyt Regular No. 1211118, ANID, Chile. Dr. Sisubalan N acknowledges the Bishop Heber College (Autonomous) and the management for providing physical facilities. Conflict of Interest There is no conflict of interest to declare.

References Abeylath SC, Ganta S, Iyer AK, Amiji M (2011) Combinatorial-designed multifunctional polymeric nanosystems for tumor-targeted therapeutic delivery. Acc Chem Res 44(10):1009–1017 Alsehli M (2020) Polymeric nanocarriers as stimuli-responsive systems for targeted tumor (cancer) therapy: recent advances in drug delivery. Saudi Pharm J 28(3):255–265 Arulmozhi S, Matchado MS, Snijesh VP, Kumar A, Singh S (2019) An insight into anti-arthritic property of C25H34O7 for rheumatoid arthritis using molecular modelling and molecular dynamics approach. Inf Med Unlocked 16:100145 Bai Y, Wang Y, Zhang X, Fu J, Xing X, Wang C, Gao L, Liu Y, Shi L (2019) Potential applications of nanoparticles for tumor microenvironment remodeling to ameliorate cancer immunotherapy. Int J Pharm 30(570):118636 Bhardwaj M, Singh S, Kumar A (2021) Health informatics-A vital strategy to tackle pandemic diseases. Biomed J Sci Tech Res 39(4):31497–31498 Buabeid MA, Arafa ES, Murtaza G (2020) Emerging prospects for nanoparticle-enabled cancer immunotherapy. J Immunol Res 3:2020 Cai H, Dai X, Wang X, Tan P, Gu L, Luo Q, Zheng X, Li Z, Zhu H, Zhang H, Gu Z (2020) A nanostrategy for efficient imaging-guided antitumor therapy through a stimuli-responsive branched polymeric prodrug. Adv Sci 7(6):1903243 Cappuccitti A, Daniels B, Galloro C, Kung K, Ly K, Mohammad AM, Dahman Y (2022) Nanotechnology for biomedical devices: cancer treatment. In: Nanotechnology for electronic applications. Springer, Singapore, pp 207–251 Chauhan G, Chopra V, Tyagi A, Rath G, Sharma RK, Goyal AK (2017) “Gold nanoparticles composite-folic acid conjugated graphene oxide nanohybrids” for targeted chemo-thermal cancer ablation: in vitro screening and in vivo studies. Eur J Pharm Sci 1(96):351–361 Cheng R, Meng F, Deng C, Klok HA, Zhong Z (2013) Dual and multi-stimuli responsive polymeric nanoparticles for programmed site-specific drug delivery. Biomaterials 34(14):3647–3657 Ding Q, Niu T, Yang Y, Guo Q, Luo F, Qian Z (2014) Preparation of curcumin-loaded poly (ester amine) nanoparticles for the treatment of anti-angiogenesis. J Biomed Nanotechnol 10(4):632– 641 Du Q, Lv F, Huang J, Tang X, Zhao Z, Chen J (2021) A multiple environment-sensitive prodrug nanomicelle strategy based on chitosan graftomer for enhanced tumor therapy of gambogic acid. Carbohyd Polym 1(267):118229

100

N. Sisubalan et al.

Franiak-Pietryga I, Ziemba B, Messmer B, Skowronska-Krawczyk D (2018) Dendrimers as drug nanocarriers: the future of gene therapy and targeted therapies in cancer. Dendrimers: fundamentals and applications 25:7 Gelperina S, Kisich K, Iseman MD, Heifets L (2005) The potential advantages of nanoparticle drug delivery systems in chemotherapy of tuberculosis. Am J Respir Crit Care Med 172(12):1487– 1490 Gong Z, Zhou B, Liu X, Cao J, Hong Z, Wang J, Sun X, Yuan X, Tan H, Ji H, Bai J (2021) Enzymeinduced transformable peptide nanocarriers with enhanced drug permeability and retention to improve tumor nanotherapy efficacy. ACS Appl Mater Interfaces 13(47):55913–55927 Gote V, Sharma AD, Pal D (2021) Hyaluronic acid-targeted stimuli-sensitive nanomicelles coencapsulating paclitaxel and ritonavir to overcome multi-drug resistance in metastatic breast cancer and triple-negative breast cancer cells. Int J Mol Sci 22(3):1257 Huang Y, Shi Y, Wang Q, Qi T, Fu X, Gu Z, Zhang Y, Zhai G, Zhao X, Sun Q, Lin G (2019) Enzyme responsiveness enhances the specificity and effectiveness of nanoparticles for the treatment of B16F10 melanoma. J Control Release 28(316):208–222 Kannan RM, Nance E, Kannan S, Tomalia DA (2014) Emerging concepts in dendrimer-based nanomedicine: from design principles to clinical applications. J Intern Med 276(6):579–617 Karthikeyan C, Sisubalan N, Sridevi M, Varaprasad K, Basha MH, Shucai W, Sadiku R (2021) Biocidal chitosan-magnesium oxide nanoparticles via a green precipitation process. J Hazard Mater 411:124884 Kesharwani P, Jain K, Jain NK (2014a) Dendrimer as nanocarrier for drug delivery. Prog Polym Sci 39(2):268–307 Kesharwani P, Tekade RK, Jain NK (2014b) Formulation development and in vitro–in vivo assessment of the fourth-generation PPI dendrimer as a cancer-targeting vector. Nanomedicine 9(15):2291–2308 Li HY, Jing FK, Hao JF (2020) GSH-responsive polyglutamic acid nanocarriers for dual targeted cancer therapy. J Appl Polym Sci 137(43):49339 Li J, Li X, Liu H, Ren T, Huang L, Deng Z, Yang Y, Zhong S (2019) GSH and light dual stimuli-responsive supramolecular polymer drug carriers for cancer therapy. Polym Degrad Stab 1(168):108956 Liu T, Tong L, Lv N, Ge X, Fu Q, Gao S, Ma Q, Song J (2019) Two-stage size decrease and enhanced photoacoustic performance of stimuli-responsive polymer-gold nanorod assembly for increased tumor penetration. Adv Func Mater 29(16):1806429 Luo L, Xu F, Peng H, Luo Y, Tian X, Battaglia G, Zhang H, Gong Q, Gu Z, Luo K (2020) Stimuliresponsive polymeric prodrug-based nanomedicine delivering nifuroxazide and doxorubicin against primary breast cancer and pulmonary metastasis. J Control Release 1(318):124–135 Mura S, Nicolas J, Couvreur P (2013) Stimuli-responsive nanocarriers for drug delivery. Nat Mater 12(11):991–1003 Najafipour A, Mahdavian AR, Aliabadi HS, Fassihi A (2020) Dual thermo-and pH-responsive poly (N-isopropylacrylamide-co-(2-dimethylamino) ethyl methacrylate)-g-PEG nanoparticle system and its potential in controlled drug release. Polym Bull 77(6):3129–3142 Nitta SK, Numata K (2013) Biopolymer-based nanoparticles for drug/gene delivery and tissue engineering. Int J Mol Sci 14(1):1629–1654 Qian Q, Niu S, Williams GR, Wu J, Zhang X, Zhu LM (2019) Peptide functionalized dual-responsive chitosan nanoparticles for controlled drug delivery to breast cancer cells. Colloids Surf, A 5(564):122–130 Saeedi S, Murjan S, Nabid MR (2021) Redox and pH dual sensitive folate-modified star-like amphiphilic copolymer based on castor oil for controlled doxorubicin delivery. J Drug Deliv Sci Technol 1(62):102391 Sahoo SK, Labhasetwar V (2003) Nanotech approaches to drug delivery and imaging. Drug Discovery Today 8(24):1112–1120 Singh S, Jannet Vennila J, Kant R (2014) Gene interaction map: a paradigm for identifying significant pathways responsible for rheumatoid arthritis. Netw Model Anal Health Inf Bioinform 3:1–7

Recent Advancements in the Field of Stimuli-Responsive Polymeric …

101

Tomalia DA (2005) Birth of a new macromolecular architecture: dendrimers as quantized building blocks for nanoscale synthetic polymer chemistry. Prog Polym Sci 30(3–4):294–324 Varaprasad K, Karthikeyan C, Yallapu MM, Sadiku R (2022) The significance of biomacromolecule alginate for the 3D printing of hydrogels for biomedical applications. Int J Biol Macromol Xia Y, Tian X, Cui J, Zhang X, Yang Q, Zhao T, Lin Y, Cen J, Duan S, Construction of a multi-bioresponsive drug delivery system for a hyaluronic acid-geraniol polymeric prodrug and evaluation of its antitumor effect. https://doi.org/10.21203/rs.3.rs-1587453/v1 Yang K, Feng L, Liu Z (2016) Stimuli responsive drug delivery systems based on nano-graphene for cancer therapy. Adv Drug Deliv Rev 1(105):228–241 Zhao J, Zheng D, Tao Y, Li Y, Wang L, Liu J, He J, Lei J (2020) Self-assembled pH-responsive polymeric nanoparticles based on lignin-histidine conjugate with small particle size for efficient delivery of anti-tumor drugs. Biochem Eng J 15(156):107526

Nanopoxia: Antimonene-Based Nanoplatform Targeting Cancer Hypoxia for Precision Cancer Therapy Shikha Srivastava, Sarita Singh, Sanchalika Mishra, Manju Pandey, and Md. Yaqub Khan

Abstract Majority of anticancer drugs possess a wide range of toxicities that are routinely administered to cancer patients. Further hypoxic condition too reduces the drug distribution in tumours affected their therapeutic efficacies. As a result, a major portion of cancerous drugs is distributed into nearby healthy tissues causing ill effect. In cancer without overdosing of drugs states as one of the most tedious issues for their treatment. Thus, the current chapter focuses on the novel concept of nanopoxia based on tumour hypoxia that utilizes photodynamic nanotherapy for the effective release of various therapeutic agents to accomplish accuracy in cancer therapy. Later, focuse on antimonene (Sb) therapy has been made that functions by switching to cytotoxic trivalent form to kill cancerous cells under hypoxic environment. Further, the study provides clarity of hypoxia grounded accurate tumour therapy by the development of nanotherapy that gives enormous opportunity to eradicate cancer. Keywords Nanopoxia · Antimonene · Nanotherapy · Cancer therapy

1 Introduction The majority of anticancer medications with wide-ranging toxicities are routinely given to cancer-affected patients, because hypoxia reduces their distribution in tumours, and their therapeutic efficacies are jeopardized. As a result, a predominant part of cancer medications is dispersed into healthy tissues that aren’t their intended target, which frequently results in serious side effects (Singh et al. 2017). One of the most difficult problems in cancer therapy is how to use precision cancer therapy without giving patients an excessive amount of medication. Different nanoprecision medicines have been illustrated in (Fig. 1). To overcome this ice-breaking S. Srivastava · S. Singh · S. Mishra · M. Pandey (B) Institute of Pharmacy, Shri Ramswaroop Memorial University, Barabanki 225003, India e-mail: [email protected] Md. Y. Khan Biomedical Engineering, Chung Yuan Christian University, Taoyuan 32023, Taiwan © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_6

103

104

S. Srivastava et al.

Fig. 1 Programmable nanoparticles targeting various mechanisms used in cancer therapy

concept for nanopoxia came into consideration by researchers for precise therapy by reducing toxicity. Nanopoxia, being a novel photodynamic nanoplatform that utilizes the concept of tumour hypoxia for the release of therapeutic drugs to accomplish a précisedtarget cancer therapy. Recent eras have observed a significant increase in the study of two-dimensional (2D) nanomaterials for a variety of biomedical purposes, including gene and medication delivery, bio-imaging, photonic nanotherapy and biosensing, which have been discussed in Sect. 2 in detail. Researchers have explored phosphorus (black), a typical developing material, having a wide range of uses, particularly in the biological disciplines. Similarly, as black phosphorus and other 2D materials of group VA such as arsenene, antimonene and bismuthene share morphological properties; hence, it is envisaged that these 2D materials will have a wide range of uses (Tang et al. 2020; Zhao et al. 2022). One of these emerging nanosystems is 2D antimonene, which has a structure similar to graphene and is made up

Nanopoxia: Antimonene-Based Nanoplatform Targeting Cancer …

105

of sheets of sp3-hybridized antimonene atoms. Antimonene is typically produced via liquid exfoliation of bulk antimony, which has unique thermoelectric, chemically stable, optically absorbent, and superconducting properties (Wu et al. 2018). Antimonene’s honeycomb structure allows easy uploading of drugs, genes and immunosuppressant’s delivery, which points to its potential use in cancer chemotherapy, gene photothermal therapy and photothermal/immune combination therapy. The potency of Sb is further enhanced by coupling it with nanosystems and specifically with cell membrane as studied by Lu et al. (2019). Similarly, Sb-based nanomaterials display favourable photothermal characteristics and contrast-enhanced photoacoustic (PA) imaging capabilities, in particular, near-infrared (NIR) absorption. As a result, it has been anticipated that 2D antimonene might produce outstanding photothermal effects comparable to those of newly discovered 2D nanomaterials (Chen et al. 2021). Qiu et al. demonstrated that under hypoxic environment, laser irradiation causes the polymer outer shell to break down, producing anticancer reactive oxygen species. It also converts 2D antimonene (Sb) nanomaterials into cytotoxic-trivalent antimony, which together can kill cancers. Delivery of Sb nanoparticles to mice in preclinical cancer models virtually eradicates tumour development without causing any observable negative consequences (Qiu et al. 2021).

2 Different Approaches to Treat Cancer Recently, numerous treatment strategies followed for cancerous therapy for fulling different biomedical purposes, such as photodynamic, photothermal, photoacoustic, biosensing and drug–gene therapy, have been discussed below.

2.1 Photodynamic Therapy Photodynamic therapy (or PDT) uses special drug treatment, which may be called photosensitizing agents, in combination with light to kill cancerous cells (Dolmans et al. 2003). This therapy has been used to treat numerous forms of cancer as nonmelanoma skin cancer, lung, head and neck, food pipe and prostate cancers (Agostinis et al. 2011). This drug works after being activated by some specific kinds of light. PDT in terms also known as photoradiation therapy/or photochemotherapy. The photosensitizing substance is either applied to the skin or injected into the bloodstream, depending on which area of the body is being treated. The medicine is eventually taken up by the cancer cells over time. The treated area is then exposed to light which leads to a reaction with drug and forms a special form of oxygen molecule which further kills the affected cancer cells. The detailed study has been illustrated in (Fig. 2) [American Cancer Society]. Basically, effect of photodynamic therapy has been observed to destroy the cancerous cell by generating ROS by PDT which further damages vessels associated with tumours cells, then PDT also leads to the

106

S. Srivastava et al.

Fig. 2 Steps involved in photodynamic therapy

activation of immune response against tumorous cells that helps in the destruction of cancerous cells (Dolmans et al. 2003). A type photodynamic therapy called extracorporeal photopheresis treats abnormal WBCs that could cause skin-related problems in populations with cutaneous T-cell lymphoma. In ECP working, technically collects blood cells and treats them through photosensitizer, then exposed to light and administered them to body via a needle into a vein [National Cancer Institute]. Photodynamic therapy prevents damage to nearby healthy cells as the photosensitizers have a tendency to build up in abnormal cells and allows light to focus directly on them.

2.2 Photothermal Therapy Medically, in today’s era, photothermal therapy is a non-surgical, non-invasive and theoretically effective therapy procedure (Zhao et al. 2021; Xie et al. 2020). Photothermal therapy uses pulsed laser irradiation of photosensitizing chemicals to generate heat for thermal removal of cancer tumours (Chen et al. 2016). As related to traditional radiotherapy/or chemotherapy, the primary advantages of PTT include the ability for deep tissue penetration and cause minimum non-invasive effect on the surrounding healthy tissues (Jaque et al. 2014; Liu et al. 2019). A good photosensitizing agent should have a wide absorption cross section for optical wavelengths and should be low in toxicity, simple to functionalize and be readily solvable in biocompatible solutions (Jaque et al. 2014). Wang et al. (2020), demonstrated effective therapy of photothermal by coupling it with self-assembly of black phosphorous elements to improve tumour retention (Wang and Cheng 2019).

Nanopoxia: Antimonene-Based Nanoplatform Targeting Cancer …

Laser/ RF pulse

Absorption of the given energy

107

Acoustic/ ultrasonic waves

Ultrasonic detection

Thermoelastic expansion

Image formation takes place

Fig. 3 Flowchart of the photoacoustic imaging

2.3 Photoacoustic Imaging Photoacoustic imaging is a bioimaging technique grounded on the photoacoustic effect. A non-ionizing laser pulse is transmitted to living tissue and some of this energy is absorbed and converted to heat, resulting in a transient thermoelastic expansion and emission of broadband ultrasound. The generated waves are captured by an ultrasonic transducer and analysed to produce an image. It is well-known fact that light absorption is strictly related to physiological properties such as haemoglobin level and oxygen satiety (Grinvald et al. 1986; Arulmozhi et al. 2019). As a result, a physiologically distinct optical absorption contrast can be seen since the intensity of ultrasonic radiation is related to the local energy deposition. It is then possible to create a 2D or 3D image of the target area (Xu et al. 2006). Below, the flowchart shows the summary of the photoacoustic imaging formation as shown in Fig. 3.

2.4 Biosensing A biosensor’s biological receptor unit, or bioinspired receptor unit, having distinct specificities toward related specific analytes, defines the device. These analytes should be frequently biological in origin and include DNA from bacteria or viruses, proteins produced by the immune system (antibodies, antigens), and living organisms that have been infected or polluted. As biological receptor element with specificity is present, such analytes can be simple molecules like glucose or other contaminants. The effective signal capture of the biological identification event is one of the many additional difficulties in biosensor development (transduction). These transducers convert the electrochemical, electrochemiluminescent, magnetic, gravimetric or optical signals that result from the analyte’s interaction with the biological element. Similarly, an sp2 hybridized graphene oxide has been found to contain many oxygenbearing groups that aid in recognition while process of biosensing (Pumara et al. 2011). Nanomaterials are being used to raise sensitivity and lower detection limits to even single molecules (Holzinger et al. 2014).

108

S. Srivastava et al.

2.5 Drug–Gene Delivery One of the most ground-breaking medical advancements to result from the development of DNA recombination and gene cloning technology is gene therapy (Cicalese and Aiuti 2020). It is a biological procedure that modifies human genetic material. Gene therapy can directly fix or even replace the disease-causing genes at the molecular level for genetic illnesses, hence regenerating damaged protein. After decades of research and development, gene therapy has demonstrated considerable promise in the treatment of serious diseases brought on by genetic anomalies and flaws, including malignant tumours, AIDS (Acquired Immuno-Deficiency Syndrome), and cardiovascular conditions. Recent genetic therapy bioactive drugs include DNA plasmids, small-interfering RNA (Tai 2019), microRNA, short hairpin RNA and antisense oligonucleotide system. Along with the development of gene therapy and the refinement of innovative vectors, gene therapy products have received overwhelming approval from the Food and Drug Administration.

3 Role of Antimonene (AM) in Cancer Therapy Numerous methods have been explored as photodynamic therapy (PDT), photoacoustic imaging (PA), magnetic resonance imaging (MRI) and photothermal therapy (PTT), for treatment of diagnosis of cancer by the use of novel materials with specific functions. Currently, a recent element antimonene (AM) came into limelight due to its exceptional ability of effective light absorption in near-infrared region (NIR) (Fig. 4) (Yang et al. 2020a, b). This exceptional property of antimonene awards it with the potency of extraordinarily photothermal conversion efficiency, as compared to many other nanomaterials equipped photothermal agents, for example, graphene oxide, silver narods, nanoshells and quantum dots. Studies reported efficient emergence of photoacoustic imaging due to extraordinarily photothermal conversion efficiency of antimonene. These too help to generate O2 in high concentration for effective cancer hypoxia therapy.

4 Antimonene-Based Nanoplatform Targeting Cancer Hypoxia A novel photothermal treatment (PTT) agent, antimonene (AM) nanomaterial, quickly breakdowns in physiological media severely restricts its direct application. In order to overcome this issue AM have been modulated in varied form with the help of cell membrane (CM) camouflage, size management and dimension optimization. This showed noticeably better stability, higher photothermal efficacy and greater tumour-targeting ability when compared to conventional AM nanosheets. For

Nanopoxia: Antimonene-Based Nanoplatform Targeting Cancer …

109

Fig. 4 Various roles of antimonene in cancer therapy

instance, the antimonene nanosheet and quantum dots (AQDs) that were previously described had improved thermal conductivity and a high photothermal conversion effectiveness of (41.8%) and (45.5%), respectively. Additionally, Sb nanocrystals even displayed good photothermal properties. A recently discovered 2D elemental layered substance is called antimonene. Using 2D PEGylated AM nanosheets, a novel photonic drug-delivery platform is created in this study. The platform provides a number of benefits such as photothermal properties; improved drug-loading capacity; controlled release initiated by near-infrared (NIR) light or moderate acidity; accumulation at sites of tumour and deep tumour penetration by both extrinsic stimulus and intrinsic stimulus as pH) (Tao et al. 2018; Sao et al. 2021). Tao et al. reported that polyethylene glycol 2D antimonene quantum dots showed photothermal conversion efficiency of 45.5%. These also possessed a unique feature of NIR-mediated rapid degradation that aids in tumour excision ability. Thus, different works promoted biomedical applications through introduction of an entirely novel platform for PTT (Tao et al. 2017). Many more implications have been illustrated in Table 1.

5 Conclusion The current chapter focuses on the recent findings and progress on antimonene-based nanoplatforms and their uses in hypoxia-based cancer therapy. Reviewed the most promising platforms of antimonene-based nanoformulations used in various biomedical applications such as PTT, PDT, biosensing and other bioinformatics and AI (Artificial Intelligence) applications. In addition to this, the development of various antimonene-based nanoformulations such as antimonene nanosheets, antimonene

110

S. Srivastava et al.

Table 1 Implications of antimonene-based nanoformulations S. Nanoformulation Active no constituent

Targeting

Antimonene Synergistic cancer therapy

Remark

References

1

Antimonene nanosheets

This adaptable Jin et al. nanoplatform improved (2021) therapeutic efficiency through synergistic phototherapy and may be used in fluorescence and photoacoustic imaging (dual) for diagnostics and cancer treatment

2

Antimonene Antimonene Cancer two-dimensional theranostics (2D) nanomaterials

Numerous benefits Wei et al. include: outstanding (2018) photothermal properties, efficient drug loading capacity, spatiotemporally regulated drug release by near-infrared (NIR) light and mild acidic environment, superior accumulation at tumour locations and deep tumour penetration by both extrinsic as NIR light and intrinsic stimuli as NIR light

3

2D-Ultrathin nanomaterials

Antimonene Non-invasive cancer therapeutics

2D nanomaterials related to biomedical interests were introduced initially, further followed by nanoplatforms for cancer determination and treatment

4

2D Antimonene quantum dots

Antimonene Tumour cell The modified G/A Wu et al. thermo-resistance Calcium carbonate (2022) polyethylene glycol nanocatalyst exhibits prolonged blood circulation, stable at pH (neutral) but quickly degraded in acidic tumour microenvironment and causes fast drug cargo release into the tumour cells

Yang et al. (2020a, b)

(continued)

Nanopoxia: Antimonene-Based Nanoplatform Targeting Cancer …

111

Table 1 (continued) S. Nanoformulation Active no constituent

Targeting

Remark

References

5

Antimonene Xene

Antimonene Biosensors, bio-imaging, therapeutic delivery and theranostics

Xenes are thought to be Tao et al. promising agents for (2019) biosensors, bio-imaging, therapeutic delivery, theranostics and some novel bioapplications

6

Ultrathin antimonene nanoparticles

Antimonene Cancer therapy

AMNPs (ultrathin Duo et al. antimonene (2020) nanoparticles) are shown to be radiosensitizers that accomplish an efficient radio-chemotherapeutic activity by inducing a robust oxidative stress response and having very high In Vivo radiotoxicity

2D nanomaterials, 2D-ultrathin nanomaterials, 2D-antimonene quantum dots, antimonenexene and ultrathin antimonene nanoparticles have been focussed. Future studies need to be performed on antimonene nanoparticles. Acknowledgements The authors are thankful to the Director, Institute of Pharmacy, Shri Ramswaroop Memorial University, for providing an opportunity to compile a chapter.

References Agostinis P, Berg K, Cengel KA, Foster TH, Girotti AW, Gollnick SO, Hahn MR, Kessel AD, Korbelik M (2011) Photodynamic therapy of cancer: an update. CA: A Cancer J Clinicians 61(4):250–281 Arulmozhi S, Matchado MS, Snijesh VP, Kumar A, Singh S (2019) An insight into anti-arthritic property of C25H34O7 for rheumatoid arthritis using molecular modelling and molecular dynamics approach. Inf Med Unlocked 16:100145 Chen Q, Xu L, Liang C, Wang C, Peng R, Liu Z (2016) Photothermal therapy with immune adjuvant nanoparticles together with checkpoint blockade for effective cancer immunotherapy. Nat Commun 21; 7(1):1–3 Chen Y, Wang M, Zheng K, Ren Y, Xu H, Yu Z, Zhou F, Liu C, Qu J, Song J (2021) Antimony nanopolyhedrons with tunable localized surface plasmon resonances for highly effective photoacoustic-imaging-guided synergistic photothermal/immunotherapy. Adv Mater 33:2100039 Cheng L, Wang C, Feng L, Yang K, Liu Z (2014) Functional nanomaterials for phototherapies of cancer. Chem Rev 114:10869–939

112

S. Srivastava et al.

Cicalese MP, Aiuti A (2020) New perspectives in gene therapy for inherited disorders. Pediatr Allergy Immunol 31:5–7 Dolmans DE, Fukumura D, Jain RK (2003) Photodynamic therapy for cancer. Nat Rev Cancer 3:380–387 Duo Y, Huang Y, Liang W, Yuan R, Li Y, Chen T, Zhang H (2020) Ultra effective cancer therapy with an antimonene-based X-ray radiosensitizer. Adv Func Mater 30:1906010 Grinvald A, Lieke E, Frostig RD, Gilbert CD, Wiesel TN (1986) Functional architecture of cortex revealed by optical imaging of intrinsic signals. Nature 324:361–364 Holzinger M, Le Goff A, Cosnier S (2014) Nanomaterials for biosensing applications: a review. Front Chem 27:63 Jaque D, Maestro LM, del Rosal B, Haro-Gonzalez P, Benayas A, Plaza JL, Rodríguez EM, Sole JG (2014) Nanoparticles for photothermal therapies. Nanoscale 6:9494–9530 Jin X, Xu C, Hu J, Yao S, Hu Z, Wang B (2021) A biodegradable multifunctional nanoplatform based on antimonene nanosheets for synergistic cancer phototherapy and dual imaging. J Mater Chem B 9:9333–9346 Lu G, Lv C, Bao W, Li F, Zhang F, Zhang L, Wang S, Gao X, Zhao D, Wei W, Xie HY (2019) Antimonene with two-orders-of-magnitude improved stability for high performance cancer theranostics. Chem Sci 10:4847–4853 Liu Y, Bhattarai P, Dai Z, Chen X (2019) Photothermal therapy and photoacoustic imaging via nanotheranostics in fighting cancer. Chem Soc Rev 48:2053–2108 Photodynamic therapy to treat cancer – NCI, www.cancer.gov Photodynamic Therapy, www.cancer.org Pumera M (2011) Graphene in biosensing. Mater Today 14:308–315 Qiu M, Duo Y, Liang W, Yang Y, Zhang B, Xie Z, Yang X, Wang G, Xie N, Nie G, Alhartomy OA (2021) Nanopoxia: targeting cancer hypoxia by antimonene-based nanoplatform for precision cancer therapy. Adv Func Mater 31(42):2104607 Sao P, Chand Y, Kumar A, Singh S (2021) Potential drug target identification in porphyromonas gingivalis using in-silico subtractive metabolic pathway analysis. Bangladesh J Med Sci 20(4):887–896 Singh SK, Singh S, Lillard JW Jr, Singh R (2017) Drug delivery approaches for breast cancer. Int J Nanomed 24(12):6205–6218 Singh SK, Singh S (2019) Breast cancer: current knowledge and future perspective. Acta Sci Women’s Health 1:50–51 Tai W (2019) Chemical modulation of siRNA lipophilicity for efficient delivery. J Control Release 10:98–107 Tang Z, Kong N, Ouyang J, Feng C, Kim NY, Ji X, Wang C, Farokhzad OC, Zhang H, Tao W (2020) Phosphorus science-oriented design and synthesis of multifunctional nanomaterials for biomedical applications. Matter 5:297–322 Tao W, Ji X, Xu X, Islam MA, Li Z, Chen S, Saw PE, Zhang H, Bharwani Z, Guo Z, Shi J (2017) Antimonene quantum dots: synthesis and application as near-infrared photothermal agents for effective cancer therapy. Angew Chem 18:12058–12062 Tao W, Ji X, Zhu X, Li L, Wang J, Zhang Y, Saw PE, Li W, Kong N, Islam MA, Gan T (2018) Two-dimensional antimonene-based photonic nanomedicine for cancer theranostics. Adv Mater 30:1802061 Tao W, Kong N, Ji X, Zhang Y, Sharma A, Ouyang J, Qi B, Wang J, Xie N, Kang C, Zhang H (2019) Emerging two-dimensional monoelemental materials (Xenes) for biomedical applications. Chem Soc Rev 48:2891–2912 Wang X, Cheng L (2019) Multifunctional two-dimensional nanocomposites for photothermal-based combined cancer therapy. Nanoscale 11:15685–15708 Wu J, Cai X, Williams GR, Meng Z, Zou W, Yao L, Hu B, Chen Y, Zheng Y (2022) 2D antimonene-integrated composite nanomedicine for augmented low-temperature photonic tumor hyperthermia by reversing cell thermoresistance. Bioactive Mater 1:295–305 Xu M, Wang LV (2006) Photoacoustic imaging in biomedicine. Rev Sci Instrum 17:041101

Nanopoxia: Antimonene-Based Nanoplatform Targeting Cancer …

113

Yang W, Lyu Q, Zhao J, Cao L, Hao Y, Zhang H (2020a) Recent advance in near- infrared/ultrasoundsensitive 2D-nanomaterials for cancer therapeutics. Sci China Mater 63:2397–2428 Wang Y, Xie J, Kang J, Choi W, Jangili P, Zhang B, Xie N, Nie G, He J, Zhang H, Liu L (2020) Smart acid-activatable self-assembly of black phosphorous as photosensitizer to overcome poor tumor retention in photothermal therapy. Adv Func Mater 30:2003338 Wei Z, Yin X et al (2018) Antitumor effect of a Pt-loaded nanocomposite based on graphene quantum dots combats hypoxia-induced chemoresistance of oral squamous cell carcinoma. Int J Nanomed 13:1505 Xie Z, Fan T, An J, Choi W, Duo Y, Ge Y, Zhang B, Nie G, Xie N, Zheng T, Chen Y (2020) Emerging combination strategies with phototherapy in cancer nanomedicine. Chem Soc Rev 49:8065–8087 Yang X, Wu R, Xu N, Dong N, Ling G, Liu Y, Zhang P (2020b) Application and prospect of antimonene: a new two-dimensional nanomaterial in cancer theranostics. J Inorg Biochem 1:111232 Zhao L, Zhang X, Wang X, Guan X, Zhang W, Ma J (2021) Recent advances in selective photothermal therapy of tumor. J Nanobiotechnol 19:1–5 Zhao Y, Wang SB, Chen AZ, Kankala RK (2022) Nanoarchitectured assembly and surface of twodimensional (2D) transition metal dichalcogenides (TMDCs) for cancer therapy. Coord Chem Rev 1:214765

Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials Aashna Srivastava, Dharmendra Prajapati, Sachidanand Singh, and Tanvi Jain

Abstract Hypoxia has always been an obstacle to therapeutic effectiveness of radiotherapy. The hypoxic microenvironment is recognized by the presence of decreased oxygen levels. The development of methods to enhance the efficacy of radiotherapy for the treatment of hypoxia using nanomaterials is a promising strategy to tackle the issue of hypoxia. This chapter intends to provide an overview of all the innovative and novel strategies which have been developed in recent years to enhance radiotherapy using nanomaterials to reduce hypoxia. The major approaches to tackle hypoxia: (1) direct delivery of oxygen in the hypoxia region, (2) in situ generation of oxygen in the hypoxia region using catalyzed decomposition of H2 O2, (3) nanocomposites acting as radiosensitizers, and (4) delivery nanoparticles to alleviate the hypoxia region using radiations are discussed in detail in the following chapter. Keywords Hypoxia · Radiotherapy · Nanomaterials · Therapeutics

1 Introduction Hypoxia is generally known to be a decreased oxygen partial pressure (also known as oxygen tension, or pO2 ) lower than a physiological degree. However, assigning a single number to this notion is more challenging: In the human body, physiological pO2 ranges from 100 mmHg in the lung alveoli to below 1 mmHg in the mitochondria. The “oxygen cascade,” or pressure gradient, directs oxygen from the “supply” (alveolar gas) to the “sinks,” ensuring its diffusion throughout the body (the cells). Physiologically, all pO2 values in this cascade might be termed “normoxic.” A. Srivastava · T. Jain (B) Faculty of Biotechnology, Institute of Biosciences and Technology, Shri Ramswaroop Memorial University, Barabanki 225003, India e-mail: [email protected] D. Prajapati · S. Singh Department of Biotechnology, Smt. S.S. Patel Nootan Science and Commerce College, Sankalchand Patel University, Visnagar-384315, Mehsana, Gujarat, India © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_7

115

116

A. Srivastava et al.

Solid tumors need the development of a neovascular system in order to grow and survive. Tumor angiogenesis is induced by the hypoxic microenvironment produced by rapidly multiplying tumor cells. The rapidly expanding blood vessels that make up the neovascular network are frequently abnormal, disordered, and dysfunctional, which results in a lack of oxygen delivery that worsens tumor hypoxia (Baluk et al. 2005; Nagy et al. 2010). On the other hand, oxygen demand frequently becomes uncontrolled after tumor-specific metabolic alterations. When assembled together, these factors provide a dynamic, promptly evolving tumor microenvironment with areas of nutrition and oxygen deprivation (Wilson et al. 2011). Tumor hypoxia was first discovered in the 1950s, when Gray and Thomlinson established, using mathematical calculations subjected to histological segments of human tumors, that areas around necrotic sites exhibit a hypoxic oxygen gradient (Telarovic et al. 2021). It has recently been demonstrated that a wide range of variable oxygen tension values (ranging from slight oxygen shortage to anoxia) are heterogeneously distributed throughout the tumor microenvironment, in contrary to the early stance at a radial hypoxia gradient largely develops from the necrotic tumor center approaching to the tumor margin. Regardless of the tumor stage, histological grade, or lymph node status, hypoxia and necrosis have each been independently shown to be predictive of poor clinical outcomes (Vaupel 2009). Additionally, hypoxia has been linked to genomic instability, immune system repression (Facciabene et al. 2011), the many stages of the metastatic cascade (such as annexation, migration, intravasation, and extravasation, creation, and sustenance of the premetastatic niche), (Rankin et al. 2016) and an increase in chemotherapy and radiotherapy resistance (Telarovic et al. 2021). Radiotherapy is a popular and efficient cancer treatment method. The science of radiotherapy had its start when Nobel Prize winner Marie Curie learned how radiation affected human cells. Ionizing radiation is used as a therapeutic strategy due to its ability to produce varied DNA damage and lead to cellular death in desired regions (clinical and/or subclinical lesions). The ability of cancer cells to divide uncontrollably makes them more exposed to radiation-instigated DNA damage. Presently, above 60% of cancer patients undergo radiotherapy as part of their anti-cancer therapeutic. This is done using a variety of methods, such as brachytherapy and external beam (photons, protons, and electrons) (internal radioactive source). The method of administration is determined by the clinical implications (Mi et al. 2016). More than 50% of cancer patients with locally advanced solid tumors continue to get their primary care as radiotherapy. Poor tumor oxygenation (hypoxia), which is present in a variety of solid tumor forms, can, nevertheless, result in tumor resistance to radiation, incomplete tumor eradication, and local tumor recurrence (Barker et al. 2015; Horsman et al. 2012). In several cancer types, hypoxia has repeatedly been demonstrated to be a potent predictive clinical indication of radioresistance and shortened disease-free survival (Vaupel et al. 2007). For instance, tumorregions in nearly 40% of locally advanced breast cancers have oxygen concentrations below what is necessary for half-maximal radiosensitization, making radiotherapy ineffective when used alone. This has prompted research into radiotherapy combinations with other therapies and treatment options (Abbasi et al. 2016). The efficacy of radiotherapy

Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials

117

is vitally dependent on the relative quantity of oxygen in the tumor at the time of irradiation because oxygen can accelerate the creation of DNA double-strand breaks (DSB) induced directly by ionization of DNA and indirectly by free radicals generated by radiolysis of water (Bristow et al. 2008). Several studies have demonstrated that hypoxic cells can be two to three times more resistant to a single fragment of ionizing radiation than cells that are exposed to radiation in the presence of normal oxygen levels. In many different malignancies, hypoxia has also been associated with the emergence of an aggressive phenotype, primarily as a result of the overexpression of hypoxia-inducible factors (Abbasi et al. 2016). When compared to oxygenated healthy tissues, hypoxic tumors frequently exhibit high amounts of reactive oxygen species (ROS), low pH, and altered metabolism (Sharma et al. 2019). The likelihood of tumor spread is often increased by hypoxia because it causes intratumoral heterogeneity and inhibits innate and adaptive immune responses (Kumar et al. 2014; Palazón et al. 2012). It is also known that some tumor cells may live in low oxygen environments (Masson et al. 2014), and what’s worse, these tumor cells are more resistant to conventional treatments for them, such as radiation therapy, chemotherapy, and photodynamic therapy (PDT) (Höckel et al. 2001; Lu et al. 2019; Shannon et al. 2003). Recently, several studies have been conducted with the objective of improving the hypoxic microenvironment, for instance, by using inspiratory hyperoxia (Zou et al. 2019; Arulmozhi et al. 2019). Unfortunately, due to the severe anatomical abnormality of microvessels in tumors and the negative results of decompression oxygen therapy, it is difficult to adapt this technology to clinical therapy (Liu et al. 2018). Therefore, it is essential to develop effective treatments for hypoxic tumors. Over the last few decades, there has been a successful union of modern medicine with nanotechnology that nurtures to initiate the creation of individualized “nanomedicine” established on multifunctional nanomaterials (Lin et al. 2018; Wolfram et al. 2019; Mi et al. 2020; Wang et al. 2020). Recently, nanomaterials have been used to boost RT while reducing radiotoxicity by enhancing radiation reactions and overcoming radiation resistance (Kunz-Schughart et al. 2017; Yang et al. 2020). Nanomaterials could function as radiosensitizers independently or as drug carriers to transfer additional radiosensitive components for improved RT. For instance, radiosensitizers can be used directly in nanomaterials incorporating high-atomic number elements (such as Au, Bi, Hf, and W) to escalate radiation absorption and accumulation of radiation energy inside tumors during EBRT. Additionally, nanoparticles could effectively target tumors by active targeting and passive targeting (also known as the improved permeability and retention effect, or EPR effect) to deliver radiosensitizers for EBRT16 or therapeutic radioisotopes for RIT17. Additionally, multifunctional nanomaterials make it possible to combine RT with other therapies in order to influence the TME (e.g., alleviate hypoxia, regulate angiogenesis, and regulate immunosuppression), repress radiation resistance, and achieve synergistic RT (Zeng et al. 2021). This chapter discusses the various strategies which have utilized nanomaterials to increase the efficacy of radiotherapy to reduce hypoxia.

118

A. Srivastava et al.

2 Nanotechnology in Radiotherapy To overcome the low tolerance of normal tissue and obtain a high therapeutic ratio, new radiotherapy (RT) methods have been created. Despite the use of advanced techniques, tumor responses are only partially effective in hypoxic tumor situations. Hypoxia is a critical regulator of tumor growth and is a fundamental contributor to RT resistance. Furthermore, RT can cause hypoxia cycling in tumor areas. Following RT, the lack of oxygen reduces the formation of reactive oxygen species (ROS), which ultimately stops tumor cells from suffering irreparable DNA damage and leading to tumor cell death (Barker et al. 2015). Hypoxia also causes the hypoxia-inducible factor 1 (HIF-1) to be upregulated, which independently increases radioresistance (Semenza 2004). The use of nanoparticles in RT is one method of overcoming RT’s limitations. Antitumor therapies utilize nanoparticles for a number of functions, including drug administration (Zhao et al. 2014), diagnostics (Perrault et al. 2009; Qian et al. 2008), gene therapy (Conde et al. 2013), and RT (Kim et al. 2016). When nanoparticles are coupled with RT, the absorbed radiation dose increases. An overview of the applications of nanotechnology in improving the efficacy of radiotherapy against tumors is illustrated in Fig. 1.

2.1 Perfluorocarbons (PFCs) as O2 Carriers Over the last decade, various remarkable O2 nanoplatforms for sustained release have been produced. A perfluorooctane composite was loaded into hollow microparticles by Lee et al. (2015) to create a scaffolding arrangement for well-timed delivery of oxygen. An in vitro investigation showed that tissues neovascularized up to 4 mm, instead of necrosis and cell survival under hypoxic circumstances was extended by 10 days. To overcome hypoxia-related radioresistance, Cheng et al. (2015) illustrated more efficient particle accumulation and targeted O2 release in tumors. Briefly, they used a simple technique to easily encapsulate perfluorohexane within liposomes to create 100 nm nanoparticles (NPs), which they subsequently injected into tumorbearing animals before photodynamic therapy or radiation. It should be noted that this outcome was attained with no addition of extra oxygen since the oxygen packed in the core of the NPs was adequate to neutralize hypoxia, owing to the elevated permeability and retention (EPR) effect generated by a significant nanoparticle size and free oxygen dispersion in the core of NPs. The creation of innovative O2 delivery technologies is unquestionably advantageous for therapeutic modalities like radiation and photodynamic therapy. Reoxygenation and radiosensitizers may work well together in vivo. A sort of hollow Bi2 Se3 NPs with PFC loaded on them was created by Song et al. (2016) as an oxygen delivery system. In addition to bismuth’s radiosensitizing properties, this arrangement has strong near-infrared (NIR) absorbance that could be used for photothermal therapy and the targeted stimulation of swift O2

Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials

119

Fig. 1 Major strategies to target hypoxia

distribution. In in vivo, a synergistic therapeutic effect was found. They then added PFCs to tantalum oxide (TaOx) NPs to further improve radiotherapy (Li et al. 2018). Nonetheless, given the restricted penetration limit of NIR light and retention by the skin and major organs, a more versatile and responsive system to initiate the activity of these composite NPs at safe laser magnitude to prevent hyperthermia in regular tissues may be necessary. The Food and Drug Administration for ophthalmological treatments and percutaneous transluminal coronary angiography approved the PFC composites (such as Fluosol-DA, 20%) as early as 1989. Hartzler et al. (1988), Teicher et al. (1995) gave tumor-bearing mice a perflubron emulsion (8 mL/kg) with carbogen breathing (95% O2 /5% CO2 ) just after chemo-/radiotherapy; astonishingly, the hypoxic zone (PO2 5 mmHg) dropped from 85 to 27% as a result of reoxygenation. Similar outcomes for GL261 intracranial tumors were recently obtained by Feldman et al. (2017). Using a different method, Yao et al. (2015) created a poly(lactic-coglycolic acid) (PLGA)-(polyethylene glycol) (PEG/PFC) composite to attain very efficient and quick reoxygenation in cells and organisms. It was also mentioned that the possibility of hypoxia–reoxygenation injury might lead to a large amount of reactive oxygen species (ROS) to be produced and destroy healthy tissues. Due to this, the

120

A. Srivastava et al.

PEG/PFC composite dose is now under 0.2 mg/mL. Furthermore, tumor responses to radiotherapy are controlled by intermediate oxygen levels (0.5–20 mmHg) (Wouters et al. 1997).

2.2 Red Blood Cell (RBC)-Based O2 Carriers RBCs, also known as erythrocytes, have been exploited as an effective endogenous delivery system for drug administration since 1970 because of their exceptional biocompatibility, lengthy circulation durations (around 120 days), and distinctive loading volume (high surface-to-volume ratio). Gao et al. (30) created PFC@PLGARBCM NPs in order to take use of these properties of RBCs for O2 delivery and enhance the effectiveness of radiotherapy. In addition, the NP half-life was amplified to 13.93 h, which considerably raised oxygenation magnitudes from 1.6% to 24%. The key findings of this research presented that the nanosized RBC replica was capable to perforate the vasculature and diffuse deeply inside the hypoxic zones in solid tumors (Li et al. 2018).

2.3 H2 O2 Catalyst-Based Reoxygenation Nanoplatforms Strong evidence shows that major solid tumors contain endogenous H2 O2 . The “Warburg effect,” which causes tumor cells to create excessive amounts of lactic acid, also contributes to the TME’s low pH (Kuang et al. 2011). MnO2 has been used as a crucial constituent of various nanoplatforms designed to reoxygenate hypoxic tumors and increase therapeutic efficiency because it offers good catalytic properties to decompose H2 O2 into O2 and H2 O in the presence of acidic conditions (Cho et al. 2017; Tian et al. 2017). Fan et al. used a single nanoplatform to perform both stimuli-responsive imaging and O2 upregulation treatment (Fan et al. 2015a; b). They used an easy non-chemical technique to successfully anchor upconversion NPs (UCNPs) to 2D MnO2 nanosheets. Under normal circumstances, upconversion photoluminescence was quenched by MnO2 , but in the tumor region, upconversion photoluminescence was noticeably enhanced, which can be interpreted by the escalation of the MnO2 –H2 O2 redox reaction at a low pH.(5.5 vs. 7.4). The synchronized photodynamic therapy and radiation were considerably improved by the huge oxygen created during this technique, which was later proved in in vivo and in vitro experiments. Recent researches have concentrated on coupling a catalyst and a high-atomic number element, resulting in a series of inventive nanoplatforms combining H2 O2 catalysts with high-Z elements like tungsten (WS2-IO/S@MO-PEG), (Yang et al. 2018) gold (Au@MnO2), (Yi et al. 2016) or hafnium (BM@NCP (DSP)-PEG), (Liu et al. 2017) for multimodal photodynamic therapy/radiation therapy/chemotherapy. Tian et al. (2017) created 131I-labeled human serum albumin (HSA)-bound MnO2 NPs for internal radiotherapy. By adjusting the particle size, these NPs demonstrated

Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials

121

more EPR-derived tumor retention than that of free 131I-HSA. Moreover, the NP size could be successively decreased to 10 nm by progressive MnO2 deterioration in the acidic TME. The HSA-based nanocarrier’s intratumoral diffusion enhanced noticeably, and endogenous H2 O2 was transformed into H2 O and O2 , which greatly enhanced radiotherapy’s effectiveness. Yongtian Liu et al. (2021a, b) suggested a procedure for creating a BCHN (BiOCl/Cu2+ -H2 O2 @PVP) nanocomposite that alters the tumor microenvironment (TME) by providing H2 O2 , producing O2 , by the consumption of GSH, and producing •OH. Under slightly acidic conditions, BCHN can release self-carried H2 O2 to produce BCN (BiOCl/Cu@PVP) in tumor cells. Then, under the influence of overexpressed GSH in tumor cells, BCN goes through biodegradation through the reaction of Bi3+ and GSH to take up GSH and the reduction of Cu2+ to Cu+ by GSH, which upsets intracellular redox equilibrium. Self-supplied and endogenous H2 O2 combine with Cu2+ and Cu+ to create •OH and O2 . CDT is facilitated by the ongoing generation of •OH and consumption of GSH, which jointly regulate intracellular oxidative stress levels. The produced O2 also alleviates hypoxia in tumors. This approach improves the effectiveness of RT by combining it with the radiosensitizing effect of bismuth under X-ray irradiation. The synergistic anti-tumor actions of CDT and RT are improved when the amount of oxidative stress in the TME is raised. According to the results, the biodegradable nanomaterial BCHN will form the base for future advancements in CDT-enhanced RT of tumors.

2.4 Magnetic Nanoparticles Magnetic nanoparticles have been exploited as drug delivery systems for tumor therapy due to their good biocompatibility and biodegradability (Wu et al. 2019). Using polyacrylic acid-coated nanoscopic super-paramagnetic iron oxide nanoparticles (PAA@USPIOs), Yang et al. recently created a cisplatin-packed, poly dopaminecoated, and GE11 peptide-amalgamated multifunctional theranostic arrangement (GE11-PDA-Pt@USPIOs). By effectively delivering packed cisplatin to EGFRpositive tumor cells, GE11-PDA-Pt@USPIOs can improve tumor tissue hypoxia and boost the anti-tumor effectiveness of chemoradiation. Yang et al. (2019) created a drug-carrying arrangement using versatile graphene oxide in a different recent study (GO). This medication delivery method has the ability to simultaneously administer radiosensitizers like FePt magnetic nanoparticles and chemotherapeutic medicines (5-Fu). These nanocomposites’ cytotoxicity and radiosensitization were demonstrated in vitro (Liu et al. 2021).

122

A. Srivastava et al.

2.5 High Atomic Number Elements-Based Nanoparticles Zhang et al. created a new BiPt-folic acid-modified amphiphilic polyethylene glycol nanocomposite (BiPt-PFA). A Pt element in the BiPt-PFA nanocomposite has an effective capacity for NIR assimilation, which can result in photothermal results. Additionally, this fabricated nanocomposite has the capacity to boost tumor tissues’ X-ray absorption, intake glutathione, and catalyze the breakdown of peroxides into oxygen to treat hypoxia in TME. In order to moderate the microenvironment and enhance the reactivity of tumors to photothermal therapy and radiotherapy, BiPt-PFA may be used as the effective material (Zhang et al. 2020). Zhou et al. created an effective radiosensitizer known as bismuth heteropolytung state (BiP5W30) nanobundles. The BiP5W30 nanobundles, which contain the elements Bi and W, can improve tumor tissue hypoxia, escalate X-ray deposition in tumor tissues, and boost glutathione consumption, all of which can increase the effectiveness of radiation. As an NIR optical absorbance agent, reduced graphene oxide (rGO) can increase tumor blood perfusion in vivo and have a photothermal impact. Combining rGO with BiP5W30 nanoclusters may provide a brand-new sensitizer for synergistic thermo-radiotherapy (Zhou et al. 2019). Bao et al. integrated hafnium (Hf) bundles and manganese (III)– porphyrin ligands into a nanoscale metal–organic substructure to create a novel nanocomposite (fHMNM). The catalase-like Mn(III)–porphyrin ligand found in fHMNM can alleviate tumor tissue hypoxia, reduce hydrogen peroxide to oxygen, and increase PTT and RT sensitivity. Synergistic thermo-radiotherapy regulated by fHMNM effectively decreased tumor tissue growth in mice cancer models, with no apparent toxic reaction (Bao et al. 2020).

2.6 Nanospheres Yong et al. created gadolinium-containing polyoxometalate-linked chitosan nanospheres (GdW10@CS). These nanospheres can induce and mediate HIF-1a siRNA to downregulate HIF-1 expression and block DNA self-healing. Additionally, GdW10@CS nanospheres increased tumor tissue hypoxia and glutathione uptake, increasing the anti-tumor effectiveness of radiotherapy (Yong et al. 2017).

2.7 Salmonella Typhi Ty21A GNPs According to a study, attenuated strains of the anaerobic bacterium Salmonella typhi Ty21A have considerable advantages as radiosensitizers and as vehicles for delivering GNPs into hypoxic tumor volumes. This study investigated the effects of combining radiation with several types of modified nanoformulations, including “Citrate-GNPs, Gelatin-GNPs, BSA-GNPs, FA-GNPs, Glutamine-citrate-GNPs, Glut-BSA-GNPs,

Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials

123

and Glut-BSA-GNPs” by Salmonella typhi Ty21a. Among these, FA-GNPs are generally regarded as the prime option for synthesizing golden bacteria that may increase GNP delivery across anoxic tumor volume. This delivery method consequently reduced radioresistance throughout the tumor microenvironment. Delivery of nanoparticles by bacteria results in an effective photothermal therapy across the tumor volume (Luo et al. 2016). The in vivo biodistribution of NPs has a major impact on radiotherapy efficacy and tumor hypoxia; this biodistribution is solely dependent on various parameters including size, shape, charge, and other surface properties (Zhou et al. 2022).

2.8 Nanocarriers with Hypoxia-Responsive Prodrugs Recent studies have revealed that, when paired with radiation, the distribution of hypoxia-responsive prodrugs (tirapazamine (TPZ)) encapsulated in nanocarriers can play a significant role in regulating the hypoxia situation in tumor tissues. TPZ is an anti-tumor drug that exhibits toxicity to tumor cells at comparatively low oxygen tensions (Brown 1993). In order to avoid the oxygen demand during radiotherapy, a multifunctional nanoradiosensitizer (TPZ@UCHMs) was successfully synthesized to encapsulate the TPZ. In vitro and in vivo models were used to determine the effectiveness of these nanoparticles in order to determine how TPZ modulates radiosensitivity. A colony-forming assay was used to determine the involvement of oxygen and TPZ@UCHMs in radiation-induced cytotoxicity (Shuryak et al. 2009; Sao et al. 2021). When measured to hypoxic cells co-cultured with separate therapeutic classes of TPZ or UCHMs, the survival rate of cancer cells medicated with TPZ@UCHMs in integration with RT significantly declined. The blend of RT and TPZ@UCHMs demonstrated therapeutic effectiveness in tumor-bearing nude mice by inhibiting hypoxia and counteracting hypoxia-mediated metastasis (Liu et al. 2015).

2.9 Oxygen-Carrying Nanobubbles Oxygen nanobubbles with a stabilizing monolayered shell and the ability to carry oxygen have been studied for tumor oxygenation and tumor therapy effectiveness (Bhandari et al. 2017; Song et al. 2019). These oxygen nanobubbles were made of a dextran, lipid, polymer, and gas vesicle nanolayered shell surrounding an oxygen core (Song et al. 2018). Depending on the medications’ hydrophobic or hydrophilic qualities, drugs can be packed into the nanobubbles namely by encapsulating them in the core or by plating the outside covering in a covalent or non-covalent manner. When coupled with other therapies, oxygen nanobubbles have been employed to decrease the expression of hypoxia-inducible factor-1 (HIF-1) to improve the therapeutic benefits of radiotherapy and chemotherapy (Kheir et al. 2012; Song et al. 2020). To avoid premature oxygen delivery and lessen negative effects, oxygen may

124

A. Srivastava et al.

also respond by responsively releasing in tumors in response to external stimuli. In order to enable the release of oxygen to treat tumor hypoxia in the tumor microenvironment, Song et al. employed acetylated dextran, a polymer that responds to pH (Zou et al. 2021).

2.10 Haemoglobin-Based Oxygen Carriers Current research studies have demonstrated the potential of hemoglobin (Hb)-based oxygen delivery systems for treating tumor hypoxia, as these carriers have a high O2 -loading ability (Yang et al. 2018). Hb can be used as an effective component for oxygen delivery, but its use is restricted due to nephrotoxicity and immunogenicity (Jain et al. 2017). To get around this problem, Xia D. et al. created Au-Hb@PLT and examined its radiosensitization effectiveness; Au-Hb@PLT reduced hypoxia in tumor tissue by ensuring appropriate oxygen release during radiotherapy (Xia et al. 2020). The clinical outcomes of radiotherapy were improved by Au-Hb@PLT since it could deliver oxygen molecules directly and worked well as a radiosensitizer throughout the tumor tissues. The tumor targeting efficacy of Au-Hb@PLT when coupled with nanoparticles was tested in HeLa cells and tumor-bearing nude mice (Zhou et al. 2022).

2.10.1

Gas-Generating Nanomaterials

NO, a messenger molecule, is crucial for immunological response, angiogenesis, and cardiovascular homeostasis, among other physiological processes (Bogdan 2015; Moncada et al. 2002). In tumor therapy, NO is a concentration-dependent “double-edged sword,” though. For tumor inhibition, NO directly kills tumor cells at high concentrations (>1 mM). NO not only reduces tumor hypoxia and multidrug resistance-associated proteins expression at relatively low concentrations (1 M– 1 mM) (Guo et al. 2017) but also inhibits P-glycoprotein and multidrug resistanceassociated protein expression (Jin et al. 2017). The processes by which NO relieves a hypoxic tumor include modifying blood vessel relaxation to enhance blood flow, speeding the metabolism of intracellular GSH, and lowering the oxygen consumption rate of the tumor to oxygenate it (Riganti et al. 2005). Shi’s group developed an X-ray-activated synergistic NO gas/radiotherapy system (PEG-USMSs-SNO) to treat hypoxic tumors by grafting mesoporous silica with S-nitrosothiol (R-SNO). After the S–N link was selectively broken by X-ray radiation, NO was released. This released NO might then promote radiosensitization of hypoxic tumors and enhance the radiotherapeutic actions against these tumors (Fan et al. 2015a; b).

Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials

125

3 Conclusion The chapter intended to summarize the novel methods in radiotherapy which use nanomaterials that have been created to reduce hypoxia. With the development of technology, the field of radiation therapy in the treatment of cancer is rapidly evolving. These advancements include the ability to deliver high dosages to more accurate volumes and moving targets. The most rational answer is that radiotherapy efficacy is constrained by normal tissue damage, tumor resistance, and precise radiotherapy administration. Therefore, extensive studies into the contribution that nanotechnology can play in overcoming these restraints could be beneficial for radiation oncology. Nanotechnology has the potential to improve the transport and/or concentration of radiosensitizers or radioisotopes, hence increasing their anti-tumor effectiveness. Recent research on the impact of radiotherapy on tumor microenvironments has also sparked interest in alternative radiotherapy combination therapies, particularly those that combine radiotherapy and immunotherapy. Tumor hypoxia might be alleviated by delivering oxygen to tumors via oxygen-packed nanoparticles, oxygen-creating nanomaterials, or oxygen-budgeting nanomaterials, which sensitized or improved the therapeutic benefits of oxygen-dependent tumor therapy. Due to the entanglement of hypoxia tumors, a lot of research has also concentrated on hypoxic tumor treatment methods using nanomaterials that are less oxygendependent. Because of their distinctive physical and chemical characteristics, nanomaterials have been extensively used to improve RT. As it is greatly said by the great physicist and Nobel Prize laureate, Richard Feynman, “There’s Plenty of Room at the Bottom” when he introduced the world to the concept of nanotechnology in 1959, it is believable that there are a lot of exploration to be done and advances to be made in medical science using nanotechnology.

References Abbasi AZ, Gordijo CR, Amini MA, Maeda A, Rauth AM, DaCosta RS, Wu XY (2016) Hybrid manganese dioxide nanoparticles potentiate radiation therapy by modulating tumor hypoxia. Cancer Res 76(22):6643–6656 Arulmozhi S, Matchado MS, Snijesh VP, Kumar A, Singh S (2019) An insight into anti-arthritic property of C25H34O7 for rheumatoid arthritis using molecular modelling and molecular dynamics approach. Inf Med Unlocked 16:100145 Baluk P, Hashizume H, McDonald DM (2005) Cellular abnormalities of blood vessels as targets in cancer. CurrOpin Genet Dev. 15(1):102–111 Bao J, Zu X, Wang X, Li J, Fan D, Shi Y, Xia Q, Cheng J (2020) Multifunctional Hf/Mn-TCPP metal-organic framework nanoparticles for triple-modality imaging-guided PTT/RT synergistic cancer therapy. Int J Nanomed 15:7687–7702 Barker HE, Paget JT, Khan AA, Harrington KJ (2015) The tumour microenvironment after radiotherapy: mechanisms of resistance and recurrence. Nat Rev Cancer 15:409–425 Bhandari PN, Cui Y, ElzeyBD et al (2017) Oxygen nanobubblesrevert hypoxia by methylation programming. Sci Rep 7:9268

126

A. Srivastava et al.

Bogdan C (2015) Nitric oxide synthase in innate and adaptive immunity: an update. Trends Immunol 36:161–178 Bristow RG, Hill RP (2008) Hypoxia and metabolism. Hypoxia, DNA repair and genetic instability. Nat Rev Cancer 8:180–192 Brown JM (1993) SR 4233 (Tirapazamine): a new anticancer drug exploiting hypoxia in solid tumours. Br J Cancer 67:1163–1170 Cheng Y, Cheng H, Jiang C et al (2015) Perfluorocarbon nanoparticles enhance reactive oxygen levels and tumour growth inhibition in photodynamic therapy. Nat Commun 6:8785 Cho MH, Choi E, Kim S, Goh S, Choi Y (2017) Redox-responsive manganese dioxide nanoparticles for enhanced MR imaging and radiotherapy of lung cancer. Front Chem 5:109 Conde J, Rosa J, de la Fuente JM, Baptista PV (2013) Gold-nanobeacons for simultaneous gene specific silencing and intracellular tracking of the silencing events. Biomaterials 34:2516–2523 Facciabene A, Peng X, Hagemann IS, Balint K, Barchetti A, Wang L-P et al (2011) Tumour hypoxia promotes tolerance and angiogenesis via CCL28 and T reg cells. Nature 475(7355):226–230 Fan W, Bu W, Shen B et al (2015a) Intelligent MnO2 nanosheets anchored with upconversionnanoprobes for concurrent pH-/H2O2-responsive UCL imaging and oxygen-elevated synergetic therapy. Adv Mater 7(28):4155–4161 Fan W, Bu W, Zhang Z et al (2015b) X-ray radiation-controlled NO-release for on-demand depthindependent hypoxic radiosensitization. AngewChemInt Ed 54:14026–14030 Feldman LA, Fabre MS, Grasso C et al (2017) Perfluorocarbon emulsions radiosensitise brain tumors in carbogen breathing mice with orthotopic GL261 gliomas. PLoS ONE 12(9):e0184250 Guo R,Tian Y, Wang Yet al (2017) Near-infrared laser-triggered nitric oxide nanogenerators for the reversal of multidrug resistance in cancer. Adv Funct Mater 27:1606398 Hartzler GO, Rutherford BD, Mcconahay DR, Johnson WL, Giorgi LV (1988) “High-risk” percutaneous transluminal coronary angioplasty. Am J Cardiol 61(14):33G-37G Höckel M, Vaupel P (2001) Tumor hypoxia: definitions and current clinical, biologic, and molecular aspects. J Natl Cancer Inst 93:266–276 Horsman MR, Mortensen LS, Petersen JB, Busk M, Overgaard J (2012) Imaging hypoxia to improve radiotherapy outcome. Nat Rev ClinOncol 9:674–687 Jain S, Bose A, Bastia B, Sharma H, Sachdeva R, Jain AK, Pal R (2017) Oxidized hemoglobin is antigenic and immunogenic in Lupus. Front Immunol 8:732 Jin Z, Wen Y, Hu Y et al (2017) MRI-guided and ultrasound-triggered release of NO by advanced nanomedicine. Nanoscale 9:3637–3645 Kheir JN, Scharp LA, Borden MA et al (2012) Oxygen gas-filled microparticles provide intravenous oxygen delivery. Sci Transl Med 4:140ra88 Kim MS, Lee EJ, Kim JW, Chung US, Koh WG, Keum KC, Koom WS (2016) Gold nanoparticles enhance anti-tumor effect of radiotherapy to hypoxic tumor. Radiat Oncol J 34(3):230–238 Kuang Y, Balakrishnan K, Gandhi V, Peng X (2011) Hydrogen peroxide inducible DNA crosslinking agents: targeted anticancer prodrugs. J Am Chem Soc 133(48):19278–19281 Kumar V, Gabrilovich DI (2014) Hypoxia-inducible factors in regulation of immune responses in tumour microenvironment. Immunology 143:512–519 Kunz-Schughart LA, Dubrovska A, Peitzsch C, Ewe A, Aigner A, Schellenburg S, Muders MH, Hampel S, Cirillo G, Lemma F, Tietze R, Alexiou C, Stephan H, Zarschler K, Vittorio O, Kavallaris M, Parak WJ, Madler L, Pokhrel S (2017) Biomaterials 120:155–184 Lee HY, Kim HW, Lee JH, Oh SH (2015) Controlling oxygen release from hollow microparticles for prolonged cell survival under hypoxic environment. Biomaterials 53:583–591 Lin H, Chen Y, Shi JL (2018) Chem Soc Rev 47:1938–1958 Li J, Shang W, Li Y, Fu S, Tian J, Lu L (2018) Advanced nanomaterials targeting hypoxia to enhance radiotherapy. Int J Nanomed 13:5925–5936 Liu J, Chen Q, Zhu W et al (2017) Nanoscale-coordination-polymer-shelled manganese dioxide composite nanoparticles: a multistage redox/pH/H2O2-responsive cancer theranosticnanoplatform. Adv Funct Mater 27(10):1605926

Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials

127

Liu W, Chen B, Zheng H, Xing Y, Chen G, Zhou P, Qian L, Min Y (2021a) Advances of nanomedicine in radiotherapy. Pharmaceutics 13(11):1757 Liu Y, Jiang Y, Zhang M et al (2018) Modulating hypoxia via nanomaterials chemistry for efficient treatment of solid tumors. Acc Chem Res 51:2502–2511 Liu Y, Liu Y, Bu W, Xiao Q, Sun Y, Zhao K, Fan W, Liu J, Shi J (2015) Radiation-/hypoxia-induced solid tumor metastasis and regrowth inhibited by hypoxia-specific upconversionnanoradiosensitizer. Biomaterials 49:1–8 Liu Y, Zhang J, Du J, Song K, Liu J, Wang X, Li B, Ouyang R, Miao Y, Sun Y, Li Y (2021b) Biodegradable BiOCl platform for oxidative stress injury–enhanced chemodynamic/radiation therapy of hypoxic tumors. Acta Biomaterialia 129:280–292, 1742–706 Lu J, Yang L, Zhang W et al (2019) Photodynamic therapy for hypoxic solid tumors via Mn-MOF as a photosensitizer. Chem Commun 55:10792–10795 Luo C-H, Huang C-T, Su C-H, Yeh C-S (2016) Bacteria-mediated hypoxia-specific delivery of nanoparticles for tumors imaging and therapy. Nano Lett 16:3493–3499 Masson N, Ratcliffe PJ (2014) Hypoxia signaling pathways in cancer metabolism: the importance of co-selecting interconnected physiological pathways. Cancer Metab 2:3 Mi Y, Shao Z, Vang J et al (2016) Application of nanotechnology to cancer radiotherapy. Cancer Nano 7:11 Mi P, Cabral H, Kataoka K (2020) Adv Mater 32:e1902604 Moncada S, Erusalimsky JD (2002) Opinion-does nitric oxide modulate mitochondrial energy generation and apoptosis? Nat Rev Mol Cell Biol 3:214–220 Nagy JA, Chang S-H, Shih S-C, Dvorak AM, Dvorak HF (2010) Heterogeneity of the tumor vasculature. SeminThromb Hemost. 36(03):321–331 Palazón A, Aragonés J, Morales-Kastresana A et al (2012) Molecular pathways: hypoxia response in immune cells fighting or promoting cancer. Clin Cancer Res 18:1207–1213 Perrault SD, Walkey C, Jennings T, Fischer HC, Chan WC (2009) Mediating tumor targeting efficiency of nanoparticles through design. Nano Lett 9:1909–1915 Qian X, Peng XH, Ansari DO et al (2008) In vivo tumor targeting and spectroscopic detection with surface-enhanced Raman nanoparticle tags. Nat Biotechnol 26:83–90 Rankin EB, Giaccia AJ (2016) Hypoxic control of metastasis. Science 352(6282):175–180 Riganti C, Miraglia E, Viarisio D et al (2005) Nitric oxide reverts the resistance to doxorubicin in human colon cancer cells by inhibiting the drug efflux. Cancer Res 65:516–525 Sao P, Chand Y, Kumar A, Singh S (2021) Potential drug target identification in porphyromonas gingivalis using in-silico subtractive metabolic pathway analysis. Bangladesh J Med Sci 20(4):887–896 Semenza GL (2004) Intratumoral hypoxia, radiation resistance, and HIF-1. Cancer Cell 5:405–406 Shannon AM, Bouchier-Hayes DJ, Condron CM et al (2003) Tumour hypoxia, chemotherapeutic resistance and hypoxia-related therapies. Cancer Treat Rev 29:297–307 Sharma A, Arambula JF, Koo S et al (2019) Hypoxia-targeted drug delivery. Chemsoc Rev 48:771– 813 Shuryak I, Hahnfeldt P, Hlatky L, Sachs RK, Brenner DJ (2009) A new view of radiation-induced cancer: Integrating short- and long-term processes. Part I: Approach Radiat Environ Biophys 48:263–274 Song G, Liang C, Yi X et al (2016) Perfluorocarbon-loaded hollow Bi2Se3 nanoparticles for timely supply of oxygen under near-infrared light to enhance the radiotherapy of cancer. Adv Mater 28(14):2716–2723 Song L, Wang G, Hou X et al (2020) Biogenic nanobubbles for effective oxygen delivery and enhanced photodynamic therapy of cancer. Acta Biomater 108:313–325 Song R, Hu D, Chung HY et al (2018) Lipid-polymer bilaminar oxygen nanobubbles for enhanced photodynamic therapy of cancer. ACS Appl Mater Inter 10:36805–36813 Song R, Peng S, Lin Q et al (2019) pH-responsive oxygen nanobubbles for spontaneous oxygen delivery in hypoxic tumors. Langmuir 35:10166–10172

128

A. Srivastava et al.

Teicher BA, Holden SA, Ara G, Dupuis NP, Goff D (1995) Restoration of tumor oxygenation after cytotoxic therapy by a perflubron emulsion/carbogen breathing. Cancer J Sci Am 1(1):43–48 Telarovic I, Wenger RH, Pruschy M (2021) Interfering with tumor hypoxia for radiotherapy optimization. J Exp Clin Cancer Res 40:197 Tian L, Chen Q, Yi X et al (2017) Albumin-templated manganese dioxide nanoparticles for enhanced radioisotope therapy. Small 13(25):1700640 Vaupel P (2009) Prognostic potential of the pretherapeutic tumor oxygenation status. Adv Exp Med Biol 645:241–246 Vaupel P, Mayer A (2007) Hypoxia in cancer: significance and impact on clinical outcome. Cancer Metastasis Rev 26:225–239 Wang W, Tang Z, Zhang Y, Wang Q, Liang Z, Zeng X (2020) Macromol Biosci 20:e2000222 Wilson WR, Hay MP (2011) Targeting hypoxia in cancer therapy. Nat Rev Cancer 11(6):393–410 Wolfram J, Ferrari M (2019) Nano Today 25:85–98 Wouters BG, Brown JM (1997) Cells at intermediate oxygen levels can be more important than the “hypoxic fraction” in determining tumor response to fractionated radiotherapy. Radiat Res 147(5):541–550 Wu K, Su D, Liu J, Saha R, Wang JP (2019) Magnetic nanoparticles in nanomedicine: a review of recent advances. Nanotechnology 30:502003 Wu Q, Wu W, Kuca K (2020) From hypoxia and hypoxia-inducible factors (HIF) to oxidative stress: a new understanding of the toxic mechanism of mycotoxins, Jan 2020. Food Chem Toxicol 135:110968 Xia D, Hang D, Li Y, Jiang W, Zhu J, Ding Y, Gu H, Hu Y (2020) Au–hemoglobin loaded platelet alleviating tumor hypoxia and enhancing the radiotherapy effect with low-dose X-ray. ACS Nano 14:15654–15668 Yang X, Gao L, Guo Q, Li Y, Ma Y, Yang J, Gong C, Yi C (2020) Nano Res 13:2579–2594 Yang G, Zhang R, Liang C et al (2018) Manganese dioxide coated WS2@ Fe3O4/sSiO2 nanocomposites for pH-responsive MR imaging and oxygen-elevated synergetic therapy. Small 14(2) Yang C, Mi X, Su H, Yang J, Gu Y, Zhang L, Sun W, Liang X, Zhang C (2019) GE11-PDAPt@USPIOs nano-formulation for relief of tumor hypoxia and MRI/PAI-guided tumor radiochemotherapy. Biomater Sci 7:2076–2090 Yang J, Li W, Luo L, Jiang M, Zhu C, Qin B, Yin H, Yuan X, Yin X, Zhang J et al (2018) Hypoxic tumor therapy by hemoglobin-mediated drug delivery and reversal of hypoxia-induced chemoresistance. Biomaterials 182:145–156 Yao Y, Zhang M, Liu T et al (2015) Perfluorocarbon-encapsulated PLGA-PEG emulsions as enhancement agents for highly efficient reoxygenation to cell and organism. ACS Appl Mater Interfaces 7(33):18369–18378 Yi X, Chen L, Zhong X et al (2016) Core–shell Au@MnO2 nanoparticles for enhanced radiotherapy via improving the tumor oxygenation. Nano Res 9(11):3267–3278 Yong Y, Zhang C, Gu Z, Du J, Guo Z, Dong X, Xie J, Zhang G, Liu X, Zhao Y (2017) Polyoxometalate-based radiosensitization platform for treating hypoxic tumors by attenuating radioresistance and enhancing radiation response. ACS Nano 11:7164–7176 Zou M-Z, Liu W-L, Chen H-S, Bai X-F, Gao F, Ye J-J, Cheng H, Zhang X-Z (2021) Advances in nanomaterials for treatment of hypoxic tumor. Natl Sci Rev 8:2 Zeng L, Cao Y, He L, Ding S, Bian X, Tian G (2021) Metal–ligand coordination nanomaterials for radiotherapy: emerging synergistic cancer therapy. J Mater Chem B 9(2):208–227 Zhang J, Liu Y, Wang X, Du J, Song K, Li B, Chang H, Ouyang R, Miao Y, Sun Y et al (2020) Nanozyme-incorporated biodegradable bismuth mesoporousradiosensitizer for tumor microenvironment-modulated hypoxic tumor thermoradiotherapy. ACS Appl Mater Interfaces 12:57768–57781 Zhao Y, Yu B, Hu H, Hu Y, Zhao NN, Xu FJ (2014) New low molecular weight polycationbased nanoparticles for effective codelivery of pDNA and drug. ACS Appl Mater Interfaces 6:17911–17919

Novel Strategies in Radiotherapy to Reduce Hypoxia Using Nanomaterials

129

Zhou R, Zhao D, Beeraka NM, Wang X, Lu P, Song R, Chen K, Liu J (2022) Novel implications of nanoparticle-enhanced radiotherapy and brachytherapy: Z-effect and tumor hypoxia. Metabolites 12(10):943 Zhou R, Wang H, Yang Y, Zhang C, Dong X, Du J, Yan L, Zhang G, Gu Z, Zhao Y (2019) Tumor microenvironment -manipulated radiocatalytic sensitizer based on bismuth heteropoly- tungstate for radiotherapy enhancement. Biomaterials 189:11–22

Nanoproteomics: An Approach for the Identification of Molecular Targets Associated with Hypoxia J. Deepa Arul Priya, Sumira Malik, Mohammad Khalid, and Akash Gautam

Abstract A thorough examination of every protein’s array in the human proteome is necessary to understand the causes, pathophysiology, and possible treatment targets of any disease or disorder at the molecular level. The study of this total proteome in any organism, i.e. proteomics, has been augmented by nanotechnology applications. This new field of nanoproteomics has several technical advantages over the traditional method of proteomics, as it helps in the proteomics analyses of small as well as the specific population of cells in living organisms. In the last decade, advanced techniques of nanoproteomics have assisted scientists in processing biological samples, their MS detection and analysis of the data, even from a single cell. This chapter discusses the different approaches of nanoproteomics in identifying several cellular proteins, their interactions and their modifications during hypoxia. Moreover, a summary of the impacted proteins and their interactions during hypoxic conditions has been stated. Finally, this chapter mentions the critical factors and challenges for nanoproteomics which are crucial to successfully detecting molecular targets for hypoxia and associated disorders. Keywords Proteomics · Nanotechnology · Biomarkers · Hypoxia · Tumour microenvironment

J. Deepa Arul Priya Department of Biotechnology, Sri Paramakalyani College, Tirunelveli, India S. Malik Amity Institute of Biotechnology, Amity University Jharkhand, Ranchi, India M. Khalid Graphene & Advanced 2D Materials Research Group, School of Engineering and Technology, Sunway University, No. 5, Jalan Universiti, Bandar Sunway, 47500 Petaling Jaya, Selangor, Malaysia A. Gautam (B) Centre for Neural and Cognitive Sciences, School of Medical Sciences, University of Hyderabad, Hyderabad, India e-mail: [email protected] © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_8

131

132

J. Deepa Arul Priya et al.

1 Introduction The term proteome (coined in the 1990s) defines the complete set of proteins encoded by a genome. The proteome closely reflects the biological and chemical processes occurring in a biological system (De Angelis and Calasso 2014; Zipes 2019). Therefore, distinguishing the proteins expressed by a cell provides important hints to the function, organization and responsiveness inherent in a cell (Garrels 2001). The tool to study the proteome of an organism (or cell line or tissue) is called proteomics (De Angelis and Calasso 2014). The concept of proteomics is based on protein separation, identification and data analysis of molecular interactions (Westergren-Thorsson et al. 2006) which involves an inclusive study of protein structures, abundance, modifications, functions and interactions in an organism at definite time and conditions (Jia et al. 2013). Beforehand, the conventional proteomic techniques were performed through lowthroughput methods like (Fig. 1); gel-based methods (Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), Two-dimensional gel Electrophoresis (2DE), 2D-Difference Gel Electrophoresis (2D-DIGE)) for separation of multiplex proteins (Dunn and Burghes 1986; Marouga et al. 2005; Issaq and Veenstra 2008), chromatography-based methods (Ion Exchange (IEC), Size Exclusion (SEC), Affinity (AC)) for purification (Jungbauer and Hahn 2009; Voedisch and Thie 2010; Hage et al. 2012) and antibody-based methods (Enzyme-Linked Immunosorbent Assay (ELISA), western blot) for evaluation (Lequin 2005; Kurien and Scofield 2006). However, with the advancements in technologies, proteomics studies can now be performed through high-throughput methods including protein microarrays (Sutandy et al. 2013) and mass spectrometry (Yates 2011). The state-of-the-art proteomics tools are capable of identifying proteins, quantifying proteins, locating different

Fig. 1 A general idea of various proteomics techniques (Source Aslam et al. 2017)

Nanoproteomics: An Approach for the Identification of Molecular …

133

proteins, deciphering the structure and function of proteins, revealing the protein– protein interactions, as well as finding out the post-translation modifications (PTMs) in these proteins. Therefore, the proteomics research had been extended with a focus on biomedical applications to study disease aetiology, finding molecular markers for diagnosis and observation of disease, interpretation of drug action and possible drug targets (Hanash 2003; Moseley et al. 2007). There are several current studies of using different proteomics techniques for identifying biomarkers for various diseases. For example, Olfactomedin-4 was identified for the detection of ‘colorectal cancer’ (Karagiannis et al. 2014), Myeloperoxidase (MPO) was found as molecular marker for ‘acute bacterial meningitis’ (ABM) and Lactotransferrin (LTF) for ‘cerebral malaria’ (CM) (Njunge et al. 2017), Intelectin2, Moesin, Neuroserpin, Neogenin and Secretogranin-2 were discovered as new biomarkers for Human African trypanosomiasis (HAT) (Bonnet et al. 2019). In a row, studies on hypoxia with proteomics approaches were also performed. Li et al. (2019) worked on proteomic characterization of human Periodontal Ligament Cells (hPDLCs) during hypoxia for analysing hypoxia-based therapeutic strategies for periodontal diseases. Du´s-Szachniewicz et al. (2021) reported that the response of DLBCL (diffuse large B-cell lymphoma) phenotypes after hypoxia in lymphoproliferative malignancies was intricate and highly diversified. Hypoxia or low-oxygen tension initiates a sequence of string-like reactions on gene expression cascade leading to cellular function alterations which can be irreversible too (Pugh and Ratcliffe 2017) (Fig. 2). HIF (Hypoxia-inducible factor) has a significant role in the regulation of hypoxic responses. This factor consists of 3α and 1β subunits (Koyasu et al. 2018). The other transcription factors involved are Nuclear factor (NF-κB), CREB (cAMP-responsive transcription factor), Nrf2 (Nuclear factor erythroid 2-related factor 2), STAT (Signal transducer and activator of transcription) and Myc (Lee et al. 2006; Kim et al. 2011; Nakayama 2013; Wong et al. 2013). Hence, these factors play a critical role in diseases like cancer, metabolic diseases and chronic heart and kidney diseases. This chapter highlights the current advances in the integration of nanotechnology and proteomics to identify different proteins which get impacted during hypoxia and possibly pave the way for the development of therapeutic target in the near future.

Fig. 2 Gene expression machinery under hypoxia (Source Nakayama and Kataoka 2019)

134

J. Deepa Arul Priya et al.

2 Proteomics and Nanotechnology (NT) Protein molecules in biological samples at low concentrations of 10–12 –10–15 M generally synchronize with molecules at high concentrations of 10–3 –10–4 M. This makes detection of low-concentration molecules more difficult. The other limitations faced in proteome research were sensitivity, dynamic range, detection time, biological complexities, etc. Such limitations challenge both top-down and bottomup approaches (Ray et al. 2011). In clinical proteomics, the identification and characterization of low-range molecules as molecular markers is an effectual tool for early diagnosis and evaluation of the efficacy of therapeutic interventions for fatal diseases (Ramachandran et al. 2008). So, the development of ultrasensitive and powerful technology for speedy detection of low-concentration molecules in samples has been a top priority in proteomic methodology (Bell et al. 2009) (Fig. 3). Biomedical engineers have integrated proteomics with emerging disciplines such as NT to overcome these complexities. This led to the emergence of a unique research field known as ‘Nanoproteomics’ to focus mainly on the varying concentration range of mixed proteins present in biological samples. The benefits of NT compared with traditional methods are real-time assays for multiple samples, utilization of less sample and reagent, high sensitivity and specificity, less duration for processing and nanoscale analytical systems (Jia et al. 2013).

Fig. 3 Schematic diagram to overcome challenges in top-down proteomics through novel strategies (Source Melby et al. 2021b)

Nanoproteomics: An Approach for the Identification of Molecular …

135

3 Advantages of Nanoproteomics Over Traditional Proteomics Tools Proteomics studies are based on two different mass spectrometry-related approaches: bottom-up and top-down proteomics (Gregorich et al. 2014) (Fig. 4). The former approach is well acknowledged for analyses of peptides by protein digestion, but it shows insignificance in picturizing PTMs and sequence variants (Chait 2006). In contrast, the latter approach analyses intact proteins to depict PTM codes along with mutations and alternative splicing (Chen et al. 2018). In the past decade, nanoproteomics research has developed rapidly through technological advances (Tiambeng et al. 2020) for understanding biological events, unravelling mechanism of action for diseases and targeting molecular markers (Toby et al. 2019; Melby et al. 2021a).

3.1 Protein Solubility in Sample Preparation The traditional workups before MS analysis need some technological improvement. Sample isolation techniques: ‘Fluorescence-activated cell sorting’ (FACS) and ‘laser

Fig. 4 Schematic diagram of the top-down and bottom-up proteomics approaches (Source Gregorich et al. 2014)

136

J. Deepa Arul Priya et al.

capture microdissection’ (LCM) were coupled with ‘Nanodroplet processing in onepot for trace samples’ (nanoPOTS) (Zhu et al. 2018a) (Fig. 5). LCM was found to be a useful alternative in comparison with FACS in resolving the loss of spatial information by preincubating nanowells with dimethyl sulfoxide (DMSO) (Zhu et al. 2018b). Also, sample preparation in nanoPOTS increased protein concentration by enhancing enzymatic digestion and efficient sample recovery, which was not achieved through online processing systems like strong cation exchange (SCX) processed in proteomic reactor (Ethier et al. 2006) and simplified nanoproteomics platform (SNaPP) processed in immobilized enzyme reactor (IMER) (Huang et al. 2016). In some cases, according to samples, surfactants are essentially used for solubilizing proteins in biological samples. One such surfactant is SDS which was found incompatible with the process due to the loss of sample (Wi´sniewski et al. 2009). Some surfactants, like octyl β-D-glucopyranoside (OGP) and n-dodecyl βD-maltoside (DDM) with mild, non-ionic activity were found to have limited solubilization ability (Laganowsky et al. 2013). Moreover, acid-cleavable surfactants

Fig. 5 Proteomic sample preparation with nanoPOTS (Source Zhu et al. 2018a)

Nanoproteomics: An Approach for the Identification of Molecular …

137

Fig. 6 Protein solubility challenge overcome by a novel photocleavable surfactant (Source Melby et al. 2021b)

such as RapiGest (Yu et al. 2003), ProteaseMax (Saveliev et al. 2013), and MScompatible slowly degradable surfactant (MaSDeS) (Chang et al. 2015; Arulmozhi et al. 2019), had solubilization and digestion efficiency but were not suitable for top-down proteomics. To overcome this, a novel photocleavable surfactant called 4hexylphenylazosulfonate or Azo was identified for high-throughput proteomics (Fig. 6). Azo facilitated robust extraction and rapid enzymatic digestion followed by subsequent MS analysis. It was degraded upon UV irradiation, thus enabling both approaches in proteomics (Brown et al. 2019). Azo also enhanced access to proteins from cellular components and extracellular regions. Extracellular matrix proteins mainly contribute to pathologies and are nearly insoluble. A new decellularization/extraction method was done with Azo to minimize sample clean-up before MS analysis (Knott et al. 2020).

3.2 Protein Sampling Sample processing is more challenging than instrumentation. The development of ‘single tube’ strategies like stage-tip (Rappsilber et al. 2007), filter-aided and (Wi´sniewski et al. 2009) and single-pot solid-phase (Hughes et al. 2014) sample processing has made significant progress in nanoproteomics. These strategies are optimized for digestion in a single tube to reduce labware-related sample loss. With the advantages of Rapigest SF Surfactant (acid-cleavable detergent), a nanoproteomic workflow was optimized and developed, known as ‘Nanogram TMT Processing in One Tube’ (NanoTPOT) (Fig. 7). The workflow started with evaluating enzymatic digestion, continued by labelling of tandem mass tag (TMT), followed by online and offline fractionation methods. Overall the quantitative one-pot workflow was a high-throughput nanoproteomic analysis for biomedical applications (Wu et al. 2020a). Recently, a facile one-pot method known as ‘SOPs-MS’ (surfactant-assisted one-pot sample-MS) was developed for convenient robust proteome profiling (Fig. 8). This combines all steps in low-bind one tube or multi-well plates (Martin et al. 2021). Sampling in the presence of nanoparticles (NPs) is yet another breakthrough. When NPs are exposed to biological fluid or lysate samples, a biological complex

138

J. Deepa Arul Priya et al.

Fig. 7 Trypsin digestion optimization in the NanoTPOT workflow (Source Wu et al. 2020a)

called protein corona (PC) is cast on the surface of NPs (Monopoli et al. 2011). This complex is affected by several factors like properties of NP (size, charge, surface properties and configuration of the particle core), varying properties of protein (isoelectric points, molecular weight, structure and folding), temperature, time and concentration of samples (Kalantari et al. 2016). Thus, the comparative study of the PCs with proteomic data analysis is favourable for identifying novel molecular targets (Lai et al. 2012).

3.3 High Dynamic Range In the present scenario, appropriate protein enrichment techniques are required to concentrate low-range proteoforms before MS analysis (Xie et al. 2009). Antibodybased affinity purification was preferred for intact protein targeting (Lollo et al. 2014). But some significant limitations like high cost, limited availability, variability in subsequent batches and low stability of antibodies urged the need for better affinity components (Baker 2015). To overcome this, surface-functionalized multivalent superparamagnetic NPs were developed. NPs are produced by simple, fast and scalable synthesis with better

Nanoproteomics: An Approach for the Identification of Molecular …

139

Fig. 8 Schematic diagram of the SOPs-MS workflow at the standard sample processing volume (∼50 μL) (Source Martin et al. 2021)

penetration, multiple flexible binding sites and suitable protein interactions (Hwang et al. 2015). A coherent workflow for phosphoprotein analysis by cobalt ferrite (CoFe2 O4 ) NPs coupled with LC–MS/MS online mode analysis was established (Chen et al. 2017). Surface-silanized magnetite (Fe3 O4 ) NPs were synthesized for the enrichment of phosphoproteome (Roberts et al. 2019). A peptide-functionalized NP was developed using a nanoproteomics strategy for low-abundance proteins with high specificity and reproducibility (Tiambeng et al. 2020).

140

J. Deepa Arul Priya et al.

This top-down nanoproteomics platform is efficient for low-abundance proteoforms. In future, advances in aptamer developments may enable substituted affinity components with novel NPs for analysis by top-down approach (Wang et al. 2019).

3.4 Proteome Complexity Proteome complexity in terms of protein molecular weights in the varying range is a key challenge in the proteomic approach (Picotti et al. 2009). This requires separation before MS analysis to reduce signal-to-noise ratio fluctuations (Compton et al. 2011). To resolve this, an offline technique called ‘Gel-eluted liquid fraction entrapment electrophoresis’ (GELFrEE) was developed to bin proteins into size-selected fractions (Tran and Doucette 2009). Another technique for proteome fractionation based on size was called ‘Passively Eluting Proteins from Polyacrylamide gels as Intact species’ for MS (PEPPI-MS) (Fig. 9). This was done by separating protein bands from SDS-PAGE (Takemori et al. 2020). The size-based proteome fractionization can also be done through ‘serial Size exclusion chromatography’ (sSEC) in continuation with RPLC-MS/MS (Fig. 10). In the sSEC method, the separation of high molecular weight proteins becomes effective due to varying porosity columns and mobile phases suiting both MS modes (online and offline). This method detects up to 223 kDa of proteoform by employing a Q-TOF/MS (Cai et al. 2017). The growing interest in native separations has led to the origination of a non-denaturing technique called ‘Capillary electrophoresis-MS’ (CE-MS) (Shen et al. 2021) (Fig. 11). This proved to be a useful alternative for analysing the

Fig. 9 Schematic view of polyacrylamide-gel-based prefractionation (Source Takemori et al. 2020)

Nanoproteomics: An Approach for the Identification of Molecular …

141

Fig. 10 Overview of the complex protein mixture separation by serial size-exclusion chromatography (sSEC) (Source Melby et al. 2021b)

samples processed by the ‘Nanoparticle-aided nanoreactor for the nanoproteomics’ (Nano3) method. Nano3 was developed for analysing complex proteome samples through bottom-up proteomics (Yang et al. 2021) (Fig. 12). A gas-phase technique called ‘Ion-mobility spectrometry’ (IMS), separated molecules on the basis of their working phenomenon inside the drift tube. Also, high-resolution IMS involves rapid proteoforms separation with high throughput (Dodds and Baker 2019). For a finer depiction of larger proteoforms, a ‘2D-Liquid chromatography’ (2DLC) technique integrating sSEC (by size) and RPLC-MS (by hydrophobicity)

Fig. 11 Protein analysis by native capillary zone electrophoresis–mass spectrometry (CZE-MS) method (Source Shen et al. 2021)

142

J. Deepa Arul Priya et al.

Fig. 12 An overview of nanoparticle-aided nanoreactor (Source Yang et al. 2021)

was constructed for separation (Cai et al. 2017). Then, further, a ‘Three dimensionalLC’ (HIC-IEX-RPC) technique which runs in both offline and online modes was also developed. This method had great potential for intact protein separation with deep proteome coverage (Valeja et al. 2015). In the coming years, the fascination on automated mechanical techniques will pave the way for many new separation method combos.

3.5 Data Analysis Due to complexity and labour-intensiveness, a key barrier in proteomic approaches is data analysis. Thus, developing software solutions for predicting spectral data and comparing them against databases can be promising (Chen et al. 2018). The database search algorithms like MS-Align, TopPIC, pTOP, Mascot and Proteoform Suite (Karabacak et al. 2009; Liu et al. 2012; Kou et al. 2016; Sun et al. 2016; Cesnik et al. 2018) have been validated as beneficial tools for data analysis. A software called ‘Informed Proteomics’ was designed with Promex algorithm (LC–MS feature-finding) and MSPathfinder algorithm (database search) for data analysis (Park et al. 2017a). A user-friendly interface software ‘MASH Suite Pro’ was developed with combined data analysis tools, but this showed limited access to raw data files (Cai et al. 2016; Bhardwaj et al. 2021). An improved version of ‘MASH Explorer’ was created with flexibility in targeted and discovery modes (Fig. 13). The main features were high-resolution spectral data, proteoform analysis, graphical data representation, data validation and automated workflow. This software package can become an integral part of advancing proteomics research in biomedical field (Wu et al. 2020b). A machine learning technique combo to process all resultant peak lists from various deconvolution algorithms was developed. This showed more accuracy in

Nanoproteomics: An Approach for the Identification of Molecular …

143

Fig. 13 Schematic diagram of proteomics data processing by various MASH explorer functions (Source Wu et al. 2020b)

database searches on protein formulations. Hence, combining this machine learning technique and MASH Explorer for proteoform analysis in wide range has been in preference (McIlwain et al. 2020). The deconvolution algorithm ‘FlashDeconv’ has already been integrated into MASH Explorer to outpace other deconvolution tools (Fig. 14). This works on the basis of logarithmic transformation to match a spectral pattern with high throughput (Jeong et al. 2020). Incorporating ‘UniDec’ into MASH Explorer is expected as a promising feature for extracting data on native protein with protein fragment ions imaging parallelly. UniDec works on Bayesian algorithm to separate a complex spectrum’s mass and charge dimension by incorporating ion mobility–mass spectra (Marty et al. 2015). A new algorithm ‘ClipsMS’ has been developed for analysing terminal and internal fragments of protein sequence. These are generated from MS fragmentation and can be used to locate alterations in the sequence (Fig. 15). Combined with ion mobility analysis, this can give a comprehensive outcome (Lantz et al. 2021).

144

J. Deepa Arul Priya et al.

Fig. 14 FLASHDeconv technique for top-down proteomics (Source Jeong et al. 2020)

Comparative analysis of MS/MS data with search algorithms showed that Sequest HT/INFERYS combo benefitted with the highest proteome coverage for sample loads below 1 ng (Stejskal et al. 2021).

4 Application of Nanoproteomics for Hypoxia 4.1 Molecular Markers of Hypoxia Hypoxia is defined as insufficient oxygen levels due to a lack of oxygen supply or excessive oxygen consumption for maintaining normal cellular function. In normal conditions, oxygen exchange starts in the lung’s alveoli, with more oxygen (95%) diffusing into the capillary vessels and then binding to haemoglobin. This oxygenated blood returns to the heart (left atrium). From there, it is pumped out through the left ventricle to all parts to maintain the proper function of every cell. If the state of continuous lack of oxygen is for a short period (within minutes), it is termed acute

Nanoproteomics: An Approach for the Identification of Molecular …

145

Fig. 15 The workflow of the algorithm and how it matches peaks input by the user (Source Lantz et al. 2021)

hypoxia, which is a response to alterations in pre-existing proteins. If it is for an extended period (lasting for hours) it is termed chronic hypoxia, which is a response to alterations in gene expression (Keeley and Mann 2019). Blood flow hindrance can cause high-risk effects on organ structure and function. This may be seen in cases of cerebral ischaemia, myocardial ischaemia and regulating tumour growth and metastasis (Michiels 2004). The gene expression is mostly controlled by HIF or NF-κB. There are three HIFα’s (1α, 2α and 3α) and one HIF-1β (aryl hydrocarbon nuclear translocator, ARNT) subunits. These subunits are relatively expressed to form a heterodimeric functional unit. Among them, HIF-1α is expressed in most human tissues, but HIF-2α and HIF3α are expressed in specific tissues only at developmental stages. So, HIF-1α plays an essential role in regulating transcription of all cells during hypoxia. HIF-2α and HIF-3α play much-confined roles in oxygen homeostasis (Semenza 2000). Under normoxia (Fig. 16), HIF-α’s protein gets hydroxylated by two oxygendependent enzymes—prolyl hydroxylases (PHDs) and factor-inhibiting HIF (FIH). That is, in normoxia, these are activated and, in hypoxia, these are suppressed via distinct mechanisms. The action of PHDs starts with catalysing the proline hydroxylation of HIF-α’s followed by E3 ubiquitin ligase, von Hippel–Lindau (VHL) binding to HIF-α’s protein to promote the degradation via the ubiquitin–proteasome degradation pathway. The action of FIH starts with catalysing the asparagine hydroxylation in the C-terminal transactivation domain of HIF-α’s. This prevents the recruitment of transcriptional coactivator CREB-binding protein (CBP) and its homolog, p300 which inhibits HIF-α’s transcription activity. In addition, PHDs and FIH can also inactivate NF-κB by targeting the inhibitor of the κB kinase (IKK) complex through direct hydroxylation (Ke and Costa 2006).

146

J. Deepa Arul Priya et al.

Fig. 16 Schematic diagram of HIF-1α and NF-κB regulation under normoxic and hypoxic conditions (Source Chen et al. 2020)

Under hypoxia (Fig. 16), oxygen deficiency interrupts the activity of PHDs. This prevents HIF-α’s from VHL-dependent protein degradation. In yet another condition, mitogen-activated protein kinases (MAPKs) phosphorylate HIF-α’s, which increases the stability of the ‘α’ subunit. These phosphorylated HIF-α proteins travel to the nucleus and are associated with HIF-1β. This forms a HIF-α/1β heteroduplex which binds to the hypoxia-responsive element (HRE) of target genes. The FIH activity under hypoxia gets suppressed and increases CBP/p300 recruitment to enhance the transcription of HIF target genes (Semenza 2012). In NF-κB, the hypoxic condition prevents hydroxylation of IKK. Non-hydroxylated IKK complex promotes the phosphorylation, ubiquitination and degradation of the inhibitor. Therefore, NF-κB once released, travels to the nucleus to regulate the target gene transcription (D’Ignazio and Rocha 2016).

4.2 Hypoxia and Related Diseases 4.2.1

Cancer

The main feature of hypoxia in the tumour microenvironment (TME) is impaired oxygen delivery and consumption (Fig. 17). Hypoxic cellular response leads to the existence and reproduction of cancer cells, cell transition, cell differentiation and reduction in drug effectiveness. This concludes that the HIF regulatory pathway plays a dominant role in cancer biology (Schito and Semenza 2016). Hypoxic molecular responses stabilize HIFs and mediate cells to adapt hypoxic stress by overexpression

Nanoproteomics: An Approach for the Identification of Molecular …

147

Fig. 17 Hypoxia-regulated cancer progression (Source Chen et al. 2020)

of downstream genes (Semenza 2012). This mechanism is reported in different cancer types like colon, lung, breast, gastric, pancreatic, prostrate (Luo et al. 2022) and thyroid (García-Vence et al. 2020). HIF-1α and HIF-2α are found in advanced stages in patients with low survival rate (Schito and Semenza 2016).

4.2.2

Cardiovascular Diseases

Hypoxia is a common feature that causes physiological and functional changes in cardiovascular-related diseases (atherosclerosis, arrhythmia, cardiomyopathy, congenital heart disease and pulmonary hypertension) (Luo et al. 2022). The factors which play essential roles in these disorders are HIF-1α and NF-κB. The mechanism starts with the upregulation of NF-κB by HIF-1α, which in turn activates the transcription of HIF-1α, thus worsening the disease (Eltzschig and Carmeliet 2011).

4.2.3

Kidney Diseases

The occurrence of hypoxia is also significant in Acute Kidney Injury (AKI). HIF-1α and TGF-β1 signalling during kidney fibrosis are excessively expressed, which can slowly lead to chronic kidney disease (CKD) over time (LeBleu et al. 2013) (Fig. 18).

4.2.4

Neurodegenerative Diseases

Impaired oxygen supply (hypoxia) to the brain causes many metabolic changes in neuronal and non-neural cells. The HIF signalling response in brain tissues seems to be complex (Fig. 19). In neurospheres HIF-2 regulates Vascular endothelial growth factor (VEGF) and so, HIF-regulated growth factors play a critical role in cognitive

148

J. Deepa Arul Priya et al.

Fig. 18 Schematic diagram of the kidney-related diseases under hypoxic conditions (Source Chen et al. 2020)

functions (Koester-Hegmann et al. 2019). These factors are linked with Amyotrophic lateral sclerosis and several neurodegenerative disorders (Parkinson’s disease and Alzheimer’s disease) (Luo et al. 2022). Hence, HIF-2 might be a valuable target for neuroregenerative therapy (Wakhloo et al. 2020). Hypoxia’s role is also seen in many disorders like Preeclampsia (hypertension featured during pregnancy) (Duley 2009) and Endometriosis (Wu et al. 2019). HIF1α is found to be upregulated in preeclamptic placenta and is the main driving force in endometriosis. Identifying novel sensors and functional modulators can provide insights for developing potential diagnostic and therapeutic approaches (Chen et al. 2020).

4.3 Therapeutics with Molecular Markers Nanoproteomics was considered impracticable for biomedical applications for a long time. But now, it is a reality enabling complete characterization of cellular responses and structural mapping of proteins in pathological tissues (Zhu et al. 2018b; Rangisetty et al. 2023). This approach has helped us to understand the need for finding new strategies for multi-target drug discovery by characterizing molecular events (Vinaiphat et al. 2021). In this section, nanoproteomics approach for identification of molecular targets for hypoxia will be detailed.

Nanoproteomics: An Approach for the Identification of Molecular …

149

Fig. 19 HIF-1 and HIF-2 regulation in hypoxia/ischaemia causing brain damage (Source Schneider Gasser et al. 2021)

4.3.1

Nanotherapeutics

Many solid tumours are characterized by hypoxia TME. TME is a prominent barrier to oxygen-dependent therapy such as radiotherapy, chemotherapy, photodynamic therapy and immunotherapy (Graham and Unger 2018). A novel therapeutic approach for reversing TME by nanotechnology has emerged (Fig. 20). The main strategies are (1) external oxygen linked to hypoxic TME, (2) nanotechnology-based oxygen generation, (3) structural regulation of TME and (4) decreasing oxygen consumption in TME (Wu et al. 2022). The recent advancements in these strategies reveal that (1) external oxygen strategy can be executed by oxygen carriers. Different carriers used are artificial Red Blood Cells Substitutes (RBCs) (Jia et al. 2016; Gao et al. 2017), PFC-based oxygen carriers (Song et al. 2017), Metal–Organic Frameworks (MOFs) (Gao et al. 2018; Ren et al. 2020) and oxygen microcapsules (Wu et al. 2021). (2) Oxygen generation can be carried out by chemical decomposition method (Liu et al. 2020; Liang et al. 2020), H2 O2 breakdown by catalase (Phua et al. 2019) and by water splitting (Zhao et al. 2020). (3) Structural regulation can be made by increasing blood flow (Shih et al. 2021). (4) Decreasing oxygen consumption can inhibit oxidative respiration through an ‘O2 economizer’ (Li et al. 2020; Yuan et al. 2021). A self-calibrated activatable nanoprobe (Cy7-1/PG5-Cy5@LWHA) was designed to detect nitro-reductase activity, which is expressed more in hypoxic tumour cells. This nanoprobe allows for ratiometric calibration due to dual fluorescence emission and eliminates target-independent interference. Researchers demonstrated that this

150

J. Deepa Arul Priya et al.

Fig. 20 Schematic diagram of the nanotechnology strategies for overcoming tumor hypoxia microenvironment (Source Wu et al. 2022)

imaging property of the Cy7-1/PG5-Cy5@LWHA probe will be a promising tool to track tumour hypoxia and to explore novel anti-hypoxia therapeutics (Feng et al. 2022).

4.3.2

Targeted Therapeutics

HIF-1α is marked as the main element in the hypoxia signalling pathway. This has led researchers to study each step in the pathway from the onset of diseases. This helped in designing target therapeutics for hypoxia-associated diseases (Moyer 2012). The advancement in multiplexed quantitative techniques has supported research groups in assessing accurate modifications in protein interactions and abundances in biological complexes (Han et al. 2008; Singh et al. 2014). Different MS ionization techniques have revolutionized protein identification and analysis of protein expression, structure, interaction, etc. (Fig. 21). This has given us a better understanding of complex biological processes and diseases (Li et al. 2017).

Thyroid Cancer Recently, García-Vence et al. (2020) studied thyroid cancer with a nanoparticleassisted proteomics approach. The challenges in processing were overcome with some techniques (Fig. 22). The Formalin-Fixed Paraffin-Embedded (FFPE) tissue samples were collected for analysis. Deparaffinization was an important step in protein extraction where water was used instead of xylene solvent (Kalantari et al. 2016). The extracted proteins were incubated with respective NPs (Au, Ag and Fe) to form protein corona. PCs were then separated by SDS-PAGE, digested with trypsin, and analysed by LC–MS/MS. The data files were processed using bioinformatic tools meant for database search and data grouping such as ProteinPilot 5.0.1 and

Nanoproteomics: An Approach for the Identification of Molecular …

151

Fig. 21 An overview of techniques adopted in different components of MS-based nanoproteomics (Source Yi et al. 2017)

Progroup (Shilov et al. 2007). Functional interaction networks and protein ontology classification were analysed using STRING and PANTHER (Szklarczyk et al. 2015). The differentially expressed proteins were compiled as novel molecular targets for therapeutics. The data analysis showed that HIF-1α, commonly overexpressed in thyroid cancer (anaplastic (undifferentiated) thyroid carcinoma (ATC)), plays a decisive role in processes like glycolysis activation and OxPhos inhibition. It was also identified in all follicular cell-derived tumours. This glycolysis process generally occurs in high proliferating cancer cells where glucose consumption is increased with subsequent production of lactate in place of oxygen. Thus, mitochondrial OxPhos is suppressed and induces growth of cancer cells as a part of hypoxic response (Denko 2008).

Prostrate Cancer Tumour treatments are dominated by therapies mediated with reactive oxygen species (ROS). In hypoxia-related tumours, ROS-related treatments are resisted by strong antioxidant defence mechanism and protective autophagy. To overcome this (Fig. 23), an X-ray-triggered nitrite (NO2 ) was used in prostate cancer therapy. This inhibited autophagy and increased nitrosative stress based on an electrophilic zeolitic imidazole framework (ZIF-82-PVP). The workflow was that, after incorporation of pHresponsive ZIF-82-PVP nanoparticles, electrophilic ligands and Zn2+ were delivered into cancer cells. Firstly, electrophilic ligands consumed Glutathione (GSH) and

152

J. Deepa Arul Priya et al.

Fig. 22 Flow chart depicting tissue extract pre-treatment (Source García-Vence et al. 2020)

then, low-energy electrons derived from X-rays were captured to generate NO2 . This was followed by the inhibition of autophagy and nitrosative stress level enhancement. Then Zn2+ through ion interference particularly limited the progression of cancer cells. In vitro and in vivo studies indicated that ZIF-82-PVP nanoparticles can stimulate the apoptosis of cancer cells with X-ray irradiation effectively. Overall, this nitrosative stress-mediated tumour therapy provided a new insight for targeting hypoxic tumours (Li et al. 2021).

Cardiovascular Diseases Tiambeng et al. (2020) had come up with an approach to resolving proteoforms for analysing low-abundance proteins directly from serum. Until then, this was an unresolved issue because of the presence of high dynamic proteome range in blood. This study was carried out by pairing peptide-functionalized superparamagnetic NPs (NP-Pep) and MS for analysing cardiac troponin (cTnI), a gold-standard biomarker for cardiovascular diseases (Fig. 24). cTnI is first released after cardiac injury into the blood. So, the detection becomes difficult because of its low abundance (Park et al. 2017b). The binding of NP-Pep with cTnI was favourable for detection through top-down LC/MS. This revealed unique cTnI proteoform fingerprints in each human

Nanoproteomics: An Approach for the Identification of Molecular …

153

Fig. 23 Schematic diagram of ZIF-82-PVP synthesis and its mechanism of action in hypoxic prostate cancer therapy (Source Li et al. 2021)

serum sample with distinct pathophysiology. Since cTnI was heavily modified, its proteoforms from PTMs provided a new perception of the molecular mechanisms significant in cardiovascular diseases (Soetkamp et al. 2017). The use of peptides for protein enrichment offered indicative advantages over antibodies.

5 Conclusion Biomarkers or molecular markers are identified for detecting a disease, early diagnosis and drug discovery. Among all kinds of biomarkers, proteins are very sensitive and accomplish labour-intensive workups (Alharbi 2020). These limitations faced in traditional proteome research like sensitivity, dynamic range, detection time, biological complexities, etc., were overcome by a novel approach known as ‘Nanoproteomics’. The integration of nanotechnology with proteomics has proved to be advantageous in designing nanomaterial miniatures, nano-level separation media and channels for biomedical research.

154

J. Deepa Arul Priya et al.

Fig. 24 Proteoform-resolved analysis of low-abundance cardiac troponin I in human serum by nanoproteomics (Source Melby et al. 2021b)

Nanoproteomics: An Approach for the Identification of Molecular …

155

Earlier studies on hypoxia gave us an overview of the fundamental concept. But, recent studies illustrate an in-depth view of the signalling pathways triggered in hypoxic conditions (Chen et al. 2020). Although more research on targeting hypoxia has been conducted, clinical applications are lagging. Many nanotechnology strategies to reverse hypoxic TME are not approved by FDA (Wu et al. 2022). The biocompatibility and safety issues of nanoparticles are of present concern. So, further research is being looked forward for a suitable candidate biomarker for biomedical applications. In future, several single molecule or single particle MS techniques in trend can be combined with nanomechanical systems-based analysis with parallel developments in mass analysers, sampling processes, separation methods and data analysis for fast evolution of potential therapeutic approaches (Tamara et al. 2022).

References Alharbi RA (2020) Proteomics approach and techniques in identification of reliable biomarkers for diseases. Saudi J Biol Sci 27:968–974 Arulmozhi S, Matchado MS, Snijesh VP, Kumar A, Singh S (2019) An insight into anti-arthritic property of C25 H34 O7 for rheumatoid arthritis using molecular modelling and molecular dynamics approach. Inf Med Unlocked 16:100145 Aslam B, Basit M, Nisar MA, Khurshid M, Rasool MH (2017) Proteomics: technologies and their applications. J Chromatogr Sci 55:182–196 Baker M (2015) Blame it on the antibodies. Nature 521:274 Bell AW, Deutsch EW, Au CE, Kearney RE, Beavis R, Sechi S, Nilsson T, Bergeron JJ (2009) A HUPO test sample study reveals common problems in mass spectrometry–based proteomics. Nat Methods 6:423–430 Bhardwaj M, Singh S, Kumar A (2021) Health informatics-A vital strategy to tackle pandemic diseases. Biomed J Sci Tech Res 39(4):31497–31498 Bonnet J, Garcia C, Léger T, Couquet MP, Vignoles P, Vatunga G, Ndung’u J, Boudot C, Bisser S, Courtioux B (2019) Proteome characterization in various biological fluids of Trypanosoma brucei gambiense-infected subjects. J Proteomics 196:150–161 Brown KA, Chen B, Guardado-Alvarez TM, Lin Z, Hwang L, Ayaz-Guner S, Jin S, Ge Y (2019) A photocleavable surfactant for top-down proteomics. Nat Methods 16:417–420 Cai W, Guner H, Gregorich ZR, Chen AJ, Ayaz-Guner S, Peng Y, Valeja SG, Liu X, Ge Y (2016) MASH suite pro: a comprehensive software tool for top-down proteomics. Mol Cell Proteomics 15:703–714 Cai W, Tucholski T, Chen B, Alpert AJ, McIlwain S, Kohmoto T, Jin S, Ge Y (2017) Top-down proteomics of large proteins up to 223 kDa enabled by serial size exclusion chromatography strategy. Anal Chem 89:5467–5475 Cesnik AJ, Shortreed MR, Schaffer LV, Knoener RA, Frey BL, Scalf M, Solntsev SK, Dai Y, Gasch AP, Smith LM (2018) Proteoform suite: software for constructing, quantifying, and visualizing proteoform families. J Proteome Res 17:568–578 Chait BT (2006) Mass spectrometry: bottom-up or top-down? Science 314:65–66 Chang YH, Gregorich ZR, Chen AJ, Hwang L, Guner H, Yu D, Zhang J, Ge Y (2015) New mass-spectrometry-compatible degradable surfactant for tissue proteomics. J Proteome Res 14:1587–1599 Chen B, Brown KA, Lin Z, Ge Y (2018) Top-down proteomics: ready for prime time? Anal Chem 90:110–127

156

J. Deepa Arul Priya et al.

Chen B, Hwang L, Ochowicz W, Lin Z, Guardado-Alvarez TM, Cai W, Xiu L, Dani K, Colah C, Jin S, Ge Y (2017) Coupling functionalized cobalt ferrite nanoparticle enrichment with online LC/MS/MS for top-down phosphoproteomics. Chem Sci 8:4306–4311 Chen PS, Chiu WT, Hsu PL, Lin SC, Peng I, Wang CY, Tsai SJ (2020) Pathophysiological implications of hypoxia in human diseases. J Biomed Sci 27:1–19 Compton PD, Zamdborg L, Thomas PM, Kelleher NL (2011) On the scalability and requirements of whole protein mass spectrometry. Anal Chem 83:6868–6874 D’Ignazio L, Rocha S (2016) Hypoxia Induced NF-Kb. Cells 5:10 De Angelis M, Calasso M (2014) Molecular biology – proteomics. In: Encyclopaedia of food microbiology, 2nd ed. Academic Press, pp 793–802 Denko NC (2008) Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat Rev Cancer 8:705–713 Dodds JN, Baker ES (2019) Ion mobility spectrometry: fundamental concepts, instrumentation, applications, and the road ahead. J Am Soc Mass Spectrom 30:2185–2195 Duley L (2009) The global impact of pre-eclampsia and eclampsia. In: Seminars in perinatology. WB Saunders, pp 130–137 Dunn MJ, Burghes AH (1986) High resolution two-dimensional polyacrylamide-gel electrophoresis. In: Gel electrophoresis of proteins. Butterworth-Heinemann, pp 203–261 Du´s-Szachniewicz K, Gdesz-Birula K, Zduniak K, Wi´sniewski JR (2021) Proteomic-based analysis of hypoxia-and physioxia-responsive proteins and pathways in diffuse large B-cell lymphoma. Cells 10:2025 Eltzschig HK, Carmeliet P (2011) Hypoxia and inflammation. N Engl J Med 364:656–665 Ethier M, Hou W, Duewel HS, Figeys D (2006) The proteomic reactor: a microfluidic device for processing minute amounts of protein prior to mass spectrometry analysis. J Proteome Res 5:2754–2759 Feng X, Li Y, Zhang S, Li C, Tian J (2022) Quantitative hypoxia mapping using a self-calibrated activatable nanoprobe. J Nanobiotechnology 20:1–15 Gao M, Liang C, Song X, Chen Q, Jin Q, Wang C, Liu Z (2017) Erythrocyte-membrane-enveloped perfluorocarbon as nanoscale artificial red blood cells to relieve tumor hypoxia and enhance cancer radiotherapy. Adv Mater 29:1701429 Gao S, Zheng P, Li Z, Feng X, Yan W, Chen S, Guo W, Liu D, Yang X, Wang S, Liang XJ (2018) Biomimetic O2 -evolving metal-organic framework nanoplatform for highly efficient photodynamic therapy against hypoxic tumor. Biomaterials 178:83–94 García-Vence M, Chantada-Vázquez MD, Cameselle-Teijeiro JM, Bravo SB, Núñez C (2020) A novel nanoproteomic approach for the identification of molecular targets associated with thyroid tumors. Nanomaterials 10:2370 Garrels JI (2001) Proteome. In: Encyclopaedia of genetics. Academic Press, pp 1575–1578 Graham K, Unger E (2018) Overcoming tumor hypoxia as a barrier to radiotherapy, chemotherapy and immunotherapy in cancer treatment. Int J Nanomed 13:6049 Gregorich ZR, Chang YH, Ge Y (2014) Proteomics in heart failure: top-down or bottom-up? Pflügers Arch 466:1199–1209 Hage DS, Anguizola JA, Bi C, Li R, Matsuda R, Papastavros E, Pfaunmiller E, Vargas J, Zheng X (2012) Pharmaceutical and biomedical applications of affinity chromatography: recent trends and developments. J Pharm Biomed Anal 69:93–105 Han X, Aslanian A, Yates JR III (2008) Mass spectrometry for proteomics. Curr Opin Chem Biol 12:483–490 Hanash S (2003) Disease proteomics. Nature 422:226–232 Huang EL, Piehowski PD, Orton DJ, Moore RJ, Qian WJ, Casey CP, Sun X, Dey SK, BurnumJohnson KE, Smith RD (2016) SNaPP: simplified nanoproteomics platform for reproducible global proteomic analysis of nanogram protein quantities. Endocrinology 157:1307–1314 Hughes CS, Foehr S, Garfield DA, Furlong EE, Steinmetz LM, Krijgsveld J (2014) Ultrasensitive proteome analysis using paramagnetic bead technology. Mol Syst Biol 10:757

Nanoproteomics: An Approach for the Identification of Molecular …

157

Hwang L, Ayaz-Guner S, Gregorich ZR, Cai W, Valeja SG, Jin S, Ge Y (2015) Specific enrichment of phosphoproteins using functionalized multivalent nanoparticles. J Am Chem Soc 137:2432– 2435 Issaq HJ, Veenstra TD (2008) Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE): advances and perspectives. Biotechniques 44:697–700 Jeong K, Kim J, Gaikwad M, Hidayah SN, Heikaus L, Schlüter H, Kohlbacher O (2020) FLASHDeconv: ultrafast, high-quality feature deconvolution for top-down proteomics. Cell Syst 10:213–218 Jia L, Lu Y, Shao J, Liang XJ, Xu Y (2013) Nanoproteomics: a new sprout from emerging links between nanotechnology and proteomics. Trends Biotechnol 31:99–107 Jia Y, Duan L, Li J (2016) Hemoglobin-based nanoarchitectonic assemblies as oxygen carriers. Adv Mater 28:1312–1318 Jungbauer A, Hahn R (2009) Ion-exchange chromatography. Methods Enzymol 463:349–371 Kalantari N, Bayani M, Ghaffari T (2016) Deparaffinization of formalin-fixed paraffin-embedded tissue blocks using hot water instead of xylene. Anal Biochem 507:71–73 Karabacak NM, Li L, Tiwari A, Hayward LJ, Hong P, Easterling ML, Agar JN (2009) Sensitive and specific identification of wild type and variant proteins from 8 to 669 kDa using top-down mass spectrometry* S. Mol Cell Proteomics 8:846–856 Karagiannis GS, Pavlou MP, Saraon P, Musrap N, Xie A, Batruch I, Prassas I, Dimitromanolakis A, Petraki C, Diamandis EP (2014) In-depth proteomic delineation of the colorectal cancer exoproteome: mechanistic insight and identification of potential biomarkers. J Proteomics 103:121–136 Ke Q, Costa M (2006) Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol 70:1469–1480 Keeley TP, Mann GE (2019) Defining physiological normoxia for improved translation of cell physiology to animal models and humans. Physiol Rev 99:161–234 Kim TH, Hur EG, Kang SJ, Kim JA, Thapa D, Lee YM, Ku SK, Jung Y, Kwak MK (2011) NRF2 blockade suppresses colon tumor angiogenesis by inhibiting hypoxia-induced activation of HIF-1α. Cancer Res 71:2260–2275 Knott SJ, Brown KA, Josyer H, Carr A, Inman D, Jin S, Friedl A, Ponik SM, Ge Y (2020) Photocleavable surfactant-enabled extracellular matrix proteomics. Anal Chem 92:15693–15698 Koester-Hegmann C, Bengoetxea H, Kosenkov D, Thiersch M, Haider T, Gassmann M, Schneider Gasser EM (2019) High-altitude cognitive impairment is prevented by enriched environment including exercise via VEGF signaling. Front Cell Neurosci 12:532 Kou Q, Xun L, Liu X (2016) TopPIC: a software tool for top-down mass spectrometry-based proteoform identification and characterization. Bioinformatics 32:3495–3497 Koyasu S, Kobayashi M, Goto Y, Hiraoka M, Harada H (2018) Regulatory mechanisms of hypoxiainducible factor 1 activity: two decades of knowledge. Cancer Sci 109:560–571 Kurien BT, Scofield RH (2006) Western blotting. Methods 38:283–293 Laganowsky A, Reading E, Hopper JT, Robinson CV (2013) Mass spectrometry of intact membrane protein complexes. Nat Protoc 8:639–651 Lai ZW, Yan Y, Caruso F, Nice EC (2012) Emerging techniques in proteomics for probing nano–bio interactions. ACS Nano 6:10438–10448 Lantz C, Zenaidee MA, Wei B, Hemminger Z, Ogorzalek Loo RR, Loo JA (2021) ClipsMS: an algorithm for analyzing internal fragments resulting from top-down mass spectrometry. J Proteome Res 20:1928–1935 LeBleu VS, Taduri G, O’connell J, Teng Y, Cooke VG, Woda C, Sugimoto H, Kalluri R (2013) Origin and function of myofibroblasts in kidney fibrosis. Nat Med 19:1047–1053 Lee MY, Joung YH, Lim EJ, Park JH, Ye SK, Park T, Zhang Z, Park DK, Lee KJ, Yang YM (2006) Phosphorylation and activation of STAT proteins by hypoxia in breast cancer cells. Breast 15:187–195 Lequin RM (2005) Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA). Clin Chem 51:2415–2418

158

J. Deepa Arul Priya et al.

Li M, Shao Y, Kim JH, Pu Z, Zhao X, Huang H, Xiong T, Kang Y, Li G, Shao K, Fan J (2020) Unimolecular photodynamic O2 -economizer to overcome hypoxia resistance in phototherapeutics. J Am Chem Soc 142:5380–5388 Li Q, Luo T, Lu W, Yi X, Zhao Z, Liu J (2019) Proteomic analysis of human periodontal ligament cells under hypoxia. Proteome Sci 17:1–11 Li X, Wang W, Chen J (2017) Recent progress in mass spectrometry proteomics for biomedical research. Sci China Life Sci 60:1093–1113 Li Y, Gong T, Gao H, Chen Y, Li H, Zhao P, Jiang Y, Wang K, Wu Y, Zheng X, Bu W (2021) ZIF-based nanoparticles combine X-ray-induced nitrosative stress with autophagy management for hypoxic prostate cancer therapy. Angew Chem Int Ed Engl 133:15600–15609 Liang JH, Zheng Y, Wu XW, Tan CP, Ji LN, Mao ZW (2020) A Tailored multifunctional anticancer nanodelivery system for ruthenium-based photosensitizers: tumor microenvironment adaption and remodeling. Adv Sci 7:1901992 Liu J, Gao J, Zhang A, Guo Y, Fan S, He Y, Yang K, Wang J, Cui D, Cheng Y (2020) Carbon nanocage-based nanozyme as an endogenous H2 O2 -activated oxygenerator for real-time bimodal imaging and enhanced phototherapy of esophageal cancer. Nanoscale 12:21674–21686 Liu X, Sirotkin Y, Shen Y, Anderson G, Tsai YS, Ting YS, Goodlett DR, Smith RD, Bafna V, Pevzner PA (2012) Protein identification using top-down spectra. Mol Cell Proteomics 11 Lollo B, Steele F, Gold L (2014) Beyond antibodies: new affinity reagents to unlock the proteome. Proteomics 14:638–644 Luo Z, Tian M, Yang G, Tan Q, Chen Y, Li G, Zhang Q, Li Y, Wan P, Wu J (2022) Hypoxia signaling in human health and diseases: implications and prospects for therapeutics. Signal Transduct Target Ther 7:1–30 Marouga R, David S, Hawkins E (2005) The development of the DIGE system: 2D fluorescence difference gel analysis technology. Anal Bioanal Chem 382:669–678 Martin K, Zhang T, Lin TT, Habowski AN, Zhao R, Tsai CF, Chrisler WB, Sontag RL, Orton DJ, Lu YJ, Rodland KD (2021) Facile one-pot nanoproteomics for label-free proteome profiling of 50–1000 mammalian cells. J Proteome Res 20:4452–4461 Marty MT, Baldwin AJ, Marklund EG, Hochberg GK, Benesch JL, Robinson CV (2015) Bayesian deconvolution of mass and ion mobility spectra: from binary interactions to polydisperse ensembles. Anal Chem 87:4370–4376 McIlwain SJ, Wu Z, Wetzel M, Belongia D, Jin Y, Wenger K, Ong IM, Ge Y (2020) Enhancing topdown proteomics data analysis by combining deconvolution results through a machine learning strategy. J Am Soc Mass Spectrom 31:1104–1113 Melby JA, de Lange WJ, Zhang J, Roberts DS, Mitchell SD, Tucholski T, Kim G, Kyrvasilis A, McIlwain SJ, Kamp TJ, Ralphe JC (2021a) Functionally integrated top-down proteomics for standardized assessment of human induced pluripotent stem cell-derived engineered cardiac tissues. J Proteome Res 20:1424–1433 Melby JA, Roberts DS, Larson EJ, Brown KA, Bayne EF, Jin S, Ge Y (2021b) Novel strategies to address the challenges in top-down proteomics. J Am Soc Mass Spectrom 32:1278–1294 Michiels C (2004) Physiological and pathological responses to hypoxia. Am J Pathol 164:1875– 1882 Monopoli MP, Walczyk D, Campbell A, Elia G, Lynch I, Baldelli Bombelli F, Dawson KA (2011) Physical−chemical aspects of protein corona: relevance to in vitro and in vivo biological impacts of nanoparticles. J Am Chem Soc 133:2525–2534 Moseley FL, Bicknell KA, Marber MS, Brooks G (2007) The use of proteomics to identify novel therapeutic targets for the treatment of disease. J Pharm Pharmacol 59:609–628 Moyer MW (2012) Targeting hypoxia brings breath of fresh air to cancer therapy. Nat Med 18:636– 638 Nakayama K (2013) cAMP-response element-binding protein (CREB) and NF-κB transcription factors are activated during prolonged hypoxia and cooperatively regulate the induction of matrix metalloproteinase MMP1. J Biol Chem 288:22584–22595

Nanoproteomics: An Approach for the Identification of Molecular …

159

Nakayama K, Kataoka N (2019) Regulation of gene expression under hypoxic conditions. Int J Mol Sci 20:3278 Njunge JM, Oyaro IN, Kibinge NK, Rono MK, Kariuki SM, Newton CR, Berkley JA, Gitau EN (2017) Cerebrospinal fluid markers to distinguish bacterial meningitis from cerebral malaria in children. Wellcome Open Res 2 Park J, Piehowski PD, Wilkins C, Zhou M, Mendoza J, Fujimoto GM, Gibbons BC, Shaw JB, Shen Y, Shukla AK, Moore RJ (2017a) Informed-proteomics: open-source software package for top-down proteomics. Nat Methods 14:909–914 Park KC, Gaze DC, Collinson PO, Marber MS (2017b) Cardiac troponins: from myocardial infarction to chronic disease. Cardiovas Res 113:1708–1718 Phua SZ, Yang G, Lim WQ, Verma A, Chen H, Thanabalu T, Zhao Y (2019) Catalase-integrated hyaluronic acid as nanocarriers for enhanced photodynamic therapy in solid tumor. ACS Nano 13:4742–4751 Picotti P, Bodenmiller B, Mueller LN, Domon B, Aebersold R (2009) Full dynamic range proteome analysis of S. cerevisiae by targeted proteomics. Cell 138:795–806 Pugh CW, Ratcliffe PJ (2017) New horizons in hypoxia signaling pathways. Exp Cell Res 356:116– 121 Rangisetty PT, Kilaparthi A, Akula S, Bhardwaj M, Singh S (2023) RSAD2: an exclusive target protein for Zika virus comparative modeling, characterization, energy minimization and stabilization. Int J Health Sci 17(1):12–17 Ramachandran N, Srivastava S, LaBaer J (2008) Applications of protein microarrays for biomarker discovery. Proteomics Clin Appl 2:1444–1459 Rappsilber J, Mann M, Ishihama Y (2007) Protocol for micro-purification, enrichment, prefractionation and storage of peptides for proteomics using StageTips. Nat Protoc 2:1896–1906 Ray S, Reddy PJ, Choudhary S, Raghu D, Srivastava S (2011) Emerging nanoproteomics approaches for disease biomarker detection: a current perspective. J Proteomics 74:2660–2681 Ren SZ, Wang B, Zhu XH, Zhu D, Liu M, Li SK, Yang YS, Wang ZC, Zhu HL (2020) Oxygen self-sufficient core–shell metal–organic framework-based smart nanoplatform for enhanced synergistic chemotherapy and photodynamic therapy. ACS Appl Mater Interfaces 12:24662–24674 Roberts DS, Chen B, Tiambeng TN, Wu Z, Ge Y, Jin S (2019) Reproducible large-scale synthesis of surface silanized nanoparticles as an enabling nanoproteomics platform: enrichment of the human heart phosphoproteome. Nano Res 12:1473–1481 Saveliev SV, Woodroofe CC, Sabat G, Adams CM, Klaubert D, Wood K, Urh M (2013) Mass spectrometry compatible surfactant for optimized in-gel protein digestion. Anal Chem 85:907– 914 Schito L, Semenza GL (2016) Hypoxia-inducible factors: master regulators of cancer progression. Trends Cancer 2:758–770 Schneider Gasser EM, Gassmann M, Thiersch M (2021) HIF-2: an important player in neuronal response to ischemia. Pflügers Arch 473:1175–1176 Semenza GL (2000) HIF-1: mediator of physiological and pathophysiological responses to hypoxia. J Appl Physiol Semenza GL (2012) Hypoxia-inducible factors in physiology and medicine. Cell 148:399–408 Shen X, Liang Z, Xu T, Yang Z, Wang Q, Chen D, Pham L, Du W, Sun L (2021) Investigating native capillary zone electrophoresis-mass spectrometry on a high-end quadrupole-time-of-flight mass spectrometer for the characterization of monoclonal antibodies. Int J Mass Spectrom 462:116541 Shih CY, Wang PT, Su WC, Teng H, Huang WL (2021) Nanomedicine-based strategies assisting photodynamic therapy for hypoxic tumors: state-of-the-art approaches and emerging trends. Biomedicines 9:137 Shilov IV, Seymour SL, Patel AA, Loboda A, Tang WH, Keating SP, Hunter CL, Nuwaysir LM, Schaeffer DA (2007) The Paragon algorithm, a next generation search engine that uses sequence temperature values and feature probabilities to identify peptides from tandem mass spectra. Mol Cell Proteomics 6:1638–1655

160

J. Deepa Arul Priya et al.

Singh S, Jannet Vennila J, Kant R (2014) Gene interaction map: a paradigm for identifying significant pathways responsible for rheumatoid arthritis. Netw Model Anal Health Inf Bioinf 3:1–7 Soetkamp D, Raedschelders K, Mastali M, Sobhani K, Bairey Merz CN, Van Eyk J (2017) The continuing evolution of cardiac troponin I biomarker analysis: from protein to proteoform. Expert Rev Proteomics 14:973–986 Song G, Ji C, Liang C, Song X, Yi X, Dong Z, Yang K, Liu Z (2017) TaOx decorated perfluorocarbon nanodroplets as oxygen reservoirs to overcome tumor hypoxia and enhance cancer radiotherapy. Biomaterials 112:257–263 Stejskal K, Op de Beeck J, Dürnberger G, Jacobs P, Mechtler K (2021) Ultrasensitive nanoLC-MS of subnanogram protein samples using second generation micropillar array LC technology with orbitrap exploris 480 and FAIMS PRO. Anal Chem 93:8704–8710 Sun RX, Luo L, Wu L, Wang RM, Zeng WF, Chi H, Liu C, He SM (2016) pTop 1.0: a high-accuracy and high-efficiency search engine for intact protein identification. Anal Chem 88:3082–3090 Sutandy FR, Qian J, Chen CS, Zhu H (2013) Overview of protein microarrays. Curr Protoc Protein Sci 72:27–31 Szklarczyk D, Franceschini A, Wyder S, Forslund K, Heller D, Huerta-Cepas J, Simonovic M, Roth A, Santos A, Tsafou KP, Kuhn M (2015) STRING v10: protein–protein interaction networks, integrated over the tree of life. Nucleic Acids Res 43:447–452 Takemori A, Butcher DS, Harman VM, Brownridge P, Shima K, Higo D, Ishizaki J, Hasegawa H, Suzuki J, Yamashita M, Loo JA (2020) PEPPI-MS: polyacrylamide-gel-based prefractionation for analysis of intact proteoforms and protein complexes by mass spectrometry. J Proteome Res 19:3779–3791 Tamara S, den Boer MA, Heck AJ (2022) High-resolution native mass spectrometry. Chem Rev 122:7269–7326 Tiambeng TN, Roberts DS, Brown KA, Zhu Y, Chen B, Wu Z, Mitchell SD, Guardado-Alvarez TM, Jin S, Ge Y (2020) Nanoproteomics enables proteoform-resolved analysis of low-abundance proteins in human serum. Nat Commun 11:1–12 Toby TK, Fornelli L, Srzenti´c K, DeHart CJ, Levitsky J, Friedewald J, Kelleher NL (2019) A comprehensive pipeline for translational top-down proteomics from a single blood draw. Nat Protoc 14:119–152 Tran JC, Doucette AA (2009) Multiplexed size separation of intact proteins in solution phase for mass spectrometry. Anal Chem 81:6201–6209 Valeja SG, Xiu L, Gregorich ZR, Guner H, Jin S, Ge Y (2015) Three dimensional liquid chromatography coupling ion exchange chromatography/hydrophobic interaction chromatography/reverse phase chromatography for effective protein separation in top-down proteomics. Anal Chem 87:5363–5371 Vinaiphat A, Low JK, Yeoh KW, Chng WJ, Sze SK (2021) Application of advanced mass spectrometry-based proteomics to study hypoxia driven cancer progression. Front Oncol 11:559822 Voedisch B, Thie H (2010) Size exclusion chromatography. In: Antibody engineering. Springer, pp 607–612 Wakhloo D, Scharkowski F, Curto Y, Javed Butt U, Bansal V, Steixner-Kumar AA, Wüstefeld L, Rajput A, Arinrad S, Zillmann MR, Seelbach A (2020) Functional hypoxia drives neuroplasticity and neurogenesis via brain erythropoietin. Nat Commun 11:1–12 Wang T, Chen C, Larcher LM, Barrero RA, Veedu R (2019) Three decades of nucleic acid aptamer technologies: lessons learned, progress and opportunities on aptamer development. Biotechnol Adv 37:28–50 Westergren-Thorsson G, Marko-Varga G, Malmström J, Larsen K (2006) Proteome. In: Encyclopedia of respiratory medicine. Elsevier Inc., pp 527–532 Wi´sniewski JR, Zougman A, Nagaraj N, Mann M (2009) Universal sample preparation method for proteome analysis. Nat Methods 6:359–362 Wong WJ, Qiu B, Nakazawa MS, Qing G, Simon MC (2013) MYC degradation under low O2 tension promotes survival by evading hypoxia-induced cell death. Mol Cell Biol 33:3494–3504

Nanoproteomics: An Approach for the Identification of Molecular …

161

Wu B, Sun Z, Wu J, Ruan J, Zhao P, Liu K, Zhao CX, Sheng J, Liang T, Chen D (2021) Nanoparticlestabilized oxygen microcapsules prepared by interfacial polymerization for enhanced oxygen delivery. Angew Chem Int Ed Engl 60:9284–9289 Wu J, Song J, Yin X, Tang J, Zhang J, Wang X, Ji Y, Zhao Y, Chen D, Sheng J, Bai X (2022) Recent advancements of nanotechnology-based strategies for overcoming tumor microenvironment hypoxia. Front Biosci-Landmark 27:145 Wu MH, Hsiao KY, Tsai SJ (2019) Hypoxia: the force of endometriosis. J Obstet Gynaecol Res 45:532–541 Wu R, Pai A, Liu L, Xing S, Lu Y (2020a) NanoTPOT: enhanced sample preparation for quantitative nanoproteomic analysis. Anal Chem 92:6235–6240 Wu Z, Roberts DS, Melby JA, Wenger K, Wetzel M, Gu Y, Ramanathan SG, Bayne EF, Liu X, Sun R, Ong IM (2020b) MASH explorer: a universal software environment for top-down proteomics. J Proteome Res 19:3867–3876 Xie S, Moya C, Bilgin B, Jayaraman A, Walton SP (2009) Emerging affinity-based techniques in proteomics. Expert Rev Proteomics 6:573–583 Yang Z, Zhang Z, Chen D, Xu T, Wang Y, Sun L (2021) Nanoparticle-aided nanoreactor for nanoproteomics. Anal Chem 93:10568–10576 Yates JR III (2011) A century of mass spectrometry: from atoms to proteomes. Nat Methods 8:633–637 Yi L, Piehowski PD, Shi T, Smith RD, Qian WJ (2017) Advances in microscale separations towards nanoproteomics applications. J Chromatogr A 1523:40–48 Yu YQ, Gilar M, Lee PJ, Bouvier ES, Gebler JC (2003) Enzyme-friendly, mass spectrometrycompatible surfactant for in-solution enzymatic digestion of proteins. Anal Chem 75:6023–6028 Yuan P, Deng FA, Liu YB, Zheng RR, Rao XN, Qiu XZ, Zhang DW, Yu XY, Cheng H, Li SY (2021) Mitochondria targeted O2 economizer to alleviate tumor hypoxia for enhanced photodynamic therapy. Adv Healthc Mater 10:2100198 Zhao C, Chen Z, Shi R, Yang X, Zhang T (2020) Recent advances in conjugated polymers for visible-light-driven water splitting. Adv Mater 32:1907296 Zhu Y, Dou M, Piehowski PD, Liang Y, Wang F, Chu RK, Chrisler WB, Smith JN, Schwarz KC, Shen Y, Shukla AK (2018a) Spatially resolved proteome mapping of laser capture microdissected tissue with automated sample transfer to nanodroplets. Mol Cell Proteomics 17:1864–1874 Zhu Y, Piehowski PD, Zhao R, Chen J, Shen Y, Moore RJ, Shukla AK, Petyuk VA, CampbellThompson M, Mathews CE, Smith RD (2018b) Nanodroplet processing platform for deep and quantitative proteome profiling of 10–100 mammalian cells. Nat Commun 9:1–10 Zipes DP (2019) Biomarkers and use in precision medicine. In: Braunwald’s heart disease: a textbook of cardiovascular medicine. Elsevier Inc., pp 73–82

Nanomaterial-Mediated Theranostics for Vascular Diseases Tejaswini Divanji, Krisha Desai, Bhupendra Prajapathi, and Saritha Shetty

Abstract Vascular diseases are widely known to be the leading cause of death and disabilities across the world. Although the conventional treatment methods have shown success there is a need for advancement in order to enable early detection, screening and diagnosis. Nanotechnology is such a unique branch of science that enables not only effective treatment of malfunctioning cells in particular vessel lesions and evaluation of disease progression but also aids in screening, diagnosis and ultimately prevention of vascular diseases. There have been many developments in the field of nanotechnology which make combining therapeutic and diagnostic moieties possible. This review focuses on the use of nanotechnology-based theranostics for a myriad of vascular diseases, such as coronary artery disease, atherosclerosis, neurovascular diseases and thrombosis to name a few, and highlights their advantages, drawbacks and future scope of advancements. Keywords Nanotechnology · Theranostic · Nanoparticle · Coronary artery disease · Aneurysm · Nanoparticles · Atherosclerosis · Aneurysm · Inflammation

Abbreviations AAA ACPP ALA AuNPs CAA

Abdominal aortic aneurysm Activatable cell penetrating peptide Aminolevulinic acid Gold nanoparticles Cerebral Amyloid Angiopathy

T. Divanji · K. Desai · S. Shetty (B) Shobhaben Pratapbhai Patel School of Pharmacy & Technology Management, SVKM’s NMIMS, V.L. Mehta Road, Vile Parle [W], Mumbai 400056, India e-mail: [email protected] B. Prajapathi (B) S.K. Patel College of Pharmaceutical Education and Research, Ganpat University, Mehsana, Gujarat, India e-mail: [email protected] © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_9

163

164

CAD CEST CVD DOX DVT EaRASMC EC ECM EMMPRIN FTP HMGB-1 IONPs MFNPs MI MMP-2 NCD PDT PS PVD ROS SLN SMCs SOD SPIONPs TAA TLRs VEGF

T. Divanji et al.

Coronary artery disease Chemical exchanger saturation transfer Cardiovascular disease Doxycyline Deep vein thrombosis Aneurysmal smooth muscle cells Endothelial cells Extracellular matrix Extracellular matrix metalloproteinase inducer Fibrin-binding peptide High mobility group box Iron oxide nanoparticles Magnetic fluorescent nanoparticles Myocardial infarction Metalloproteases Non-communicable diseases Photodynamic therapy Photosensitiser Peripheral vascular disease Reactive oxygen species Solid lipid nanoparticles Smooth muscle cells Superoxide dismutase Superparamagnetic iron oxide nanoparticles Thoracic aortic aneurysm Toll-like receptors Vascular endothelial growth factors

1 Introduction The global health network has made leaps in advancements to protect and maintain human health. Nevertheless, the world today is witnessing an increasing incidence in the rate of non-communicable diseases (NCDs). The NCDs include cardiovascular diseases (CVDs), diabetes, respiratory diseases and cancer among others. Since the NCDs would account for about 70% of global death by the year 2025, they have proven to be a global health issue. Among them, 84% of the global death will be by the occurrence of CVDs (48%), cancer (21%), respiratory diseases (12%) and diabetes (3%) (Pala et al. 2021). Since CVDs have the highest incidence rate, it is the most prominent NCD with a high-risk factor. CVDs include ailments such as atherosclerosis, myocardial infarction, aneurysms, peripheral vascular disease, etc. It is estimated that by the year 2030, the global mortality rates due to CVDs will rise up to 22 million (from 17.7 million in 2015). Due to this reason, there is a need arising

Nanomaterial-Mediated Theranostics for Vascular Diseases

165

for advances in technology and therapeutic substitutes. Although immunotherapy, chemotherapy and other conventional methods are being used as front-line treatment, there is a significant effort being made to upgrade or curate diagnostic and therapeutic alternatives (Zumla et al. 2016).

1.1 Nanotheranostics A combinatorial modality termed ‘theranostics’ was put forth by John Funkhouser in 2002 involving both diagnosis and therapeutics. It includes the association of diagnostic tools and efficient therapeutic measures under a single platform. A series of sequential steps are followed beginning with early detection and diagnosis of the disease, disease prognosis, selecting the therapy and precisely monitoring the therapeutic efficacy. Theranostic tools have helped develop a customised, target-guided therapy platform for the treatment of chronic infectious diseases, inflammatory diseases and cancer therapy. Some of its salient benefits also include a non-invasive platform, low toxicity, target selectivity and specificity and tunability. ‘Nanotheranostics’ is an advanced concept wherein nanoparticles of different classes and physicochemical properties are combined with theranostic modalities to optimise disease diagnosis and therapies for the same. Recently, nanomaterials are being explored as prolific contrasting agents. This proves them instrumental in the early diagnosis and prognosis of diseases and, hence, effective to be used as bioimaging modalities (Pala et al. 2021). On the other hand, features like ease of synthesis, good biocompatibility and stability, bimodal applications, non-immunogenicity, large surface area and pore volume provide a unique platform for redefining therapeutic measures used for various chronic health conditions (Deng et al. 2020). Commonly used nanomaterials for drug delivery include lipid-forming micelles or liposomes, polymeric nanoparticles, dendrimers, carbon nanotubes and metallic nanoparticles such as crystalline iron oxide and gold nanoparticles as shown in Fig. 1 (Matoba et al. 2017). Commonly used diagnostic tools and therapeutics in nanotheranostics are depicted in Fig. 2. A lot of progress has been made to treat cardiovascular diseases with the use of nanotechnology. For example, to treat hypertriglyceridemia, non-formulations of fenofibrate are currently being used to overcome difficulties associated with drug solubility and absorption (Flores et al. 2019).

1.2 Vascular Diseases: Pathophysiology, Diagnosis and Treatment Vascular disease is a collective term used to describe conditions that affect the circulatory system, i.e. arteries, veins, capillaries and lymph vessels. Vascular diseases include but are not limited to inherited conditions, degenerative processes, trauma

166

T. Divanji et al.

Fig. 1 Various nanomaterials in drug delivery systems

Fig. 2 Diagnostics and therapeutics modalities of nanotheranostics

and autoimmune and infectious diseases. Vascular diseases impact one or all layers of the blood vessel such as adventitia, media and intima, and sometimes even the blood in the lumen of the vessel (Stone 2016). Ath1erosclerosis is a condition characterised by lipid accumulation, fibrous elements and calcification of arterial walls. Increasing plasma cholesterol levels

Nanomaterial-Mediated Theranostics for Vascular Diseases

167

leads to an increase in endothelial permeability. The endothelium’s ability to maintain homeostasis which makes the arterial wall susceptible to vasoconstriction and migration of LDL-C is also seen. The lipoidal migration into the vessel is followed by the adhesion of monocytes to endothelial cells (EC). The monocyte adhesion is succeeded by the formation of foamy macrophages, platelet activation and high levels of oxidative stress which triggers the inflammatory response including the release of high mobility group box (HMGB-1), toll-like receptors (TLRs), interleukin (Il-6, IL-β) and tumour necrosis factor (TNF-α) (Agrawal et al. 2020; Jebari-Benslaiman et al. 2022). The progression of atherosclerosis is shown in Fig. 3. Atherosclerosis manifests clinically into angina pectoris, myocardial infarction and ultimately into coronary artery disease (CAD) depending upon the level of obstruction (JebariBenslaiman et al. 2022). Lipoprotein modification is one of the most commonly employed treatment strategies which includes the use of cholesterol absorption inhibitors like ezetimibe,3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitors or statins and bile acid sequestrants like colestipol. Lifestyle modifications like dietary changes, stimulation of physical activity and cessation of smoking are used as risk mitigation measures. Angioplasty is used as a surgical intervention in more serious cases (Bergheanu et al. 2017). Peripheral vascular disease (PVD) presents itself as a manifestation of progressing systemic atherosclerosis (Fig. 3) characterised by narrowing of blood vessels distal to the aortal arch leading to partial or complete vascular obstruction and reduced blood flow to visceral organs ultimately resulting in end-organ ischaemia (Sontheimer 2006; Peripheral Vascular Disease—StatPearls—NCBI Bookshelf 2022). Smoking, diabetes mellitus, hypertension, hyperlipidemia and a sedentary lifestyle are the usual risk factors for PVD (Sontheimer 2006). Acute limb ischaemia is the typical symptom along with intermittent claudication (Sontheimer 2006). Diagnosis is complicated as many patients present similar comorbid and asymptomatic/atypical

Fig. 3 Schematic representation of progression in atherosclerosis

168

T. Divanji et al.

conditions. Physical and neurological examination is required to distinguish the symptoms of PVD from other neurological and vascular disorders (Peripheral Vascular Disease—StatPearls—NCBI Bookshelf 2022). Anti-platelet drugs such as Clopidogrel and Aspirin are used as treatment options. Other treatment strategies include cardiovascular risk factor modification (smoking cessation), exercise therapy, revascularisation, endovascular stenting, angioplasty and intra-arterial thrombolytic drug (Urokinase) for acute limb ischaemia (Sontheimer 2006; Peripheral Vascular Disease—StatPearls—NCBI Bookshelf 2022).

2 Nanomaterials as Theranostics for Vascular Diseases Nanoparticle mediated theranostics in vascular diseases are listed in Table 1. A few disease conditions along with theranostics are detailed below.

2.1 Atherosclerosis Atherosclerosis, widely known as the underlying pathology leading to myocardial infarction and stroke is a combination of hyperlipidemia and inflammation. The formation of atherosclerotic plaque is modulated by not only the innate but also the adaptive immune systems. Over the years, highly advanced techniques such as cytometry by time of flight, and single-cell RNA sequencing are useful in revealing viable therapeutic targets (MacRitchie et al. 2021). Nanotechnology has proven to be highly useful as a diagnostic tool as well as a therapeutic one. Polymeric nanoparticles, dendrimers, liposomes, micelles, carbon nanotubes and metallic nanoparticles like gold, silver and silica are commonly used for the treatment of cardiovascular diseases. Generally, small particles like radionucleotides (PET scan), iodinated compounds (CT scan) or microbubbles are used in diagnostic tests both of which have their own set of limitations. Therefore, nanoparticles are used due to their myriad advantages (MacRitchie et al. 2021). The therapeutic targets for atherosclerosis are depicted in Fig. 4. Gold nanoparticles (AuNPs) are used as carriers of photosensitisers (PS) for photodynamic therapy (PDT) which is a noninvasive treatment approach for atherosclerosis and cancer. PS molecules bind to the atherosclerotic plaque forming plaque destroying reactive oxygen species (ROS) upon the action of light and irradiation. Aminolevulinic acid (ALA) was incorporated into AuNPs and administered in adult white rabbits (New Zealand species) which leads to an increase of protoporphyrin IX in tissues as a result of uncontrolled cellular proliferation seen in atheromatous plaque. This coupled with the high stability of the conjugate leads to easy drug administration making ALA: AuNPs suitable for the diagnosis and treatment of atherosclerosis (de Oliveira Gonçalves et al. 2015). Iron oxide nanoparticles (IONPs) are another type of nanoparticle used as a theranostic due to their large surface area providing ease of functionalisation and their magnetic properties

Gold nanoparticles (AuNPs) conjugated with polyethylene glycol (PEG)

Polymer lipid nanoparticles Magnetic imaging resonance (MRI)

Iron-perfluorohexane (PFH)-PLGA/Chitosan (CS) nanoparticles

Iron oxide nanoparticles (IONPs)

Protoporphyrin IX (PPIX) synthesis pathway leading to uncontrolled cellular proliferation in atheromatous plaque

Collagen IV expressed on atherosclerotic plaque

Class A scavenger receptors (SR-A)

Reactive oxygen species (ROS)

MRI

Ultrasound imaging and MRI

Fluorescence

Photosensitisers

Gold nanoparticles (AuNPs)

Reactive oxygen species (ROS)

Imaging modality

Nanoparticle

Target

Cerium oxide

Dextran sulphate (DS)

Iron oxide–Paclitaxel conjugated with collagen specific peptide: C11

5-aminolevulinic acid

Photodynamic therapy

Therapeutic modality

Table 1 List of various targets along with nanomaterials mediated theranostics used in vascular diseases Atherosclerosis

Applicati on

(continued)

ROSs are made Atherosclerosis non-reactive due to the action of cerium oxide to allow treatment, whereas Iron oxide aids in improving MRI contrast

The NPs demonstrated Atherosclerosis binding capacity with macrophages leading to their apoptosis

Improved MRI contrast Atherosclerosis and anti-atheros clerotic efficacy

Endogenous porphyrins Atherosclerosis were synthesised and PPIX pathway is overloaded Thus, particle can be used to diagnose and treat atherosclerosis

Photosensitiser molecules bind to the atherosclerotic plaque leading to destroying the plaque

Outcome

Nanomaterial-Mediated Theranostics for Vascular Diseases 169

MRI

Micro computed tomography (CT)

Aneurysmal smooth PLGA NPs with muscle cells (EaRASMCs) supermagnetic iron oxide

Gold nanoparticles

Paramagnetic fluorescent micellar nanoparticles

Degraded elastin fibres in aneurysmal tissue

Extracellular matrix metalloproteinase inducer (EMMPRIN)

MRI

MRI

HDL mimicking particles

High-density lipoprotein (HDL): hyperlipidemia

Imaging modality

Nanoparticle

Target

Table 1 (continued)

EMMPRIN-binding peptide NAP9

Elastin

Doxycyline

Statins

Therapeutic modality

Applicati on

Reperfusion damage and apoptosis progression of myocytes is reduced

(continued)

Myocardia l infarction

The EL-GNPs Aneurysm accumulated around the degraded elastin fibres; used as effective diagnostic tool

The synthesis of Aneurysm metalloproteases (MMP-2; MMP-9) is inhibited; elastic regeneration of SMCs improved Acts as a non-surgical intervention for treating AAAs

The lipophilic nature of Atherosclerosis statins enabled them to lower the excess levels of cholesterol and slowed the progression of inflammation in atherosclerotic mice

Outcome

170 T. Divanji et al.

Magnetic microbubbles with silicon oxide nanoparticles

Paramagnetic liposomes

Phosphatidylcholine and PEG-ylated phosphatidylehtanolamine liposomes

Ischaemic tissues

Plaque in peripheral vessels

Superoxide dismutase–superoxide anion

MRI

MRI, PET, CT

Ultrasound imaging

Fluorescence imaging

Fluorescent silica nanoparticles

Ischaemic smooth muscle cells

Imaging modality MRI

Nanoparticle

Tissue factors expressed in Perfluorocarbon smooth muscle cells nanoparticles

Target

Table 1 (continued)

M4041

Glucocorticoids and anti-inflammatory drugs

VEGF

Vascular endothelial growth factor (VEGF)

Paclitaxel and Doxirubicin

Therapeutic modality

Neuroprotec tive effect and improvement in mouse brain

Improved signal intensity was observed along the inflamed vessel along with atherosclerotic activity

NPs demonstrated strong bioluminescences and improved angiogenesis in ischaemic mice

Ischaemia targeted imaging of tissues was possible owing to the improved permeability of blood vessels VEGF improved perfusion and angiogenesis

Smooth cell proliferation was inhibited by the drug moieties

Outcome

(continued)

Cerebral ischaemia

Peripheral vascular diseases

Limb ischaemia

Hindlimb ischaemia

Peripheral Vascular Disease

Applicati on

Nanomaterial-Mediated Theranostics for Vascular Diseases 171

Phase transition liposome

Polymeric hybrid micelle

Thrombus

Active coagulation factor FXIIIa

Near-infrared imaging

LIFY and acoustic droplet vaporisation

CEST MRI

Fluroescence imaging

Liposomes

Perfluorocarbon nanoparticles

Cloth-bound thrombin

Near-infrared imaging

Ischaemic tissues

Immunoliposome

PirB

MRI and fluorescence imaging

Imaging modality

Magnevist®

Immunoliposomes

HSP72 biomarker on peri-infarct tissue

Amyloid β protein deposits Polymeric nanoparticles

Nanoparticle

Target

Table 1 (continued)

Fibrinolytic drug lumbrokinase

ACPP and FTP

Citicoline

Cyclophosphamide

Thrombin inhibitors like bivalirudin

sPirB

Citicoline

Therapeutic modality

Applicati on

Thrombolysis

Nanoexcavation of thrombus due to microbubble formation

A self-guided platform for delivery of diamagnetic drugs which has neuroprotective effects

Amyloid deposits were targeted

Inhibition of thrombin activity and platelet deposition

Improved neurite growth and motor activities

Carotid thrombosis

Deep vein thrombosis

Cerebral ischaemia

Cerebral amyloid angiopathy (CAA)

Cerebralthrombosis

Cerebral stroke

Reduced lesion volume Cerebral stroke in animals with citicoline

Outcome

172 T. Divanji et al.

Nanomaterial-Mediated Theranostics for Vascular Diseases

173

making them suitable for diagnosis. There are many approaches to administering iron oxide nanoparticle-based theranostic. One approach is to combine the MRI ability of IONPs with drug payloads within encapsulating materials leading to the formation of nanocapsules (Talev and Kanwar 2020). Following this approach, solid lipid nanoparticles (SLNs) containing IONPs contrast agents with prostacyclin were developed and these had better MRI properties for atherosclerosis (Matuszak et al. 2018). Another approach is conjugating nanocapsules by targeting ligands, particularly for biomarkers that are overexpressed in atherosclerosis (Talev and Kanwar 2020). C11, a collagen-specific peptide was added onto the exterior of iron oxide– paclitaxel polymer lipid nanoparticles to target the collagen IV expressed on the atherosclerotic plaque. The resultant molecule exhibited improved MRI contrast and anti-atherosclerotic therapeutic efficacy in an animal model (Rabbit) (Ye et al. 2019). The last approach involves blocking the molecular activity of reactive oxygen species (ROS) (Talev and Kanwar 2020). Iron oxide–cerium oxide nanoparticles were developed: the iron oxide aids in MRI imaging of the plaque and the cerium oxide reacts with ROS to make them non-reactive working on treating atherosclerosis (Bietenbeck et al. 2016). Since hyperlipidemia is also a cause of atherosclerotic plaque, high-density lipoprotein (HDL) mimicking nanoparticles are also used. In 2003, the Tuzcu group tested the anti-atherosclerotic efficacy of HDL-mimicking particles with acute coronary disease. Mulder and workers prepared statin-loaded HDL particles to deliver statins locally which slowed the progression of inflammation in atherosclerotic mice (Duivenvoorden et al. 2014). Fig. 4 Various therapeutic targets for atherosclerosis

174

T. Divanji et al.

2.2 Aneurysm An aneurysm is a vascular condition referring to the expansion/bulging/distention of arteries as a result of the weakening of the arterial walls resulting in fatalities post rupture at the late stage of the disease. Aneurysms are classified based on the location of arterial distention into three types: Aortic aneurysms refer to the aorta carrying blood through the chest and abdomen from the left ventricle and are further divided into abdominal aortic aneurysms (AAA) and thoracic aortic aneurysms (TAA); Cerebral aneurysms affect blood vessels supplying blood to the cerebrum; and Peripheral aneurysms are those that affect peripheral blood vessels. AAA has acquired widespread scrutiny from academia and research circles as it is the chief reason for mortality and morbidity in adults (Pala et al. 2021). Early diagnosis is of utmost importance in aneurysms as there is a high risk of rupture similarly curative treatments are also crucial and antibiotics like doxycycline and tetracycline have proven to be efficacious when administered orally but it is low due to the absence of targeted delivery of the active drug moiety. Thus, nanoparticles prove to be highly useful (Yin et al. 2021). AAA is characterised by the chronic upregulation of metalloproteases (MMPs)-2 and 9, which degrades the elastin matrix of the aortic wall leading to the increasing loss of vascular elasticity. Doxycycline (DOX), a tetracycline antibiotic is known to slow down the growth of AAAs but its oral administration has systemic side effects and does not contribute to the deposition of new elastin fibres in the AAA tissue (Sivaraman and Ramamurthi 2013; Jennewine et al. 2017). Therefore, DOX-loaded poly-lactic-co-glycolic acid (PLGA) nanoparticles were developed to provide the sustained and controlled delivery of DOX in small dosages. Surface functionalisation of the DOX-PLGA NPs with cationic amphiphiles resulted in enhanced aortic uptake of DOX as well as elastin binding through hydrophobic interactions as exhibited in smooth muscle cell (SMCs) cultures (Sivaraman and Ramamurthi 2013). To improve the targetability and specificity of the DOX-PLGA NPs were incorporated with superparamagnetic iron oxide nanoparticles (SPIONPs) which do so under an external magnetic field. When administered to aneurysmal smooth muscle cell cultures (EaRASMC), the DOX-SPIONPs, inhibit the synthesis of MMP-2 and -9 while improving the elastic regeneration and can serve as a non-surgical intervention for treating small growing AAAs (Sivaraman et al. 2017). Cationic amphiphile (DMAB) modified sub-micron particles (SMPs) of DOX were also prepared and these improve elastogenesis while reducing proteolysis. The DOX-SMPs were conjugated with an antibody against Cathepsin-K, a lysosomal protease that is overexpressed in an aneurysm which enhances the pro-elastogenic and anti-proteolytic effects while maintaining a steady release of DOX (Jennewine et al. 2017). Angiotensin II (AngII) was used to induce aneurysms in mice that were injected with elastin-gold nanoparticles (EL-GNPs) retro-orbitally. Micro-computed tomography (CT) technique was used to determine the distribution of the EL-GNPs which were seen to accumulate around the degraded elastin fibres and not around the healthy, intact elastin. Thus, EL-GNPs can be used as an effective diagnostic tool to estimate the elastin damage and the potential of a rupture of an aneurysm (Wang et al.

Nanomaterial-Mediated Theranostics for Vascular Diseases

175

2019). Similarly, Ang II was infused in apoE−/− mice leading to the development of an aneurysm. Human ferritin NPs (HFn) are functionalised with arginine–glycine– aspartic acid (Arg-Gly-Asp) which impacts vascular inflammation enhancing molecular imaging by magnetic resonance imaging (MRI) (Golestani and Sadeghi 2014). Iron oxide NPs were administered to porcine pancreatic elastase-induced AAA mice which act on the macrophage accumulation and homing allowing imaging of AAA using bioluminescence (BLI) and MRI (Yin et al. 2021).

2.3 Myocardial Infarction Derived from the Latin words Infarctus myocardii, myocardial infarction is defined as cardiomyocyte cell death as a result of an ischaemic insult (Frangogiannis 2015). Some common risk factors include old age, smoking tobacco, increased levels of low-density lipoprotein, elevated blood pressure levels, diabetes, sedentary lifestyle, obesity, chronic kidney disease and alcohol and drug abuse among others. MI manifests itself as chest pain, shortness of breath, sweating, nausea, anxiety, fatigue, etc. (Zhang et al. 2022). Diagnosis of MI is dependent on clinical findings, electrocardiograph findings, imaging studies and biomarkers (Frangogiannis 2015). Nanocarriers present a unique and effective opportunity for diagnosing and treating MI. Nanocarriers are loaded with therapeutic agents and target the specific site of disease either actively or passively. In active targeting, the nanocarrier is conjugated with the sitespecific molecule whereas in passive targeting nanocarrier reaches the target site by making use of highly perfusable cells (Manners et al. 2022). It is known that microRNA 199a-3p provides protection against MI. Macrophage membrane-coated nanoparticles (MMNPs) containing miR199a-3p were prepared and administered to MI mice and they were shown to prevent hypoxia-induced apoptosis of myocytes, reducing cardiac inflammation, fibrosis and ventricular modelling while improving cell proliferation. Thus, MMNPs represent an effective therapeutic approach to treating MI as it targets the three most important factors in MI: myocyte apoptosis, inflammation and fibrosis (Xue et al. 2021). Cardiac remodelling is preceded by extracellular matrix (ECM) degradation and it occurs as a result of ischaemia and reperfusion deriving from the occlusion of the coronary artery. Extracellular matrix metalloproteinase inducer (EMMPRIN), a highly expressed molecule in MI was targeted by conjugating EMMPRIN with fluorescent paramagnetic micellar nanoparticles. The nanoparticles were developed and administered to mice with acute myocardial infarction and the NPs conjugated with EMMPRIN binding peptide (NAP9) reduced the damage to myocytes from reperfusion as well as the progression of myocyte apoptosis (Cuadrado et al. 2016). Another unique therapy targeting inflammation induced by monocytes which leads to myocardial ischaemia–reperfusion (IR) injury was developed using Pioglitazone (peroxisome proliferator-activated receptor agonist). Lactic acid/glycolic acid–Pioglitazone nanoparticles were designed and administered to the mouse IR model. The NPs inhibit the Ly6Chigh, reduce inflammatory gene expression in

176

T. Divanji et al.

IR and reduce mortality in MI. In porcine IR models, Pioglitazone NPs also exhibit a cardioprotective effect and reduce cardiac remodelling (Tokutome et al. 2018). Oleate Adenosine prodrug (Ade-OA) and Anti-natriuretic peptide (ANP)distearoylphosphatidylethanolamine-polyethylene glycol were synthesised. ANPmodified Ade-loaded lipid nanocarriers (ANP Ade/LNCs) were formed using the solvent evaporation method and were then administered to rats and these nanosystems inhibited infarct size in vivo (Yu et al. 2018).

2.4 Peripheral Vascular Diseases Peripheral vascular disease (PVD), a slow and escalating disorder of the arterial vessels in the lower extremities is characterised by narrowing of the vessels resulting in their obstruction ultimately leading to the loss of functional capacity. Ankle Brachial Index (ABI) and Toe Brachial Index (TBI) are the typical methods of diagnosing PVD, whereas medication, surgical bypass, endovascular therapies and cellbased therapies are the conventional methods of treating PVD (Noukeu et al. 2018). Nanoparticles are an effective option to diagnose and treat PVD. There are four strategies by which nanoparticles are employed in the treatment of PVD as shown in Fig. 5: as protein carriers for delivering angiogenic growth factors to ischaemic tissues; as gene carriers, they are able to deliver the drug molecules directly into the cell after bypassing the cell membrane; as a facilitator of existing cell therapies to improve angiogenesis; and as tissue engineered vascular scaffold (Tu et al. 2015). Hydrogen peroxide-activatable carbon dioxide bubble generating indocyanine green-loaded boronated maltodextrin (ICG-BM) nanoparticles were developed and they amplified fluorescence and ultrasound signals. ICG-BM NPs also demonstrated anti-inflammatory and pro-angiogenic properties in mouse models with hindlimb ischaemia. The NPs target hydrogen peroxide since they are the most common ROS and they have a longer half-life than all the other ROS making them a promising therapeutic biomarker. Thus, ICG-BM NPs are an effective way of accurately diagnosing lesion sites (Jung et al. 2019). Perfluorocarbon nanoparticles (NPs) were loaded onto paclitaxel and doxorubicin to target tissue factors that are expressed postangioplasty. MRI was used to determine the uptake of NPs within the smooth muscle cells and the drugs exhibited anti-proliferative properties (Noukeu et al. 2018). Hyperlipidemic rats were administered with paramagnetic nanoparticles (PMNPs) loaded with statins and functionality is restored in occluded vessels (Noukeu et al. 2018). In another study, vascular endothelial growth factor (VEGF)-loaded fluorescent silica nanoparticles were injected intravenously into the ischaemic hindlimb of mice which resulted in improved blood perfusion by 93% as compared to the controls. The NPs accurately performed both proangiogenic therapy as well as diagnosis of ischaemic tissues in the mice (Kim et al. 2011). Poly (β amino esters) nanoparticles (PBAE NPs) were designed and used to improve the expression of VEGF and CXC-chemokine receptor type 4 (CXCR4) in ischaemic mice allowing the site of administration as well as the pharmacokinetics and efficacy of the drug to be monitored effectively. It

Nanomaterial-Mediated Theranostics for Vascular Diseases

177

Fig. 5 Therapeutic strategies for peripheral vascular disease (PVD)

also improved perfusion and muscle regeneration in the limbs (Deveza et al. 2016). Magnetic microbubbles are now being used as an imaging agent by ultrasound (US) imaging. Magnetic microbubbles with silicon oxide-coated nanoparticles (SO-Mag) were developed in order to administer VEGF to ischaemic mice which resulted in improved angiogenesis within 72 h, the NPs also exhibited strong bioluminescence making it easy to determine uptake (Heun et al. 2017). Anti-inflammatory drugs have also been used to treat PVD and glucocorticoid-loaded paramagnetic liposomes were developed as a nanotheranostic. MRI, PET and CT were used as imaging modalities to image the plaque, and an increase in signal intensity was seen 2 days after IV administration along the entirety of the inflamed vessel. Quantitative information was obtained using PET scans and the liposomes showed efficacious results over a course of 7 days (Lobatto et al. 2012).

2.5 Neurovascular Diseases Neurological diseases refer to atypical physical condition of the nervous system which impacts the central as well as peripheral nervous system. They manifest themselves as structural, biochemical as well as electrical abnormalities of the brain, spinal

178

T. Divanji et al.

cord and peripheral nerves in the body. The current imaging methods for neurological diseases have many drawbacks such as increased time consumption, reduced sensitivity in MRI, high tissue scattering of fluorescence-based imaging and exhibition of undesirable effects in the surrounding healthy tissues. Similarly, conventional therapies also have disadvantages: difficulty crossing blood brain barrier (BBB), low biocompatibility, solubility and half-life (Sharma et al. 2019). Shahzeeb et al. developed M4041 encapsulated in phosphatidylcholine and PEGylated phosphatidylethanolamine liposomes which catalysed superoxide dismutase. It is widely known that endogenous manganese-based superoxide (Mn-SOD) protects cerebral tissue against damage due to superoxide anion (O2 − ). A synthetic enzyme mimicking SOD with a higher catalytic activity than the native SOD enzyme called M4041 was used. The liposomes were administered to mouse systemically and the presence of Mn (II), a common contrasting agent in neuroimaging in M4041 allowed for imaging using MRI. The nanoparticles also led to region-specific improvement in mouse brain. Thus, M4041 liposomes have proven to be an effective nanotheranostic for MRI-mediated tracking of therapeutic delivery (Shazeeb et al. 2014). In order to develop nanoplatforms for the diagnosis and treatment of cerebral stroke, immunoblotting, immunohistological and proteonomic studies were conducted to determine the expression of molecular biomarkers on ‘peri-infarct’ tissue. A selectively expressed protein on the peri-infarct tissue called HSP72 was selected. In order to separate peri-infarct tissues in vivo, antiHSP72 bearing stealth immunoliposomes with probes for MRI and fluorescence imaging were developed. The liposomes were encapsulated with drug citicoline and led to 30% smaller lesion volume in animals compared to those animals which were administered with non-encapsulated drug (Agulla et al. 2014). A novel theranostic system targeting ischaemic stroke was developed by labelling an anti-PirB with NIR probe. PirB, an immunoglobulin-like receptor B is expressed in neurons and have shown to negatively impact neurite outgrowth. Soluble ectodomain (sPirB) protein was used as a therapeutic agent and the resultant sPirB immunoliposome was shown improve motor abilities due to accumulation in cerebral ischaemic tissue model of mice (Wang et al. 2017). In the case of inflammatory atherosclerosis, magnetic fluorescent nanoparticles (MFNPs) conjugated with dextran coated magnetic nanoparticles and near-infrared dye (NIR) dye were developed. The resulting nanoparticles were photosensitive and upon exposure to light radiation at 646 nm, a singlet oxygen species was released which aided in killing macrophages while protecting the neighbouring healthy cells. Thus, these nanoparticles eradicated inflammatory macrophages and stabilised lesions (McCarthy et al. 2010). A commonly employed strategy in suppressing thrombosis is restoring antithrombotic surfaces which has proven to be an effective alternate to systemic anticoagulation which has a significant risk of bleeding. Keeping this strategy in mind, perfluorocarbon nanoparticles (PFCNPs) functionalised with thrombin inhibitors like bivalirudin were administered to C57BL6 mice. The NPs targeted clot bound thrombin by binding to active clotting sites resulting in inhibition of thrombin activity and platelet deposition (Agyare et al. 2014; Myerson et al. 2014). A neurovascular disease, Cerebral Amyloid Angiopathy (CAA) characterised by the deposition of amyloid beta protein in the cerebral vascular walls

Nanomaterial-Mediated Theranostics for Vascular Diseases

179

followed by aggressive vascular inflammation and ultimately resulting in recurrent hemorrhagic strokes was also diagnosed and treated by nanotheranostics. For CAA, the nanotheranostic consists the following: a polymeric core made up of an MRI contrasting agent Magnevist® conjugated with chitosan and cyclophosphamide (CYC), an immunosuppressant. To further target the cerebrovascular amyloid, the nanocore surface was functionalised by F(ab, )2 fragment (F(ab, )24.1) of a novel antiamyloid antibody. In vitro studies were conducted in polarised human microvascular endothelial cell monolayers (hCMEC/D3), whereas in vivo studies were conducted in mice. These studies confirmed the capability of the nanoparticle in targeting and imaging the amyloid using MRI and single photon computed tomography (Agyare et al. 2014). Cytidine-5’-diphosphocholine or Citicoline (CDPC), a natural precursor of phosphatidylcholine (Ptd-Cho) is widely known to have neuroprotective properties. The intrinsic chemical exchanger saturation transfer (CEST) property of CDPC owing to the presence of exchangeable amine and hydroxyl protons allows its use as tracing tool for drug delivery in cerebral ischaemia. Targeted delivery was ensured by preparing encapsulating citicoline within liposomes and their effect was tested in a rat brain model with temporary ischaemia. Intra-arterial administration of the liposome demonstrated a detectable CEST MRI contrast at 2 ppm. The nanoparticle made it possible to detect diamagnetic drug delivery and thus, a self-guided nanotheranostic platform was developed (Liu et al. 2016).

2.6 Thrombosis Thrombotic events are inter-related with the occurrence of cardiovascular diseases as well as unhealthy life choices such as poor diet, lack of exercise, alcohol and drug use. Over the last 2 years, COVID-19 has emerged as a cause of thrombosis and it has gained significant attention from the scientific and medical communities after reports emerged that the vaccines developed by AstraZeneca (ChAdOx1-S) and Johnson and Johnson (AD26.COV2. S) lead to thromboembolisms as a side effect in people. All of these only highlight the need for timely diagnosis and treatment to prevent high rates of morbidity and mortality. Following the rupture of unstable atherosclerotic plaque (Yang et al. 2020), the blood becomes highly susceptible to procoagulant factors leading to the aggregation of platelets, fibrin and red blood cells (RBCs) resulting in the formation of a clot or thrombi. Coagulation and platelet cascade play an important role in the process of thrombus formation specifically the conversion of fibrinogen to fibrin by thrombin which is involved in both the cascades. The activation of glycoprotein IIb/IIIa allows binding with fibrin and leads to burst release of thrombin and exacerbates thrombus growth (Russell et al. 2022). The detection and dissolution of deep venous thrombosis (DVT) non-invasively present a unique challenge to scientists, especially in areas where the thrombus blockage and thickness is particularly large. In order to combat these challenges, a thrombin-responsive complete thickness infiltration nonpharmaceutical nanoplatform theranostic platform was developed. The theranostic is a thrombin-responsive

180

T. Divanji et al.

phase transition liposome consisting of a liquid perfluoro pentane (PFP) core and two binding peptides: activatable cell-penetrating peptide (ACPP) and fibrin-binding ligand (FTP) which assists in targeting and accumulation within the thrombus. The liposome functions as a nanoexcavator: it uses low intensity-focused ultrasound (LIFU) and acoustic droplet vaporisation to dig out the thrombus. Microbubbles are formed due to the droplet vaporisation which allows the monitoring of the therapeutic process in real time (Yang et al. 2020). Self-assembling, functionalised polymeric hybrid micelle was developed from polycaprolactone-polyethlyene glycol (PCLPEG) and amphiphilic polycaprolactone-polyethlenimine (PCL-PEI). Near-infrared imaging nanoparticles targeting activated coagulation factor XII (FXIIIa), a protein which controls fibrin cross-linking called IR780/FPHM/LK NPs were developed. These nanoparticles were loaded with fibrinolytic drug lumbrokinase (LK) and were administered to a mouse carotid thrombosis model (Wang et al. 2021). Table 1 represents the various targets and nanomaterials along with their therapeutic modality with applications.

3 Advantages and Limitations of Nanoparticle-Mediated Theranostics As is evident, nano-based theranostics was discovered to have an impact on the diagnosis of atherosclerosis, aneurysms and other CVDs by accurately identifying pathophysiological conditions through sensitive detection and determining the most suitable therapeutic approaches (Deb et al. 2015). The site-specific targeted drug delivery proves to be fast and accurate which increases its biological efficacy and bioavailability. By improving drug stability and water solubility, increasing drug uptake and cycle time and reducing degradations by enzymes, nanomaterial-mediated drug delivery improves the safety and efficacy of the therapy. This proves to reduce offtarget effects, systemic effects, and drug resistance (Agrawal et al. 2020). There were encouraging results from the use of silica–gold nanoparticles for atheroprotective management of plaques in a first-in-man trial (the NANOM FIM trial NCT01270139) with three patient groups: (1) nano-intervention with the delivery of silica–gold NP in a bioengineered on-artery patch (n = 60 nano group), or (2) nano-intervention with the delivery of silica–gold iron-bearing NP with targeted micro-bubbles and stem cells using a magnetic navigation system (n = 60 ferro group) versus (3) stent implantation (n = 60 stenting group), which showed a significantly lower risk of cardiovascular death in the first group compared with others (91.7% vs. 81.7% and 80% respectively; p < 0.05) (Kharlamov et al. 2015). The advantages of nanoparticle-mediated theranostics are summarised in Table 2. Nanotheranostics seem to be an effective approach; however, there are challenges in its implementation. The heterogeneous composition of nanoparticles poses difficulties in large-scale production while complying with pharmaceutical GMP guidelines (e.g., sterility, stability, and purity). Even the slightest bit of contamination

Nanomaterial-Mediated Theranostics for Vascular Diseases Table 2 Advantages and disadvantages of nanoparticles in theranostics

181

Advantages

Disadvantages

• Targeted drug delivery • Site-specific • Diagnosis, therapeutic and theranostic applications • Increased permeability • Faster and accurate drug delivery • Lower systemic effects • Reduced degradation by enzymes • Reduced off-target effects • Reduced drug resistance • Increased biological efficacy • Increased bioavailability • Improved drug stability • Improved water solubility • Increased drug uptake • Increased safety and efficacy

• Complex synthesis • Possibilities of contamination • Nanoparticle related toxicity • Therapy cannot be discontinued • Difficulty in evaluating blood distribution • Challenges with biodegradability • Low sensitivity • Expensive

during the manufacture of nanotubes, nanospheres, etc., may lead to extreme adverse effects in-patient. Hence, maintaining aseptic conditions during manufactures is a must. For successful large-scale production, it is of vital importance to employ techniques that are highly reproducible, affordable and time-effective (Ambesh et al. 2017; Flores et al. 2019). The translation of in-laboratory settings to a large-scale clinical production and marketed-oriented approaches poses challenges in costeffectiveness and ethical issues (Nakhlband et al. 2018). Additionally, there is a major gap in the evaluation of safety of these nanomedicines in vivo since some nanoparticles undergoing preclinical development have proved to be cytotoxic or immunogenic (Wolfram et al. 2015). The toxicity is generally due to the chemical makeup of the nanoparticles which directly causes an adverse effect on the cell (Ambesh and Angeli, 2015). One study involving embryonic stem cells of the mouse shows that the frequency of mutation doubles, causing DNA damage, after injecting multi-walled carbon nanotubes (Zhu et al. 2007). More intensive and integrative approaches need to be developed with possible toxicity profiles and other regulatory issues (Tang et al. 2012). The limitations of nanoparticle-mediated theranostic are listed in Table 2.

4 Recent Advancements and Future Scope Strides in advancements have been made in the past few decades in developing nanomaterial-mediated theranostics used in CVS. Gold nanorods impregnated within UV-cross-linkable gelatin methacrylate-based cardiac patches were

182

T. Divanji et al.

patented (US20170143871 A1). This showed improved physicochemical properties such as increased surface area and conductivity. Nanoparticular stem cell conjugates were developed and patented as a treatment for post-infarction period (JP5495215B2). Solid lipid nanoparticles were combined with traditional Chinese medicine and patented (CN103027981B). Additionally, Magnetic nanoparticles (MNP) were also developed and patented for the theranostics of coronary heart disease (CN102085380A) (Fan et al. 2020). MNP-based therapy has proved to produce circumferential re-endothelialisation of blood vessels (Haldar et al. 2016). Current state-of-the-art nanoparticle technologies are being developed wherein instead of giving nanoparticles intravenously, heart-targeted agents like MMP-2 and MMP-9 targeting peptides are administered (Nguyen et al. 2015). Cyclic peptides CSTSMLKAC and CRSWNKADNRSC have also shown to have selective targeting to the ischaemic heart (Fan et al. 2020). These help in improving the efficacy and the side effects profile by increasing the retention of the particles at the infarct site (Nguyen et al. 2015). It has also been demonstrated that the naturally generated atrial natriuretic peptide (ANP) exhibits cardioprotective characteristics via cGMPdependent signalling involving guanylyl cyclase A (GC-A) receptors (Potter et al. 2009).

5 Conclusion To conclude, strides in technology and health care have been made to develop and design one or more ways to diagnose and treat CVDs. Nanomaterial-mediated theranostics being one such venture for the same. This dual-faced modality can have a significant impact on both diagnosis and therapies of CVD in a single platform. Nanotheranostics have revolutionised and shone a light on not only the pathophysiology of CVDs but also the appropriate therapeutic measures. The use of theranostic nanoparticles for the treatment of CVDs has increased significantly in laboratory settings over the past few years. They have exhibited promising results in the development of atherosclerosis, aneurysms, myocardial infarction and peripheral vascular diseases. Nanomedicine can effectively solve the issues involved with targeted drug delivery, drug resistance and bioavailability. However, commercial, cost-effective, large-scale production is a major challenge for these theranostic nanoparticles. Scaling up the synthesis of the nanoparticles is more complicated since they must be optimised for their effectiveness, long-term stability, purity and sterility. Choosing the right chemical structures for the materials, carriers and surface modifiers are all equally related to the efficacy of the nanoparticles. Formulating new drug technologies requires a lot of manpower, material resources and time compared to continuing the existing technologies. So to summarise, nanotheranostics used in CVDs are still in their very early preclinical stages of development. Therefore, nanomaterial-mediated theranostics show a highly promising scope in the future but to translate them into clinical therapies a lot more development needs to be done to deem them fit for human use.

Nanomaterial-Mediated Theranostics for Vascular Diseases

183

References Agrawal S et al (2020) Nanomaterial-mediated theranostics for vascular diseases. J Nanotheranostics 2(1):1–15 Agulla J et al (2014) In vivo theranostics at the peri-infarct region in cerebral ischemia. Theranostics 4(1):90–105 Agyare EK et al (2014) Engineering theranostic nanovehicles capable of targeting cerebrovascular amyloid deposits. J Control Release 185(1):121–129 Ambesh P et al (2017) Nanomedicine in coronary artery disease. Indian Heart J (Elsevier B.V.) 244–251 Ambesh P, Angeli DG (2015) Nanotechnology in neurology: genesis, current status, and future prospects. Ann Indian Acad Neurol (Medknow Publications) 382–386 Bergheanu SC, Bodde MC, Jukema JW (2017) Pathophysiology and treatment of atherosclerosis: current view and future perspective on lipoprotein modification treatment. Neth Hear J 25(4):231–242 Bietenbeck M et al (2016) Remote magnetic targeting of iron oxide nanoparticles for cardiovascular diagnosis and therapeutic drug delivery: where are we now? Int J Nanomed (Dove Medical Press Ltd.) 3191–3203. https://doi.org/10.2147/IJN.S110542 Cuadrado I et al (2016) EMMPRIN-targeted magnetic nanoparticles for in vivo visualization and regression of acute myocardial infarction. Theranostics 6(4):545–557 Deb S et al (2015) Nanoimaging in cardiovascular diseases: current state of the art Deng Y et al (2020) Application of the nano-drug delivery system in treatment of cardiovascular diseases. Front Bioeng Biotechnol (Frontiers Media S.A.) Deveza L et al (2016) Polymer-DNA nanoparticle-induced CXCR4 overexpression improves stem cell engraftment and tissue regeneration in a mouse hindlimb ischemia model. Theranostics 6(8):1176–1189 Duivenvoorden R et al (2014) A statin-loaded reconstituted high-density lipoprotein nanoparticle inhibits atherosclerotic plaque inflammation. Nat Commun 5 Fan C et al (2020) Nanoparticle-mediated drug delivery for treatment of ischemic heart disease. Front Bioeng Biotechnol (Frontiers Media S.A.) Flores AM et al (2019) Nanoparticle therapy for vascular diseases. In: Arteriosclerosis, thrombosis, and vascular biology. NLM (Medline), pp 635–646 Frangogiannis NG (2015) Pathophysiology of myocardial infarction. Compr Physiol 5(4):1841– 1875 Golestani R, Sadeghi MM (2014) Emergence of molecular imaging of aortic aneurysm: implications for risk stratification and management. J Nucl Cardiol 21(2):251–267 Haldar R et al (2016) Contemporary social network sites: relevance in anesthesiology teaching, training, and research. J Anaesthesiol Clin Pharmacol (Medknow Publications) 382–385 Heun Y et al (2017) Targeting of magnetic nanoparticle-coated microbubbles to the vascular wall empowers site-specific lentiviral gene delivery in vivo. Theranostics 7(2):295–307 Jebari-Benslaiman S et al (2022) Pathophysiology of atherosclerosis. Int J Mol Sci (MDPI) Jennewine B, Fox J, Ramamurthi A (2017) Cathepsin K-targeted sub-micron particles for regenerative repair of vascular elastic matrix. Acta Biomater 52:60–73 Jung E et al (2019) Stimulus-activatable echogenic maltodextrin nanoparticles as nanotheranostic agents for peripheral arterial disease. Biomaterials 192:282–291 Kharlamov AN et al (2015) Silica-gold nanoparticles for atheroprotective management of plaques: results of NANOM-FIM trial on behalf of the team of NANOM FIM study Kim J et al (2011) Targeted delivery of nanoparticles to ischemic muscle for imaging and therapeutic angiogenesis. Nano Lett 11(2):694–700 Liu H et al (2016) Label-free CEST MRI detection of citicoline-liposome drug delivery in ischemic stroke. Theranostics 6(10):1588–1600

184

T. Divanji et al.

Lobatto ME et al (2012) Imaging the efficacy of anti-inflammatory liposomes in a rabbit model of atherosclerosis by non-invasive imaging. In: Methods in enzymology. Academic Press Inc., pp 211–228 MacRitchie N et al (2021) Nanoparticle theranostics in cardiovascular inflammation. In: Seminars in immunology. Academic Press Manners N et al (2022) Theranostic nanomedicines for the treatment of cardiovascular and related diseases: current strategies and future perspectives. Pharmaceuticals (MDPI) Matoba T et al (2017) Nanoparticle-mediated drug delivery system for atherosclerotic cardiovascular disease. J Cardiol (Japanese College of Cardiology (Nippon-Sinzobyo-Gakkai)) 206–211 Matuszak J et al (2018) Drug delivery to atherosclerotic plaques using superparamagnetic iron oxide nanoparticles. Int J Nanomed 13:8443–8460 McCarthy JR et al (2010) A light-activated theranostic nanoagent for targeted macrophage ablation in inflammatory atherosclerosis. Small 6(18):2041–2049 Myerson JW et al (2014) Thrombin-inhibiting nanoparticles rapidly constitute versatile and detectable anticlotting surfaces. Nanotechnology 25(39) Nakhlband A et al (2018) Combating atherosclerosis with targeted nanomedicines: recent advances and future prospective. BioImpacts 8(1):59–75 Nguyen MM et al (2015) Enzyme-responsive nanoparticles for targeted accumulation and prolonged retention in heart tissue after myocardial infarction. Adv Mater 27(37):5547–5552 Noukeu LC et al (2018) Nanoparticles for detection and treatment of peripheral arterial disease. Wiley-VCH Verlag, Small de Oliveira Gonçalves K et al (2015) Aminolevulinic acid with gold nanoparticles: a novel theranostic agent for atherosclerosis. Analyst 140(6):1974–1980 Pala R et al (2021) Nanomaterials as novel cardiovascular theranostics. Pharmaceutics (MDPI AG) Peripheral Vascular Disease—StatPearls—NCBI Bookshelf (2022) Potter LR et al (2009) Natriuretic peptides: their structures, receptors, physiologic functions and therapeutic applications. In: Handbook of experimental pharmacology, pp 341–366 Russell P et al (2022) Theranostic nanoparticles for the management of thrombosis. Theranostics (Ivyspring International Publisher) 2773–2800 Sharma M et al (2019) Nanotheranostics, a future remedy of neurological disorders. Expert Opin Drug Deliv 16(2):113–128 Shazeeb MS, Feula G, Bogdanov A (2014) Liposome-encapsulated superoxide dismutase mimetic: theranostic potential of an MR detectable and neuroprotective agent. Contrast Media Mol Imaging 9(3):221–228 Sivaraman B et al (2017) Magnetically-responsive, multifunctional drug delivery nanoparticles for elastic matrix regenerative repair. Acta Biomater 52:171–186 Sivaraman B, Ramamurthi A (2013) Multifunctional nanoparticles for doxycycline delivery towards localized elastic matrix stabilization and regenerative repair. Acta Biomater 9(5):6511–6525 Sontheimer DL (2006) Peripheral vascular disease: diagnosis and treatment Stone JR (2016) Diseases of small and medium-sized blood vessels. In: Cardiovascular pathology, 4th ed, pp 125–168 Talev J, Kanwar JR (2020) Iron oxide nanoparticles as imaging and therapeutic agents for atherosclerosis. Semin Thromb Hemost 46(5):553–562 Tang J et al (2012) Nanomedical theranostics in cardiovascular disease. Current Cardiovasc Imaging Rep (current Medicine Group LLC) 1:19–25 Tokutome M et al (2018) PPARγ-targeting nanomedicine promotes cardiac healing after acute myocardial infarction by skewing monocyte/macrophage polarization in preclinical animal models Tu C et al (2015) Nanoscale strategies: treatment for peripheral vascular disease and critical limb ischemia. ACS Nano 9(4):3436–3452 Wang J et al (2017) Neuronal PirB upregulated in cerebral ischemia acts as an attractive theranostic target for ischemic stroke

Nanomaterial-Mediated Theranostics for Vascular Diseases

185

Wang X et al (2019) Gold nanoparticles that target degraded elastin improve imaging and rupture prediction in an AngII mediated mouse model of abdominal aortic aneurysm. Theranostics 9(14):4156–4167 Wang Y et al (2021) Functionalized polymeric hybrid micelles as an efficient nanotheranostic agent for thrombus imaging and thrombolysis. Acta Biomater 122:278–290 Wolfram J et al (2015) Safety of nanoparticles in medicine HHS public access this mini-review addresses the safety considerations for nanoparticles in medicine. Curr Drug Targets Xue Y et al (2021) Engineered macrophage membrane-enveloped nanomedicine for ameliorating myocardial infarction in a mouse model. Bioeng Transl Med 6(2) Yang A et al (2020) Thrombin-responsive engineered nanoexcavator with full-thickness infiltration capability for pharmaceutical-free deep venous thrombosis theranostics. Biomater Sci 8(16):4545–4558 Ye M et al (2019) SR-A-targeted phase-transition nanoparticles for the detection and treatment of atherosclerotic vulnerable plaques. ACS Appl Mater Interfaces 11(10):9702–9715 Yin L et al (2021) Nanoparticle-assisted diagnosis and treatment for abdominal aortic aneurysm. Front Med (Frontiers Media S.A.) Yu J, Li W, Yu D (2018) Atrial natriuretic peptide modified oleate adenosine prodrug lipid nanocarriers for the treatment of myocardial infarction: in vitro and in vivo evaluation. Drug Des Dev Ther 12:1697–1706 Zhang P et al (2022) Theranostic nanoparticles with disease-specific administration strategies Zhu L et al (2007) DNA damage induced by multiwalled carbon nanotubes in mouse embryonic stem cells. Nano Lett 7(12):3592–3597 Zumla A et al (2016) Host-directed therapies for infectious diseases: current status, recent progress, and future prospects. Lancet Infect Dis (Lancet Publishing Group) e47–e63

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors in Tumor Diagnosis Sudha Srivastava, Namita Sharma, and Manisha Singh

Abstract This chapter describes the early detection of cancer employing nanosensors as a diagnostic tool. The changes in the tumor microenvironment like hypoxia, acidosis, nutrient deprivation, and high interstitial pressure are indicative of a malignant tumor irrespective of the stages. Two distinctive features of a cancerous environment—acidosis and hypoxia can be employed as a biomarker for the identification of radio/chemotherapy-resistant tumors. Here, we present technological advancements and limitations thereof of various nanomaterials/dye-based hypoxia estimation techniques. Further, we describe various fluorescence-based imaging techniques and their sensitivity toward the detection of changes in pH. This is followed by a discussion on the development of various pH-sensitive nanomaterials overcoming drawbacks of existing imaging techniques. This review highlights optical biosensors, utilizing different types of nanomaterials, for the early detection of the tumor. Keywords Acidosis · Hypoxia · Nanosensors · Tumor

1 Introduction Worldwide statistics show alarmingly high incidences of cancer, with approximately 10 million deaths reported in 2020 (Ferlay et al. 2020; de Martel et al. 2020). Research efforts focused towards cancer diagnosis and therapeutics have led to a 29% decrease in the cancer death rate in the last two decades. Cancer cases diagnosed at stage I (early diagnosis) report as high as 90–95% survival rates, while delayed diagnosis is directly proportional to decreasing survival rates. In other words, early diagnosis of cancer plays a decisive role in improving cancer prognosis and survival rates. However, early diagnosis is difficult due to asymptomatic presentation of most of the cancers at the initial stage and/or technological limitations of diagnostic techniques to detect small-sized, early-stage tumors (Ferlay et al. 2020; de Martel et al. 2020). S. Srivastava (B) · N. Sharma · M. Singh Department of Biotechnology, Jaypee Institute of Information Technology, Noida, Uttar Pradesh, India e-mail: [email protected] © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_10

187

188

S. Srivastava et al.

Cancer diagnosis relies on diagnostic tests like histopathological, cytological, and imaging exploiting morphological and (bio)chemical changes in the cells/tissue in addition to biomarker analysis in body fluids (blood, urine, saliva, etc.). Cancer diagnostics have come a long way, but still there are numerous problems/restrictions based on the degree of invasiveness, early detection, and diagnosis accuracy of a given method/tool. Choice of the diagnostic method varies for different cancer types affecting different organs. It can be quite challenging since no two tumors are same even for cancer kinds that attack the same organs owing to variability in genetic makeup, growth rate, capacity for metastasis, malignancy, etc. Diagnostic methods for cancer can be broadly categorized as (a) imaging-based, (b) solid biopsy, and (c) liquid biopsy. Imaging methods, like ultrasound, X-ray-computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET) scans, are noninvasive in nature. These are effective for identification of tumor’s physical characteristics, such as its exact position and size, but they are unable to provide information on the tumor’s development and spread. Recent technological advancements in radiomics have enabled early identification of cancerous tumors and could differentiate between slow-growing and aggressive tumors (Frangioni 2008). However, these techniques suffer from major issues like increased radiation exposure for the patient, are expensive, and frequently provide false-positive results. Solid biopsy involves aspiration of tumor tissue sample that is then sent to a laboratory for physical and (bio)chemical analysis. Biopsy results are generally used for confirmation of cancerous growth that remains inconclusive from imaging-based techniques. The invasive nature of this diagnostic method restricts its use in the diagnosis of high-risk individuals and tracking cancer progression. In addition to this, the amount of sample taken might vary depending on the technique used, handling expertise, and sometimes there aren’t enough tissues available, which might make it difficult to identify and classify cancer (Palmirotta et al. 2018). Liquid biopsy term was coined for the identification of tumor DNA from cell-free samples of body fluids. Broadly speaking, liquid biopsy involves cancer diagnosis based on the identification of cancer biomarkers in body fluids like blood, urine, saliva, and sweat. The major drawback of biomarkers identified for various cancer types is the lack of specificity. Till date, none of the identified biomarkers are unique to a cancer type—either they are up/down-regulated in more than one type of cancer or other disease conditions also and/or are present in healthy individuals as well. Hence, it is always recommended to analyze a panel of biomarkers rather than one single biomarker. However, compared to solid tissue extraction and testing, liquid biopsy is considerably a less intrusive form of cancer diagnosis. Hence, research efforts are on for identification of a panel of biomarkers that would increase the specificity and sensitivity of cancer diagnosis (Lone et al. 2022). Cancer biomarkers, or tumor-specific signature markers produced on the cell surface, can aid in the early diagnosis of cancer, however, major hurdle is posed by the detection limit of the diagnostic technique and specificity of the proposed biomarker (Sarhadi and Armengol 2022).

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors …

189

Recently, hypoxia and acidosis have been identified as both diagnostic as well as prognostic indicators of cancer. Hypoxia is characteristic of most of the solid tumors. This is attributed to the fact that diffusion length scales of oxygen is 100–200 μm only while tumors are of much greater size (>1–2 mm), this limits the supply of oxygen and nutrients needed for solid tumor growth from nearby blood capillaries (Godet et al. 2022). This, in turn, promotes unregulated neovascularization (formation of new blood vessels) directed towards the tumor (Vaupel and Harrison 2004) with decreased oxygen pressure that is directly proportional to the distance from blood arteries. In some of the solid tumors, oxygen pressures can be as low as 0 mmHg pO2 as opposed to greater than 30 mmHg pO2 in healthy tissues. Breast tumors show partial pressure of oxygen (pO2 ) from 2.5 to 28 mmHg, with an average value of 10 mmHg (~1% O2 ), while normal human breast tissue has a value of 65 mmHg (8% O2 ). Oxygen percentage in different healthy tissues and the corresponding tumor are shown in Table 1. As observed, oxygen percentage in healthy tissues ranges from 3.9 to 9.5%, while cancerous tissues are clearly demarcated by a much lower range of 0.3–1.8% in different tissue types. In addition to this, decreased oxygen percentage or increased hypoxia further leads to an increase in reductive stress, and hence elevated levels of reductive enzymes are observed in cancer (Mathejczyk et al. 2012; McNeel et al. 2019; Fan et al. 2017). Irrespective of the stage of the tumor or the tumor type, solid tumors typically have a slightly acidic pH as compared to healthy tissue (Zhang et al. 2010). As shown in Table 1, the pH decreases in cancerous tissue as compared to healthy counterparts with the exception of breast cancer where an increase in pH in cancerous tissue has been reported. pH changes by approximately 0.5–1.0 pH unit in cancer tissue as compared to normal tissue. Though the number looks smaller, the change in hydrogen ion concentration is much higher (tenfold), rendering it significant for other biochemical processes/changes (Hao et al. 2018; Lee and Shanti 2021). Conventional imaging techniques, such as ultrasound, single-photon emission tomography (SPECT), CT, PET (Mirabello et al. 2018), MRI, electron paramagnetic resonance imaging (EPRI), surface-enhanced Raman (SER) spectroscopy, magnetic particle-based imaging, optical imaging, and mass spectrometry-based imaging, can be used to detect pH and hypoxic conditions of a tumor (Chen and Pagel 2015; Subasinghe et al. 2022). However, these techniques are qualitative in nature. Electrochemical probes (Liu et al. 2017a, b), EPR microdialysis, and MRI can be used to monitor tissue oxygen levels in real time (Marland et al. 2020; Wisniewski et al. 2017), but are time-consuming and require big expensive equipment. Optical imaging seems to be the most suitable technique that offers simplicity at the same time has high sensitivity for quantification of hypoxia and acidosis (Liu et al. 2017a, b). However, the major problem encountered by imaging techniques due to background signal from auto-fluorescence, varying tumor depths, and decreased phosphorescence/fluorescence intensity by tissue absorption or quenching of signal by nonspecific binding (photobleaching) needs to be addressed. Fluorescent nanosensors can overcome the above issues by using nanomaterials as reference dye to take care of the background signal. Furthermore, these nanosensors could also take care of varying tumor depth by choosing nanocomplexes having

190

S. Srivastava et al.

Table 1 Oxygen percentage and pH levels in normal and cancerous tissue for various cancer tissue Cancer

Oxygen percentage in cancer/normal tissue

pH range in cancer/normal tissue

Reference(s)

Brain tumor

1.7/4.6

6.4–7.0/7.2–7.5

Muz et al. (2015), Hao et al. (2018)

Breast cancer

1.5/8.5

7.4/7.2

Muz et al. (2015), Lee et al. (2021)

Cervical cancer

1.2/5.5

4.6–5.0/3.8–4.5

Muz et al. (2015), Teng and Hao (2020)

Renal cancer

1.3/9.5

/6.3–7.0

Muz et al. (2015), Minhas et al. (2020)

Liver cancer

0.8/4.0–7.3

6.6–6.8/7.3–7.4

Muz et al. (2015), Tang et al. (2020)

Non-small-cell lung cancer

2.2/5.6

6.0–6.5/7.4

Muz et al. (2015), Pang et al. (2020)

Pancreatic tumor

0.3/7.5

6.5–6.9/7.2–7.4

Muz et al. (2015), Kimbrough et al. (2015)

Rectal carcinoma

1.8/3.9

8.02/7.2–12.1

Muz et al. (2015), Newmark and Lupton (1990), Hua (2019)

fluorescence with improved penetration depth. The next section describes different types of nanomaterials and fluorescent dyes employed for fabrication of fluorescent nanoprobes for quantitative estimation of hypoxia (Shamsipur et al. 2019), acidosis (Tian et al. 2019), and hypoxia-acidosis for cancer diagnosis (Chen et al. 2021a, b) and the technological development thereof.

2 Hypoxia Responsive Fluorescent Nanosensors for Cancer Diagnosis Schematic representation of hypoxia-responsive fluorescent nanosensor fabrication is shown in Fig. 1. Hypoxia nanosensor comprises of (1) a recognition molecule (MO) that undergoes chemical/structural changes in response to hypoxic environment leading to increased fluorescence and (2) a nanomaterial (NM), that may or may not be fluorescent molecule, that acts as a carrier of hypoxia-sensitive recognition molecule (MO-NM nanoconjugate) for targeted delivery/distribution to the tumor site(s). Recognition molecules used for hypoxia-based sensors are either sensitive to oxygen or hypoxia-sensitive enzymes like nitroreductase (Luo et al. 2017), quinone reductase, azoreductase, and Hypoxia-Inducible Factor (HIF)-1 protein. Nitroreductase enzyme under hypoxic conditions reduces aromatic nitro compounds (like

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors …

191

Fig. 1 Schematic representation of hypoxia-responsive fluorescent nanosensor using nanomaterial and oxygen sensing moiety

nitroimidazole derivative, nitrobenzene, and nitrofuran), to primary amine. These aromatic nitro compounds are well known as fluorescence quenching agents owing to their electron-accepting nature. The reduced product being primary amine is an electron donor, hence results in increased fluorescence. Similarly, azoreductase reduces azo compounds into primary amines and quinone reductase reduces quinone to hydroquinone (Gebremedhin et al. 2019). Table 2 shows a list of hypoxia-sensitive (Zheng et al. 2015) fluorescent nanosensors. Nanomaterials used for the development of fluorescent nanosensors are either (1) Quantum dots (QD) having intrinsic fluorescence characteristics such as CdSe semiconductor nanocrystals, or (2) Upconversion nanomaterials (UCNPs) which can convert near-infra-red (NIR) excitation into visible or ultraviolet light emission, or (3) Non-fluorescent nanomaterials like gold nanoparticles (AuNPs), that have capability to quench the fluorescence of conjugated compounds, or (4) Nanomaterials that are neither fluorescent nor quenchers but are used as carriers of fluorescent molecules, for example, polymeric nanoparticles, meso/nanoporous silica, etc. (Smith and Gambhir 2017). Liu and coworkers exploited stronger fluorescence emission, less photobleaching, broad absorption band, and narrow emission peak characteristics of QDs, to develop a QD and Iridium complex-based luminous hypoxia-sensing nanoprobe (Liu et al. 2017a, b). They developed glycerol monoolein nanoparticles (GMNPs) having oil/water channel walls and used them as a vehicle for segregated loading of QDs and hypoxia-sensitive dye. This design of GMNPs took care of two issues—Firstly, provide an optimum stablizing microenvironment for QDs which are very reactive, highly unstable, and susceptible to small fluctuation in salt, pH, temperature, etc., that eventually leads to agglomeration of QDs and quenching of the fluorescence signal. Secondly, the problem of florescence resonance energy transfer (FRET)

542 nm

Gold nanoparticles with β-cyclodextrins (βCD-AuNPs)

480–440 nm/520

800 nm

–

pRF-GQDs (pH-responsive fluorescent graphene quantum dots)

Mn2+ -doped CaP nanoparticles with a poly(ethylene glycol) (PEG) shell (PEGMnCaP) –

575 ± 15 nm)

613 nm

Mechanism

Fluorescence

pH

Mice

Fluorescence

pH

(continued)

Mi et al. (2016)

Fan et al. (2017)

McNeel et al. (2019)

Oxygen Li et al. (2020)

Oxygen Zhu et al. (2019)

Oxygen Liu et al. (2014)

Sensing Reference analyte

Tumor xenografts and Upconversion pH cells derived from photoluminescence HeLa, HepG2, PANC-1 (UCPL) cancer, A549 and U87MG cancer cells

Normal human breast Fluorescence fibroblast, human breast carcinoma

HeLa cell lines, MDA-MB-231 cell lines and NSCLC, CH27 cell lines, zebrafish

Fluorescence

U87MG cells, Zebrafish Luminescence resonance energy transfer (LRET), confocal laser scanning

Test model

560–20 nm (TAMRA-red) HepG2 cells and 625–675 nm (Cy5-green)

400–500 nm/550–650 nm

Emission

NanoGUMBOS (fluorescein, FL, rhodamine 535 ± 15 nm B, RhB, and tetradecyltrihexyl phosphonium ions, P66614)

pH nanosensors

bio-MOF-100-[Ru(dpp)3 ]2+ Cl2 nanocrystals 980 nm (BMU-Ru nanosensors)

980 nm

Excitation

Upconversion nanoparticles (UCNP@hmSiO2 ) UCNPs: NaYF4 :Yb/Tm@NaYF4

Hypoxia nanosensors

Nanoparticles

Table 2 Fluorescent-based hypoxic, pH and hypoxia and pH nanobiosensors

192 S. Srivastava et al.

–

455 nm, 650 nm

690–770 nm and 820–880 nm, 692 nm

–

Extremely small iron oxide nanoparticles (ESIONs)

Fe3 O4 nanoparticles (Cy5.5, ANNA, and Folic Acid with Fe3 O4 as nanoprobe)

Ultrathin Mn-oxides [MnOx] nanosheet–semiconducting polymer nanoparticles [SPNs]

Superparamagnetic iron oxide (SPIO) particles

AuNR@mSiO2 loaded with Rho-TP and azobenzene

–

–

–

–

–

Emission Fluorescence

Mechanism

Female athymic nude mice

Mouse breast cancer 4T1 cells

Mice

Fluorescence

Fluorescence

Fluorescence

Fluorescence

CT26-tumor-bearing Fluorescence mice and homogeneous HCT116-tumor-bearing mice

Mice

Test model

580 nm/559 nm/580 nm 600–700 nm MCF-7 cells, mice /570–650 nm/620–700 nm

–

Superparamagnetic iron oxide nanoparticles (SPIONs)

Hypoxia and pH nanosensors

Excitation

Nanoparticles

Table 2 (continued)

pH and oxygen

pH

pH

pH

pH

pH

Chen et al. (2021a, b)

Wang et al. (2015)

Lu et al. (2020)

Ma et al. (2018)

Ling et al. (2014)

Yang et al. (2019)

Sensing Reference analyte

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors … 193

194

S. Srivastava et al.

Fig. 2 Quantum dots and Iridium complex as fluorescent nanosensor showing fluorescence response with increasing hypoxic environment in a cells, b mouse with tumor, c emission spectra of the nanosensor, and d the ratiometric calibration plot. Reprinted (adapted) with permission from {Liu, J., Wu, Y., Yu, Y., Li, K., Ji, Y., & Wu, D. (2017). Quantitative ratiometric phosphorescence hypoxia-sensing nanoprobes based on quantum dots/Ir (III) glycerol monoolein cubic-phase nanoparticles. Biosensors and Bioelectronics, 98:119–125} Copyright {2017}

(Chen et al. 2013) when QDs and hypoxia-responsive dye are conjugated directly or loaded onto same vehicle leading to poor signal/sensitivity of nanosensor. The QDs are used as reference dye, while Iridium acted as the hypoxia-sensitive element. It was observed that the intensity of QD emission displayed no change in hypoxic and normoxic conditions, while Iridium emission intensity exhibited greater than a fourfold increase under hypoxic conditions (Fig. 2a–b). The nanosensor emission spectra (Fig. 2c) and corresponding calibration plot displayed a linear correlation between the ratio of emission intensity of iridium and QDs and oxygen concentration (Fig. 2d). The nanosensor was evaluated under in vitro and in vivo conditions and exhibited linear calibration plot for ratiometric estimation/imaging of hypoxic environment (Liu et al. 2017a, b). Li and coworkers (Liu et al. 2020) developed the fluorescent nanosensor activated by near-infrared (NIR) light to minimize the detrimental effect of long UV or visible light exposure, at the same time exploited the advantage of increased penetration depth of tissue, less harmful, and excellent spatial resolution of NIRs. Nanosensor was built on Lanthanide-doped up conversion-based nanoparticles (UCNPs) to detect the hypoxic condition in vitro and in vivo. UCNPs based nanosensor comprised of a UCNP core/hollow mesoporous silica shell-structured nanoparticles for energy donation for luminescence resonance energy transfer (LRET) process and oxygen indicator tris (4, 7-diphenyl-1,10-phenanthroline) ruthenium (II) dichloride as [Ru(dpp)3 ]2+ Cl2 for detection of hypoxia, since UCNPs themselves are insensitive to variation in oxygen concentration. Ru(dpp)3 ]2+ Cl2 indicator was employed owing to its quenching capability—having absorption maxima 463 nm that overlaps with the emission peaks of UCNPs at 450 and 475 nm. Fluorescence response of the developed nanoprobe displays quenching of the red emission under normoxic circumstances while a high-intensity emission was regained under oxygen-deficient or hypoxic conditions (Liu et al. 2014). The LRET efficiency was found to be around 90.1%. Advantages of using this nanosensor lie in its high selectivity towards change

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors …

195

Fig. 3 Hypoxia nanosensors showing change in luminescence under hypoxic conditions and corresponding ratiometric calibration plot as a function of dissolved oxygen. Reprinted (adapted) with permission from {Liu, J., Liu, Y., Bu, W., Bu, J., Sun, Y., Du, J., & Shi, J. (2014). Ultrasensitive nanosensors based on upconversion nanoparticles for selective hypoxia imaging in vivo upon nearinfrared excitation. Journal of the American Chemical Society, 136(27), 9701–9709} Copyright {2014} American Chemical Society

in oxygen and its unchanged behavior towards acidic microenvironment in cancer cells. Furthermore, the high penetration depth of the nanosensor makes it suitable for in vivo sensing of tumor and its progression. As shown in Fig. 3, the calibration plot of ratiometric emission shows an exponential increase with increasing hypoxia or decreasing dissolved oxygen concentration (Zhang et al. 2020). Though UCNPs have high penetration depth, the sensitivity of NIR-excited nanosensors is not very good. Li and coworkers developed biological metal–organic framework-based nanomaterials (Bio-MOF) having anionic cavities suitable for improving the sensitivity of fluorescent dyes (Li et al. 2020). Controlled anionic core atmosphere avoids aggregation as well as steric hindrance of [Ru(dpp)3 ]2+ Cl2 dye in the Bio-MOFs pores. A core/satellite nanostructure was formed by loading the oxygen-sensitive dye in mesoporous Bio-MOFs and, on its surface, core–shell UCNPs (NaYF4 :Yb,Tm@NaYF4 ) were attached. The multiple UCNPs as NIR antenna (donor) in close proximity of fluorescent dye [Ru(dpp)3 ]2+ Cl2 (acceptor) under controlled anionic core atmosphere lead to high FRET efficiency.

3 pH-Responsive Fluorescent Nanosensors for Cancer Diagnosis As discussed in Sect. 1, during tumor growth the extracellular pH changes. The flourescent nanomaterial-based pH sensing can be used not only for extracellular pH monitoring but also for intracellular pH as well since the membrane porosity of

196

S. Srivastava et al.

cancerous cells is high and nanomaterials can enter the cells (Brasuel et al. 2001; Chen et al. 2015, 2016). Furthermore, the nanomaterials should not agglomerate under in vivo conditions and their fluorescence should remain unaffected by other intra/extracellular environmental factors like salts, etc. Major hurdle faced by pH nanosensors is the variation in fluorescent intensity due to difference in path length because of different positions of the cells/tissue or varied tumor size (Shaked et al. 2009). This leads to inaccuracy in pH estimation and demands a reference sensor to take care of calibration issues and background fluorescence. In 1993, to overcome the problems associated with single fluorescence intensity-based sensors, Agayan and Walt (1993) developed a ratiometric fluorescent sensor, using a pH-sensitive fluorophore (acryloylflourescien) immobilized on polyhydroxyethyl methacrylate. Acryloylflourescien on excitation with a wavelength of 530 nm emits two fluorescence peaks at 430 and 485 nm, ratio of these peaks correlates well with the change in pH. However, the lifetime of these sensors is reduced due to extensive photobleaching. Offenbacher and coworkers (Offenbacher et al. 1986) observed no photobleaching in fluorescent sensors developed using 7-hydroxycoumarin-3 carboxylic acid (HCC that is insensitive to oxygen fluctuations) and 1-hydroxypyren-3,6,8-trisulphonate (HPTS that displayed 0.02% variation with oxygen). In addition to this, HPTS fluorophore emits two peaks—(1) for acid at 405 nm and (2) for conjugate base at 457 nm, and ratio of the peaks takes care of the calibration problem. Furthermore, quenchers present in blood like chloride, bromide, iodide, phosphate, sulfate, and salicylate exhibit none or negligible interference. Hence, ratiometric fluorescent sensor using HPTS seems to be an apt candidate for in vivo measurements. However, the orientation of fluorophores at the tip of the sensor affects the fluorescence intensity. Wang and coworkers (Wang et al. 2013) addressed calibration issue by taking two different fluorophores immobilized on core–shell silica nanoparticles with fluorescein (FL), the pH-sensitive probe on the surface of silica nanoparticles, while pH-insensitive fluorophore that served as reference (5,10,15,20- tetrakis(pentafluorophenyl)porphyrin (TFPP)) was encapsulated in the core. McNeel and coworkers (McNeel et al. 2019) synthesized nanomaterial using FL, Rhodamine B (RhB), and tetradecyltrihexyl phosphonium (P66614) to develop a cost-effective and highly sensitive pH nanosensor. Among the three ions, FL absorbance shows pH sensitivity while RhB is known for its insensitivity towards change in pH. On formation of the nanomaterial P66614-RhB-FL, the emission peak intensity of FL decreased, while that of RhB increased as a result of FRET due to the closer proximity of the two. After incubating with nano P66614-RhB-FL, cancer cells fluoresced brightly throughout the cell except the nucleus, while normal cells had scattered fluorescence that was significantly less fluorescence distribution than the cancer cells. FL dye fluoresced in both normal and cancer cells and RhB also demonstrated the nuclear fluorescence after incubation in normal and cancer cells. With the exception of pH 6, two bands of FL and RhB exhibited improved values with increasing pH due to increased absorbance in more anionic forms of FL and increased FRET. Due to the ease and speed required for visual diagnostic analyses, P66614-RhB-FL nanomaterial could be useful for imaging cancer cells (McNeel et al. 2019).

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors …

197

A research team led by Fan (Fan et al. 2017) developed pH-responsive fluorescent graphene quantum dots (pRFGQDs) by electrolysis of graphite in sodium p-toluenesulfonate acetonitrile solution. The synthesized pRFGQDs on excitation with 800 nm wavelength, display an emission peak at 520 nm. These graphene quantum dots have stable photoluminescence, good biocompatibility, and photobleaching resistance as compared to other traditional imaging materials. They created a new class of pRF-GQDs with a sharp fluorescence transformation at pH 6.8 that can detect solid tumors of various origins at an early stage (see Fig. 4). Tumor detection was accomplished using five distinct xenografts derived from five different cancer cells. Since there is a sharp variation in fluorescence emission, pRF-GQDs can be used to distinguish tumors from normal tissue. Choi and Co-workers (Choi et al. 2018) exploited fluorescent nanosensors for pH sensing as well as targeted drug delivery of drugs having poor solubility or high toxicity, e.g. Paclitaxel. developed pH and redox-active fluorescent nanoparticles (FNPs) for targeted delivery of Paclitaxel drug. To create smart nanoparticles for multi-stimuli pH response sensors, two FNPs were synthesized and crosslinked along with MnO2 . (1) Polydopamine was covalently conjugated with hyaluronic acid followed by acid treatment to yield FNP(HA-D), and (2) Polyethylene glycolg–poly(dimethylamino)ethyl methacrylate was treated with acid to get FNP(B-PgD). Both FNP(HA-D) and FNP(B-PgD) crosslinked with each other to form carbonized fluorescent nanoparticle C-FNP. Self-assembled MnO2 nanosheets could very well self-assemble into the C-FNP core(see Fig. 5). MnO2 nanosheets quench the fluorescence emission of C-FNPs (fluorescence “switch off” state). The MnO2 nanosheets are redox-sensitive hence, in presence of glutathione Mn2+ ions are generated resulting in a “switch on” state of the fluorescence. In another study by Mi and coworkers (Mi et al. 2016), MnO2 was encapsulated in calcium phosphate nanoparticles to develop pH-responsive nanosensor to be used

Fig. 4 Diagrammatic representation of pH-responsive fluorescent GQDs (pRFGQDs) a pH < 6.8, pH > 6.8, b tumor imaging. Reprinted (adapted) with permission from Fan Z, Zhou, S., Garcia, C., Fan, L., & Zhou, J. (2017). pH-Responsive fluorescent graphene quantum dots for fluorescenceguided cancer surgery and diagnosis. Nanoscale, 9:4928–4933. Copyright {2017}

198

S. Srivastava et al.

Fig. 5 Schematic representation of MnO2 /PTX-loaded C-FNP. Reprinted (adapted) with permission from Choi, C. A., Lee, J. E., Mazrad, Z. A. I., In, I., Jeong, J. H., & Park, S. Y. (2018). Redoxand pH-responsive fluorescent carbon nanoparticles–MnO2 -based FRET system for tumor-targeted drug delivery in vivo and in vitro. Journal of Industrial and Engineering Chemistry, 63, 208–219. Copyright {2018}

in MRI imaging. The release of Mn2+ in acidic pH conditions increased the contrast of cancerous tissues by increasing the relaxivity. Ling et al. (2014) created pH-responsive magnetic nanogrenades (PMNs) consisting of iron oxide magnetic nanoparticles and pH-sensitive ligands. They synthesized iron oxide nanoparticles in the size range of 3 nm. Ligand poly(ethylene glycol)–poly(β-benzyl-L-aspartate) (PEG–PBLA) was modified with Chlorin e6 (Ce6) to facilitate the binding with primary amines. Finally, imidazole group was attached to the Ce6-modified ligand that acted as a pH-sensitive element since it is easily ionizable and has pKa 6.8 (compatible with TME) (see Fig. 6). Magnetic nanoparticles and ligands form a core–shell micellar structure in water. These PMNs when come in contact with tumor cells (pH < 6.8), the charge on surface of the PMNs becomes positive facilitating easy internalization by the cells. Once inside, the pH further lowers and the PMNs break down and the relaxation time (R2) decreases as compared to intact PMNs. This leads to increased relaxivity and hence better contrast is observed in Magnetic resonance imaging. Wang et al. (2015) showed the possibility of using hybrid nanogels for magnetic resonance contrast. SGM (SPIO@GCS/acryl/biotin@Mn-gel) and there clusters were made by combining superparamagnetic iron oxide particles with polysaccharide nanomaterial. The dual-mode SGM’s in vitro MRI results revealed an intriguing pH responsiveness, with both T1 and T2 relaxivities turning “ON” in acidic pH conditions. On similar lines, Yanga and his colleagues (Yanga et al. 2019) developed a tumor-responsive magnetic nanobomb (HTAMN), for diagnostic testing and photodynamic therapy. The HTAMN was created through the self-assembly of a chlorin

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors …

199

Fig. 6 pH-sensitive magnetic nanogrenades (PMNs) fluorescence. Ling, D., Park, W., Park, S. J., Lu, Y., Kim, K. S., Hackett, M. J., Hyeon, T. (2014). Reprinted (adapted) with permission from Multifunctional tumor pH-sensitive self-assembled nanoparticles for bimodal imaging and treatment of resistant heterogeneous tumors. Journal of the American Chemical Society, 136:5647–5655. Copyright {2014}

e6 (Ce6)-functionalized polypeptide ligand, methoxy poly (ethyleneglycol)-blockpoly dopamine-ethylenediamine-2,3-dimethylmaleic anhydride)-L-glutamate-Ce6 [mPEG-b-P (Dopa-Ethy-DMMA)LG-Ce6] and superparamagnetic ion oxide nanoparticles (SPIONs) (as shown in Fig. 7a). At the tumor location, under acidic tumor environment (pH < 6.8), the surface charge of the negatively charged HTAMNs is reversed from slightly negative to slightly positive. This resulted in tumor deposition and cellular uptake of this complex. Once inside the cell, HTAMs surface charge becomes more positive at pH 5.0 and it disintegrates, thereby generating a high fluorescence signal inside the cancerous cell (see Fig. 7b). It was observed that, on excitation with 600 nm, the absorption peak of free Clorin e6 at 410 nm displayed a faster decaying behavior as compared to HTANs peak, suggesting better photostability of HTANs. Both in vitro and in vivo findings show that HTAMNs have effective tumor accumulation, internalization, diagnostic sensitivity, and superior photodynamic therapy effect. It has been reported that cancer invasion, progression, and metastasis are strongly associated with dysregulated expression levels of proteases like MMP-9, along with decreased extracellular pH. In 2018 (Ma et al. 2018), a novel dual-ratiometric fluorescent probe for mapping tumor marker MMP-9 activity along with pH sensitivity. The pH-sensitive dye ANNA (N-carboxyhexyl derivative of 3-amino-1,2,4-triazole fused 1,8-naphthalimide) was linked with Fe3 O4 nanoparticles via a peptide linker. The covalent linkage of dye with the nanoparticles led to quenching of the fluorescence intensity of dye (see Fig. 8). The nanoconjugate of Fe3 O4 -ANNA gets cleaved when exposed to tumor microenvironment containing MMP-9 thereby restoring the fluorescence intensity of free ANNA dye.

200

S. Srivastava et al.

Fig. 7 a Synthesis of hierarchical tumor acidity-responsive magnetic nanobomb (HTAMN) and b HTAMNs pH response under different acidic conditions. Reprinted (adapted) with permission from Yang, H. Y., Jang, M. S., Li, Y., Fu, Y., Lee, J. H. Lee, D. S. (2019). Hierarchical tumor acidityresponsive self-assembled magnetic nanotheranostics for bimodal bioimaging and photodynamic therapy. Journal of Controlled Release, 301, 157–165. Copyright {2019}

4 Dual (Acidosis–Hypoxia)-Mode Fluorescent Nanosensors for Cancer Diagnosis As discussed in previous sections many hypoxia-based nanosensors displayed poor sensitivities due to photobleaching or nonspecific adsorption by blood/cellular components or change in pH. The same limitations were reported for pH-based nanosensor showing calibration issues or non-reproducible results due to fluctuations in oxygen levels. Hence, there is a need to develop nanosensor for the diagnosis of tumor employing both hypoxic and pH sensing. To detect and measure oxygen and pH levels inside the cells simultaneously, Wang and his team (Wang et al. 2012), developed a dual fluorescent nanosensor.

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors …

201

Fig. 8 a Formation of fluorescent nanoprobe consists of Cy5.5, FA, ANNA, and Fe3 O4 nanoparticle and b Process of quenching ANNA, ratiometric pH dye under acidic conditions. Reprinted (adapted) with permission from Ma, T., Hou, Y., Zeng, J., Liu, C., Zhang, P., Jing, L, Gao, M. (2018). Dualratiometric target-triggered fluorescent probe for simultaneous quantitative visualization of tumor microenvironment protease activity and pH in vivo. Journal of the American Chemical Society, 140: 211–218. Copyright {2018}

Sensing of pH and oxygen at the same site was achieved by using oxygensensitive luminescent probe and reference dye comprised of platinum(II) mesotetraphenyl tetrabenzoporphyrin (PtTPTBP) and the (inert) reference dye 5,10,15,20tetrakis(pentafluorophenyl) porphyrin (TFPP). Pegylated Pluronic F-127 polymer was tagged with fluorescent dye fluorescein on both the ends. Modified Pluronic F-127 as well as both active and inert hypoxic dye PtTPTBP and TFPP were mixed with silica precursor solution during silica nanoparticle synthesis. This resulted in formation of core–shell silica nanoparticles with oxygen-sensitive dye in the core and pH-sensitive dye on the surface. Response towards oxygen was monitored through quenching of probe PtTPTBP by oxygen. The green fluorescence was obtained in response to pH. The ratio of the intensity in oxygen-saturated and oxygen-free solution was reported to be as high as 10.6 (Wang et al. 2012).

202

S. Srivastava et al.

Fig. 9 Response of silica-coated gold nanorods under hypoxic and acidic pH conditions

Recently, Chen and colleagues (Chen et al. 2021a, b) developed another nanobiosenor, employing mesoporous silica-coated gold nanorods (AuNR@mSiO2 ). The presence of azobenzene/β-cyclodextrin polymer and Rho-TP led to dual mode sensing of pH and oxygen. The double-mode responsive fluorescent nanosensor comprised of a nanocarrier-cum-nanoquencher, AuNR@mSiO2 loaded with the pHsensitive fluorescent reporter (Rho-TP). Under hypoxic conditions, Azo/ß-CDP was considered as first lock opened in the presence of highly expressed “azoreductase” and reduced to amines, while Rho-TP as the second lock, opened in the presence of acidic pH (Chen et al. 2021a, b; Zhu et al. 2019) leading to reestablishment of fluorescence signal (Fig. 9).

5 Conclusion Diagnosis of tumor or cancer is still a big challenge worldwide. Developing improved technologies could play a crucial role in detecting tumor in early stages to increase survival rates. Generally, protein, DNA, RNA, etc., are used as tumor markers, however, their detectable levels in blood or other bodily fluids appear at a much later stage of cancer. Tumor microenvironment can be alternatively considered as an important primary source of diagnosis at early stage. Two reported parameters for sensing the tumor at early stage are hypoxia and pH level inside and outside cancer/tumor cells. This chapter highlights different fluorescent nanosensors developed for diagnosis of tumor. Sensing and imaging of the tumor cells/tissue using

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors …

203

nano-based fluoroprobes can be done in vitro as well as under in vivo conditions. The choice of nanomaterial as well as the dye should be based on their biocompatible characteristics and toxicity, for in vivo sensing applications. We have also discussed different types of nanomaterials used, i.e.—intrinsically fluorescent, quenchers, upconversion nanomaterials, or simply nanocarrieers of fluorescent molecules. The limitations of fluorescent nanosensors such as concentration dependence, probe labeling, requirement of other conjugates or molecules, stability of fluorescence emission and its quenching ability, background fluorescence, variable depth of tumors, and photobleaching have been discussed and design of nanosensors to take care of these limitations for pH and hypoxia sensing. Finally, we have discussed nanosensor design that can detect both the hypoxia as well as pH without any calibration hassles in tumor cells/tissue.

References Agayn VI, Walt DR (1993) Fiber-optic sensor for continuous monitoring of fermentation pH. Biotechnology 72:6–9 Brasuel M, Kopelman R, Miller TJ, Tjalkens R, Philbert MA (2001) Fluorescent nanosensors for intracellular chemical analysis: decyl methacrylate liquid polymer matrix and ion-exchangebased potassium PEBBLE sensors with real-time application to viable rat C6 glioma cells. Anal Chem 73:2221–2228 Chen S, Chen M, Yang J, Zeng X, Zhou Y, Yang S, Yang R, Yuan Q, Zheng J (2021a) Design and engineering of hypoxia and acidic pH dual-stimuli-responsive intelligent fluorescent nanoprobe for precise tumor imaging. Small 17:1–11 Chen G, Song F, Xiong X, Peng X (2013) Fluorescent nanosensors based on fluorescence resonance energy transfer (FRET). Ind Eng Chem Res 52:11228–11245 Chen LQ, Pagel MD (2015) Evaluating pH in the extracellular tumor microenvironment using CEST MRI and other imaging methods. In: Advances in radiology Chen Q, Liu X, Chen J, Zeng J, Cheng Z, Liu Z (2015) A self-assembled albumin-based nanoprobe for in vivo ratiometric photoacoustic pH imaging. Adv Mater 27:6820–6827 Chen Q, Liu X, Zeng J, Cheng Z, Liu Z (2016) Albumin-NIR dye self-assembled nanoparticles for photoacoustic pH imaging and pH-responsive photothermal therapy effective for large tumors. Biomaterials 98:23–30 Chen S, Chen M, Yang J, Zeng X, Zhou Y, Yang S, Zheng J (2021b) Design and engineering of hypoxia and acidic pH dual-stimuli-responsive intelligent fluorescent nanoprobe for precise tumor imaging. Small 17:2100243 Choi CA, Lee JE, Mazrad ZAI, In I, Jeong JH, Park SY (2018) Redox-and pH-responsive fluorescent carbon nanoparticles–MnO2-based FRET system for tumor-targeted drug delivery in vivo and in vitro. J Ind Eng Chem 63:208–219 de Martel C, Georges D, Bray F, Ferlay J, Clifford GM (2020) Global burden of cancer attributable to infections in 2018: a worldwide incidence analysis. Lancet Glob Health 8(2):e180–e190 Fan Z, Zhou S, Garcia C, Fan L, Zhou J (2017) pH-Responsive fluorescent graphene quantum dots for fluorescence-guided cancer surgery and diagnosis. Nanoscale 9:4928–4933 Ferlay J, Ervik M, Lam F, Colombet M, Mery L, Pineros M et al (2020) Global cancer observatory: cancer today. Lyon: International Agency for Research on Cancer (https://gco.iarc.fr/today. Accessed February 2021) Frangioni JV (2008) New technologies for human cancer imaging. J Clin Oncol Am Soc Clin Oncol 2624:4012–4021

204

S. Srivastava et al.

Gebremedhin KH, Li Y, Yao Q, Xiao M, Gao F, Fan J, Peng X (2019) Development of a red-light emission hypoxia-sensitive two-photon fluorescent probe for in vivo nitroreductase imaging. J Mater Chem B 7:408–414 Godet I, Doctorman S, Wu F, Gilkes DM (2022) Detection of hypoxia in cancer models: significance, challenges, and advances. Cells 11:686 Hao G, Xu ZP, Li L (2018) Manipulating extracellular tumour pH: an effective target for cancer therapy. RSC Adv 8:22182–22192 Hua S (2019) Physiological and pharmaceutical considerations for rectal drug formulations. Front Pharmacol 10:1196 Kimbrough CW, Khanal A, Zeiderman M, Khanal BR, Burton NC, McMasters KM, Vickers SM, Grizzle WE, McNally LR (2015) Targeting acidity in pancreatic adenocarcinoma: multispectral optoacoustic tomography detects pH-low insertion peptide probes in vivo. Clin Cancer Res off J Am Assoc Cancer Res 21:4576–4585 Lee S, Shanti A (2021) Effect of exogenous pH on cell growth of breast cancer cells. Int J Mol Sci 22:9910 Ling D, Park W, Park SJ, Lu Y, Kim KS, Hackett MJ, Kim BH, Yim H, Jeon YS, Na K, Hyeon T (2014) Multifunctional tumor pH-sensitive self-assembled nanoparticles for bimodal imaging and treatment of resistant heterogeneous tumors. J Am Chem Soc 136:5647–5655 Liu J, Liu Y, Bu W, Bu J, Sun Y, Du J, Shi J (2014) Ultrasensitive nanosensors based on upconversion nanoparticles for selective hypoxia imaging in vivo upon near-infrared excitation. J Am Chem Soc 136:9701–9709 Liu J, Wu Y, Yu Y, Li K, Ji Y, Wu D (2017a) Quantitative ratiometric phosphorescence hypoxia–sensing nanoprobes based on quantum dots/Ir (III) glycerol monoolein cubic-phase nanoparticles. Biosens Bioelectron 98:119–125 Liu L, Zhao F, Liu W, Zhu T, Zhang JZH, Chen C, Dai Z, Peng H, Huang JL, Hu Q, Bu W, Tian Y (2017b) An electrochemical biosensor with dual signal outputs: toward simultaneous quantification of pH and O2 in the brain upon ischemia and in a tumor during cancer starvation therapy. Angew Chem 56:10471–10475 Li Y, Liu J, Wang Z, Jin J, Liu Y, Chen C, Tang Z (2020) Optimizing energy transfer in nanostructures enables in vivo cancer lesion tracking via near-infrared excited hypoxia imaging. Adv Mater 32:1907718 Lone SN, Nisar S, Masoodi T, Singh M, Rizwan A, Hashem S, El-Rifai W, Bedognetti D, Batra SK, Haris M, Bhat A, Macha MA (2022) Liquid biopsy: a step closer to transform diagnosis, prognosis and future of cancer treatments. Mol Cancer 21:79 Lu C, Zhang C, Wang P, Zhao Y, Yang Y, Wang Y, Yuan H, Qu S, Zhang X, Song G, Pu K (2020) Light-free generation of singlet oxygen through manganese-thiophene nanosystems for pH-responsive chemiluminescence imaging and tumor therapy. Chem 6:2314–2334 Luo S, Zou R, Wu J, Landry MP (2017) A probe for the detection of hypoxic cancer cells. ACS Sensors 2:1139–1145 Ma T, Hou Y, Zeng J, Liu C, Zhang P, Jing L, Gao M (2018) Dual-ratiometric target-triggered fluorescent probe for simultaneous quantitative visualization of tumor microenvironment protease activity and pH in vivo. J Am Chem Soc 140:211–218 Marland JRK, Grayb ME, Dunare C, Blair EO, Tsiamis A, Sullivan P, González-Fernándezd E, Greenhalgh SN, Gregson R, Clutton RE, Parys MM, Dyson A, Singer M, Kunkler IH, Potter MA, Mitra S, Terry JG, Smith S, Mount AR, Underwood I, Walton AJ, Argyle DJ, Murray AF (2020) Real-time measurement of tumour hypoxia using an implantable microfabricated oxygen sensor. Sens Bio-Sens Res 30 Mathejczyk JE, Pauli J, Dullin C, Resch-Genger U, Alves F, Napp J (2012) High-sensitivity detection of breast tumors in vivo by use of a pH-sensitive near-infrared fluorescence probe. J Biomed Opt 17:076028 McNeel KE, Siraj N, Bhattarai N, Warner IM (2019) Fluorescence-based ratiometric nanosensor for selective imaging of cancer cells. ACS Omega 4:1592–1600

Tissue Oxygenation and pH-Responsive Fluorescent Nanosensors …

205

Minhas AS, Sharkey J, Randtke EA, Murray P, Wilm B, Pagel MD, Poptani H (2020) Measuring kidney perfusion, pH, and renal clearance consecutively using MRI and multispectral optoacoustic tomography. Mol Imag Biol 22:494–503 Mirabello V, Cortezon-Tamarit F, Pascu SI (2018) Oxygen sensing, hypoxia tracing and in vivo imaging with functional metalloprobes for the early detection of non-communicable diseases. Front Chem 6:27 Mi P, Kokuryo D, Cabral H, Wu H, Terada Y, Saga T, Aoki I, Nishiyama N, Kataoka K (2016) A pH-activatable nanoparticle with signal-amplification capabilities for non-invasive imaging of tumour malignancy. Nat Nanotechnol 11:724–730 Muz B, de la Puente P, Azab F, Azab AK (2015) The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia 3:83 Newmark HL, Lupton JR (1990) Determinants and consequences of colonic luminal pH: implications for colon cancer. Nutr Cancer 14:161–173 Offenbacher H, Wolfbeis OS, Furlinger E (1986) Fluorescence optical sensors for continuous determination of near-neutral pH values. Sens Actuators 9:73–84 Palmirotta P, Lovero D, Cafforio P, Felici C, Mannavola F, Pelle E, Quaresmini D, Tucci M, Silvestris F (2018) Liquid biopsy of cancer: a multimodal diagnostic tool in clinical oncology. Ther Adv Med Oncol 10:1–24 Pang J, Xing H, Sun Y, Feng S, Wang S (2020) Non-small cell lung cancer combination therapy: hyaluronic acid modified, epidermal growth factor receptor targeted, pH sensitive lipid-polymer hybrid nanoparticles for the delivery of erlotinib plus bevacizumab. Biomed Pharmacother 125:109861 Sarhadi VK, Armengol G (2022) Molecular biomarkers in cancer. Biomolecules 12:1021 Shaked NT, Rinehart MT, Wax A (2009) Dual-interference channel quantitative-phase microscopy of live cell dynamics. Opt Lett 34:767–769 Shamsipur M, Barati A, Nematifar Z (2019) Fluorescent pH nanosensors: design strategies and applications. J Photochem Photobiol C 39:76–141 Smith BR, Gambhir SS (2017) Nanomaterials for in vivo imaging. Chem Rev 117:901–986 Subasinghe SAAS, Pautler RG, Samee AH, Yustein JT, Allen MJ (2022) Dual-mode tumor imaging using probes that are responsive to hypoxia-induced pathological conditions. Biosensors 12:478 Tang Y, Xiao G, Shen Z, Zhuang C, Xie Y, Zhang X, Yang Z, Guan J, Shen Y, Chen Y, Lai L, Chen Y, Chen S, Dai Z, Wang R, Wu R (2020) Noninvasive detection of extracellular pH in human benign and malignant liver tumors using CEST MRI. Front Oncol 10:578985 Teng P, Hao M (2020) A population-based study of age-related associations between vaginal pH and the development of cervical intraepithelial neoplasia. Cancer Med 9:1890–1902 Tian Q, Li Y, Jiang S, An L, Lin J, Wu H, Huang P, Yang S (2019) Tumor pHresponsive albumin/polyaniline assemblies for amplified photoacoustic imaging and augmented photothermal therapy. Small 15:1902926 Vaupel P, Harrison L (2004) Tumor hypoxia: causative factors, compensatory mechanisms, and cellular response. Oncologist 5:4–9 Wang XD, Stolwijk JA, Lang T, Sperber M, Meier RJ, Wegener J, Wolfbeis OS (2012) Ultra-small, highly stable, and sensitive dual nanosensors for imaging intracellular oxygen and pH in cytosol. J Am Chem Soc 134:17011–17014 Wang Xd, Meier RJ, Wolfbeis OS (2013) Fluorescent pH sensitive nanoparticles in an agarose matrix for imaging of bacterial growth and metabolism. Angew Chem 125:424–427 Wang X, Niu D, Wu Q, Bao S, Su T, Liu X, Zhang S, Wang Q (2015) Iron oxide/manganese oxide co-loaded hybrid nanogels as pH-responsive magnetic resonance contrast agents. Biomaterials 53:349–357 Wisniewski NA, Nichols SP, Gamsey SJ, Pullins S, Au-Yeung KY, Klitzman B, Helton K L (2017) Tissue-integrating oxygen sensors: continuous tracking of tissue hypoxia. In: Oxygen transport to tissue XXXIX: advances in experimental medicine and biology. Springer, Cham, vol 977

206

S. Srivastava et al.

Yang HY, Jang MS, Li Y, Fu Y, Wu TP, Lee JH, Lee DS (2019) Hierarchical tumor acidity-responsive self-assembled magnetic nanotheranostics for bimodal bioimaging and photodynamic therapy. J Control Release 301:157–165 Zhang X, Lin Y, Gillies RJ (2010) Tumor pH and its measurement. J Nucl Med 51:1167–1170 Zhang Z, Feng Q, Yang M, Tang Y (2020) A ratiometric fluorescent biosensor based on conjugated polymers for sensitive detection of nitroreductase and hypoxia diagnosis in tumor cells. Sens Actuators, B Chem 318:128257 Zheng X, Wang X, Mao H, Wu W, Liu B, Jiang X (2015) Hypoxia-specific ultrasensitive detection of tumours and cancer cells in vivo. Nat Commun 6:1–12 Zhu C, Zou Z, Huang C, Zheng J, Liu N, Li J, Yang R (2019) Highly selective imaging of lysosomal azoreductase under hypoxia using pH-regulated and target-activated fluorescent nanoprobes. Chem Commun 55:3235–3238

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis Saroj Kumar Das, Nishant Ranjan Chauhan, and Subhash Mehto

Abstract Neurological manifestations in diverse forms are rising in recent decades and are supposed to be the leading cause of mortality across the globe. Cerebral ischemia (ischemic stroke) is considered as one of the life threatening cerebral disorders caused by “thrombosis”, blood clot formation in the arteries which supply blood to the vital brain regions. The early catastrophic change following cerebral ischemia is associated with inflammatory processes leading to neuronal cell death and resulting in loss of functions of the affected brain regions. The severity of these structural and functional problems depends on the affected areas, the size of the clot and the time taken to recover from the ischemic stroke following treatment. The principal method to treat cerebral ischemia is reperfusion and thrombectomy. The reperfusion method which involves the use of tissue plasminogen activator (tPA) to remove thrombosis is associated with several side effects such as an upsurge in the generation of reactive oxygen species (ROS)/reactive nitrogen species (RNS), heightened proinflammatory response and leukocyte infiltration in the brain. Correspondingly, the use of mechanical thrombectomy is associated with limited access to trained neurosurgeon,technical difficulty with the navigating wire in intracranial vessels, may aggravate the stroke. Therefore, a prognostic marker for early diagnosis of cerebral ischemia to improve the efficacy of treatment strategies is the paramount requisite in recent days. The compensatory measure for the reduction in CBF, oxygen extraction factor (OEF) in the affected brain region is augmented during the early stage of cerebral ischemia to sustain oxidative metabolism near the basal level. The use S. K. Das (B) P.G. Department of Life Sciences, Sri Krushna Chandra Gajapati (Autonomous) college, Paralakhemundi, Gajapati 761200, Odisha, India e-mail: [email protected] Neurobiology Laboratory, Centre for Biotechnology, School of Pharmaceutical Sciences, Siksha ‘O’ Anusandhan (Deemed to Be University), Bhubaneswar 751003, Odisha, India N. R. Chauhan Cell Biology and Infectious Disease Biology Division, Institute of Life Sciences, NALCO Square, Bhubaneswar 751023, Odisha, India S. Mehto Division of Infectious Diseases and Immunology, University of Massachusetts-Chan Medical School, Worcester, MA, USA © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_11

207

208

S. K. Das et al.

of nanotechnology has recently emerged as an innovative means towards the diagnosis and treatment of several life threatening diseases including cerebral ischemia. The development of hypoxia responsive nanomaterials towards early diagnosis of cerebral ischemia is gaining significant attention. In this chapter, a comprehensive pathology of cerebral ischemia and its related maladies, a brief emphasis towards available diagnosis methods, an introduction of the concept of nanotechnology as a novel diagnostic tool towards early identification of cerebral ischemia and finally available treatment strategies are discussed. Keywords Cerebral ischemia · Thrombosis · Diagnosis · Hypoxia · Stroke therapy · Nanoparticles · Hypoxia responsive nanoparticles

Abbreviations AM BCAO DOPE DPPC DPPG DSPC DSPE-PEG 2000 eNOS iNOS LCAO MCAO MPP MPP/SCB MSNs-TQ NMDAR NPs PEG PLGA PS ROS SLNs SOD TIMP-1 TNF-α WGA-NPs

Encapsulated agonistic micelle Bilateral common carotid artery occlusion Dioleoyl phosphatidylethanolamine Dipalmitoyl-phosphatidylcholine 1,2-Dipalmitoyl-sn-glycero-3-phospho Distearoylphosphatidylcholine Distearoylphosphoethanolamine-polyethyleneglycol-2000 Endothelial nitric oxide synthase Inducible nitric oxide synthase Left common carotid artery occlusion Middle cerebral artery occlusion PH-sensitive polymeric nanovehicle with a 4T1 cell membrane SCB-loaded pH-sensitive polymeric nanovehicle with a 4T1 cell membrane Mesoporous silica nanocarriers-Thymoquinone N-methyl-D-aspartate receptor Nanoparticles Polyethylene glycol Poly lactic-co-glycolic acid nanoparticles Phosphatidylserine Reactive oxygen free radicals Solid lipid nanoparticles Superoxide dismutase Tissue inhibitor of matrix metalloproteinases Tumor necrosis factor-alpha Wheat germ agglutinin-modified nanoparticle

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

209

1 Introduction The global burden of neurological disorders is increasing at an alarming rate in the last few decades and thus stance a significant challenge for the sustainability of health systems of both developed and developing nations. However, developing countries are facing a more pronounced upsurge in neurological burden in recent decades due to rapid changeover in demography and epidemiology. In this regard, a thorough understanding of neurological disorders with respect to their prevalence, occurrence, risk factors and health care planning is of paramount requisite in low-income and middleincome countries. More than 1000 neurological disorders have been identified till date, but the correct diagnosis, treatment, prevention and rehabilitation are restricted to only a few of its type. Among these, cerebrovascular diseases (CVDs) make a class of neurological disorders that includes a wide range of anomalies involving vertebral stenosis, carotid stenosis, intracranial stenosis, stroke, aneurysm, embolism and vascular malformations. With the available data, CVDs represent the most common global life threatening neurological disorders. As per the report, stroke represents the second leading cause of both disability and death worldwide and accounts for 6 × 106 death annually (Woodruff et al. 2011). The stroke is a medical condition that primarily occurs when there is an interruption of blood flow to the brain or its parts generating a hypoxic ailment leading to transient loss of function or permanent damage to the site. Medically, strokes can be broadly classified into five types such as transient ischemic stroke, ischemic stroke, hemorrhagic stroke, brain stem stroke and cryptogenic stroke. Out of these strokes, ischemic stroke/cerebral ischemia accounts for 85% of strokes (Beal 2010; Sacco et al. 2013) (Table 1). Cerebral ischemia: pathophysiology, epidemiology and risk factors Cerebral ischemia/ischemic stroke occurs when there is an obstruction in blood flow to the vital parts of the brain. The obstruction due to cerebral ischemia is residing with respect to clot formation in arteries (atherosclerosis). In a broad sense, cerebral ischemia is of two types such as global and focal. In global cerebral ischemia, a large brain region or entire brain is restricted or cut off from blood supply and this usually leads to cardiac arrest (Zhang et al. 2020). Focal cerebral ischemia is primarily confined to a specific area of the brain when a clot has blocked an artery of the brain, and usually, this may result in embolus or thrombus. In this regard, ischemic stroke can be embolic (embolism) when blood clot travels from other parts of the body to the brain through cerebral circulation causing the stroke and accounts for 75% of all cerebral ischemia. On the other hand, thrombotic cerebral ischemia is considered as an ischemic stroke when there is clot formation in the blood vessel of the brain itself and it accounts for the remainder of cerebral ischemia (Mergenthaler et al. 2004; Ahad et al. 2020). However, the fundamental pathophysiology of cerebral ischemia includes cognitive dysfunction, neural signaling failure, energy failure, cell-ion homeostatic loss, augmented intracellular calcium, complement activation, cytokine-instigated cytotoxicity, excitotoxicity, blood–brain barrier (BBB) disruption, leucocytes infiltration and oxidative stress (Woodruff et al. 2011; Neumann

210

S. K. Das et al.

Table 1 Classification of Cerebrovascular diseases and their etiology with available treatment strategies Sl. no Cerebrovascular diseases

Etiology

Available treatment strategies

References

1

Ischemic stroke A. Thrombotic B. Embolic

A. Thrombus blocking • Alteplase (Tissue Brekenfeld et al. an artery to the brain plasminogen (2012), Smith activator), tPA and et al. (2016) B. Embolus (thrombus tenecteplase or plaque that moves in • Intraarterial bloodstream until it thrombolysis blocks an artery • Carotid downstream) endarterectomy or stenting of the intracranial vessels • Mechanical thrombectomy by The Merci Retriever and the Penumbra System

2

Hemorrhagic stroke A. Intracerebral hemorrhage B. Subarachnoid hemorrhage

(i) High blood pressure • Surgery to relieve Feldstein (2014), Muehlschlege (hypertension), intracranial (2018) ruptured brain pressure, surgery aneurysm, abnormal to repair damaged development of blood blood vessel, vessels (vascular catheter and stent malformation), (tiny platinum anticoagulation coils), medication clipping/vessel complications/bleeding bypass, disorders/cerebral craniotomy, amyloid angiopathy hyperventilation, (CAA)/tumors stereotactic clot aspiration (ii) Head trauma, • Endovascular ruptured brain techniques: coiling aneurysm and/or stenting/flow diversion

3

Transient ischemic attack

Temporary blockage of • Treatment of Renner et al. an artery to the brain Carotid artery (2021) disease or cardiac problems • Anti-platelet therapy (aspirin, clopidogrel and the combination of aspirin plus dipyridamole and cilostazol)

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

211

Table 2 Stages of Cerebral ischemia and available treatment strategies Sl. Cerebral Stages no ischemia

Syndromes (symptoms)

Etiology

Treatment strategies

References

1

Focal Specific • Frailty in one • Blockage • Carotid Artery ischemia region arm or leg of Surgery • Frailty in one • Computerof the cerebral entire side of Assisted Surgery brain vessel body (CAS) affected due to blood clot • Stenting • Double vision • Due to • Dizziness • Endovascular thromNeurosurgery • Vertigo bosis or • Frailty on both • Microsurgery embolism sides of the body • Dysarthria • Ataxia

Smith (2004)

2

Global Wide • Most of the • Cardiac ischemia areas of symptoms are arrest brain similar to focal tissue ischemia affected • Unconsciousness, weakness in both sides of the body, focal brain infarction

Brekenfeld et al. (2012)

• Alteplase (Tissue plasminogen activator: tPA) • Treatment as per the requirement described above

et al. 2013; Manzanero et al. 2013). Several experimental animal modelings have shown the recurrence of multiple mechanisms of ischemic brain injury. Oxidative stress following cerebral ischemia is associated with a reduction in the bioavailability of nitric oxide (NO) in endothelial cells (Lubos et al. 2008). As NO primarily prevents thrombosis by inhibiting leukocyte aggregation and platelet adhesion, it leads to heightened ischemic stroke through a reduction in CBF (Table 2). A chain of biochemical events occurs soon after the occlusion of the cerebral artery following cerebral ischemia. This effect leads to restriction in nutrient and oxygen supply to affected brain regions and thus there is a failure of aerobic glycolysis in neurons. Followed by there is an augmented influx of sodium and calcium into the affected neurons leading to calcium overload. Afterwards, local acidification occurs following surplus formation and accumulation of lactate. Neuronal excitotoxicity is another sequential event that occurs following increased secretion of excitatory neurotransmitters and augmented generation of free radicals. As a final assault, protease and lipase hyperactivity leads to the activation of apoptotic signal and thus promotes apoptotic neuronal death in the region of cerebral ischemia. Therefore, a sustained reduction in CBF leading to edema following cerebral ischemia is detrimental to the brain regions and associated functions and if persists for a long duration may lead to permanent disability or death (Gu et al. 2022). The abrupt disruption of CBF is associated with necrosis and consequently leads to cerebral edema in the principal zone of cerebral ischemia which may lead to the obliteration of the

212

S. K. Das et al.

blood–brain barrier (BBB) (Yemisci et al. 2015). With the release of necrotic cells, there is substantial upregulation in the level of inflammatory cytokines and apoptotic signaling cascades leading to the death of nearby cells in the infarcted areas and thus aggravating the cerebral ischemia-persuaded brain injury (Li et al. 2018a, b). The prevalence of cerebral ischemia is rising at an alarming rate in both developed and developing countries in recent decades. The changing life styles involving physical inactivity, food habit, working environment and gross stress might aggravate the occurrence of cerebral ischemia. In gross, cerebral ischemia is the third leading cause of death in USA alone. Available data suggested that cerebral ischemia accounts for eight of ten stroke cases. Further, cerebral ischemia represents the most general reason for disability, third leading root of dementia and fourth leading cause of death in developing and developed countries. With this prevalence, cerebral ischemia is cited with a number of documented risk factors. Even if the contribution of these risk factors towards the pathophysiology of cerebral ischemia is different, the outcomes fall within the symptoms of cerebral ischemia. The risk factors for cerebral ischemia are classified into the following two types based on whether they are influenced (modifiable) or not (non-modifiable) (Fig. 1). The non-modifiable risk factors mainly involved age, gender, ethnicity, genetics and previous history of ischemic stroke (Sveinsson et al. 2014). Age is the prime nonmodifiable risk factor for cerebral ischemia, that’s why it is very uncommon below the age of 40. However, the prevalence of ischemic stroke upsurge with increasing age, and the incidence gets double in each successive decade after the age of 55. Hence, the risk of ischemic stroke for people older than 80 years is almost double that of individuals 60–79 years of age. Further, the incidence of cerebral ischemia is moderately higher in men in comparison with women up to the age of 75. The ethnic population-based study showed that blacks have a higher prevalence of ischemic stroke than whites (Sacco et al. 1998; White et al. 2005). In addition, a family history of ischemic stroke increases the incidence and this could be attributed to the existence of a family history of other risk factors such as high blood pressure, diabetes and hypercholesteremia. The modifiable or treatable risk factors of cerebral ischemia include living practice, atherogenic traits (high blood pressure, diabetes mellitus, obesity and hypercholesteremia), smoking and a few diseases. These risk factors primarily predispose the individual towards the occurrence of ischemic stroke, and among these, hypertension is the most common modifiable risk factor of cerebral ischemia (Rigaud et al. 2000). High blood pressure (hypertension) substantially increases the risk of a cerebral stroke three to four times. On the other hand, diabetes mellitus is associated with a high risk towards the occurrence of cerebral ischemia and it is independent of age and other related cardiovascular factors (Burchfiel et al. 1994). Considering dyslipidaemia (hypercholesteremia), the direct association between cerebral ischemia and cholesterol level is not established. However, smoking is considered as an independent modifiable risk factor of most subtypes of cerebral ischemia (Whisnant et al. 1996). The report also cited that passive smoking is associated with an increased risk of cerebral ischemia (Lee et al. 2006; You et al. 1999). Several other modifiable risk factors are also reported to be associated with the increasing occurrence of ischemic stroke.

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

213

Fig. 1 Diagrammatic representation showing risk factors (modifiable and non-modifiable) of cerebrovascular diseases

2 Hypoxia and Cerebral Ischemia The clinical symptoms and manifestations of cerebral ischemia are reported to be highly assorted and further subjected to the location and extent of the damage. The first and foremost cause of cerebral ischemia resides with respect to a reduction in cerebral blood flow (CBF) to the affected brain area or multiple brain locations following blockage of cerebral arteries which leads to a reduced supply of oxygen (hypoxia) and nutrients (Fig. 2). The impaired oxygen supply to the brain leading to the hypoxic condition following ischemic stroke is associated with profound changes in metabolic signaling in both neural and non-neural cells. Following hypoxic condition, the response in cytosolic level is reflected in the form of rapid change in membrane lipid composition and enzyme activities and further leads to change in transcription and translation of vital cellular entities. However, the central nervous system (CNS) can endure the ischemia or cerebral hypoxia for a restricted period of time, a phenomenon called ischemic tolerance. Thus, hypoxia is considered as the critical factor for neuronal survival or death in cerebral ischemia; however, the pathophysiology of cerebral ischemia-hypoxia is not completely understood. The cellular hypoxia following ischemic stroke is followed by production of inflammatory cytokines and subsequent activation of both pro-survival and proapoptotic signaling pathways. In the beginning, following ischemic stroke-induced reduction in CBF, the regional oxygen extraction fraction is augmented as a compensatory measure to maintain cerebral metabolic rate of oxygen close to the normal

214

S. K. Das et al.

Fig. 2 Schematic illustration showing gross etiology of cerebral ischemia

cell (Baron 2011; Adhami et al. 2006). Earlier report following comprehensive of investigation of energy metabolism of cells, cortical neurons were found out to be the most susceptible neurons to reduced oxygen supply (Lukyanova and Kirova 2015). At cellular level, hypoxia persuades reprogramming of respiratory chain function, and swapping from NAD oxidation to succinate oxidation is required for instant acclimatization to persistent cellular hypoxia (Lukyanova and Kirova 2015). The pathophysiological effects associated with cerebral ischemia-induced hypoxia involve multifaceted signaling cascades. One pathophysiological effect is hypoxia-persuaded fibrin deposition primarily caused by the transformed anticoagulant property of endothelial cells (Adhami et al. 2006). Additionally, a reduction in CBF in association with hypoxia might persuade unprompted thrombosis and this may lead to a further reduction in blood perfusion. In this regard, cerebral ischemia/hypoxia-persuaded microvascular thrombosis might also prevent cerebral reperfusion even after the release/removal of thrombotic occlusion from large artery. However, reperfusion of the affected brain area following clearance of the artery during ischemic stroke is not beneficial rather it involves more detrimental consequences in ischemic brain tissues called cerebral ischemia/reperfusion (I/R) injury (Liang et al. 2021; Zhao et al. 2022). Recent reports advocated that I/R injury is associated with several forms of cell death such as apoptosis, pyroptosis, necrosis, necroptosis and autophagy (Wu et al. 2018; Zhao et al. 2022). As cerebral ischemia is associated with heightened neuroinflammation and oxidative stress, the detrimental effect of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

215

Fig. 3 Illustrative presentation of cellular pathophysiological mechanism involved in ischemic stroke

inflammasome activation and dysfunctional autophagy can’t be ignored. A detailed of neuropathological events that occurred during ischemic stroke and the cytosolic response is enumerated in Fig. 3. Therefore, the regulatory role of autophagy and inflammasome activation could play a pivotal role in the pathological manifestation of cerebral ischemia-persuaded I/R injury (Lv et al. 2021). Existing information on the pathophysiology of cerebral ischemia also focused towards regulatory role of long non-coding RNAs (lncRNAs) and their direct role towards controlling the expression pattern of protein-coding genes and associated signaling cascades of diseases (Ren and Yang 2018; Wang et al. 2019a, b, c). Therefore, lncRNAs can be considered as potential biomarkers for diagnosis and therapeutic intervention for cerebral ischemia.

3 Cerebral Ischemia Diagnosis Practices and Recent Advances As cerebral ischemia is associated with sudden reduction in CBF due to thrombosis (embolism), necrosis, cerebral edema and disruption of BBB, there exist a limited time window for correct diagnosis of thrombosis. Primarily, two different imaging tools are frequently utilized for the diagnostic and observation of individuals assumed of acute cerebral ischemia such as computed tomographic (CT) scan and magnetic resonance imaging (MRI) (Wang et al. 2016; Fernandes et al. 2018). Clinically, CT scan is preferred for the detection of infarction; however, it is not sensitive

216

S. K. Das et al.

enough and also not able to differentiate between new events of cerebral ischemia (Fig. 4). Further, ischemic stroke-like symptoms are also represented in other nonvascular maladies such as migraine and epilepsies, thus a correct diagnosis is essential for early-stage administration of thrombolytic drugs. In this regard, MRI is a gold standard imaging tool which provides an advantage for the early assessment of acute ischemic stroke. However, the effectiveness of MRI in detecting ischemic stroke within 3 h of symptoms onset is around 70% (Chalela et al. 2007). Considering this clinical fact, in present days, a more precise diagnosis of ischemic stroke is achieved through the collective use of CT scan and MRI (Brazzelli et al. 2009). Still, there is a boundless requirement for a more powerful and sensitive technique for early and accurate detection of ischemic stroke and thus several advancements have been done to meet the short coming of conventional principles of CT scan and MRI (Fig. 4). In this situation, a vast number of clinical and preclinical studies have been conducted to identify novel serum biomarkers, new diagnostic-imaging technique and nanoprobes for imaging application in nanotechnology (Mouhieddine et al. 2015; Wang et al. 2016).

Fig. 4 Diagram showing etiology of diagnostic criteria of cerebral ischemia

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

217

4 Available Treatment Strategies for Cerebral Ischemia Preclinical validation of a number of drugs against cerebral ischemia-persuaded I/R injury has been used and demonstrated the mechanism of neuroprotection (Ruan et al. 2021). Studies have demonstrated that targeting neuroinflammatory response especially NLRP3 inflammasome activation by the therapeutic intervention of novel compounds (natural and synthetic) provides substantial neuroprotection following ischemic stroke through regulation of diverse cellular signaling (Qiu et al. 2016; Ma et al. 2019a, b; Peng et al. 2020; Bian et al. 2021; Liu et al. 2021; Ran et al., 2021a, b). Earlier findings advocated that Sinomenine, a natural anti-inflammatory and anti-apoptotic molecule, provides substantial neuroprotection against middle cerebral artery occlusion (MCAO) in mice model by reducing cerebral infraction, cerebral edema and neural apoptosis through suppression of NLRP3 inflammasome activation (Qiu et al. 2016) (Table 3). Presently, clinical measure for the treatment of cerebral ischemia to restore the blood flow in the brain is available by intravenous administration of tissue plasminogen activator (tPA) or by the surgery involving mechanical thrombectomy (Fig. 5). The tPA administration is a gold standard clinical treatment for cerebral ischemia; however, tPA administration is done within 4.5 h of experiencing ischemic stroke symptoms and clinically there is an alleviation of symptoms (Yoo et al. 2011). Despite being one of the important treatment strategies available in recent days, tPA administration is associated with severe secondary complications in the form of intracerebral hemorrhage leading to increase mortality (Kurth et al. 2007). Alternatively, when cerebral ischemia is associated with large artery occlusion or if the treatment window for tPA administration crosses the suitable time range, mechanical thrombectomy is an alternative clinical strategy for the removal of clots by microcatheter from clotting blood vessels following ischemic stroke. However, restoration of cerebral blood perfusion is associated with a neuroinflammatory response following the generation of reactive oxygen species (ROS) by the sudden influx of oxygen. In this regard, several drugs have been discovered to provide neuroprotection against I/R injury, but none were clinically approved due to failure at multiple sites such as unproductive BBB permeability, poor stability, toxicity, short circulation time and difficulty in choosing the right drug due to heterogeneity of cerebral ischemia (Dong et al. 2020) (Table 4).

5 Nanoparticles as Effective Drug Delivery System for the Treatment of Cerebrovascular Diseases As most of the available treatment strategies are associated with poor stability in the blood vascular system, short blood circulation time and ineffective BBB permeability, a novel drug delivery vehicle with control release is a paramount requisite to improve the treatment of cerebral ischemia. In this regard, nanotechnology plays a pivotal role

Name of compound

Astragaloside IV

Ascovertin

Buckwheat polyphenol

Crataegus flavanoids

Curcumin

Gastrodin

Sl. no

1

2

3

4

5

6

LCAO

MCAO

Type of ischemia

Gastrodia elata

Curcuma longa Linn

Crataegus Pinnalifida

MCAO

MCAO and BCAO

MCAO and BCAO

Fagopyrum esculentum TIA

Complex of ascorbic acid and dihydroquercetin

Astragalus membranaceus

Source

Table 3 Medicinal compounds used for the treatment of cerebral ischemia

Rats

Rats

Rat

Animal model

100 mg/kg, i.p

30 mg/kg, i.p

Rats

Mongolian gerbils

0.5 and Mongolian 2.5 mg/ml, oral gerbils

600 mg/kg, oral

70 mg/kg, oral

20 mg/kg, i.p

Treatment mode and dosage

Wang et al. (2005)

Zhang et al. (2004)

Pu et al. (2004)

Logvinov et al. (2001)

Zhang et al. (2019)

References

(continued)

Reduces inflammatory Wang et al. response, decreases (2018a) apoptosis, high rate of revascularization, reduces neurological symptoms

Suppress mitochondrial-mediated apoptosis

Scavenge superoxide anion

Inhibit excessive release of glutamate and restored production of NO2

Prevent brain Na+ K+ -ATPase activity

Inhibits apoptosis by promoting p62-LC3-mediated autophagy; reduces neurological symptoms

Mechanism of action

218 S. K. Das et al.

Ginsenoside Rg1 Panax ginseng

Lecithin and α tocopherol

Safranal

Salidroside

8

9

10

11

Rhodiola rosea

Crocus sativus L

Soybean, Nuts, seeds and vegetable oil

Ginkgo biloba

Ginkgolide B

7

Source

Name of compound

Sl. no

Table 3 (continued)

MCAO/Reperfusion

Four vessel occlusion

BCAO

MCAO

Thrombotic cerebral ischemia induced by photochemical

Type of ischemia

Rats

Rats

Tree shrews

Animal model

2.5, 5, 10, 20 mg/kg, i.v

Mice

72.5 mg/kg, i.p Rats

300 mg/kg (Lecithin), 200 mg/kg (tocopherol), oral

10, 20, 40 mg/kg, i.p

5 mg/kg, i.v

Treatment mode and dosage

Controls microglial polarization, diminishes cerebral infarction, upregulates M2 macrophage marker

Antioxidant activity

Antioxidant activity

Downregulation of IkBα phosphorylation and NF-kB nuclear translocation, reduces infarct area and neurological symptoms

Suppress pathological manifestation of platelet activating factors such as calcium overload toxicity and edema in brain

Mechanism of action

(continued)

Liu et al. (2018)

Hosseinzadeh et al. (2005)

Aabdallah et al. (2004)

Zheng et al. (2019)

Li et al. (1999)

References

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis 219

Flavonoids (SSF) Scutellaria baicalensis BCAO Georgi

Thymoquinone

14

Nigella sativa

Cerebral reperfusion injury

MCAO

13

Erigeron breviscapus

Scutellarin

Type of ischemia

12

Source

Name of compound

Sl. no

Table 3 (continued)

2.5, 5 and 10 mg/kg, i.p

35 mg/kg, oral

50 and 75 mg/kg, oral

Treatment mode and dosage

Rats

Rats

Rats

Animal model

Suppress lipid peroxidation

Blocks the pathological process in vascular dementia. Provides protection against neuronal injury and modulates energy metabolites

Increase eNOS and decrease iNOS levels

Mechanism of action

Hosseinzadeh et al. (2007)

Shang et al. (2005)

Hu et al. (2005)

References

220 S. K. Das et al.

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

221

Fig. 5 Diagram illustrating various treatment strategies for cerebral ischemia

to design nanomaterials with potential biomedical applications for several neurodegenerative diseases and cerebral ischemia (Saraiva et al. 2016). These nanoparticles may be natural or synthetic polymers with varied sizes ranging from 1 to 1000 nm. However, nanoparticles with a size range of 1–500 nm in diameter are effortlessly taken up by the cells. The increased efficiency of nanoparticles to interact with cells depends upon the surface area to volume ratio. Therefore, it can be inferred that the smaller the nanoparticles, the large the surface area to volume ratio (Salatin et al. 2015). However, the half-life of nanoparticles in biological systems depends upon the shape, size and surface modification (Mohammadi et al. 2017). The structural modification of nanoparticles in the form of PEGylation is linked with the biocompatibility of nanoparticles, by which antioxidants are brought to affected/targeted brain tissues. Additionally, PEGylation in combination with polyethylene glycol (PEG) and nanoparticles can enhance the hydrophilicity and steadiness of nanoparticles (Suk et al. 2016). Nanoparticles of specific sizes are used for packaging or encapsulating of drugs to augment their water solubility, membrane permeability and tissue targeting; thus, nanoparticle drug delivery tools will solve most of the existing challenges in cerebral ischemia treatment and management. In this line, we have discussed novel nanoparticles-based drug delivery systems and their application towards the treatment of ischemic stroke.

T7 and SHp coupled PEGylated liposomes

Basic fibroblast growth factor (bFGF) conjugated liposomes

PEGylated liposome nanoparticles (tPA/fasudil)

3

4

5

PEGylated liposomes (Asialo-erythropoietin)

T7-conjugated liposome nanoparticles (ZL006)

Liposomes nanoparticles

1

Name of nanomedicines

2

Broad Classification of Nanoparticles

Sl. no

Table 4 Nanomedicines in the treatment of cerebral ischemia

i.v

i.n

i.v

i.v

i.v

Drug delivery mode

Rats

Rats

Rats

Rats, brain capillary endothelial cells

Rats

Modeling

References

Treating fasudil-lip prior to tPA reduced the danger of tPA-mediated brain hemorrhage and thus increase the therapeutic time period of tPA

bFGF aggregation in brain tissues ameliorated spontaneous locomotion in animals

Augmented aggregation of NPs in brain improved ischemic damage

Augment transportation of liposomes across blood–brain barrier and T7-PLPs/ZL006 decreases infarct area and protects from neurological symptoms

(continued)

Fukuta et al. (2017)

Zhao et al. (2016b)

Zhao et al. (2016a)

Wang et al. (2015)

Increase aggregation of AEPO Ishii et al. (2012) at the site of ischemia and improved brain ischemic reperfusion injury

Treatment effects

222 S. K. Das et al.

DOPE, DSPE-PEG 2000

Polymorphonuclear leucocyte membrane-resultant nanovesicles (Resolvin D2)

9

10

Biomimetic nanoparticles

FK506

8

i.v

i.v

i.v

DPPC, Egg phosphocholine, i.v PEG2000, DPPG, cholesterol (Xenon)

i.v

Drug delivery mode

7

Name of nanomedicines

DSPE-PEG2000 liposome

Broad Classification of Nanoparticles

6

Sl. no

Table 4 (continued)

Mouse

Rats

Rats

Rats

Mouse

Modeling

References

Targets the ischemic area of endothelium and releases Resolvin D2 to impede MPO activity and reduce inflammatory response at ischemic damaged area

Recovering of infarct area, brain edema, neurological symptoms; suppress inflammatory signaling like myeloperoxidase (MPO) activity and TNF-α levels

Diminishes infarct area, Suppresses leucocyte infiltration and downregulates the TNF-α levels

Inhibits apoptosis in neurons and lowered their mortality

(continued)

Dong et al. (2019)

Partoazar et al. (2017)

Fukuta et al. (2015)

Miao et al. (2018)

Liposomes aggregated in Al-Ahmady et al. ischemic area of brain at early (2019) phase (0.5 h) and late stage (48 h)

Treatment effects

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis 223

Unknown, miR-195

C (RGDyK) peptide-ligated exosomes

13

14

Curcumin, Poly (ethylene glycol)-b-poly (D,L-lactide)

Polymeric nanoparticles

12

Name of nanomedicines

Platelets membrane covered-γ-Fe2 O3 magnetic nanoparticles (L-arginine)

Broad Classification of Nanoparticles

11

Sl. no

Table 4 (continued)

Stereotaxic injection

i.v

i.v

i.v

Drug delivery mode Li et al. (2020)

References

Anti-apoptotic for damaged Cheng et al. (2019) neurons by inhibiting Sema3A/Cdc42/JNK signaling cascade; neurogenesis by stimulating neuronal stem cell propagation and migration; anti-inflammatory action by obstructing NF-kB pathway

Suppressed the rise in MMP-9 Wang et al. (2019a, levels; maintenance of BBB b, c) Integrity; decrease activated M1 microglial cells and lessen upsurge in TNF-α and IL-1β levels

L-arginine at ischemic lesion area disturbed platelets accumulation and improves cerebral blood flow

Treatment effects

(continued)

Bone marrow-derived Targets infarct area of Tian et al. (2018) MSCs, Mice ischemic stroke and microglia to inhibit the inflammation and guard the affected part of the brain

Rats

Mice

Mice

Modeling

224 S. K. Das et al.

SHp conjugated, RBC membrane shelled-polymer (NR2B9C)

T7-ligated RBS-covered Mn3 O4 NPs

OX26-PEGylated-Senanoparticles

18

19

20

Inorganic nanoparticles

Cationic BSA-conjugated PEG-PLA (Tanshinone IIA)

16

Name of nanomedicines

Poly (ethylene glycol)-block-poly (D, L-lactide) loaded with C3 siRNA

Broad Classification of Nanoparticles

15

Sl. no

Table 4 (continued)

i.p

i.v

i.v

i.v

i.v

Drug delivery mode

Rats

Rats

Rats

Rats

Mice

Modeling

References

Suppress action of jak2/stat3 signalling pathway and decrease the transcriptional levels of inflammatory mediator Adamts1

The NPs scavenged free radicals and restoration of the oxygen levels

The nanomedicine, SHp-RBC-NP targeted ischemic area and improved neuro-scores and infarct volume

(continued)

Amani et al. (2019)

Shi et al. (2020)

Lv et al. (2018)

The formulated nanomedicine, Liu et al. (2013a, b) CBSA-PEG-TIIA-NPs lessened infarct volume and neural symptoms

Reduce C3 expression in Wang et al. (2018b) microglial cells as well as ischemic brain areas; decrease the infiltration of inflammatory cells and level of proinflammatory mediators

Treatment effects

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis 225

Functionalized carbon nanotubes (f-CNT)

Allotropic dissimilarity of carbon atom (Fullerenol)

24

25

Platinum nanoparticle (nPt)

23

Carbon-Based NPs

PEGylated-ceria nanoparticles

22

Name of nanomedicines

PLGA functionalized magnetic Fe3 O4 NP

Broad Classification of Nanoparticles

21

Sl. no

Table 4 (continued)

Stereotaxic injection

i.p

i.v

i.v

–

Drug delivery mode

Treatment effects

Rats

Rats

Mice

Rats

Kim et al. (2012)

Lu et al. (2021)

References

Knockdown of Caspase-3 gene by f-CNT in neuronal tissues of brain to produce neuroprotective effect

Al-Jamal et al. (2011)

Decrease infarct area; decrease Sarami Foroshani IL-6 and MMP-9 et al. (2018) transcriptional levels to protect blood–brain barrier integrity; prevention of brain edema post cerebral I/R injury

nPt decreased ROS generation Takamiya et al. and enhanced neurological (2011), Takamiya symptoms, et al. (2012) nPt prevents ischemic damage to brain by decreasing MMP-9 activity and neurovascular unit disruption

ROS scavenging and decrease ischemic brain injury

Human fibroblast cell Effective drug loading rate; line control release efficacy of nanoparticles

Modeling

226 S. K. Das et al.

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

227

5.1 Emerging Role of Nanoparticles for Ischemic Stroke Therapy As most of the available treatment strategies are associated with poor stability in the blood vascular system, formulated nanoparticles with their potential applications and compensate for the limitations associated with available treatment strategies. The most important approach towards the development of nanoparticles-based drug delivery systems for the treatment of cerebral ischemia resides by inhibiting proinflammatory cytokines production, lipid peroxidation, immune cell infiltration and apoptosis (Kyle and Saha 2014; Sarmah et al. 2017). Earlier reports have described four important nanoparticles-based drug delivery systems for the treatment and management of cerebral ischemia such as liposome, polymeric, biomimetic, inorganic and carbon-based nanoparticles (Lin et al. 2022) (Fig. 5). Besides, several other nanoparticles-based drug delivery systems are available for treatment and management of cerebral ischemia such as antioxidant drug-loaded and excitotoxicity inhibitors-loaded nanoparticles (Guan et al. 2018; Hou et al. 2019; Li et al. 2019a, b; Zhong et al. 2019; Yuan et al. 2021). These nanoparticles are primarily used as drug delivery vehicles for the shipment of drugs spatiotemporally and in a controlled manner to the site of ischemic stroke for the restoration of biological functions.

5.1.1

Liposome Nanoparticles

Lipid-based (liposome) nanoparticles were developed as the first generation of nanoparticles-based drug delivery systems with an objective to increase the delivery load and decrease the ejection load of targeted drugs. Further, liposome nanoparticles are provided with the added advantage of being biodegradable and biocompatible to protect drugs from enzymatic degradation, thus can effortlessly cross BBB with an high loading capacity of drugs (Joshi et al. 2019). Due to improved permeability and retaining effect, liposome nanoparticles can easily seepage through brain parenchyma to get accumulated in the ischemic area during the acute phase of cerebral ischemia. The neuroprotective agents such as FK506 and cyclosporin A were tested for liposome nanoparticles-based drug delivery and were found to be effective in reducing infarct size, leucocyte infiltration and TNF-α expression in animal models (Fukuta et al. 2015; Partoazar et al. 2017). Further report also demonstrated the use of asialoerythropoietin (AEPO)-modified PEGylated liposomes (AEPO-liposomes) for the treatment and management of cerebral I/R injury. The AEPO-liposomes were found to be retained and accumulated in the ischemic area over 24 h after administration and associated with reduced infarct size and neural apoptosis in MCAO rat model (Ishii et al. 2012). Another finding also demonstrated that the blood–brain permeability of highly effective neuroprotectant ZL006 for the treatment of ischemic stroke was increased by loading to T7-conjugated PEGylated liposomes (Zhou et al. 2010; Wang et al. 2015). As tPA treatment is associated with a narrow therapeutic time

228

S. K. Das et al.

window of 4.5 h, a recent study was conducted on rat model by combining tPA and liposomal fasudil (fasudil-Lip) to upsurge the therapeutic advantage of tPA. The findings showed that joint therapy upsurges the neuroprotective efficacy as compared to independent administration of tPA or fasudil-Lip against cerebral I/R injury. Additionally, the study also demonstrated that fasudil-Lip administration decreases the cerebral hemorrhage and thus protracted the therapeutic window of tPA (Fukuta et al. 2017).

5.1.2

Polymeric Nanoparticles

Polymeric nanoparticles as drug delivery vehicles represent a promising option for the treatment of brain-related pathophysiological outcomes due to their essential properties such as biocompatibility, biodegradability, non-toxicity, controlled drug release and regulated architecture (Masserini 2013; Li et al. 2021). The polymers used to make polymeric nanoparticles involve both natural and synthetic polymers. The natural polymers (e.g., chitosan) mostly include amino acids, proteins and polysaccharides whereas the most common synthetic polymers include polyester lactic acid (PLA), poly-n-butylcyanoacrylate (PBCA), polyethylene glycol (PEG)-PLA and poly lactic-co-glycolic acid (PLGA). The polymeric nanoparticles having wellmarked pharmacokinetics and great surface modification potential are the preferable nanoparticles for the development of drug delivery systems (El-Say and El-Sawy 2017). Among these, PGLA is the most commonly used for the preparation of polymeric nanoparticles-based drug delivery tools. An earlier report demonstrated that cationic bovine serum albumin-conjugated tanshinone IIA PEGylated nanoparticles (CBSA-PEG-TIIA-NPs) are effective for the treatment of cerebral I/R injury in rat model by reduction of inflammatory response, infarction size, neutrophil infiltration and neural apoptosis (Liu et al. 2013a, b; Li et al. 2021). Earlier reports also demonstrated that polymeric nanoparticles were modulated with reactive oxygen species (ROS)-reactive application to regulate drug release in the affected area of the brain following cerebral ischemia (Lv et al. 2018). This study showed that NR2B9C, a potent neuroprotectant conjugated to nanoparticle (SHp-RBC-NPs) having ROS responsive boronic ester, triggers the release of NR2B9C in response to high ROS levels in ischemic brain regions and found to be effective in reducing infarction volume, oxidative damage and neuronal apoptosis.

5.1.3

Biomimetic Nanoparticles

Having the exogenous characteristics, most of the nanoparticles used for therapeutic purposes are always at risk of being cleared by the reticuloendothelial system. To negate this limitation, biomimetic nanoparticles-based drug delivery systems such as cell itself, cell membrane vesicles and exosomes could be possible alternatives (Chen et al. 2022). Additionally, biomimetic nanoparticles pose an advanced drug delivery cum targeting ability and biosafety. Natural nanocarriers found in biological

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

229

systems can be modified to deliver targeted drugs through biomimetic nanoparticles (Sabu et al. 2018). Among these, direct administration of exogenous stem cells is found to be a biomimetic nanoparticle-based therapy for stroke models. Several stem cell types have been documented to be used through intravenous administration in stroke (Tan et al. 2018; Shi et al. 2020). On the other hand, cell membrane coating has currently arisen as a hopeful biomimetic method for bioengineered nanoparticles for effective drug delivery. Recent reports advocated that the delivery of nanoparticles to the ischemic brain area is upsurged by surface coating with the cell membrane of neural stem cells (Ma et al. 2019a, b). Alternatively, exosomes derived from stem cells loaded with biologically active molecules can be used for cerebral ischemia-instigated infarction. In this regard, exosomes generated from adipose-derived mesenchymal stem cells (ADSCs) administration are associated with substantial neurological recovery following brain injury (Chen et al. 2022).

5.1.4

Inorganic Nanoparticles

Inorganic metallic nanoparticles mostly play their pivotal role as ROS scavengers. ROS is primarily involved in oxidative damage of tissue and organ and is the basis of brain injury following ischemic stroke. Silica and iron oxides are the most preferred inorganic materials used for the preparation of inorganic nanoparticles and can be used both for diagnosis and therapy as being able to track by MRI (Chen et al. 2022). Earlier reports showed that Se-nanoparticles provide substantial protection in the stroke model of murine by reducing inflammation and apoptosis (Amani et al. 2019). Platinum nanoparticles are innovative ROS scavengers for their bulky surface area and electron density which helps in neutralizing ROS (Watanabe et al. 2009). Similarly, ceria nanoparticles have been found to provide substantial neuroprotection against cerebral ischemia-persuaded ROS elevation and apoptosis (Kim et al. 2012).

5.1.5

Carbon-Based Nanoparticles

The wide and diverse range of medical usage of carbon-based nanomaterials resides with respect to its unique physicochemical properties and diverse structural possibilities of nanostructures (Loh et al. 2018). These properties are mainly attributed to the special electron configuration and electrons movement at discrete energy levels in nanostructures. Carbon-based nanoparticles are characterized into the following types based on their unique structures such as carbon nanotubes, nanodiamonds, graphene, fullerenes and mesoporous carbon (Zhang et al. 2017). With remarkable multifunctional surface area, these nanoparticles exemplified enhanced biocompatibility, advanced drug payload capacity and minimal immunological reactions (Mohajeri et al. 2018). The biological applications of carbon-based nanoparticles can be enhanced by chemical modifications of specific moieties. However, most of the nanoparticles come up with several side effects and toxicity, thus limiting their potential applications.

230

S. K. Das et al.

6 Novel Tools Towards Early Diagnosis of Cerebral Ischemia and Implication of Nanotechnology Proper diagnosis is considered as the first step towards the formulation of the right treatment strategies and effective recovery from a number of life threatening diseases including cerebral ischemia. With the currently available diagnostic tools for cerebral ischemia in the form of tPA administration and mechanical thrombectomy, the narrow time window limits the usage of the right treatment and thus increases mortality. However, few advances have been prescribed with respect to CT scan for specific types of cerebral ischemia. For correct diagnosis of arterial stenosis and occlusion, CT angiography is considered as an extremely accurate tool (Koelemay et al. 2004). On the other hand, CT perfusion is a much better radiological diagnostic tool for ischemic stroke than that of CT angiography and non-contrast CT (van Seeters et al. 2013). Despite this advancement, these diagnostic techniques have some limitations, e.g., CT perfusion and CT angiography scanning involve the requirement of iodine contrast agents, which were reported as high-risk factors for acute kidney injury in addition to several other side effects (Davenport et al. 2009; Tsai et al. 2014; McCullough et al. 2016). On the other hand, MRI poses an innate benefit over CT scan by providing several options to evaluate diverse structural and functional aspects of different tissues and regions of the brain through perfusion imaging, diffusion imaging, etc. For correct diagnosis of acute cerebral ischemia, diffusion-weighted imaging (DWI) is considered as the most sensitive imaging tool as it records the actual change in water diffusion in affected brain regions (Alegiani et al. 2017). MRI also works on a similar principle to a CT scan in that it utilizes intravenous gadolinium as a contrast agent during tracking through brain circulation. An obstructed perfusion or hypoperfused brain regions will generate a scattered, reduced and/or delayed time series of signal intensity of MRI in comparison to a healthy brain region or tissue. Despite having the accuracy of acute cerebral ischemia diagnosis, MRI also has limitation and disadvantages, such as expensive, slow scanning-examination procedure and inclined to generate artifacts due to movements of body parts during scanning (Kim et al. 2019). Taking into account the available tools, it can be inferred that they are not equipped with paramount requirements associated with the diagnosis of cerebral ischemia such as early detection, BBB permeability, precise binding, clear difference and simultaneous monitoring of ischemic treatment. Further, few contrast agents are difficult or not able to pass through BBB and thus pose a significant limitation to MRI (Zhang et al. 2021). Therefore, revitalization and improvement of these orthodox imaging tools is a paramount requirement in recent decades. An active surveillance of ischemic stroke severity is clinically important and thus a real-time imaging of thrombus provides several advantages to clinicians such as visual identification of thrombus load, their localization and leads for undergoing immediate mechanical thrombectomy (Lin et al. 2022). In this regard, nanotechnology emerge as a ground-breaking technology towards diagnosis and drug delivery for the treatment of a wide range of diseases. Nanomaterials could possibly be used for the effective delivery of drugs across BBB, site-specific drug release, prolonged

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

231

drug circulation time and augmented drug accumulation at the ischemic site. Further, nanomaterials can also be used as an effective diagnostic tool to detect several biomarkers for early-stage cerebral ischemia. Recent advancements in nanotechnology are primarily pursued for the accurate detection of thrombolysis and recanalization in the affected brain regions/tissues after cerebral ischemia. Firstly, the structured nanoparticles bind to the thrombus site and thus real-time tracking of thrombolysis is possible by imaging in the early stage of thrombosis. In this regard, polymorphonuclear leucocytes (neutrophils) are the initial and furthermost abundant inflammatory cells that appear on site towards a thrombogenic response to ischemic stroke. Thus, neutrophils are considered as the vital constituent of the thrombogenic process, thus adhering nanoparticles to neutrophils can be utilized for thrombus targeting for early diagnosis of cerebral ischemia (Tang et al. 2019). As neutrophils are considered as the primary player towards the pathophysiology of acute cerebral ischemia by their unchecked infiltration to cerebral ischemic regions/tissues leading to the generation of reactive oxygen species (ROS) and later act as the major contributor to cerebral reperfusion injury, neutrophil targeting for early diagnosis, treatment and subsequent prognosis will be considered as the major site for the management of cerebral ischemia. Therefore, checking the adherence of inflammatory neutrophils to the endothelial cells is considered as an beneficial therapeutic strategy by reducing neutrophils infiltration and inflammation against acute cerebral ischemia. In this regard, β2-integrin, a complex WBCs adhesion molecule required for leukocyte trafficking, endothelial adhesion and infiltration, phagocytosis, ROS generation and T-cell activation, plays a vital role in neutrophils infiltration during acute ischemic stroke. Targeting β2-integrin is the paramount requisite to make sustained inflammation under check during the early stage of ischemic stroke. Piceatannol, a selective spleen tyrosine kinase inhibitor, has been demonstrated to block β2-integrin signaling in neutrophils, thereby preventing the adhesion of neutrophils to endothelial cells; however, limitation resides with respect to the selective delivery of piceatannol to inflammatory neutrophils. Therefore, biodegradable nanoparticles might act as important therapeutic and diagnostic targets to be used as carrier systems for the delivery of drugs to the target cells in the affected area of ischemic stroke. Due to specific cell–cell recognition that happens between blood platelets and neutrophils, platelet-mimetic nanoparticles (PTNPs) have been developed and shown promising benefits as direct recognition, inhibition and real-time monitoring of inflammatory neutrophils in the treatment of acute ischemic stroke. For selective identification and therapeutic intervention of adherent inflammatory neutrophils, PTNPs were combined with piceatannol and T2 contrast agent, i.e., superparamagnetic iron oxide (SPIO). This combination is effective in the selective delivery of piceatannol to adherent inflammatory neutrophils and thus detached them from endothelial cells into the circulation, thereby plummeting neutrophil infiltration and infarct size after acute cerebral ischemia. Further, utilizing the properties of SPIO, a real-time monitoring of inflammatory neutrophils migration and associated therapeutic effects of acute cerebral ischemia is possible with the application of MRI (Tang et al. 2019).

232

S. K. Das et al.

On the other hand, ultrasonography is a cost-effective diagnosis mode with medical applications; however, it shortages specificity for analysis and diagnosis of early ischemic stroke. However, advancement in the field of nanotechnology has overcome these short comings of ultrasound with the usage of specifically designed nanomaterials. Earlier findings utilized platelet membrane-derived biomimetic nanobubbles (PNBs) for early diagnosis and timely perfusion intervention in acute ischemic stroke (Li et al. 2018a, b). As a part of treatment, PNBs exhibit accurate infarct targeting capability due to the presence of natural protein and lipid components derived from the membrane of platelets and thus enhanced the process of microvascular recanalization of obstructed cerebral vessels. As PNBs get accumulated on the site of ischemic stroke areas, a real-time monitoring is possible by utilizing ultrasound imaging. Additionally, a recent report showed that Xe-encapsulated nanobubbles (Xe-NBs) are effective in the early diagnosis of ischemic stroke by producing ultrasound contrast imaging with infarct regions in the animal model (Jin et al. 2021). However, several advancement in the field of nanotechnology is still resides mostly as preclinical validations and its utilization as a prominent diagnostic tools is limited.

7 Current Status on Hypoxia Responsive Nanomaterials for Early Diagnosis of Cerebral Ischemia Cerebral ischemia is considered as one of the most prevalent neurological diseases with a high rate of mortality and post-treatment complications. As cerebral ischemia is associated with thrombus-mediated reduction in blood flow to the affected brain areas, subsequently leading to a lack of oxygen supply (i.e., hypoxia) and nutrient supply to the brain. A sustained deprivation of food and oxygen supply to the affected brain regions following ischemic stroke leads to neuronal death by accelerated necrosis and apoptosis forming a core brain infarct area surrounded by penumbra. Therefore, hypoxia is identified as the hallmark of cerebral ischemic areas and can be considered as a prognosis factor for real-time monitoring of infarct progression. With ischemic stroke being primarily associated with a hypoxic environment, a hypoxia responsive diagnostic tool will be beneficial in the early diagnosis of various types of cerebral ischemia. The reduced level of oxygen in the cytosolic environment (hypoxic microenvironment) has resulted in the profound build-up of FADH2 and NADH and later leads to a further reduction in cytosolic oxygen level and augmented formation of ROS, thereby producing more detrimental effect following ischemic stroke. As a compensatory measure to the hypoxia following acute cerebral ischemia, anaerobic glycolysis is predominant leading to acidosis in the extracellular environment. Additionally, several cytosolic enzymes of oxidoreductive pathways were upregulated following hypoxia. Theses factors must be taken into account to target for designing of hypoxia responsive diagnostic tools for early diagnosis of cerebral ischemia. For hypoxia imaging, several quinones and nitroaromatic derivatives with

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

233

hypoxia responsive components were added in the construction of smart diagnostic agents (Kiyose et al. 2010). Out of these, 2-nitroimidazoles were the most extensively used moiety for the preparation of hypoxia responsive nanoparticles (HR-NPs) due to their high sensitivity to hypoxia (Chu et al., 2007; Takasawa et al. 2008; Okuda et al. 2012; Li et al. 2020). Mechanistically, the hydrophobic 2-nitroimidazole is converted to hydrophilic 2-aminoimidazole through sequential events of cellular bio-reduction accompanied by cascades of bio-reductases and the latter is highly reactive to macromolecules in hypoxic tissue/areas (Hodgkiss et al. 1992). Therefore, 2-nitroimidazole can be considered as an effective hypoxia responsive marker cum carrier for the diagnosis and therapeutic of cerebral ischemia. In this context, designing a nanoparticle conjugated to hypoxia responsive ligand along with the target drug can identify the ischemic stroke regions and potentiate the selective drug delivery to minimize the side effects and maximized the therapeutic effects. However, a typical approach for the development of hypoxia responsive nanomaterials for the diagnosis of ischemic stroke must be focused on the effective sensitivity of nanoparticles for hypoxia, targeting the ability for hypoxia inducible factor and good circulation time. Therefore, the basic principle to fabricate HR-NPs depends upon the ligation of moieties to NPs that are acting as substrates for overexpressed cellular enzymes following hypoxia. In this context, nitro-compounds and azobenzene compounds are the commonly used moieties for the preparation of HR-NPs against hypoxia-triggered disease conditions. To the best of our knowledge, usage of these targets is restricted to a few diseases including cancer diagnosis and therapy. Therefore, the hypoxia responsive properties of 2-nitroimidazole following its hydrophobic-hydrophilic biotransformation can be used for the generation of varieties of nanomaterial-based diagnostic cum therapeutic targets against cerebral ischemia. Additionally, exosomes as extracellular vesicles in the circulatory system act as cellular signal transducers in the body and can be targeted for the diagnosis and treatment of ischemic stroke (Li et al. 2019a, b; Lee et al. 2022). In this context, several studies have demonstrated the usage of biomimetic exosomes in effective diagnosis, therapy and prognosis of ischemic stroke in animal models. Earlier reports also advocated that plasma exosomes are typically enriched in hypoxia inducible transcription factor 1α (HIF-1α) and thus can be targeted for early diagnosis and development of an effective drug delivery system as a nanocarrier for the treatment of ischemic stroke. To completely understand the potential applications of selective targets for making up of hypoxia responsive nanomaterials towards early diagnosis of cerebral ischemia, development of innovative ligands with a quicker delivery to responsive tissue and their effective binding, effective circulation time, biodegradable ability and non-toxic to the cellular environment are central goals for forthcoming research.

234

S. K. Das et al.

8 Conclusion In past decades, several studies both in vitro and in vivo have been conducted to give effective strategies for early diagnosis, effective treatment and prognosis of cerebral ischemia. However, the available treatment strategies and diagnostic parameters utilizing the principle of nanotechnology were not clinically validated. In this context, in recent years, scientific investigation is focused on the designing of HRNPs, their toxicity to normal cells, their circulation time, penetration ability, hypoxiaprompted drug release and ischemic stroke targeting. To the best of our knowledge, currently HR-NPs are clinically validated for early diagnosis, treatment and prognosis of cerebral ischemia. Therefore, the unique biocompatibility, high sensitivity for diagnosis, high competence for targeted drug delivery and high therapeutic efficiency of HR-NPs are presently a hot research area in the field of nanomedicines for the development of the most effective diagnostic and therapeutic tools for cerebral ischemia. Acknowledgements The authors acknowledge the SOA (Deemed to be University) and Institute of Life Sciences for providing the infrastructure facility and support. The authors also acknowledge the contribution of Dr. Ritendra Mishra, Mumbai, India, who helped in the proofreading and copyediting of the manuscript. Figures were produced with the assistance of Biorender (https://app.bio render.com).

References Aabdallah DM, Eid NI (2004) Possible neuroprotective effects of lecithin and alpha-tocopherol alone or in combination against ischemia/reperfusion insult in rat brain. J Biochem Mol Toxicol 18:273–278 Adhami F, Liao G, Morozov YM, Schloemer A, Schmithorst VJ, Lorenz JN, Dunn RS, Vorhees CV, Wills-Karp M, Degen JL, Davis RJ, Mizushima N, Rakic P, Dardzinski BJ, Holland SK, Sharp FR, Kuan CY (2006) Cerebral ischemia-hypoxia induces intravascular coagulation and autophagy. Am J Patho 169:566–583 Ahad MA, Kumaran KR, Ning T, Mansor NI, Effendy MA, Damodaran T, Lingam K, Wahab HA, Nordin N, Liao P, Müller CP, Hassan Z (2020) Insights into the neuropathology of cerebral ischemia and its mechanisms. Rev Neurosci 31:521–538 Al-Ahmady ZS, Jasim D, Ahmad SS, Wong R, Haley M, Coutts G, Schiessl I, Allan SM, Kostarelos K (2019) Selective liposomal transport through blood brain barrier disruption in ischemic stroke reveals two distinct therapeutic opportunities. ACS Nano 13:12470–12486 Alegiani AC, MacLean S, Braass H, Siemonsen S, Gerloff C, Fiehler J, Cho TH, Derex L, Hermier M, Berthezene Y, Nighoghossian N, Thomalla G (2017) Comprehensive analysis of early fractional anisotropy changes in acute ischemic stroke. PLoS ONE 12:e0188318 Al-Jamal KT, Gherardini L, Bardi G, Nunes A, Guo C, Bussy C, Herrero MA, Bianco A, Prato M, Kostarelos K, Pizzorusso T (2011) Functional motor recovery from brain ischemic insult by carbon nanotube-mediated siRNA silencing. Proc Natl Acad Sci U S A 108:10952–10957 Amani H, Habibey R, Shokri F, Hajmiresmail SJ, Akhavan O, Mashaghi A, Pazoki-Toroudi H (2019) Selenium nanoparticles for targeted stroke therapy through modulation of inflammatory and metabolic signaling. Sci Rep 9:6044

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

235

Baron JC (2011) Perfusion thresholds in human cerebral ischemia: historical perspective and therapeutic implications. Cerebrovasc Dis 11(Suppl 1):2–8 Beal CC (2010) Gender and stroke symptoms: a review of the current literature. J Neurosci Nurs 42:80–87 Bian HJ, Xu SY, Li HQ, Jia JQ, Ye L, Shu S, Xia SN, Gu Y, Zhu X, Xu Y, Cao X (2021) JLX001 ameliorates cerebral ischemia injury by modulating microglial polarization and compromising NLRP3 inflammasome activation via the NF-κB signaling pathway. Int Immunopharmacol 101:108325 Brazzelli M, Sandercock PA, Chappell FM, Celani MG, Righetti E, Arestis N, Wardlaw JM, Deeks JJ (2009) Magnetic resonance imaging versus computed tomography for detection of acute vascular lesions in patients presenting with stroke symptoms. Cochrane Database Syst Rev. 4:CD007424 Brekenfeld C, Gralla J, Zubler C, Schroth G (2012) Mechanical thrombectomy for acute ischemic stroke. Rofo 184:503–512 Burchfiel CM, Curb JD, Rodriguez BL, Abbott RD, Chiu D, Yano K (1994) Glucose intolerance and 22-year stroke incidence. The Honolulu Heart Program. Stroke 25:951–957 Chalela JA, Kidwell CS, Nentwich LM, Luby M, Butman JA, Demchuk AM, Hill MD, Patronas N, Latour L, Warach S (2007) Magnetic resonance imaging and computed tomography in emergency assessment of patients with suspected acute stroke: a prospective comparison. Lancet 369:293–298 Chen J, Jin J, Li K, Shi L, Wen X, Fang F (2022) Progresses and prospects of neuroprotective agents-loaded nanoparticles and biomimetic material in ischemic stroke. Front Cell Neurosci 16:868323 Cheng HY, Wang YS, Hsu PY, Chen CY, Liao YC, Juo SH (2019) miR-195 has a potential to treat ischemic and Hemorrhagic stroke through neurovascular protection and neurogenesis. Mol Ther Methods Clin Dev 13:121–132 Chu T, Li Z, Liu X, Wang X (2007) In vivo evaluation of the nitroimidazole-based thioflavin-T derivatives as cerebral ischemia markers. Int J Biomed Imaging 2007:49791 Davenport MS, Cohan RH, Caoili EM, Ellis JH (2009) Repeat contrast medium reactions in premedicated patients: frequency and severity. Radiology 253:372–379 Dong X, Gao J, Su Y, Wang Z (2020) Nanomedicine for ischemic stroke. Int J Mol Sci 21:7600 Dong X, Gao J, Zhang CY, Hayworth C, Frank M, Wang Z (2019) Neutrophil membrane-derived Nanovesicles alleviate inflammation to protect mouse brain injury from ischemic stroke. ACS Nano 13:1272–1283 El-Say KM, El-Sawy HS (2017) Polymeric nanoparticles: promising platform for drug delivery. Int J Pharm 528:675–691 Feldstein CA (2014) Early treatment of hypertension in acute ischemic and intracerebral hemorrhagic stroke: progress achieved, challenges, and perspectives. J Am Soc Hypertens 8:192–202 Fernandes LF, Bruch GE, Massensini AR, Frézard F (2018) Recent advances in the therapeutic and diagnostic use of liposomes and carbon nanomaterials in ischemic stroke. Front Neurosci 12:453 Fukuta T, Asai T, Yanagida Y, Namba M, Koide H, Shimizu K, Oku N (2017) Combination therapy with liposomal neuroprotectants and tissue plasminogen activator for treatment of ischemic stroke. FASEB J 31:1879–1890 Fukuta T, Ishii T, Asai T, Sato A, Kikuchi T, Shimizu K, Minamino T, Oku N (2015) Treatment of stroke with liposomal neuroprotective agents under cerebral ischemia conditions. Eur J Pharm Biopharm 97:1–7 Gu Y, Zhou C, Piao Z, Yuan H, Jiang H, Wei H, Zhou Y, Nan G, Ji X (2022) Cerebral edema after ischemic stroke: Pathophysiology and underlying mechanisms. Front Neurosci 16:988283 Guan D, Li Y, Peng X, Zhao H, Mao Y, Cui Y (2018) Thymoquinone protects against cerebral small vessel disease: Role of antioxidant and anti-inflammatory activities. J Biol Regul Homeost Agents 32:225–231

236

S. K. Das et al.

Hodgkiss RJ, Middleton RW, Parrick J, Rami HK, Wardman P, Wilson GD (1992) Bioreductive fluorescent markers for hypoxic cells: a study of 2-nitroimidazoles with 1-substituents containing fluorescent, bridgehead-nitrogen, bicyclic systems. J Med Chem 35:1920–1926 Hosseinzadeh H, Nassiri Asl M, Parvardeh S (2005) The effects of carbenoxolone, a semisynthetic derivative of glycyrrhizinic acid, on peripheral and central ischemia-reperfusion injuries in the skeletal muscle and hippocampus of rats. Phytomedicine 12:632–637 Hosseinzadeh H, Parvardeh S, Asl MN, Sadeghnia HR, Ziaee T (2007) Effect of thymoquinone and Nigella sativa seeds oil on lipid peroxidation level during global cerebral ischemia-reperfusion injury in rat hippocampus. Phytomedicine 14:621–627 Hou J, Yang X, Li S, Cheng Z, Wang Y, Zhao J, Zhang C, Li Y, Luo M, Ren H, Liang J, Wang J, Wang J, Qin J (2019) Accessing neuroinflammation sites: Monocyte/neutrophil-mediated drug delivery for cerebral ischemia. Sci Adv 5:eaau8301 Hu XM, Zhou MM, Hu XM, Zeng FD (2005) Neuroprotective effects of scutellarin on rat neuronal damage induced by cerebral ischemia/reperfusion. Acta Pharmacol Sin 26:1454–1459 Ishii T, Asai T, Oyama D, Fukuta T, Yasuda N, Shimizu K, Minamino T, Oku N (2012) Amelioration of cerebral ischemia-reperfusion injury based on liposomal drug delivery system with asialoerythropoietin. J Control Release 160:81–87 Jin J, Li M, Li J, Li B, Duan L, Yang F, Gu N (2021) Xenon Nanobubbles for the imageguided Preemptive treatment of acute ischemic stroke via neuroprotection and microcirculatory restoration. ACS Appl Mater Interfaces 13:43880–43891 Joshi MD, Prabhu RH, Patravale VB (2019) Fabrication of nanostructured lipid carriers (NLC)based gels from microemulsion template for delivery through skin. Methods Mol Biol 2000:279– 292 Kim CK, Kim T, Choi IY, Soh M, Kim D, Kim YJ, Jang H, Yang HS, Kim JY, Park HK, Park SP, Park S, Yu T, Yoon BW, Lee SH, Hyeon T (2012) Ceria nanoparticles that can protect against ischemic stroke. Angew Chem Int Ed Engl 51:11039–11043 Kim H, Irimia A, Hobel SM, Pogosyan M, Tang H, Petrosyan P, Blanco REC, Duffy BA, Zhao L, Crawford KL, Liew SL, Clark K, Law M, Mukherjee P, Manley GT, Van Horn JD, Toga AW (2019) The LONI QC system: a semi-automated, web-based and freely-available environment for the comprehensive quality control of neuroimaging data. Front Neuroinform 13:60 Kiyose K, Hanaoka K, Oushiki D, Nakamura T, Kajimura M, Suematsu M, Nishimatsu H, Yamane T, Terai T, Hirata Y, Nagano T (2010) Hypoxia-sensitive fluorescent probes for in vivo real-time fluorescence imaging of acute ischemia. J Am Chem Soc 132:15846–15848 Koelemay MJ, Nederkoorn PJ, Reitsma JB, Majoie CB (2004) Systematic review of computed tomographic angiography for assessment of carotid artery disease. Stroke 35:2306–2312 Kurth T, Heuschmann PU, Walker AM, Berger K (2007) Mortality of stroke patients treated with thrombolysis: analysis of nationwide inpatient sample. Neurology 68:710 Kyle S, Saha S (2014) Nanotechnology for the detection and therapy of stroke. Adv Healthc Mater 3:1703–1720 Lee EC, Ha TW, Lee DH, Hong DY, Park SW, Lee JY, Lee MR, Oh JS (2022) Utility of exosomes in ischemic and hemorrhagic stroke diagnosis and treatment. Int J Mol Sci 23:8367 Li C, Sun T, Jiang C (2021) Recent advances in nanomedicines for the treatment of ischemic stroke. Acta Pharm Sin B 11:1767–1788 Li G, Morris-Blanco KC, Lopez MS, Yang T, Zhao H, Vemuganti R, Luo Y (2018a) Impact of microRNAs on ischemic stroke: From pre- to post-disease. Prog Neurobiol 163–164:59–78 Li M, Li J, Chen J, Liu Y, Cheng X, Yang F, Gu N (2020) Platelet membrane biomimetic magnetic Nanocarriers for targeted delivery and in situ generation of nitric oxide in early ischemic stroke. ACS Nano 14:2024–2035 Li M, Liu Y, Chen J, Liu T, Gu Z, Zhang J, Gu X, Teng G, Yang F, Gu N (2018b) Platelet bionanobubbles as microvascular recanalization nanoformulation for acute ischemic stroke lesion theranostics. Theranostics 8:4870–4883

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

237

Li R, Huang Y, Chen L, Zhou H, Zhang M, Chang L, Shen H, Zhou M, Su P, Zhu D (2019a) Targeted delivery of intranasally administered nanoparticles-mediated neuroprotective peptide NR2B9c to brain and neuron for treatment of ischemic stroke. Nanomedicine 18:380–390 Li S, Meng Q, Zhang L (1999) Experimental therapy of a platelet-activating factor antagonist (ginkgolide B) on photochemically induced thrombotic cerebral ischaemia in tree shrews. Clin Exp Pharmacol Physiol 26:824–825 Li Y, Ren C, Li H, Jiang F, Wang L, Xia C, Ji X (2019b) Role of exosomes induced by remote ischemic preconditioning in neuroprotection against cerebral ischemia. NeuroReport 30:834–841 Liang TY, Peng SY, Ma M, Li HY, Wang Z, Chen G (2021) Protective effects of sevoflurane in cerebral ischemia reperfusion injury: a narrative review. Med Gas Res 11:152–154 Lin X, Li N, Tang H (2022) Recent advances in nanomaterials for diagnosis, treatments, and neurorestoration in ischemic stroke. Front Cell Neurosci 16:885190 Liu J, Ma W, Zang CH, Wang GD, Zhang SJ, Wu HJ, Zhu KW, Xiang XL, Li CY, Liu KP, Guo JH, Li LY (2021) Salidroside inhibits NLRP3 inflammasome actiation and apoptosis in microglia induced by cerebral ischemia/reperfusion injury by inhibiting the TLR4/NF-κB signaling pathway. Ann Transl Med 9:1694 Liu X, An C, Jin P, Liu X, Wang L (2013a) Protective effects of cationic bovine serum albuminconjugated PEGylated tanshinone IIA nanoparticles on cerebral ischemia. Biomaterials 34:817– 830 Liu X, Wen S, Yan F, Liu K, Liu L, Wang L, Zhao S, Ji X (2018) Salidroside provides neuroprotection by modulating microglial polarization after cerebral ischemia. J Neuroinflammation 15:39 Liu X, Ye M, An C, Pan L, Ji L (2013b) The effect of cationic albumin-conjugated PEGylated tanshinone IIA nanoparticles on neuronal signal pathways and neuroprotection in cerebral ischemia. Biomaterials 34:6893–6905 Liu Y, Ai K, Ji X, Askhatova D, Du R, Lu L, Shi J (2017) Comprehensive insights into the multiantioxidative mechanisms of melanin nanoparticles and their application to protect brain from injury in ischemic stroke. J Am Chem Soc 139:856–862 Logvinov SV, Pugachenko NV, Potapov AV, Krasnov EA, Plotnikov MB, Maslov MY, Aliev OI, Tyukavkina NA (2001) Ischemia-induced changes in synaptoarchitectonics of brain cortex and their correction with ascovertin and Leuzea extract. Bull Exp Biol Med 132:1017–1020 Loh KP, Ho D, Chiu GNC, Leong DT, Pastorin G, Chow EK (2018) Clinical applications of carbon nanomaterials in diagnostics and therapy. Adv Mater 30:e1802368 Lu X, Zhang Y, Wang L, Li G, Gao J, Wang Y (2021) Development of L-carnosine functionalized iron oxide nanoparticles loaded with dexamethasone for simultaneous therapeutic potential of blood brain barrier crossing and ischemic stroke treatment. Drug Deliv 28:380–389 Lubos E, Handy DE, Loscalzo J (2008) Role of oxidative stress and nitric oxide in atherothrombosis. Front Biosci 13:5323–5344 Lukyanova LD, Kirova YI (2015) Mitochondria-controlled signaling mechanisms of brain protection in hypoxia. Front Neurosci 9:320 Lv S, Liu H, Wang H (2021) The interplay between autophagy and NLRP3 Inflammasome in ischemia/reperfusion injury. Int J Mol Sci 22:8773 Lv W, Xu J, Wang X, Li X, Xu Q, Xin H (2018) Bioengineered boronic ester modified dextran polymer nanoparticles as reactive oxygen species responsive nanocarrier for ischemic stroke treatment. ACS Nano 12:5417–5426 Ma C, Wang X, Xu T, Yu X, Zhang S, Liu S, Gao Y, Fan S, Li C, Zhai C, Cheng F, Wang Q (2019a) Qingkailing injection ameliorates cerebral ischemia-reperfusion injury and modulates the AMPK/NLRP3 Inflammasome Signalling pathway. BMC Complement Altern Med 19:320 Ma J, Zhang S, Liu J, Liu F, Du F, Li M, Chen AT, Bao Y, Suh HW, Avery J, Deng G, Zhou Y, Wu P, Sheth K, Wang H, Zhou J (2019b) Targeted drug delivery to stroke via chemotactic recruitment of nanoparticles coated with membrane of engineered neural stem cells. Small 15:e1902011 Manzanero S, Santro T, Arumugam TV (2013) Neuronal oxidative stress in acute ischemic stroke: sources and contribution to cell injury. Neurochem Int 62:712–718 Masserini M (2013) Nanoparticles for brain drug delivery. ISRN Biochem 2013:238428

238

S. K. Das et al.

McCullough PA, Choi JP, Feghali GA, Schussler JM, Stoler RM, Vallabahn RC, Mehta A (2016) Contrast-induced acute kidney injury. J Am Coll Cardiol 68:1465–1473 Mergenthaler P, Dirnagl U, Meisel A (2004) Pathophysiology of stroke: lessons from animal models. Metab Brain Dis 19:151–167 Miao YF, Peng T, Moody MR, Klegerman ME, Aronowski J, Grotta J, McPherson DD, Kim H, Huang SL (2018) Delivery of xenon-containing echogenic liposomes inhibits early brain injury following subarachnoid hemorrhage. Sci Rep 8:450 Mohajeri M, Behnam B, Sahebkar A (2018) Biomedical applications of carbon nanomaterials: Drug and gene delivery potentials. J Cell Physiol 234:298–319 Mohammadi MR, Nojoomi A, Mozafari M, Dubnika A, Inayathullah M, Rajadas J (2017) Nanomaterials engineering for drug delivery: a hybridization approach. J Mater Chem B 5:3995–4018 Mouhieddine TH, Itani MM, Nokkari A, Ren C, Daoud G, Zeidan A, Mondello S, Kobeissy FH (2015) Nanotheragnostic applications for ischemic and hemorrhagic strokes: improved delivery for a better prognosis. Curr Neurol Neurosci Rep 15:505 Muehlschlegel S (2018) Subarachnoid Hemorrhage. Continuum (minneap Minn) 24:1623–1657 Neumann JT, Cohan CH, Dave KR, Wright CB, Perez-Pinzon MA (2013) Global cerebral ischemia: synaptic and cognitive dysfunction. Curr Drug Targets 14:20–35 Okuda K, Okabe Y, Kadonosono T, Ueno T, Youssif BG, Kizaka-Kondoh S, Nagasawa H (2012) 2-Nitroimidazole-tricarbocyanine conjugate as a near-infrared fluorescent probe for in vivo imaging of tumor hypoxia. Bioconjug Chem 23:324–329 Partoazar A, Nasoohi S, Rezayat SM, Gilani K, Mehr SE, Amani A, Rahimi N, Dehpour AR (2017) Nanoliposome containing cyclosporine A reduced neuroinflammation responses and improved neurological activities in cerebral ischemia/reperfusion in rat. Fundam Clin Pharmacol 31:185–193 Peng J, Wang H, Gong Z, Li X, He L, Shen Q, Pan J, Peng Y (2020) Idebenone attenuates cerebral inflammatory injury in ischemia and reperfusion via dampening NLRP3 inflammasome activity. Mol Immunol 123:74–87 Pu F, Mishima K, Egashira N, Iwasaki K, Kaneko T, Uchida T, Irie K, Ishibashi D, Fujii H, Kosuna K, Fujiwara M (2004) Protective effect of buckwheat polyphenols against long-lasting impairment of spatial memory associated with hippocampal neuronal damage in rats subjected to repeated cerebral ischemia. J Pharmacol Sci 94:393–402 Qiu J, Wang M, Zhang J, Cai Q, Lu D, Li Y, Dong Y, Zhao T, Chen H (2016) The neuroprotection of Sinomenine against ischemic stroke in mice by suppressing NLRP3 inflammasome via AMPK signaling. Int Immunopharmacol 40:492–500 Ran Y, Qie S, Gao F, Ding Z, Yang S, Tian G, Liu Z, Xi J (2021a) Baicalein ameliorates ischemic brain damage through suppressing proinflammatory microglia polarization via inhibiting the TLR4/NF-κB and STAT1 pathway. Brain Res 1770:147626 Ran Y, Su W, Gao F, Ding Z, Yang S, Ye L, Chen X, Tian G, Xi J, Liu Z (2021b) Curcumin ameliorates white matter injury after ischemic stroke by inhibiting microglia/macrophage Pyroptosis through NF-κB suppression and NLRP3 Inflammasome inhibition. Oxid Med Cell Longev 2021:1552127 Ren W, Yang X (2018) Pathophysiology of long non-coding RNAs in ischemic stroke. Front Mol Neurosci 11:96 Renner C, Favilla CG, Cucchiara BL (2021) Transient ischemic attack, decision-making in adult neurology, 84–85 Rigaud AS, Seux ML, Staessen JA, Birkenhäger WH, Forette F (2000) Cerebral complications of hypertension. J Hum Hypertens 14:605–616 Ruan C, Guo H, Gao J, Wang Y, Liu Z, Yan J, Li X, Lv H (2021) Neuroprotective effects of metformin on cerebral ischemia-reperfusion injury by regulating PI3K/Akt pathway. Brain Behav 11:e2335 Sabu C, Rejo C, Kotta S, Pramod K. Bioinspired and biomimetic systems for advanced drug and gene delivery. J Control Release 287:142–155

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

239

Sacco RL, Boden-Albala B, Gan R, Chen X, Kargman DE, Shea S, Paik MC, Hauser WA (1998) Stroke incidence among white, black, and Hispanic residents of an urban community: the Northern Manhattan Stroke Study. Am J Epidemiol 147:259–268 Sacco RL, Kasner SE, Broderick JP, Caplan LR, Connors JJ, Culebras A, Elkind MS, George MG, Hamdan AD, Higashida RT, Hoh BL, Janis LS, Kase CS, Kleindorfer DO, Lee JM, Moseley ME, Peterson ED, Turan TN, Valderrama AL, Vinters HV; American Heart Association Stroke Council, Council on Cardiovascular Surgery and Anesthesia; Council on Cardiovascular Radiology and Intervention; Council on Cardiovascular and Stroke Nursing; Council on Epidemiology and Prevention; Council on Peripheral Vascular Disease; Council on Nutrition, Physical Activity and Metabolism (2013) An updated definition of stroke for the 21st century: a statement for healthcare professionals from the American Heart Association/American Stroke Association. Stroke 44:2064–89. Erratum in (2019) Stroke 50:e239 Salatin S, Maleki Dizaj S, Yari Khosroushahi A (2015) Effect of the surface modification, size, and shape on cellular uptake of nanoparticles. Cell Biol Int 39:881–890 Saraiva C, Praça C, Ferreira R, Santos T, Ferreira L, Bernardino L (2016) Nanoparticle-mediated brain drug delivery: Overcoming blood-brain barrier to treat neurodegenerative diseases. J Control Release 235:34–47 Sarami Foroshani M, Sobhani ZS, Mohammadi MT, Aryafar M (2018) Fullerenol nanoparticles decrease blood-brain barrier interruption and brain Edema during cerebral ischemia-reperfusion injury probably by reduction of interleukin-6 and matrix metalloproteinase-9 transcription. J Stroke Cerebrovasc Dis 27:3053–3065 Sarmah D, Saraf J, Kaur H, Pravalika K, Tekade RK, Borah A, Kalia K, Dave KR, Bhattacharya P (2017) Stroke Management: an emerging role of nanotechnology. Micromachines (basel) 8:262 Shang Y, Cheng J, Qi J, Miao H (2005) Scutellaria flavonoid reduced memory dysfunction and neuronal injury caused by permanent global ischemia in rats. Pharmacol Biochem Behav 82:67– 73 Shi J, Yu W, Xu L, Yin N, Liu W, Zhang K, Liu J, Zhang Z (2020) Bioinspired Nanosponge for Salvaging Ischemic Stroke via Free Radical Scavenging and Self-Adapted Oxygen Regulating. Nano Lett 20:780–789 Smith WS, Furlan AJ (2016) Brief history of Endovascular acute ischemic stroke treatment. Stroke 47:e23–e26 Smith WS (2004) Pathophysiology of focal cerebral ischemia: a therapeutic perspective. J Vasc Interv Radiol 15:S3-12 Suk JS, Xu Q, Kim N, Hanes J, Ensign LM (2016) PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Adv Drug Deliv Rev 99:28–51 Sveinsson OA, Kjartansson O, Valdimarsson EM (2014) Cerebral ischemia/infarction—epidemiology, causes and symptoms. Laeknabladid 100:271–279 Takamiya M, Miyamoto Y, Yamashita T, Deguchi K, Ohta Y, Abe K (2012) Strong neuroprotection with a novel platinum nanoparticle against ischemic stroke- and tissue plasminogen activatorrelated brain damages in mice. Neuroscience 221:47–55 Takamiya M, Miyamoto Y, Yamashita T, Deguchi K, Ohta Y, Ikeda Y, Matsuura T, Abe K (2011) Neurological and pathological improvements of cerebral infarction in mice with platinum nanoparticles. J Neurosci Res 89:1125–1133 Takasawa M, Moustafa RR, Baron JC (2008) Applications of nitroimidazole in vivo hypoxia imaging in ischemic stroke. Stroke 39:1629–1637 Tan C, Zhao S, Higashikawa K, Wang Z, Kawabori M, Abumiya T, Nakayama N, Kazumata K, Ukon N, Yasui H, Tamaki N, Kuge Y, Shichinohe H, Houkin K (2018) [18F]DPA-714 PET imaging shows immunomodulatory effect of intravenous administration of bone marrow stromal cells after transient focal ischemia. EJNMMI Res 8:35 Tang C, Wang C, Zhang Y, Xue L, Li Y, Ju C, Zhang C (2019) Recognition, intervention, and monitoring of neutrophils in acute ischemic stroke. Nano Lett 19:4470–4477

240

S. K. Das et al.

Tian T, Zhang HX, He CP, Fan S, Zhu YL, Qi C, Huang NP, Xiao ZD, Lu ZH, Tannous BA, Gao J (2018) Surface functionalized exosomes as targeted drug delivery vehicles for cerebral ischemia therapy. Biomaterials 150:137–149 Tsai TT, Patel UD, Chang TI, Kennedy KF, Masoudi FA, Matheny ME, Kosiborod M, Amin AP, Messenger JC, Rumsfeld JS, Spertus JA (2014) Contemporary incidence, predictors, and outcomes of acute kidney injury in patients undergoing percutaneous coronary interventions: insights from the NCDR Cath-PCI registry. JACC Cardiovasc Interv 7:1–9 van Seeters T, Biessels GJ, Niesten JM, van der Schaaf IC, Dankbaar JW, Horsch AD, Mali WP, Kappelle LJ, van der Graaf Y, Velthuis BK, Investigators D (2013) Reliability of visual assessment of non-contrast CT, CT angiography source images and CT perfusion in patients with suspected ischemic stroke. PLoS ONE 8:e75615 Wang J, Zhang H, Ni D, Fan W, Qu J, Liu Y, Jin Y, Cui Z, Xu T, Wu Y, Bu W, Yao Z (2019a) High-performance Upconversion Nanoprobes for multimodal MR imaging of acute ischemic stroke. Small 12:3591–3600 Wang Q, Liu X, Zhu R (2019b) Long noncoding RNAs as diagnostic and therapeutic targets for ischemic stroke. Curr Pharm Des 25:1115–1121 Wang Q, Sun AY, Simonyi A, Jensen MD, Shelat PB, Rottinghaus GE, MacDonald RS, Miller DK, Lubahn DE, Weisman GA, Sun GY (2005) Neuroprotective mechanisms of curcumin against cerebral ischemia-induced neuronal apoptosis and behavioral deficits. J Neurosci Res 82:138–148 Wang S, Nan Y, Zhu W, Yang T, Tong Y, Fan Y (2018a) Gastrodin improves the neurological score in MCAO rats by inhibiting inflammation and apoptosis, promoting revascularization. Int J Clin Exp Pathol 11:5343–5350 Wang Y, Li SY, Shen S, Wang J (2018b) Protecting neurons from cerebral ischemia/reperfusion injury via nanoparticle-mediated delivery of an siRNA to inhibit microglial neurotoxicity. Biomaterials 161:95–105 Wang Y, Luo J, Li SY (2019c) Nano-curcumin simultaneously protects the blood-brain barrier and reduces M1 microglial activation during cerebral ischemia-reperfusion injury. ACS Appl Mater Interfaces 11:3763–3770 Wang Z, Zhao Y, Jiang Y, Lv W, Wu L, Wang B, Lv L, Xu Q, Xin H (2015) Enhanced anti-ischemic stroke of ZL006 by T7-conjugated PEGylated liposomes drug delivery system. Sci Rep 5:12651 Watanabe A, Kajita M, Kim J, Kanayama A, Takahashi K, Mashino T, Miyamoto Y (2009) In vitro free radical scavenging activity of platinum nanoparticles. Nanotechnology 20:455105 Whisnant JP, Wiebers DO, O’Fallon WM, Sicks JD, Frye RL (1996) A population-based model of risk factors for ischemic stroke: rochester, Minnesota. Neurology 47:1420–1428 White H, Boden-Albala B, Wang C, Elkind MS, Rundek T, Wright CB, Sacco RL (2005) Ischemic stroke subtype incidence among whites, blacks, and Hispanics: the Northern Manhattan Study. Circulation 111:1327–1331 Woodruff TM, Thundyil J, Tang SC, Sobey CG, Taylor SM, Arumugam TV (2011) Pathophysiology, treatment, and animal and cellular models of human ischemic stroke. Mol Neurodegener 6:11 Wu MY, Yiang GT, Liao WT, Tsai AP, Cheng YL, Cheng PW, Li CY, Li CJ (2018) Current mechanistic concepts in ischemia and reperfusion injury. Cell Physiol Biochem 46:1650–1667 Yemisci M, Caban S, Gursoy-Ozdemir Y, Lule S, Novoa-Carballal R, Riguera R, Fernandez-Megia E, Andrieux K, Couvreur P, Capan Y, Dalkara T (2015) Systemically administered brain-targeted nanoparticles transport peptides across the blood-brain barrier and provide neuroprotection. J Cereb Blood Flow Metab 35:469–475 Yoo AJ, Pulli B, Gonzalez RG (2011) Imaging-based treatment selection for intravenous and intraarterial stroke therapies: a comprehensive review. Expert Rev Cardiovasc Ther 9:857–876 You RX, Thrift AG, McNeil JJ, Davis SM, Donnan GA (1999) Ischemic stroke risk and passive exposure to spouses’ cigarette smoking. Melbourne Stroke Risk Factor Study (MERFS) Group. Am J Public Health 89:572–575

Hypoxia Responsive Nanomaterials for Cerebral Ischemia Diagnosis

241

Yuan J, Li L, Yang Q, Ran H, Wang J, Hu K, Pu W, Huang J, Wen L, Zhou L, Jiang Y, Xiong X, Zhang J, Zhou Z (2021) Targeted treatment of ischemic stroke by bioactive nanoparticlederived reactive oxygen species responsive and inflammation-resolving nanotherapies. ACS Nano 15:16076–16094 Zhang B, Zhong Q, Chen X, Wu X, Sha R, Song G, Zhang C, Chen X (2020) Neuroprotective effects of Celastrol on transient global cerebral ischemia rats via regulating HMGB1/NF-κB signaling pathway. Front Neurosci 14:847 Zhang DL, Zhang YT, Yin JJ, Zhao BL (2004) Oral administration of Crataegus flavonoids protects against ischemia/reperfusion brain damage in gerbils. J Neurochem 90:211–219 Zhang Q, Wu Z, Li N, Pu Y, Wang B, Zhang T, Tao J (2017) Advanced review of graphene-based nanomaterials in drug delivery systems: synthesis, modification, toxicity and application. Mater Sci Eng C Mater Biol Appl 77:1363–1375 Zhang X, Zhou J, Gu Z, Zhang H, Gong Q, Luo K (2021) Advances in nanomedicines for diagnosis of central nervous system disorders. Biomaterials 269:120492 Zhang Y, Zhang Y, Jin XF, Zhou XH, Dong XH, Yu WT, Gao WJ (2019) The role of Astragaloside IV against cerebral ischemia/reperfusion injury: suppression of apoptosis via promotion of P62-LC3-autophagy. Molecules 24:1838 Zhao H, Yang Y, Si X, Liu H, Wang H (2022) The role of Pyroptosis and autophagy in ischemia reperfusion injury. Biomolecules 12:1010 Zhao Y, Jiang Y, Lv W, Wang Z, Lv L, Wang B, Liu X, Liu Y, Hu Q, Sun W, Xu Q, Xin H, Gu Z (2016a) Dual targeted nanocarrier for brain ischemic stroke treatment. J Control Release 233:64–71 Zhao YZ, Lin M, Lin Q, Yang W, Yu XC, Tian FR, Mao KL, Yang JJ, Lu CT, Wong HL (2016b) Intranasal delivery of bFGF with nanoliposomes enhances in vivo neuroprotection and neural injury recovery in a rodent stroke model. J Control Release 224:165–175 Zheng T, Jiang H, Jin R, Zhao Y, Bai Y, Xu H, Chen Y (2019) Ginsenoside Rg1 attenuates protein aggregation and inflammatory response following cerebral ischemia and reperfusion injury. Eur J Pharmacol 853:65–73 Zhong C, Wang G, Xu T, Zhu Z, Guo D, Zheng X, Wang A, Bu X, Peng H, Chen J, Xu T, Peng Y, Li Q, Ju Z, Geng D, He J, Zhang Y (2019) Tissue inhibitor metalloproteinase-1 and clinical outcomes after acute ischemic stroke. Neurology 93:e1675-e1685. Erratum in Neurology 93:819 Zhou L, Li F, Xu HB, Luo CX, Wu HY, Zhu MM, Lu W, Ji X, Zhou QG, Zhu DY (2010) Treatment of cerebral ischemia by disrupting ischemia-induced interaction of nNOS with PSD-95. Nat Med 16:1439–1443

Multifunctional Hypoxia Imaging Nanoparticles Preeti Sharma, Pooja Kumari, Tikam Chand Dakal, Jyotsana Singh, and Narendra Kumar Sharma

Abstract Hypoxia is a condition in which there is a scarcity of oxygen. It is mainly associated with diseased circumstances, although it can also be a trait of healthy physiology. It is a pathophysiological condition linked to numerous ailments such as hyperglycemia and hypoglycemia inflammation, wounds, tumors, and so on. These conditions are all among the most detrimental to human health. Presently Rapid acknowledgment of this condition becomes essential. In the course of the evolution of clinical and preclinical investigation for the diagnosis of multiple diseases, molecular imaging has become a potent technique. A promising way to solve this drawback is the development of multifunctional hypoxia imaging nanoparticles that enhance the efficiency, specificity, and sensitivity of molecular imaging by providing molecular and functional information in an invasive manner under in vivo conditions. The most significant feature of Nanoparticles throughout the hypoxia imaging procedure is their higher penetration and retention implication with lengthy circulation capacity and easy encapsulation capability, generating them the most influential target for utilization in the biomedical application as a hypoxia imaging technique. Here, we concentrate on a recently generated nanoparticle that has been adopted as a tissuelevel molecular imaging probe for numerous diseases. Keywords Hypoxia · Nanoparticles · Hypoxia imaging · Specificity · Sensitivity

P. Sharma · P. Kumari · N. K. Sharma (B) Department of Bioscience and Biotechnology, Banasthali Vidyapith (Deemed University), P.O. Banasthali Vidyapith Distt., Tonk 304022, Rajasthan, India e-mail: [email protected]; [email protected] J. Singh Genome and Computational Biology Lab, Department of Biotechnology, Mohanlal Sukhadia University, Udaipur, Rajasthan, India T. C. Dakal University of Texas, MD Anderson Cancer Center, Houson, TX, 77030, USA © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_12

243

244

P. Sharma et al.

Abbreviations HIF PH MRI UCNP ROS PDT O2 MTT MMC AZQ DNA Np NADPH NOS HyP-1 PET PEGylated

Hypoxia Inducible Factor Potential of Hydrogen, Magnetic Resonance Imaging Up Conversion Nanoparticle Core Reactive Oxygen Species Photodynamic Therapy Oxygen (3.(4,5-Demethylthiazolyl-2)-2,5-diphenyltetrazolium bromide Mitomycin c Aziquone Deoxy Ribo Nucleic Acid Nanoparticle Nicotinamide adenine dinucleotide Phosphate Nitric Oxide Synthase Hypoxia Probe 1 Positron emission tomography Polyethylene Gylated

1 Introduction Oxygen is a key determinant of standard aerobic metabolism in mammals. When cells suffer from lower-than-usual oxygen levels and pressure, it is called a hypoxic condition. Hypoxia is caused by poor oxygen transport, low blood oxygen levels, and decreased cellular oxygen uptake (Eltzschig and Carmeliet 2011). Mammals have their mechanism of adaptation to hypoxia by increasing respiration, blood flow, and survival response. If in any situation oxygenation is disrupted, this adaptive mechanism assists them in restoring oxygenation and helping the body adapt to hypoxia. This cellular modification mechanism is dependent on the transcription factor HIF, which is active when there is a lack of oxygen and is dormant when oxygen is present. HIF (hypoxia-inducible factor) controls other immune responses in inflammatory bowel disease, certain cancers, and infections (Lee et al. 2019). Various studies show that hypoxia signals control human organ systems such as the lungs, liver, kidneys, and heart. So we can use this hypoxic state as a therapeutic board and also as a biomarker for imaging certain health problems. Recent studies show that hypoxia emerges as a hallmark for the early detection of tumors. This interdisciplinary research should focus on the development of nanomaterials that can be used for effective and non-invasive early detection of cancer (Lledos et al. 2018). In the case of a cancer study, the hypoxia-responsive nanoparticles act vigorously in the existence of a hypoxic tumor microenvironment. The various researchers’ results suggest that its activity improves the effectiveness of radiotherapy, chemotherapy, and

Multifunctional Hypoxia Imaging Nanoparticles

245

other imaging and therapeutic strategies. Hypoxia is associated with many infections such as cancer, neurological disorders, and kidney disease. Numerous researches have been conducted to illustrate the benefit of hypoxia-activated nanoparticles and pro-drugs in tumor-directed cure. Hypoxia-responsive nanoparticles are dormant throughout perfusion and under standard physiological circumstances but become activated during hypoxia when they enter the hypoxic tumor microenvironment (Wang et al. 2021). Overcoming hypoxia is therefore a significant strategy used for the cure of dense tumors. For that purpose, researchers have attempted to enhance useful chemicals and nanoparticles that could be utilized for scanning and indulge hypoxic tumors noninvasively and efficiently. The aim is to retreat hypoxia by creating oxygen and activating nanomolecules/medicines into the hypoxic tumor microenvironment, as well as targeting tumor hypoxia biomarkers to enhance treatment efficacy. In addition, so many studies are being conducted for non-invasive imaging using nanomaterial. In recent years, these suggesting approaches get more attention in which the formation of the multifunctional platform occurs including lanthano crystals, iron oxide, polymers, and gold. These are the active Nanoprobes that can respond to the nearby environment and serve as biomarkers to enhance the effectiveness, specificity, and sensitivity of molecular imagining. During this imaging, the physiological limits that come under will be redox state, presence of enzyme, pH, and hypoxia that can be designed as stimuli for the activated probes (Liu et al. 2019). At the moment, all researchers are looking for non-invasive imaging approaches. Non-invasive hypoxia imaging technologies that are commonly used include calculated tomography and positron emission tomography, MRI, photoacoustic imaging, and optical imaging. Optical hypoxia imaging is the most economical, sensitive, and high-output imaging technology in all of them. This is due to the design of the probe, which is mostly composed of a metal complex and organic dye. The metal complex has several properties, including high oxygen sensitivity, a lengthy phosphorescent for a lifetime, and the ability to penetrate within the cell membranes. It is applied in cell culture and it can also add to animal tissue samples of tumors. To improve the precision and stop the blocking of tissue, researchers develop a ratiometric hypoxia sensor that uses the hypoxia-insensitive dye with a hypoxia sensor and uses its percentage for describing the absorption of oxygen (Ma and Xia 2021). Meanwhile, other investigators are concentrating on organelle structural hypoxia conditions like the distribution of mitochondrial hypoxia. It is essentially a sensorbased system in which the hypoxia sensor accomplished mitochondrial targeting, allowing us to detect the concentration of oxygen inside the cell. These probes and sensors could also be used in preclinical cancer diagnostics. Here, Fig. 1 depicted the basic design of nanoparticles with their vast applications. To accommodate all scanning modalities through optical hypoxia imaging, the technologies require a transporter onto which entirely agents are inserted. The nanoparticle technique would be extremely suggested for this particular reason. Nanomaterials have unique properties that make them suitable for this purpose, such as lengthy circulation and facile endocytosis. These characteristics aid the hypoxic imaging sensor in reaching a specific location. Furthermore, the diverse basic designs of nanomaterials allocate each modality and purpose to act collectively (Das et al.

246

P. Sharma et al.

Fig. 1 Applications of multifunctional hypoxia imaging nanoparticles

2020). In this case, the nanoparticle can also apply active aiming and immune leak functional group by the surface alteration. Meanwhile numerous confront and drawbacks remain in the application of multifunctional hypoxia imaging nanomaterials. Here, we are focusing on recent advancements and various kinds of multifunctional hypoxia imaging nanoparticles.

2 Types of Nanoparticles The complex architectures of multifunctional hypoxia imaging nanoparticles vary. Nanoparticles are characterized according to their structure as self-assembled nanoparticles, core–shell structure nanoparticles, micelle-liposome-like nanoparticles, and matrix dispersion nanoparticles.

2.1 Self-assembled Nanoparticles The inner core–shell of these particles is inherently hydrophobic, making bioavailability much better than any other nanoparticle. Some researchers fabricate such particles using porphyrin Pt (II) and polyfluorene-based hyperbranched complexed polyelectrolytes. Substances that adhere to these particles are with a size of about

Multifunctional Hypoxia Imaging Nanoparticles

247

10 nm. They do not have much greater encapsulation or loading capacity compared to nanocarriers. Hydrophobicity and polarity play key roles in drug development and as a hypoxia sensor. These particles exhibit van der Walls binding between molecules, giving them a relatively loose structure. Some other related data also show that strength can be increased through crosslinking activity. This makes it much stronger (Park et al 2015; Shi et al. 2014; Zhou et al. 2015).

2.2 Core–Shell Nanoparticles As the name suggests, these nanoparticles have a center and a shell. In most cases, the principal is formulated as a hydrophobic treatment and metal compound. Some nanoparticles are also formulated in the core part. They are hydrophilic in nature, making them inherently water soluble and biocompatible. It consists of biomaterials such as hyaluronic acid and many others. In one study, it was found that a researcher formed an up-conversion nanoparticle core (UCNP), the core of which interacts with hypoxia sensors in the shell. This modification ability of nanoparticles allows them to target hypoxia imaging therapy (Li et al. 2018a, b, c; Wang et al. 2016; Xu et al. 2016).

2.3 Micelle-Liposome-Like Nanoparticle Micelles and liposomes work together to form an emulsion and deliver a drug. This is one of the best nanocarriers used in optical hypoxia sensors and drug delivery. Micelles have a monolayer and liposomes have bilayers. The lipophilic agent is layered in a single phase and the hydrophilic water phase can be loaded into the bilayer liposome. According to several types of research, it has been found that the cancer cell membrane shows binding with metal ions and they further show immune evasion ability for tumor targeting. A researcher attaches a cancer cell membrane to an organometallic porphyrin scaffold that exhibits a distinctive tumor-targeting feature. Apart from this, membrane-targeting phospholipid and amphiphilic polymers can also be used in the manufacture of these particles. There show biocompatibility and much better results than other nanoparticles (Feng et al. 2017a, b; Huang et al. 2018; Luo et al. 2016; Yu et al. 2017).

2.4 Matrix-Dispersed Nanoparticles The matrix area of these particles consists exclusively of polymer. Wherein the sensors of hypoxia and the imaging agent are evenly distributed. These nanoparticles and sensors would be small by nature. A researcher fabricated polymer-based

248

P. Sharma et al.

Table 1 The general disruption of the general dispersed nanoparticles is summarized in the form of a table Nanoparticles

Structure

Diagrammatic view

References

Self-assembled nanoparticles

Have a relatively loose structure

Zhou et al. (2015), Park et al. (2015), Shi et al. (2014)

Core–shell nanoparticles

High loading capacity

Xu et al. (2016), Wang et al. (2016), Li et al. (2018a, b, c)

Matrix-dispersed nanoparticles

Small molecules distributed evenly

Luo et al. (2016), Huang et al. (2018), Yu et al. (2017), Feng et al. (2017a, b)

Micelle-liposome-like nanoparticle

Better Biocompatibility

Li et al. (2018a, b, c), Qian et al. (2016), Liu et al. (2017), Liu et al. (2014), Papkovsky (2013)

supramolecular nanoparticles used for ratiometric hypoxia imaging. In some places, small nanoparticles can be encapsulated with a matrix to grow larger. These small particles are metal clusters, dots of quantum, and UCNP, which can serve as the fluorescent functional causes. These particles protect themselves by self-extinction (Li et al. 2018a, b, c; Liu et al. 2014, 2017; Papkovsky 2013; Qian et al. 2016). The general disruption of these particles is summarized in form of a table (Table 1).

3 Hypoxia Imaging Tumor-Directed Nanoparticles Several multifunctional image-guided nanoparticles for the Therapy of cancer were evolved to construct a theranostic platform for the treatment of cancer. Hypoxia plays a debilitating role in cancer for physicians and patients. This state rendered the solid tumor resilient to radiotherapy and chemotherapy. Cancer cell mutations and

Multifunctional Hypoxia Imaging Nanoparticles

249

tumor metastasis can be caused by hypoxia. Hypoxia imaging can help localize the tumor area. Various techniques have been created by researchers to further exploit optical imaging of hypoxia to guide cancer therapy and observe therapeutic efficacy (Weissleder 2001).

3.1 Hypoxia Imaging by Phosphorescent Hypoxia Probe In this procedure, phosphorescent hypoxia probes produce ROS that eliminates cancer cells. ROS can be generated by an agent known as a photosensitizer. This technique is also known as the PDT technique and shows a better therapeutic effect than chemotherapy. It has been widely recognized that chemotherapy can also produce some side effects such as drug resistance, and toxicity at the cellular and organ levels. Compared to PDT, it is a non-invasive method that is highly accurate. It is an image-directed therapy method. In this process, triplet O2 particles respond through photosensitizer and thereafter convert to singlet form. These singlet species and reactive oxygen species are used to cure tumor cells. When the light source is applied to cancer cells, due to radiation, the photosensitizer switches oxygen into single free radical oxygen that helps kill cancer cells. Here, oxygen detection is built on the linear relationship between oxygen concentration and phosphorescence intensity. The three main factors that fall under this are oxygen, phosphorescence, and light. Hypoxiabased tumor microenvironment leads to tumor PDT resistance. This method shows high effectiveness for the diagnosis of early-stage tumor cells and also killed the cell by light irradiation. These in situ hypoxia images guide PDT techniques and results can be analyzed by confocal microscopy and MTT assay. Some more complicated ones also employ phosphorescent metals such as ruthenium, platinum, porphyrins, and iridium. This complex is used to detect the image and o2 levels in existing organisms and hence can work as a real-time monitor for o2 levels (Lv et al. 2018; Zhao et al. 2015; Zhou et al. 2015). Ruthenium HIF inhibition was utilized specifically to treat hypoxia-related illness in this manner. Some researchers created a hypoxia imaging agent based on ruthenium (II) anthraquinone complexes and HIF-1 alpha suppression. This particle shows a 40-fold change in phosphorescence intensity from 20 to 1% oxygen content, so we can say that this Ru has excellent intracellular hypoxia imaging ability. It presents effective liquid solubility and a potent phosphorescence emission in hypoxia. The binding capacity of ruthenium makes it favorable to use as a complex for particle formation. One researcher designed a UCNP mesoporous silica shell structure nanomolecules with an O2 indicator for ruthenium dichloride. This form is used as a quenchable indicator to sense oxygen. Since the oxygen concentration can be reproduced through the luminescence at 613 nm and the excitation at 980 nm, by using this, we can do a deep tissue study (Lv et al. 2015).

250

P. Sharma et al.

Platinum (II) and porphyrin Scientists have created Pt (II) porphyrin with luminous properties of photo physical like high photostability, and large stoke shift. It shows brilliant radiometric luminescence reaction to the o2 level with great consistency and full reversibility. It was also observed in other studies for the identification of early phase and therapy system porphyrin illustrate a high PDT efficacy and extended lifetime and a high signal-to-noise ratio in fluorescence recognition (Lv et al. 2016). Iridium (III) complex This complex is reduced by intracellular oxygen which is repaired during a twoelectron reduction in a hypoxic environment. A group of scientists designed an oxygen-sensing complex and covalently attach this to iridium to monitor oxygen concentration. One more scientist designed two iridium complexes that specially mark mitochondria and lysosomes in existing cells. These mitochondria are further targeted for the improvement of the PDT effect. This type of experiment and their data demonstrate that such alteration of complexes allows better imaging for hypoxia (Feng et al. 2017a, b).

4 Hypoxia Active Nanoparticles for Chemotherapy Quinones Pro-drugs are made up of quinones which could be employed as hypoxia-responsive nanoparticles. The structure of quinone could be triggered through the presence of a hypoxic situation in the cells of a tumor where it produces semiquinones radicals or some other two electron-reducing enzymes such as hydroquinones and DT diaphoresis (Zhu et al. 2012). The class of natural compounds having quinone as their core structure becomes a highly active class for biologically active agents such as vitamin K, doxorubicin (anticancer compound), and coenzyme Q. In this quinone, some more compounds also included such as MMC (mitomycin C), porfiromycin, EO9, and AZQ. The MMC was isolated from Streptomyces caespitosus. This compound showed high solubility, and it was the first used quinone-based medication documented as bioreductive hypoxia selective alkylating agent. After the uptake of cellular or intracellular, it is distributed within the nuclei. Through a reductive metabolism, the MMC and its analogs show their cytotoxic effect. It binds with the DNA through great competence and specificity at the sequence CpG while MMC is related to acute or chronic toxicities. One researcher developed an MMC-soybean- phosphatidylcholine nanoparticle. That enhances the chemical activity of MMC NP in a hypoxia environment. One more nanoparticle was developed by the same researcher with improved characteristics that show lesser size dispersal and high zeta potential with improved stability (Caramés Masana and Reijke 2017; Erdogar et al. 2014). One

Multifunctional Hypoxia Imaging Nanoparticles

251

more nanoparticle was designed by researchers in which the first two-photon fluorescent probe combining luminescent ruthenium (II) complex by a redox-active form of anthraquinone moiety was further used for the in vivo imaging of hypoxia with high-resolution spatial imaging (Pierce et al. 2010). Nitroimidazoles It is a double responsive Nanotransporter that comes as the principal method which is used to enhance the traditional efficacy of PDT ability. This composition is made up of two components one is a 2 nitroimidazole grafting conjugated polymer and the other one is doxorubicin. This Nanocarrier complex generates reactive oxygen species after a light-triggered stimulus to release a hypoxia stimulus. This complete process starts with irradiation of light, and it generates ROS that successively induces hypoxia at the tumor site by converting 2- nitroimidazole into 2 amino imidazoles. This series of reactions occurs by the release of single electron reduction catalysis through a series of nitrocellulose reductase with bio-reducing agents like NADPH that disassembles the drug transporter. This doxorubicin induces cytotoxicity via the damaging of DNA. This strategy gives an innovative pattern for the drug release system (Hou et al. 2009, 2013). Aliphatic N oxide The AQ4N drug shows a Bis N oxide Quinone structure that experiences bioreduction in the hypoxic cell. This drug has been undergoing phase I and II clinical trials. In this process, the reduction by hem proteins including cytochrome (P 450) and nitric oxide synthase (NOS) occurs, and the final product AQ4 form. This reduction product AQ4 shows high DNA affinity by targeting topoisomerase II and this topoisomerase II stops the hypoxic cell to reenter the cell cycle. The AQ4N drug considers an ideal bioreductive drug that can enter intensely into the tumor and it shows very less tissue toxicity. Scientists designed hypoxia probe 1(HyP-1) a hypoxia-responsive agent that is used for photoacoustic imaging. In vitro, it can be used as a hypoxic activator, and in cultured cells or multiple diseases model, it is used in vivo. It is used for three-dimensional visualization of intertumoral hypoxia with the best resolution (McCarthy et al. 2003; Qian et al 2016; Zhang et al. 2015). The comparative studies of before and after administration of Hyp1 were done for understanding the clear role of these particles during hypoxia imaging. Knox et al. establish a hind limb ischemia model to understand the response of Hyp1 in a hypoxic environment during in vivo conditions. Additionally, regarding the use of AQ4N in diagnosis, Feng et al. designed an AQ4N liposome-based nanodrug that relies on a commercial hydrophilic particle as a hypoxia-activated prodrug by a modified hydrophobic Ce6 as a photosensitizer which shows encapsulation with PEGylated liposomes. This liposome can use a probe for positron emission tomography (PET) after chelation with a Cu isotope. That can be used in in vivo imaging of PET, PDT, and hypoxiadependent cytotoxicity. Based on these results, Feng et al. composed AQ4N through glucose oxidase to obtain a hypoxia-activated therapy. In this process, the glucose oxidase stops the supply of glucose to the tumor and exhausts the tumor’s oxygen to produce a hypoxic enrichment. Additional treatment with AQ4N loaded with

252

P. Sharma et al.

liposome enhanced activation of hypoxia and strong synergistic antitumor properties (Feng 2017a, b; Knox et al. 2017; Mehibel et al. 2009; Nishida et al. 2008; Paoni et al. 2002).

5 Conclusion and Future Scope Even though nanotechnology has been studied for ages, while their therapeutic application has yet to be proven. The cause of this can be related to two parts: the tumor part and the nanomaterial part, with tumor heterogeneity being one of the most important. Tumor heterogeneity is defined by diverse genetic alterations and phenotypic appearance profiles at the genetic level, and by complex cells inside the tumorlike fibroblasts, other stromal cells, and inflammatory cells at the cellular level. Although the hypoxia-active nanoparticles play an important strategy to overcome tumor heterogeneity, they can also be employed to develop analytic and medicinal approaches for a wide range of tumor cells. And the nanomolecule has various disadvantages, including limited tumor retention due to its enormous size and rapid renal clearance due to its comparatively small size. Due to their positive charge, the nanoparticles have a short perfusion time, which allows them to easily pass through membranes. Scientists have created a nanoparticle with zero potential during its perfusion period that can become positively charged after being activated by a hypoxic microenvironment. Here, we summarize recent advances in improving radiation efficiency by focusing on hypoxia. Built on the relationship between hypoxia and radiation resistance, numerous new substances have been developed for the treatment of hypoxic tumors. Radioactively labeled NP is suitable for further research in this direction due to their consistent half-life and efficacy. When compared to minor molecular imaging probes, nanoprobes are reflected as a suggesting stage for designing responsive mechanisms to various stimuli such as hypoxia, acidic conditions, various enzyme redox status, and so on, because their well-built particular area provides a large room to alter their functional moieties. Despite numerous accomplishments in this field, significant obstacles remain in the construction of activated nanoprobes for in vivo imaging. Hypoxia is increasingly recognized as a significant factor in the pathophysiology of additional important causes of death, such as myocardial ischemia, metabolic disorders, reproductive disease, and cardiac disease. For this reason, more imagingrelated nanoprobes must be created, as they will provide a wide field for imaging different diseases at the molecular level.

Multifunctional Hypoxia Imaging Nanoparticles

253

References Caramés Masana F, de Reijke TM (2017) The efficacy of Apaziquone in the treatment of bladder cancer. Expert Opin Pharmacother 18(16):1781–1788. https://doi.org/10.1080/14656566.2017. 1392510 Das SS, Bharadwaj P, Bilal M, Barani M, Rahdar A, Taboada P, Bungau S, Kyzas GZ (2020) Stimuliresponsive polymeric Nanocarriers for drug delivery, imaging, and Theragnosis. Polymers 12(6):1397. https://doi.org/10.3390/polym12061397 Eltzschig HK, Carmeliet P (2011) Hypoxia and inflammation. N Engl J Med 364(7):656–665. https://doi.org/10.1056/NEJMra0910283 Erdogar N, ˙Iskit AB, Eroglu H, Sargon MF, Mungan NA, Bilensoy E (2014) Cationic core-shell nanoparticles for intravesical chemotherapy in tumor-induced rat model: safety and efficacy. Int J Pharm 471(1–2):1–9. https://doi.org/10.1016/j.ijpharm.2014.05.014 Feng L, Cheng L, Dong Z, Tao D, Barnhart TE, Cai W, Chen M, Liu Z (2017a) Theranostic liposomes with hypoxia-activated prodrug to effectively destruct hypoxic Tumors post-photodynamic therapy. ACS Nano 11(1):927–937. https://doi.org/10.1021/acsnano.6b07525 Feng Z, Tao P, Zou L, Gao P, Liu Y, Liu X, Wang H, Liu S, Dong Q, Li J, Xu B, Huang W, Wong WY, Zhao Q (2017b) Hyperbranched phosphorescent conjugated polymer dots with iridium(III) complex as the core for hypoxia imaging and photodynamic therapy. ACS Appl Mater Interfaces 9(34):28319–28330. https://doi.org/10.1021/acsami.7b09721 Hou Z, Li Y, Huang Y, Zhou C, Lin J, Wang Y, Cui F, Zhou S, Jia M, Ye S, Zhang Q (2013) Phytosomes loaded with mitomycin C-soybean phosphatidylcholine complex developed for drug delivery. Mol Pharm 10(1):90–101. https://doi.org/10.1021/mp300489p Hou Z, Wei H, Wang Q, Sun Q, Zhou C, Zhan C, Tang X, Zhang Q (2009) New method to prepare mitomycin C loaded PLA-nanoparticles with high drug entrapment efficiency. Nanoscale Res Lett 4(7):732–737. https://doi.org/10.1007/s11671-009-9312-z Huang YY, Tian Y, Liu XQ, Niu Z, Yang QZ, Ramamurthy V, ... Wu LZ (2018) Luminescent supramolecular polymer nanoparticles for ratiometric hypoxia sensing, imaging and therapy. Mater Chem Front 2(10):1893–1899.https://doi.org/10.1039/C8QM00309B Knox HJ, Hedhli J, Kim TW, Khalili K, Dobrucki LW, Chan J (2017) A bioreducible N-oxide-based probe for photoacoustic imaging of hypoxia. Nat Commun 8(1):1794. https://doi.org/10.1038/ s41467-017-01951-0 Lee JW, Ko J, Ju C, Eltzschig HK (2019) Hypoxia signalling in human diseases and therapeutic targets. Exp Mol Med 51(6):1–13. https://doi.org/10.1038/s12276-019-0235-1 Li H, Lei W, Wu J, Li S, Zhou G, Liu D, Yang X, Wang S, Li Z, Zhang J (2018a) An upconverting nanotheranostic agent activated by hypoxia combined with NIR irradiation for selective hypoxia imaging and tumour therapy. J Mater Chem B 6(18):2747–2757. https://doi.org/10.1039/c8tb00 637g Li J, Shang W, Li Y, Fu S, Tian J, Lu L (2018b) Advanced nanomaterials targeting hypoxia to enhance radiotherapy. Int J Nanomed 13:5925. https://doi.org/10.2147/2FIJN.S173914 Li SY, Xie BR, Cheng H, Li CX, Zhang MK, Qiu WX, Liu WL, Wang XS, Zhang XZ (2018c) A biomimetic theranostic O2 -meter for cancer targeted photodynamic therapy and phosphorescence imaging. Biomaterials 151:1–12. https://doi.org/10.1016/j.biomaterials.2017. 10.021 Liu H, Xie Y, Zhang Y, Cai Y, Li B, Mao H, Liu Y, Lu J, Zhang L, Yu R (2017) Development of a hypoxia-triggered and hypoxic radiosensitized liposome as a doxorubicin carrier to promote synergetic chemo-/radio-therapy for glioma. Biomaterials 121:130–143. https://doi. org/10.1016/j.biomaterials.2017.01.001 Liu J, Liu Y, Bu W, Bu J, Sun Y, Du J, Shi J (2014) Ultrasensitive nanosensors based on upconversion nanoparticles for selective hypoxia imaging in vivo upon near-infrared excitation. J Am Chem Soc 136(27):9701–9709. https://doi.org/10.1021/ja5042989

254

P. Sharma et al.

Liu J, Liu Z, Wu D (2019) Multifunctional hypoxia imaging nanoparticles: multifunctional tumor imaging and related guided tumor therapy. Int J Nanomed 14:707–719. https://doi.org/10.2147/ IJN.S192048 Lledos M, Mirabello V, Sarpaki S, Ge H, Smugowski HJ, Carroll L, Aboagye EO, Aigbirhio FI, Botchway SW, Dilworth JR, Calatayud DG, Plucinski PK, Price GJ, Pascu SI (2018) Synthesis, radiolabelling and in vitro imaging of multifunctional nanoceramics. ChemNanoMat: Chem Nanomater Energy Biol More 4(4):361–372. https://doi.org/10.1002/cnma.201700378 Luo CH, Huang CT, Su CH, Yeh CS (2016) Bacteria-mediated hypoxia-specific delivery of nanoparticles for tumors imaging and therapy. Nano Lett 16(6):3493–3499. https://doi.org/10.1021/acs. nanolett.6b00262 Lv W, Yang T, Yu Q, Zhao Q, Zhang KY, Liang H, Liu S, Li F, Huang W (2015) A Phosphorescent Iridium(III) complex-modified nanoprobe for hypoxia bioimaging via time-resolved luminescence microscopy. Adv Sci (Weinheim, Baden-Wurttemberg, Germany) 2(10):1500107. https:// doi.org/10.1002/advs.201500107 Lv W, Zhang Z, Zhang KY, Yang H, Liu S, Xu A, Guo S, Zhao Q, Huang W (2016) A mitochondriatargeted photosensitizer showing improved photodynamic therapy effects under hypoxia. Angewandte Chemie (International ed. in English) 55(34):9947–9951. https://doi.org/10.1002/anie. 201604130 Lv Z, Zou L, Wei H, Liu S, Huang W, Zhao Q (2018) Phosphorescent starburst Pt(II) porphyrins as bifunctional therapeutic agents for tumor hypoxia imaging and photodynamic therapy. ACS Appl Mater Interfaces 10(23):19523–19533. https://doi.org/10.1021/acsami.8b05944 Ma T, Xia T (2021) Nanoparticle-based activatable probes for Bioimaging. Advanced Biology 5(1):e2000193. https://doi.org/10.1002/adbi.202000193 McCarthy HO, Yakkundi A, McErlane V, Hughes CM, Keilty G, Murray M, Patterson LH, Hirst DG, McKeown SR, Robson T (2003) Bioreductive GDEPT using cytochrome P450 3A4 in combination with AQ4N. Cancer Gene Ther 10(1):40–48. https://doi.org/10.1038/sj.cgt.770 0522 Mehibel M, Singh S, Chinje EC, Cowen RL, Stratford IJ (2009) Effects of cytokine-induced macrophages on the response of tumor cells to banoxantrone (AQ4N). Mol Cancer Ther 8(5):1261–1269. https://doi.org/10.1158/1535-7163.MCT-08-0927 Nishida CR, Ortiz de Montellano PR (2008) Reductive heme-dependent activation of the n-oxide prodrug AQ4N by nitric oxide synthase. J Med Chem 51(16):5118–5120. https://doi.org/10. 1021/jm800496s Paoni NF, Peale F, Wang F, Errett-Baroncini C, Steinmetz H, Toy K, Bai W, Williams PM, Bunting S, Gerritsen ME, Powell-Braxton L (2002) Time course of skeletal muscle repair and gene expression following acute hind limb ischemia in mice. Physiol Genomics 11(3):263–272. https://doi. org/10.1152/physiolgenomics.00110.2002 Papkovsky DB, Dmitriev RI (2013) Biological detection by optical oxygen sensing. Chem Soc Rev 42(22):8700–8732. https://doi.org/10.1039/c3cs60131e Park J, Lee J, Kwag J, Baek Y, Kim B, Yoon CJ, Bok S, Cho SH, Kim KH, Ahn GO, Kim S (2015) Quantum dots in an amphiphilic polyethyleneimine derivative platform for cellular labeling, targeting, gene delivery, and ratiometric oxygen sensing. ACS Nano 9(6):6511–6521. https:// doi.org/10.1021/acsnano.5b02357 Pierce SE, Guziec LJ, Guziec FS, Brodbelt JS (2010) Characterization of aziridinylbenzoquinone DNA cross-links by liquid chromatography-infrared multiphoton dissociation-mass spectrometry. Chem Res Toxicol 23(6):1097–1104. https://doi.org/10.1021/tx1000738 Qian C, Yu J, Chen Y, Hu Q, Xiao X, Sun W, Wang C, Feng P, Shen QD, Gu Z (2016) Light-activated hypoxia-responsive nanocarriers for enhanced anticancer therapy. Adv Mater (Deerfield Beach, Fla.) 28(17):3313–3320. https://doi.org/10.1002/adma.201505869 Shi H, Ma X, Zhao Q, Liu B, Qu Q, An Z, ... Huang W (2014) Ultrasmall phosphorescent polymer dots for ratiometric oxygen sensing and photodynamic cancer therapy. Adv Funct Mater 24(30):4823–4830.https://doi.org/10.1002/adfm.201400647

Multifunctional Hypoxia Imaging Nanoparticles

255

Wang J, Huang J, Zhou W, Zhao J, Peng Q, Zhang L, Wang Z, Li P, Li R (2021) Hypoxia modulation by dual-drug nanoparticles for enhanced synergistic sonodynamic and starvation therapy. J Nanobiotechnol 19(1):87. https://doi.org/10.1186/s12951-021-00837-0 Wang XH, Peng HS, Yang W, Ren ZD, Liu YA (2016) Mitochondria-targeted theranostic nanoparticles for optical sensing of oxygen, photodynamic cancer therapy, and assessment of therapeutic efficacy. Microchim Acta 183(10):2723–2731. https://doi.org/10.1007/s00604-016-1917-1 Weissleder R (2001) A clearer vision for in vivo imaging. Nat Biotechnol 19(4):316–317. https:// doi.org/10.1038/86684 Xu S, Zhang X, Xu H, Dong B, Qu X, Chen B, Zhang S, Zhang T, Cheng Y, Xu S, Song H (2016) Silane modified upconversion nanoparticles with multifunctions: imaging, therapy and hypoxia detection. Sci Rep 6:22350. https://doi.org/10.1038/srep22350 Yu Y, Wu Y, Liu J, Liu Y, Wu D (2017) Highly efficient dual-modal phosphorescence/computed tomography bioprobes based on an iridium complex and AuNP polyiohexol composite nanoparticles. Nanoscale 9(27):9447–9456. https://doi.org/10.1039/c7nr03185h Zhang P, Huang H, Chen Y, Wang J, Ji L, Chao H (2015) Ruthenium(II) anthraquinone complexes as two-photon luminescent probes for cycling hypoxia imaging in vivo. Biomaterials 53:522–531. https://doi.org/10.1016/j.biomaterials.2015.02.126 Zhao Q, Zhou X, Cao T, Zhang KY, Yang L, Liu S, Liang H, Yang H, Li F, Huang W (2015) Fluorescent/phosphorescent dual-emissive conjugated polymer dots for hypoxia bioimaging. Chem Sci 6(3):1825–1831. https://doi.org/10.1039/c4sc03062a Zhou X, Liang H, Jiang P, Zhang KY, Liu S, Yang T, Zhao Q, Yang L, Lv W, Yu Q, Huang W (2015) Multifunctional phosphorescent conjugated polymer dots for hypoxia imaging and photodynamic therapy of cancer cells. Adv Sci (Weinheim, Baden-Wurttemberg, Germany) 3(2):1500155. https://doi.org/10.1002/advs.201500155 Zhu C, Liu L, Yang Q, Lv F, Wang S (2012) Water-soluble conjugated polymers for imaging, diagnosis, and therapy. Chem Rev 112(8):4687–4735. https://doi.org/10.1021/cr200263w

Anaerobic Bacteria Mediated Hypoxia Specific Delivery of Nanoparticles Nisha Sharma and Smriti Gaur

Abstract Local hypoxia is a major part and parcel of many solid tumours. These hypoxic regions are found to be of very low oxygen concentration and nutrition deficient leading to the reduced efficacy of anti-cancer drugs and therapies. However, the low level of oxygen in the surroundings favours the growth of certain anaerobic bacteria. Various anaerobic bacteria like Salmonella, Bifidobacterium, Escherichia coli and Clostridium perfringens have found to be therapeutically effective against cancers and other related diseases. These anaerobic bacteria have an affinity to the deep hypoxic regions of many solid tumours and can colonise these regions. Consequently, these anaerobic bacteria can be used to achieve targeted drug delivery in tissues of many tumours. Anaerobic bacteria-mediated delivery of nano-systems can improve the possible poor penetration issues associated with nanomaterials. This chapter would summarise the various ways in which anaerobic bacteria are used as carriers for nanoparticles in hypoxic regions. Keywords Anaerobic bacteria · Hypoxia · Nanoparticles · Drug delivery · Tumour · Cancer

1 Introduction Cancer is a disease in which cells show uncontrolled and abnormal growth with a tendency towards invading or spreading into other organs and body parts of the host, resulting in fatal consequences. Numerous studies have taken place to understand the nature of cancerous cells and to find a cure but none has thus far been able to crack the puzzle. In recent years, there has been an increase in the number of diagnoses related to cancer as various types of cancers and tumours, each with its own complexity and heterogeneity, have emerged. In terms of clinical interventions, chemotherapy (CT) has emerged as the most preferred and practised treatment. It can either be used solo or combined with other treatment strategies. In some cases, radiotherapy N. Sharma · S. Gaur (B) Department of Biotechnology, Jaypee Institute of Information Technology, Noida 201309, India e-mail: [email protected] © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_13

257

258

N. Sharma and S. Gaur

(RT) is utilised in addition to CT, while many oncologists prefer combining it with immunotherapy (IT), with various other such combinations also showing efficacy. Although different combinations of therapies show varied effectiveness, it is observed that their effects are impeded by a few factors, some of these are—rapid clearance in the host due to systematic circulation, tumour selectivity, lower tendency of tumours to accumulate free-drugs, impedance to drug response due to resistivity, severity of side-effects, unsatisfactory solubility in water and non-optimal bioavailability of the drugs (Kumari et al. 2020). It is found that only a decent clinical response is observed in conventional therapeutics and this calls for a novel approach towards targeted delivery of drugs through vehicles which would help in modulating and enhancing their anti-tumour efficacy. Various studies have been conducted to discover new mechanisms related to cancer management with an aim to reduce its tendency to recur. This has resulted in dedicated efforts being made to improve the transport mechanisms of drugs to the affected mass via nano drug delivery systems (DDS). This system proved to be more efficacious and secure than those conventionally used. Thus, for targeted delivery of nanomaterials and cancer therapeutics, there is a need for the development of science behind the tumour-microenvironment (TME), which shows efficiency and receptiveness towards a therapeutic DDS, with the aid of strains of bacteria which show good results in such environments (Kumari et al. 2020).

2 Hypoxia in Cancer—Occurrence and Interactions 2.1 Development of Hypoxia Cancer cells divide rapidly and demand a high amount of oxygen which results in an imbalance in the oxygen demand, therefore, the development of local hypoxia is a common occurrence in solid malignant tumours. Normal tissue exhibits a molecular O2 level of 2%e9% v/v (on average 40 mmHg pO2), whereas in a hypoxic TME level is 0.02%e2% v/v (below 10 mm HgpO2). Peripheral tissues (normal) exhibit level of oxygen (pO2) in the range of 4%-7.5% (approximate), 5% (38 mmHg) is considered as compromise value, which is usually utilised in clinical methods, in comparison, pressure of O2 present in the solid tumour lies in the range of 0.3%-4.2%, that is, below 2% (15 mmHg) in usual circumstances (Xu et al. 2020). Tumour cell proliferation happens at an alarming rate resulting in high levels of growth which puts metabolic strain and thus increases the demand for the amount of O2 and other nutrients in the core regions of the tumour, which is expanding. This might initiate formation of micro-vesicles which are irregular or defective structurally thus resulting in impairment in micro-circulation and constraining gradient of O2 diffusion in the solid mass of the tumours. The distance for O2 diffusion is said to be 200 μm in the networks of vasculature which are impaired and thus, it is limited.

Anaerobic Bacteria Mediated Hypoxia Specific Delivery …

259

Therefore, hypoxic sites are formed in the growth regions of the tumour (Kumari et al. 2020). Thus, hypoxia influences the cells’ controls over the regulation of cell-cycle, allowing an escape of apoptosis, this anomaly results in occurrence of drug-resistant tendencies in the non-cycling cells. Therefore, it is observed that hypoxia emerges as a leading cause of recurring cases of cancer and its resistant tendencies (Zhou et al. 2020). Hypoxia has been documented to feature in more than half (50–60%) of the tumours exhibiting solid mass. Herein, the cells which are found in the deep core of the tumour in the interior are deprived of optimal supply of O2 . A gradient of O2 is observed towards the periphery of the mass of the tumour. As a consequence of this, low O2 concentration zone is developed wherein the pO2 is reduced from the tumour surface to the core of the tumour, to the extent of 0–2.5 mm of Hg in a few sites, in comparison to 30–40 mm of Hg in cells which are healthy. On several occasions, it was also observed in small masses showing malignancy which are only of the size of 1 mm. It is settled through numerous studies that while O2 pressure ranges from 3–5% (20–100 mmHg) in tissues which are normal, anaerobic environments provide 90%) at up to 80 μg/ml of CQDs, MSNs, CQD-MSNs, and FA-CQD-MSNs. Meanwhile, after the release of DOX, this cell viability dropped by almost 73%. The possibility of MSNs in cancer theranostics was further supported by the greater cellular absorption of FA-linked NPs in comparison to non-folate NPs. (c) Gold Nanoparticles (AuNPs) Due to their significant qualities, including their biodegradable and non-toxic nature, ease of surface functionalization, ease of synthesis, optical and electrical properties, greater atomic number, and surface plasmon resonance effect, AuNPs have attracted a huge interest among different inorganic NPs in the biomedical field for delivering various therapeutic agents and imaging agents (Kempen et al. 2015). Furthermore, it increases light scattering by 5–6 folds more than other organic dyes, enhancing its application in cancer therapy imaging and detection (Singh et al. 2018). The protein TfR, which is found on the luminal side of the BBB, is accountable for binding and delivering substances to the brain parenchymal cells. According to a previous study, antibodies pass the BBB via changing their affinity for TfR. To target brain capillaries, Johnsen created AuNPs that are TfR- targeted (Johnsen et al. 2018). In order to conjugate thiolated antibodies, AUNPs were functionalized with 1% maleimide (i.e., IgG, Anti TfRA, anti-TfRD, anti-TfRA-BACB1). The generated AuNPs were assessed for a variety of distinctive properties, including PDI (0.016– 0.250), zeta potential (−16.7 to −40.4 mV), and particle size (35–77 nm). Using isolated brain capillary endothelial cells, the targeted AuNPs were also assessed for in-vitro cellular uptake study. Results showed that NPs were taken up by cells in large quantities while maintaining their structural integrity. In contrast, in vivo research conducted after injecting targeted AuNPs into aged female Balb/c mice revealed a significant level of growth inside the cells. Inclusive of these findings showed that the TfR-targeted AuNPs had a greater ability to penetrate the BBB. Qui explored the potential of oridonin-loaded, HA, anti-Glypican-1 (anti-GPC1) antibody, oridonin, Gd, and Cy7 dye coupled AuNPs for multimodal imaging and therapy against pancreatic cancer (Qiu et al. 2018; Rangisetty et al. 2023). The in-vitro release of the NPs was pH dependent. The targeted NPs exhibited greater cellular uptake in PANC-1, BXPC-3, and 293 cells and inhibition of PGC 1 overexpressed

New Developments in Nano-theranostics Combined with Intelligent …

361

cancerous cells at low concentrations in cell viability studies. Moreover, on the BXPC 3 orthotopic pancreatic tumor xenograft model, in vivo anticancer effectiveness was shown. These findings demonstrated the dependability of AuNPs for theranostic applications such as targeted therapy and multimodal imaging. (d) Calcium Phosphate (CaP)NPs Because of the biocompatibility, lack of toxicity, availability of the large surface for functionalization, and pH-responsive action, CaPNPs have attracted a lot of attention in recent years. These characteristics make them multifunctional NPs that can be loaded with bioactive substances, bioimaging substances, and targeting molecules. However, the difficult synthesis processes led to an increase in size, and particle aggregation limits the multifunctionality of CaP NPs. For the PDT of tumors, Haedicke synthesized the CaP NPs (Haedicke et al. 2015; Arulmozhi et al. 2019). Additionally, CaP NPs were altered in a variety of ways, such as by conjugating them with DY682-NHS fluorescent dye to facilitate near-infrared fluorescence (NIRF) optical imaging, RGDfK peptide for targeted tumor targeting, and temoporfin as a photosensitizer (in vivo). Using fluorescent DY-734-annexin V probe after two days, it was observed that the CaP NPs display tumor apoptosis. In the following few days, it was also noted that the tumor volume and vascularization had decreased. As a result, the researchers showed CaP NPs potential for PDT. Wang synthesized Au carbon/CaP core-shell NPs that are DOX-loaded, pH and NIR dual-responsive (Haedicke et al. 2015). In an initial report, the pH-responsive drug release was studied, in addition to this the hydrodynamic diameter of NPs was measured to be 160 nm, the zeta potential was −19.1 mV, and the drug loading was 92%. The presence of CaP in the NPs was validated by X-ray powder diffraction investigations and Fourier Transform Infrared microscopy. NPs notably killed cancer cells (IC50 = 7.8 × 10−4 M) in in vitro cytotoxicity experiments. Additionally, endocytosis research in HeLa cells showed substantial cellular uptake, which was supported by a rise in red fluorescence over time. Finally, chemo-PTT against cancer demonstrated the theranostic potential of CaP-based NPs. (C) Intervention of Bio-responsive Nanomaterials in Hypoxia Malignant tumors are characterized by hypoxia, which is frequently associated with worsening tumor severity and treatment results (Adrian 2002). Therefore, for efficient tumor control, early detection and killing of hypoxic tumor cells are essential. The development of useful chemicals and nanomaterials that can be utilized to non-invasively image and effectively treat hypoxic tumors has been the focus of an upsurge in interdisciplinary research. These mostly consist of treatments that target tumor hypoxia biomarkers, hypoxia-active nanoparticles, and anti-hypoxia medicines. Inactive during blood flow and other normal physiological conditions, hypoxia-responsive nanoparticles become active once they are released from the blood vessels into the blood circulation into the hypoxic tumor microenvironment. Fluorescence, photoacoustic intensity, radiotherapy, chemotherapy, and other imaging and treatment methods can all be made more effective with their utilization.

362

K. Jain et al.

The intratumor microenvironment becomes noticeably hypoxic due to the imbalance between the blood supply and the tumor’s rapid rate of growth, which reduces the oxygen levels in the tumor (Gatenby and Gillies 2008; Lewis et al. 2016; da Motta et al. 2017). Particularly, hypoxia is a condition brought on by the ability of the quick proliferation and spreading rates of tumor cells, which raises oxygen uptake. Hypoxia-inducible factor-1 alpha (HIF-1 α) can be elevated as a tumor grows, which stimulates the synthesis of Vascular Endothelial Growth Factor (VEGF) through the HIF-1 signaling pathway, which stimulates the formation of blood vessels. Because of its impact on the control of the cell cycle, evasion of apoptosis, maintenance and quiescence of stem cells, and selection of treatment-resistant noncycling cancer cells, hypoxia is a key role in cancer regression. (Keith et al. 2012; Gilkes et al. 2014). Researchers have worked to discover useful chemicals and nanomaterials that can be utilized to effectively image and treat hypoxic tumors without surgery. Anti-hypoxia drugs, hypoxia-active nanoparticles, and hypoxia-targeting agents are three common approaches. In order to increase the effectiveness of the treatments that are provided, it is necessary to produce O2 to reverse hypoxia, which can be done by the activation of the nanoparticles or agents in the hypoxic tumor microenvironment, and to target biomarkers of tumor hypoxia. Anti-hypoxia agents are substances that produce oxygen or act as oxygen carriers, such as MnO2 and hemoglobinbased O2 carriers, which can treat tumor hypoxia and boost the effectiveness of therapy or minimize the malignancy of tumor. Hypoxia-targeting drugs, on the other hand, were formulated to image and treat hypoxia due to the high levels of HIF1, the unfolded protein response, and the mTOR pathways in hypoxia. Reducing agents or light-triggered electronic transfer may be able to activate hypoxia-active prodrugs (HAPs, also known as bioreductive prodrugs) in the tumor microenvironment. Drugs that are based on the presence of quinones, nitro-groups, aromatic N-oxides, aliphatic N-oxides, and transition metals are currently been used extensively in hypoxia-responsive systems. In order to load and release HAPs and other drugs into the tumor microenvironment, nanocarriers have demonstrated to be more effective targeted delivery solutions. This results in longer circulation times, better tumor penetration, and higher drug accumulation. The low efficacy of extravasation into the distal sections of a tumor is one of the limits of hypoxia-active nanoparticles. This is because the nanoparticles are relatively large compared to small molecular prodrugs. Azobenzene, cobalt are the most used hypoxia-active fluorescence nanoparticles. Relatively cobalt is abundant and is cheaper and is also important for the metabolism of all animals. Hypoxic-active nanoparticles are also used as radiosensitizers that helps in easy killing of tumor cells. The most used are nitroimidazoles and Sanazole (SAN). Nitroimidazoles are the most often used HAPs, and they have also undergone phase II clinical trials. Since 1950s, they are utilized for cancer therapy and they also exhibit a variety of bioactivities. (Xia et al. 2019). SAN is a nitrotriazole molecule. It is an effective radiosensitizer for hypoxic cells, and phase III clinical trials with it have shown considerable improvements in local tumor control and survival (Zhang et al. 2012). Nitroimidazoles are also used as

New Developments in Nano-theranostics Combined with Intelligent …

363

hypoxia-active nanoparticles for chemotherapy. Besides this, quinones, aliphatic Noxide, Benzotriazine-N-oxide (Tirapazamine, most commonly used) are also used as hypoxia-active nanoparticles for chemotherapy.

2 Conclusion Although nanomaterials have revolutionized the theranostics and have achieved significant milestones, however there are still several hinderances before bringing it to clinical level. In the sections above, several examples of nanocarriers for theranostics have been discussed with their challenges. Advances in polymer designing, fabrication, conjugation with various modalities have paved the way to design optimized systems. Polydispersity was one of the significant challenges and systems have been designed to keep it within an optimal range. Despite advancements in systems and strategies, biocompatibility of the diagnostic agents is still a challenge. Entrapment of such diagnostic agents in organic complexes during their residence time inside the patient’s body could safeguard it. Prodrugs have the advantage over the polymers as they get cleared from the body, whereas polymers with molecular weight >40 k Da are difficult to get cleared up if non-biodegradable. Such polymeric systems can be tuned to become biodegradable by replacing the crosslinks with ester bonds which are more susceptible to enzyme as well as pH. Drug loading onto the liposomes, polymerosomes, micelles, and nanoparticles by random encapsulation is one of the most common approaches. However, the reproducibility of such systems is quite questionable and creates problems for approval from regulatory bodies. In addition to this, one must always keep the requirements for FDA approval while designing such systems.

References Abedin MR, Umapathi S, Mahendrakar H, Laemthong T, Coleman H, Muchangi D, Barua S, et al. (2018) Polymer coated gold-ferric oxide superparamagnetic nanoparticles for theranostic applications. J Nanobiotechnol 16(1):1–13 Adrian LH (2002) Hypoxia-A key regulatory factor in tumor growth. Nat Rev Cancer 2(1):38–47 Ali A, Zafar H, Zia M, ul Haq I, Phull AR, Ali JS, Hussain A (2016) Synthesis, characterization, applications, and challenges of iron oxide nanoparticles. Nanotechnol Sci Appl 9:49 Albuquerque J, Moura CC, Sarmento B, Reis S (2015) Solid lipid nanoparticles: a potential multifunctional approach towards rheumatoid arthritis theranostics. Molecules 20(6):11103–11118 Arulmozhi S, Matchado MS, Snijesh VP, Kumar A, Singh S (2019) An insight into anti-arthritic property of C25H34O7 for rheumatoid arthritis using molecular modelling and molecular dynamics approach. Inform Med Unlocked 16:100145 Bae KH, Lee JY, Lee SH, Park TG, Nam YS (2013) Optically traceable solid lipid nanoparticles loaded with siRNA and paclitaxel for synergistic chemotherapy with in situ imaging. Adv Healthcare Mater 2(4):576–584

364

K. Jain et al.

Chuang SY, Lin CH, Huang TH, Fang JY (2018) Lipid-based nanoparticles as a potential delivery approach in the treatment of rheumatoid arthritis. Nanomaterials 8(1):42 Fan D, Wang K, Gao H, Luo Q, Wang X, Li X, Shi J (2020) A 64Cu-porphyrin-based dual-modal molecular probe with integrin αvβ3 targeting function for tumour imaging. J Label Compd Radiopharm 63(5):212–221 Gatenby RA, Gillies RJ (2008) A microenvironmental model of carcinogenesis. Nat Rev Cancer 8(1):56–61 Gilkes DM, Semenza GL, Wirtz D (2014) Hypoxia and the extracellular matrix: drivers of tumour metastasis. Nat Rev Cancer 14(6):430–439 Grillone A, Riva ER, Mondini A, Forte C, Calucci L, Innocenti C, Ciofani G et al. (2015) Active targeting of sorafenib: preparation, characterization, and in vitro testing of drug-loaded magnetic solid lipid nanoparticles. Adv Healthc Mater 4(11):1681–1690 Haedicke K, Kozlova D, Gräfe S, Teichgräber U, Epple M, Hilger I (2015) Multifunctional calcium phosphate nanoparticles for combining near-infrared fluorescence imaging and photodynamic therapy. Acta Biomater 14:197–207 Johnsen KB, Bak M, Kempen PJ, Melander F, Burkhart A, Thomsen MS, Andresen TL, et al (2018) Antibody affinity and valency impact brain uptake of transferrin receptor-targeted gold nanoparticles. Theranostics 8(12):3416 Keith B, Johnson RS, Simon MC (2012) HIF1α and HIF2α: sibling rivalry in hypoxic tumour growth and progression. Nat Rev Cancer 12(1):9–22 Kelkar SS, Reineke TM (2011) Theranostics: combining imaging and therapy. Bioconjug Chem 22(10):1879–1903 Kempen PJ, Greasley S, Parker KA, Campbell JL, Chang HY, Jones JR, Jokerst JV, et al (2015) Theranostic mesoporous silica nanoparticles biodegrade after pro-survival drug delivery and ultrasound/magnetic resonance imaging of stem cells. Theranostics 5(6):631 Lee MH, Kim JY, Han JH, Bhuniya S, Sessler JL, Kang C, Kim JS (2012) Direct fluorescence monitoring of the delivery and cellular uptake of a cancer-targeted RGD peptide-appended naphthalimide theragnostic prodrug. J Am Chem Soc 134(30):12668–12674 Lewis DM, Park KM, Tang V, Xu Y, Pak K, Eisinger-Mathason TK, Gerecht S (2016) Intratumoral oxygen gradients mediate sarcoma cell invasion. Proc Natl Acad Sci 113(33):9292–9297 Li S, Goins B, Zhang L, Bao A (2012) Novel multifunctional theranostic liposome drug delivery system: construction, characterization, and multimodality MR, near-infrared fluorescent, and nuclear imaging. Bioconjug Chem 23(6):1322–1332 Madamsetty VS, Mukherjee A, Mukherjee S (2019) Recent trends of the bio-inspired nanoparticles in cancer theranostics. Front Pharmacol 10:1264 Mahmoudi M, Sant S, Wang B, Laurent S, Sen T (2011) Superparamagnetic iron oxide nanoparticles (SPIONs): development, surface modification and applications in chemotherapy. Adv Drug Deliv Rev 63(1–2):24–46 Martínez-González R, Estelrich J, Busquets MA (2016) Liposomes loaded with hydrophobic iron oxide nanoparticles: suitable T2 contrast agents for MRI. Int J Mol Sci 17(8):1209 da Motta LL, Ledaki I, Purshouse K, Haider S, De Bastiani MA, Baban D, McIntyre A (2017) The BET inhibitor JQ1 selectively impairs tumour response to hypoxia and downregulates CA9 and angiogenesis in triple negative breast cancer. Oncogene 36(1):122–132 Müller RH, Mäder K, Gohla S (2000) Solid lipid nanoparticles (SLN) for controlled drug delivery–a review of the state of the art. Eur J Pharm Biopharm 50(1):161–177 Nandwana V, Singh A, You MM, Zhang G, Higham J, Zheng TS, Dravid VP, et al (2018) Magnetic lipid nanocapsules (MLNCs): Self-assembled lipid-based nanoconstruct for non-invasive theranostic applications. J Mater Chem B 6(7):1026–1034 Naseri N, Valizadeh H, Zakeri-Milani P (2015) Solid lipid nanoparticles and nanostructured lipid carriers: structure, preparation and application. Adv Pharm Bull 5(3):305 Pan D, Sanyal N, Schmieder AH, Senpan A, Kim B, Yang X, Lanza GM (2012) Antiangiogenic nanotherapy with lipase-labile Sn-2 fumagillin prodrug. Nanomedicine 7(10):1507–1519

New Developments in Nano-theranostics Combined with Intelligent …

365

Panwar A, Yadav KL (2015) A novel one-pot synthesis of hierarchical europium doped ZnO nanoflowers. Mater Lett 142:30–34 Panwar A, Yadav KL (2022) Silver doped zinc oxide nanostructures with antibacterial properties against GFP-expressing antibiotic resistant Escherichia coli. Mater Lett 309:131469 Patil TS, Deshpande AS, Deshpande S, Shende P (2019) Targeting pulmonary tuberculosis using nanocarrier-based dry powder inhalation: current status and futuristic need. J Drug Target 27(1):12–27 Qiu W, Chen R, Chen X, Zhang H, Song L, Cui W, Wang Z, et al (2018) Oridonin-loaded and GPC1-targeted gold nanoparticles for multimodal imaging and therapy in pancreatic cancer. Int J Nanomedicine 13:6809 Rangisetty PT, Kilaparthi A, Akula S, Bhardwaj M, Singh S (2023) RSAD2: An exclusive target protein for Zika virus comparative modeling, characterization, energy minimization and stabilization. Int J Health Sci 17(1):12–17 Sao P, Chand Y, Kumar A, Singh S (2021) Potential drug target identification in porphyromonas gingivalis using in-silico subtractive metabolic pathway analysis. Bangladesh J Med Sci 20(4):887–896 Singh S, Jannet Vennila J, Kant R (2014) Gene interaction map: a paradigm for identifying significant pathways responsible for rheumatoid arthritis. Netw Model Anal Health Inform Bioinform 3:1–7 Singh P, Pandit S, Mokkapati VRSS, Garg A, Ravikumar V, Mijakovic I (2018) Gold nanoparticles in diagnostics and therapeutics for human cancer. Int J Mol Sci 19(7):1979 Strijkers GJ, Kluza E, Van Tilborg GA, van der Schaft DW, Griffioen AW, Mulder WJ, Nicolay K (2010) Paramagnetic and fluorescent liposomes for target-specific imaging and therapy of tumor angiogenesis. Angiogenesis 13(2):161–173 Sun H, Erdman III, W, Yuan Y, Mohamed MA, Xie R, Wang Y, Cheng C, et al (2020) Crosslinked polymer nanocapsules for therapeutic, diagnostic, and theranostic applications. Wiley Interdiscip Rev: Nanomedicine Nanobiotechnology 12(6):e1653 Svenson S (2013) Theranostics: Are We There Yet? Mol Pharm 10(3):848–856 Vallabani NV, Singh S (2018) Recent advances and future prospects of iron oxide nanoparticles in biomedicine and diagnostics. 3 Biotech 8(6):1–23 Vallet-Regí M (2012) Mesoporous silica nanoparticles: their projection in nanomedicine. Int Sch Res Not Xia Y, Xu C, Zhang X, Ning P, Wang Z, Tian J, Chen X (2019) Liposome-based probes for molecular imaging: from basic research to the bedside. Nanoscale 11(13):5822–5838 Zhang LW, Wen CJ, Al-Suwayeh SA, Yen TC, Fang JY (2012) Cisplatin and quantum dots encapsulated in liposomes as multifunctional nanocarriers for theranostic use in brain and skin. J Nanopart Res 14(7):1–18 Zou R, Li J, Yang T, Zhang Y, Jiao J, Wong KL, Wang J (2021) Biodegradable manganese engineered nanocapsules for tumor-sensitive near-infrared persistent luminescence/magnetic resonance imaging and simultaneous chemotherapy. Theranostics 11(17):8448

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat Tumour Hypoxia Bhupendra G. Prajapati, Amruta Desai, Pooja Desai, Aarohi Deshpande, Aarohi Gherkar, Manas Joshi, and Shama Mujawar

Abstract Nanozymes are a class of nanomaterials that mimic natural enzymes in their ability to execute catalytic processes. Due to their great stability and low cost, nanozymes are particularly attracting attention in the biosciences and biomedical fields. Numerous variables, including pH, hydrogen peroxide (H2 O2 ), glutathione (GSH) level, and metal ion chemical state, control the enzyme-mimetic activities of nanozymes, which have significant potential for use in biomedical sciences. Multifunctional nanozymes have advanced in recent years for a variety of biomedical uses. There have been certain enzymes (mostly peroxidases) that have proven to be effective against several inflammatory diseases and cancers such as malignant melanoma, etc. The fundamental cause of cancer spread, resistance to chemotherapy and tumour recurrence is hypoxic tumour microenvironment (TME). Tumour hypoxia has not yet been resolved by any recognised therapy. In this paper, we provide a comprehensive analysis of the applications and underlying difficulties of peroxidase mimicking nanozymes as a more effective treatment for hypoxic tumour microenvironment. Keywords Catalyst · Enzyme · Peroxidase · Nanozymes · Hypoxia

B. G. Prajapati Shree S.K.Patel College of Pharmaceutical Education and Research, Ganpat University, Kherva, Gujarat 384012, India A. Desai Department of Biotechnology, Mehsana Urban Institute of Sciences, MUIS, Ganpat University, Kherva, Mehsana, Gujarat, India P. Desai Cell and Molecular Biology Laboratory, ICMR National Institute of Nutrition, Hyderabad, India A. Deshpande · A. Gherkar · S. Mujawar (B) MIT School of Bioengineering Sciences and Research, MIT-Art, Design and Technology University, Loni Kalbhor, Pune 412201, India e-mail: [email protected] M. Joshi Department of Comparative Development and Genetics, Max Planck Institute for Plant Breeding Research, Köln, Germany © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_19

367

368

B. G. Prajapati et al.

Abbreviations Ce6 PEG SFO NP Au@HCNs CMS@GOx ZIF-8 ABTS@PAH-CNts ABTS CN PB-Ft NPs AgPd@BSA/DOX MIONzyme-GOx PDAC NPs

GQD-SPNs UTMD CHT FeN CDT Co9S8 NDs

Chlorin e6 Polyethylene glycol SnFe2 O4 Au–Ag@HA Gold nanoparticle core with a porous hollow carbon shell nanospheres PEGylated Zeolitic imidazolate framework-8 Dual lock-and-key type activatable nanotherapeutic platform 2,20-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) Ceria nanocubes Prussian blue-modified ferritin nanoparticles Bovine serum capped bimetallic silver palladium nanoparticles loaded with doxorubicin Iron oxide-based nanozymes loaded with glucose oxidase Polydopamine (PDA) and ammonium bicarbonate (NH4 HCO3 ) coated and doxorubicin (Dox) loaded hollow cerium oxide (CeO2 ) NPs Graphene quantum dots/semiconducting polymer nanocomposites Ultrasound-targeted microbubble destruction Chemotherapy Fe-Metal organic framework-based nanozyme Chemodynamic therapy Cobalt sulphide nanodots

1 Introduction Understanding the relevance of an enzyme in the daily operations of the human body system is essential to comprehending the features of the peroxidase enzyme and its function as a nanozyme. The rate of a chemical process is accelerated by an enzyme, a biological catalyst. It accomplishes this by reducing a reaction’s activation energy. This is the amount of energy necessary to initiate any chemical reaction. The majority of enzymes function by reducing the activation energy barrier, which brings up a different pathway for the process to occur. Without enzymes, the digestive process could take many years, which would render life unsustainable. Enzymes are categorised according to the types and functions they perform. It is a methodical approach to understanding and evaluating the kind of reaction they catalyse. The specific substrate (reactant) an enzyme is connected with is typically indicated by the name of the enzyme. For instance, lactase catalyses the hydrolysis

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat …

369

of lactose, a disaccharide, to glucose and galactose (monosaccharide). The peroxidase enzyme belongs to the class of enzymes known as oxidoreductases, which are mostly utilised in oxidation and reduction reactions. Hydrogen peroxide serves as the electron acceptor for peroxidases.

1.1 Peroxidase When guaiacol was examined with hydrogen peroxide and other plant extracts, Schonbein discovered the peroxidase activity in 1855 (Cheng et al. 2008). These enzymes are present in a wide variety of plants and animals and are crucial to all living things. Glutathione peroxidase (GPx), myeloperoxidase (MPO), eosinophil peroxidase (EPO), uterine peroxidase, lactoperoxidase (LPO), salivary peroxidase (SPO) and thyroid peroxidase (TPO) are the most common types of peroxidases in mammals (Cheng et al. 2008). These heme-containing enzymes all have a significant impact on innate immunity, the extracellular matrix and thyroid hormone synthesis, as well as various inflammatory disorders. However, glutathione peroxidase is an exception to this rule since it uses selenium in place of heme. There are eight isoenzymes in the GPx family, each having unique features and functions. Some of these enzymes have even demonstrated efficacy against a number of skin malignancies, including malignant melanoma, which affects up to 200 000 people annually. An example of such an enzyme that has shown promise as a treatment for this illness is glutathione peroxidase 3 (GPX3). Reactive oxygen species, or ROS, which in this case are melanoma cells and cause DNA damage, are known to be inactivated by it (Yi et al. 2019). When GPX3 expression is downregulated, tumours in a variety of head and neck, gastric, cervical, thyroid, and lung malignancies are effectively suppressed (Ding et al. 2022). The GPx2 enzyme, which is expressed in the digestive system and is created during squamous cell carcinoma, is another example of an enzyme from this family (Serewko et al. 2002). Another kind that is exhibited in head and neck cancer patients after chemotherapy is GPx3 (Chen et al. 2011). All of these enzymes are unquestionably excellent at therapeutic applications, but they all have significant drawbacks in terms of convenience and operating costs. For these enzymes to be the most stable, the pH and temperature must be just right. This limits their functionality and makes them expensive to work with and challenging to store. In this situation, nanozymes can help. These nanoparticles have similar catalytic activity and some characteristics of enzymes. This phrase was initially used in 2004 by Pasquato, Scrimin and colleagues to characterise the transphosphorylation reactivity of gold nanoparticles functionalized with triazacyclononane (Manea et al. 2004). Numerous uses exist for these, such as immunoassay for bioanalysis, immunohistochemical staining for disease diagnosis, in vivo imaging and disease therapy (Huang et al. 2019). Recent developments in the field of nanotechnology have sped up the creation of nanomaterials with natural enzyme-like activity (Nanozymes), which have a number of beneficial properties (Wei et al. 2013). For catalytic actions, natural

370

B. G. Prajapati et al.

enzymes need exacting physiological conditions. A few of the significant limitations of using natural enzymes are their poor stability under extreme environmental conditions, their high cost of synthesis, isolation and purification. In contrast to natural enzymes, nanozymes have unwavering biocatalytic activity even in the most severe pH, temperature and protease-resistant environments. Therefore, it was essential to create effective enzyme substitutes for artificial ones.

1.2 Tumour Hypoxia For a period of time now, patients have been experiencing the recurrence of tumours in the same or various parts of the body. This has led us to one of the largest barriers, i.e. Hypoxia to overcome. Tumour hypoxia is a condition wherein malignancies grow out rapidly as a result of lack of oxygen in some tissues, casting aside bits of tumours in those tissues which can also be seen as an advanced form of cancer spreading throughout the body. It is seen to develop due to abnormal sizes of tumour in an area restricting oxygen supply in the entire body targeting parts which were oxygen-deprived from many years. Hypoxia is exhibited to be a succession of cancer which occurs due to poor nursing of tumour patients. A number of patients are seen to relapse from tumours, and this recurrence makes them vulnerable for outgrowth of cancer leading to TME. The chief principle of Hypoxia is notably the ability of this tumour to give off resistance towards chemotherapy and radiation giving rise to uncountable dividing cells dispersed in the body. Cells of TME are lethal to many surrounding cells that are unable to settle in this cancerous environment (Muz et al. 2015). Due to this, numerous cells are taken control over and it takes almost no time for a tumour this advanced to affect multiple organs leading to organ failure. Hypoxia achieves resistance to several treatment options by maintaining some key processes like controlling the death phase of a cell, supervising autophagy (destruction of cell making it prone for abnormal proteins to enter), administering to function of mitochondria, changing drug delivery pathways, etc. Reduced oxygen being the main cause of the cascade of TME, it was proved that cells grown in hypoxic conditions are immune to apoptosis due to fewer DNA radicals and synthesising of ROS after oxygen fixation. In the present review, we put forth the probable hypoxia treatment and the mechanism acting behind it along with some of the applications of Nanozyme emerging field.

1.3 Nanozymes: Efficiency Over Natural Enzymes The potential of using nanozymes in energy, environmental and biomedical applications has been emphasised. Quantitatively and qualitatively screening of environmental contaminants such ions, molecules and organic chemicals can be done using nanozymes (Meng et al. 2020).

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat …

371

Additionally, nanozymes are being employed to treat bacterial infections because they exhibit intriguing broad-spectrum antibacterial capabilities with little cytotoxicity (Wang et al. 2020a, b). Additionally, they have been utilised in biosensors for the quick, accurate and highly subtle detection of a variety of diseases (Tao et al. 2020; Mahmudunnabi et al. 2020). Due to the crucial role that oxygen plays in the development and therapy of cancer, the use of nanomaterial-based nanozymes in cancer treatment has recently received substantial research (Jiang et al. 2019). Numerous enzyme mimicking nanomaterials, such as POD OXD, SOD and CAT mimetic nanozymes, have undergone substantial research in an effort to be used as cancer therapies (Jiang et al. 2019). The current review primarily focuses on POD mimicking nanomaterials in cancer therapeutic applications and provides a thorough analysis of recent developments and outlooks.

1.4 Antioxidant Nanozymes Antioxidants are essential in the biological system to protect the cells or tissues from harm caused by an excess of free radicals produced during the body’s regular biochemical activities. The endogenous antioxidant system in the human body is well-established and primarily made up of enzymes that scavenge free radicals, such as catalase, superoxide dismutase (SOD), glutathione peroxidase, glutathione reductase and peroxiredoxins. Myself and colleagues discovered that cerium oxide nanoparticles (CeNPs) are an example of an inorganic antioxidant that exhibits superoxide radical foraging and hydrogen peroxide breakdown in in vitro, in vivo and other animal models (Korsvik et al. 2007; Heckert et al. 2008, Hirst et al. 2009).

1.5 Pro-oxidant Nanozymes and Its Mechanism Pro-oxidant nanozymes are those that cause oxidative stress by either blocking the antioxidant system of mammalian cells or by creating free radicals in those cells. Common medications like the pain reliever paracetamol and the cancer treatment methotrexate are pro-oxidants since they are known to produce free radicals. Similar to this, it has been reported that transition metals like iron and copper, among others, go through the Fenton reaction and Haber–Weiss reaction and then produce excessive free radicals (Singh 2019). As a result, nanozymes that catalyse reactions involving the production of free radicals (such as peroxidase and oxidase) can also be referred to as pro-oxidant nanozymes.

372

B. G. Prajapati et al.

1.6 Peroxidase Mimetic Nanoparticles: Its Mode of Action Peroxidase is a biogenic enzyme that is found in a diverse range of organisms, including human bodies, flora and microorganisms (Munir et al. 2020). A broad category of catalysts referred to as natural peroxidases typically attack peroxidase targets utilising hydrogen peroxide. The fact that peroxidase enzymes serve as free radical purifying mediators deems them extremely important (e.g. glutathione peroxidase). The peroxidase enzyme’s major function is to wear down H2 O2 into non-toxic substances. The negative residue of O2 ’s oxygenation is H2 O2 . These enzymes (such as myeloperoxidase) also aid in the resistance against invasive infections (Lin et al. 2021). The horse radish peroxidase (HRP) enzyme has been well acknowledged for its applications in bioanalytical and forensic biochemistry, chiefly for its capacity to turn a neutral reagent into a bright product, which predominately improves the monitoring of sample matrix. New study shows that specific nanomaterials can have enzymatic activity mimicking those of the peroxidase enzyme. Significant work has been put on building credible alternative POD replica enzymatic methods to circumvent the restrictions of natural enzymes. Figure 1 offers a schematic illustration of the peroxidase activity which nanozymes present. Even while evidence primarily points to iron oxides having outstanding peroxidase enzyme-like ability, various nanoparticles have gotten a lot of attention here too. The original finding was reported by (Gao et al. 2007). The findings indicate that, in the environment of hydrogen peroxide and an acidic condition, various amounts of nanoparticles containing iron oxide (30, 50, and 300 nm) could convert

Fig. 1 Advantages of nanozymes over natural enzymes

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat …

373

Fig. 2 Schematic illustration demonstrating the ability of single electron donors to form superoxide anions

the neutral 3,3, ,5,5, -tetramethylbenzidine (TMB) into a turquoise product. Nonetheless, in comparison to their larger scaled predecessors, the tiny particles have the possibility to display enhanced peroxidase-like function. Researchers have found that reaction temperature and acidity seemed to have a bearing on both peroxidase activity of the iron oxide nanoparticles and normal HRP enzymes. Contrary to HRP, the nanoparticles are more resilient and continue to have enzymatic performance even after being cultured at a diverse range of temperatures (4-900C) and ph values (1–12). A greater concentration of hydrogen peroxide is speculated to attain the greatest functionality for iron oxide nanoparticles since the kinetic study also displayed that the enzyme binding (Km) number of nanoparticles containing iron oxide with hydrogen peroxide was bigger than HRP (154 and 3.7 mM, respectively). The quantity of subcellular H2 O2 and pH have a significant role in the process by which existing Peroxidase mimicking Nano molecules emit deadly ROS. The anticipated variation of internalised H2 O2 ions is (50 100 106 M) (Manea et al. 2004). Due to their limited pharmacological activity in the tumour cells, most nanozymes aren’t nearly as helpful as combination therapies when it is used alone (Fig. 2).

2 Peroxidase Mimicking Nanozyme in Tumour Hypoxia PDT is an advanced, conventional approach to treating cancer. In PDT, the activation process is started when the first photosensitizer absorbs irradiance of the correct wavelength. The primary cause of apoptosis is the accumulation of harmful ROS, which is produced when the excited electrons interact with the cellular oxygen. PDT is largely influenced by three factors: the photosensitizer, optimum and appropriate light wavelengths, and the amount of dissolved oxygen present in cells and tissue. PDT is a reaction that requires oxygen to occur. Unfortunately, due to the aggressive exponentiation of cancer cells and the constrained blood supply that neoplasms can access, oxygen saturation in malignancies are relatively low (pressure drop of O2 5 mmHg approximately to 7 M). Despite its lack of impact, low oxygen in cancers has a significant impact on PDT. The important approaches used to combat tumour

374

B. G. Prajapati et al.

hypoxia potentially increase O2 production by biocatalysts, O2 carriers, its distribution and O2 independent photosensitizers (Luo et al. 2022; Zhang et al. 2020; Wang et al. 2020a, b). To successfully regulate PDT by addressing tumour hypoxia, improved efficacy demands the creation of photosensitizers. In order to effectively treat tumour hypoxia and mediate the PDT effects by converting subcellular H2 O2 to H2 O and ROS, more work has subsequently been put into synthesising enzyme simulated materials, notably POD mimetics. Since it was learned recently that nanoparticles containing iron oxide have enzymatic behavioural traits, interest in biosensing of peroxidase-like Fe3 O4 nanostructured materials has increased (Wang et al. 2020a, b). Recently, it has been successful to catalyse cobalt-doped magnetoferritin (M-HFn) NPs (M-HFn-Cox Fe3 − x O4 ) with tunable cobalt deposition into M-HFn nuclei. The effectiveness of imaging the tumour-specific tissue is significantly improved by the possible assistance of cobalt, which dramatically increases the peroxidase-like activity (Wang et al. 2017) Cobaltdoped Fe3 O4 nanozymes have been flagged up by (Zhu et al. 2019) to have better peroxidase activity, which allows them to constructively accelerate intracellular H2 O2 at tiny concentration and showed potential in in vitro as well as in vivo anticancer potency (100-fold higher affinity) Therefore, by overcoming tumour hypoxia with POD-like behaviour, Fe-based nanoparticles were also employed in combination with photo therapies against cancer. For heterogeneous image guided photo theranostics formed multipurpose chitosan-encapsulated Fe3 O4 nanoparticles altered by CuS and porphyrin (Zhang et al. 2020; Wang et al. 2018). PtFe@Fe3 O4 nanostructures had been manufactured and discovered to possess multiple enzyme-like properties by Liu et al. in 2019 (Niu et al. 2019). Under acidic media, these nanostructures demonstrated both the CAT and POD-like behaviour that might successfully counteract hypoxia. As a matter of fact, photo-enhanced catalyst active intervention was successful in suppressing pancreatic cancer growth. Vacuous nitrogen-doped carbon nanostructures (HNCSs) and iron phthalocyanine (FePc) had been devised by Niu et al. (2019) for effective catalytic with dual radiation treatment (Feng et al. 2021). In addition to supporting the transformation of endogenous H2 O2 into ROS and O2 for tumour enzymatic treatment as well as boosting O2 -dependent PDT, FePc/HNCSs can simultaneously exhibit POD- and CAT-like activities. Alternative nanomaterials, such as platinum (Pt), palladium (Pd) and gold (Au)-based nanocomposites are also being researched for its probable use during cancer therapeutic applications in conjunction with Fe-based POD-like nanozymes. Nanoporous carbon which combines Au nanoparticles can assist POD mimicking activity and manufacture ROS for cancer cells’ internal oxidative injury. Nanosized trimetallic (Pd, Cu and Fe) alloys nanozyme (PCF-a NEs) for ultrasound- and led cancer therapy was described by Feng et al. (2021). Using a photosensitizer (Ce6 ) and Pt nanomaterials, (Xu et al. 2020) formed a three-dimensional neuronal microporous silica nanosphere (3D-dendritic MSN) substance for hypoxia that overrides PDT by transferring subcellular H2 O2 to oxidants and delivering adequate air pressure to the environment in the tumour cells for PDT.A NIR780-loaded serum albumin folate regulating gold-doped microporous carbon (OMCAPs@ rBSA-FA@IR780) nanoprobe was presented as a multimodal theranostic platform (Gao et al. 2019a,

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat …

375

b). A virus-like Fe3 O4 @Bi2 S3 nanoparticle (F-BS NCs) was manufactured using a simplified acoustic approach. POD mimetic activity was substantially boosted by the combinatorial association of POD mimetic Fe3 O4 nanoparticles with a semiconductor including a small band gap, Bi2 S3 (BS) (Wang et al. 2020a, b). In contrast to the medical platform, Sheng et al. designed hyaluronic acid insulated chlorin e6 (Ce6 ) packed into a POD substitute metal–organic structure (MOF) MIL-100 nanoparticles (CMH NPs) for symbiotic chemo-PDT (Dan et al. 2020). Under situations of circumneutral pH, PCF-a NEs showed a chain of POD and GSH peroxidase imitating activities. Furthermore, PCF-a NEs with photothermally elevated POD features encourage better tumour cell apoptosis. Biodegradable POD mimicking boron oxynitride (BON) microspheres were created by Zeng et al. (NSs). These POD imitative BON NSs have been found to be effective in the treatment of melanoma since these can catalytically induce carcinogenic OH radicals for the successful destruction of tumour cells both in vitro and in vivo (Zhao et al. 2021). Besides the ones already discussed, a handful of certain other POD mimicking nanozyme technologies have been extensively applied in antibacterial settings with relevant conclusions.

2.1 Nanozyme Implementations for the Treatment of Hypoxic Tumour Environments (TMEs) Over the past decade, nanomaterials have progressed swiftly, expanding frontiers in the clinical field, therapeutic applications and biomedical devices. The area of focus of nanomaterials in the clinical field has been biomedical imaging, targeted drug delivery system, and tumour therapy. Therapies for TME could aid from using nanomaterials that elevate oxygen saturation in cancers for higher oxygen-dependent cancer chemotherapy. Below, we have spoken about the nanomaterials’ bioactivity and medical potency (Veroniaina et al. 2021). Some of the main nanozymes expertising in this field are POD, SOD, CAT, OXD-like nanoparticles. POD-like nanozymes are frequently utilised for cancer therapies since they can conveniently help foster organic peroxides and H2 O2 , which seems to be the primary component for addressing that kind of complicated malignancies that are broadly disseminated throughout the body. Additional transition metal-containing nanomaterials, such as V, Mn, Au, Ag and Ir, could even showcase POD-like behaviour. The function of PODs could be mimicked by graphene oxide (GO) nanosheets, according to Song and colleagues. In redox processes where oxygen is lost and turned to H2 O or H2 O2 , OXDlike nanozymes are typically employed. According to a study done in 2004 by Comott et al., gold nanoparticles were held responsible for converting glucose into gluconate while reacting like GOx. It has been demonstrated that minute nanomaterials derived from metallic nanoparticles, such as Cu, Ir, Ag and Au, have the ability to simulate OXD. In a separate study, Wang and colleagues synthesised copper hexacyanoferrate (Cu-HCF), a single-site copper complex, to generate GSHOx-like nanozymes which

376

B. G. Prajapati et al.

might promote the development of H2 O2 while absorbing glutathione (GSH) (Zou et al. 2021). The biotransformation of the superoxide anion radical (O2 ) into O2 and H2 O2 is facilitated by SOD, which serves as a crucial antioxidant defence response in the body (Fig. 2). Other nanomaterials have good detailed SOD-mimicking characteristics as well. Samuel et al. reported that hydrophilic carbon colony densities might readily salvage micromolar to millimolar doses of deleterious O2 , and even that H2 O2 and O2 were the main essential catalytic products, allowing the nanosystem a potential biocompatible SOD. A lot of problems arise due to delayed detection of malignancies in the body thereafter hampering more number of cells and providing lesser time for treatment. Nanozymes have been successfully used for the detection of cancer-related genetics, compounds and cells owing to their benefits of excellent catalytic activity, relatively inexpensive, better stability and versatility. Additionally, due to the inherent features of nanoparticles, nanozymes can act as probes for precise and early imaging. Cancer biomarkers are generated which are highly specific to a particular type of cancer (Zhang et al. 2022). Nanozymes being known for their high catalytic activity and mimicking properties, they have been successfully employed in detection of biomarkers at a relatively early stage to prevent the cancer from spreading more and beginning with the appropriate course of treatment. Many such applications have been reported by researchers where study about TME and correlating it with nanozymes remains in light.

2.2 Recent Advancements and Future Scope Nanozymes that mimic PODs are currently being employed to defeat tumour hypoxia. Photothermal therapy is widely recognised to boost the therapeutic efficacy of nanozymes (Niu et al. 2019; Fan et al. 2018; Liu et al. 2019). Dual enzyme mimetic nanostructures’ synergistic effects in tumour therapy are also investigated. A Wontonlike Bismuth@poly Vinyl Pyrolidine@Gold Platinum (Bi@PVP@AuPt) Nanoparticle that exhibits dual POD and oxidase activity (Yim et al. 2020). In hypoxic tumours, the stable dual enzymatic action of nanoparticles results in a higher concentration of oxygen. Under dual photothermal and nanocatalytic treatment, applying the hyperthermia effect to the dual nanozyme also considerably promotes ROS formation, which finally leads to good therapeutic effects. For nano catalytic tumour therapy, recently described a dual inorganic nanozyme-based nanoplatform of Gold (Au) NPs and Fe3 O4 Nanoparticles co-loaded mesoporous silica materials (Gao et al. 2019a, b). Due to the fact that Au Nanoparticles mimics GOx, it will also catalyse the oxidation of -D-glucose into gluconic acid and H2 O2 , which will then be catalysed by the peroxidase mimic Fe3 O4 Nanoparticles and release highly toxic hydroxyl radicals to cause tumour-cell death by the typical Fenton-based catalytic reaction. By regulating the tumour environment created a glucose-oxidase (GOx)-loaded biomimetic Au–Ag hollow nanotriangles (Au–Ag-GOx HTNs) for NIR light-triggered tumour therapy (Xu et al. 2020). GOx in HTNs triggers the formation of gluconic acid

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat …

377

and H2 O2 . This will result in the Peroxidase mimic nanoparticles converting H2 O2 to O2 , ultimately enhancing the production of OH radicals under NIR-II light for effective tumour therapy described the tumour catalytic treatment effect of uniform Bismuth sulphide nanorods (Bi2S3 NRs) coated with dendritic mesoporous silica (Bi2S3@DMSN) material (Thangudu and su 2021). Under low pH, these synthetic nanozymes demonstrated dual enzyme mimicry, such as POD and CAT characteristics, which significantly overcame tumour hypoxia and increased oxidative stress in a hyperthermic environment. A selenium-melanin multishell nanozyme with manganese dioxide encapsulation for intracellular antioxidation, known as Se@Me@MnO2 , was recently created (Ai et al. 2021). It displayed numerous enzyme activity, including CAT, SOD and glutathione peroxidase (GPx). Through a synergistic interaction of rapid electron transfer between Se, Me and MnO2 , such multishell platforms may effectively scavenge ROS species. Future tumour therapeutic uses for these multienzyme mimic nanoplatforms seem promising as it created a zincbased single atom supported by a metal–organic framework employing the enzyme mimicking features of single atoms and observed good POD-like behaviours (Xu et al. 2019). These POD-like mechanisms also play a role in the efficient bacterial infections in vivo, and they led to the development of PEGylated single-atom Fecontaining nanocatalysts (PSAF NCs), which were used to precisely create hydroxyl radicals (OH) in acidic tumour environments (Huo et al. 2019). More recently, chlorin e6 (Ce6 ), a nanozyme similar to catalase for oxygen generation, was developed using a single atom of ruthenium as the active catalytic site anchored in the metal–organic framework Mn3 [Co(CN)6] (Wang et al. 2020a, b). For a variety of medical uses, mono nanozymes are being curated at present. By reaping the benefits of the atomic utilisation efficiency and saturation of binding site, subatomic particle metal cores that are economical and therefore are diffused atomically can considerably improve the features of enzyme analogues. As a result, frameworks with a single atom have been designed for a number of enzyme-like functionalities, including CAT, SOD, POD, etc. (Pei et al. 2020). For important potential treatment innovation, we must also take into account the existing objective evaluation markers in TME treatment in addition to a welldesigned POD nanozyme. The layout of novel and efficient nanozymes will demonstrate immense opportunities for a wide range of research for environmental quality, biomedical research and agronomic industry by overcoming all the underlying issues.

3 Application of Nanozymes 1. Agricultural technology • Nano enzymes will transform agriculture and the food business in a number of ways, including improved agronomic systems, enhanced plant nutrient uptake, disease detection and pest management.

378

B. G. Prajapati et al.

• The seed has the ability to grow on its own, but by adding nanozymes, we can boost the seed’s potential. • Smart seed is a form of seed that contains nanoencapsulations of a particular bacterial strain. • We can germinate seeds or quicken seed germination by using nano enzyme. • A wide range of possible advantages, including elevating quality and safety of food, cutting agricultural inputs, strengthening soil’s ability to absorb microelements, etc. • Since these aid in the breakdown process, the toxic effects in the compost are decreased to a much higher level 2. Biosensors technology • DNA-nanozyme applications, which integrate nanozymes with DNA, are particularly appealing for the generation of nanozyme-based biosensors, which has piqued the interest of many researchers. • The development of nanozyme-based sensors has been greatly expedited by the reports of sensors based on Genetic material or templated nanozymes for the detection of a variety of targets. • Labelling DNA can be done using nanoenzymes. • On the basis of immobilisation techniques, such as the adsorption of enzymes via van der Waals forces, ionic bonding or covalent bonding, nano-enzyme biosensors have been produced. • Nanozyme-based biosensors have been successfully created to recognise ions (Chhillar and Rana 2019). 3. Cancer therapy • For the purpose of treating cancer, harmful nanozymes can be delivered into cancer cells via nanomedicines. • Nano enzyme-based nanomedicines assist key roles in enhancing the therapeutic efficacy of treatments by altering the cancer microenvironment, including pH, glucose concentration, redox levels and heat shock protein expression. • Cerium oxide nanoparticles (NPs) were developed to take use of the special folate conjugation capacity to aid in the immunological detection of cancer cells. • The leading contributory factor to cancer patients’ deaths was cancer cell migration. Ultra-small manganese ferrite nanoparticles and the tumourtargeting peptide CREKA were combined to create the reagent known as UMFNP-CREKA, which facilitated the detection of ultra-small breast cancer metastasis. • Cu2-xTe NEs may influence the immunosuppressive tumour cells and display tunability enzymatic activity when exposed to NIR-II light (Yu et al. 2020; Sindhu et al. 2021; Sindhu et al. 2021).

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat …

379

4. Environmental engineering • MNPs, such as peroxidase, were thought to degrade the organic contaminants. Nanozymes are the best choices for real-time and/or remote environmental pollutant monitoring and cleanup because they provide stronger stability than the comparable natural enzymes. • The photocatalysis performed by nanozymes could be employed to filter UV radiation and other dangerous solar rays. • By creating the nanozymes used for product degradation, carbon fixation can be accomplished. • Nitrogen fixation can be accelerated by introducing nanozymes in the leguminous plants, yielding fertile soil that is rich in nitrogen. • Nano enzymes may be particularly important in the event of oil spills since they can facilitate the deconstruction of lipids into safe, water-soluble products (Pathakoti et al. 2018). 5. Future nanozymes in Hypoxia cancer remedial applications 1. Enzyme Activity: The discovery of new surface conjugation techniques to increase nanozyme activity is eagerly expected. Most nanozymes demonstrate lesser activity than natural enzymes due to the additional surface conjugation on NMs. 2. Nanozyme Sensitivity and selectivity: The majority of nanozymes have numerous enzyme activities that can catalyse a variety of substrates. A thorough examination of catalytic mechanisms is still required to fully comprehend the comprehensive mechanisms of nanozymes and their applicability in biomedical applications, despite the fact that multiple studies have looked at various forms of surface conjugation approaches to achieve selectivity. 3. Toxicity/biosafety: The cytotoxicity and biocompatibility of nanozymes have not yet been proven, in contrast to normal enzymes. 4. Limited to cancer therapy: The majority of nanozymes are now only used in cancer treatment. Diagnostics and treatments for diseases caused by nanozymes should be used for both environmental and agricultural problems as well as other hereditary diseases. 5. Biomedical applications: On long-term cytotoxicity, biosafety, stability and mechanisms, more research is required.

4 Conclusion With a thorough description of the principles and functions of POD resembling nanozymes in chemotherapeutic drugs, this paper summarises back in time, existing and future projects in the manufacture of nanozymes, mainly POD mimicking nanomaterials for oxygen-dependent radiation treatment. Tumour hypoxia is the greatest

380

B. G. Prajapati et al.

barrier to successful phototherapy for tumours. The majority of phototherapy modulation formats on oxygen, and the oxygen concentration in cancer microhabitats are unsatisfactory for medical utility. We have examined the advantages of POD mimetic nanoparticles that can speed up endogenous H2 O2 to H2 O and ROS and thus bypass tumour hypoxia, to get around this condition. Pairing treatment methods such as double, multienzyme techniques and moderate PTT-induced amplification of nanozyme activities can yield several medicinal benefits. To define directions for future nanozyme-based remedies and to reform inpatient settings, foreseeable ideas and concerns for the continued evolution of nanozyme application are explored. The merits, procedures and drawbacks of nanozymes are properly understood in the mentioned review hoping it would be beneficial to the medical community. We also believe that it will make it easier to develop new POD mimetic nanozymes for potential cancer phototherapy treatments. Future research should investigate nanozymes since the majority of them are restricted to only POD, CAT, OXD and SOD. For utilisation in a wide range of applications, the investigations should be undertaken in terms of other enzyme mimic activities.

References Ai Y, You J, Gao J, Wang J, Sun HB, Ding M, Liang Q (2021) Multi-shell nanocomposites based multienzyme mimetics for efficient intracellular antioxidation. Nano Res 14(8):2644–2653 Chen B, Rao X, House MG, Nephew KP, Cullen KJ, Guo Z (2011) GPx3 promoter hypermethylation is a frequent event in human cancer and is associated with tumorigenesis and chemotherapy response. Cancer Lett 309(1):37–45 Cheng G, Salerno JC, Cao Z, Pagano PJ, Lambeth JD (2008) Identification and characterization of VPO1, a new animal heme-containing peroxidase. Free Radical Biol Med 45(12):1682–1694 Chhillar AK, Rana JS (2019) Enzyme nanoparticles and their biosensing applications: a review. Anal Biochem 581:113345 Dan Q, Hu D, Ge Y, Zhang S, Li S, Gao D, et al Sheng Z (2020) Ultrasmall theranostic nanozymes to modulate tumor hypoxia for augmenting photodynamic therapy and radiotherapy. Biomater Sci 8(3):973–987 Ding M, Zhang Y, Li J, Pu K (2022). Bioenzyme-based nanomedicines for enhanced cancer therapy—nano convergence. SpringerOpen. Accessed 22 Sept 2022 Fan K, Xi J, Fan L, Wang P, Zhu C, Tang Y, Gao L, et al (2018) In vivo guiding nitrogen-doped carbon nanozyme for tumor catalytic therapy. Nat Commun 9(1):1–11 Feng L, Dong Z, Liang C, Chen M, Tao D, Cheng L, Liu Z (2018) Iridium nanocrystals encapsulated liposomes as near-infrared light controllable nanozymes for enhanced cancer radiotherapy. Biomaterials 181:81–91 Feng L, Liu B, Xie R, Wang D, Qian C, Zhou W, Zhao Y (2021) An ultrasmall SnFe2 O4 nanozyme with endogenous oxygen generation and glutathione depletion for synergistic cancer therapy. Adv Funct Mater 31(5):2006216 Gao S, Lin H, Zhang H, Yao H, Chen Y, Shi J (2019a) Nanocatalytic tumor therapy by biomimetic dual inorganic nanozyme-catalyzed cascade reaction. Adv Sci 6:1801733. https://doi.org/10. 1002/advs.201801733 Gao L, Zhuang J, Nie L, Zhang J, Zhang Y, Gu N, Yan X (2007) Intrinsic peroxidase-like activity of ferromagnetic nanoparticles. Nat Nanotechnol 2(9):577–583

Nanozymes: A Potent and Powerful Peroxidase Substitute to Treat …

381

Gao Z, Li Y, Zhang Y, Cheng K, An P, Chen F, Sun B, et al (2019b). Biomimetic platinum nanozyme immobilized on 2D metal–organic frameworks for mitochondrion-targeting and oxygen selfsupply photodynamic therapy. ACS Appl Mater Interfaces 12(2):1963–1972 Heckert EG, Karakoti AS, Seal S, Self WT (2008) The role of cerium redox state in the SOD mimetic activity of nanoceria. Biomaterials 29(18):2705–2709 Hirst SM, Karakoti AS, Tyler RD, Sriranganathan N, Seal S, Reilly CM (2009) Anti-inflammatory properties of cerium oxide nanoparticles. Small 5:2848–2856 Huang Y, Ren J, Qu X (2019) Nanozymes: classification, catalytic mechanisms, activity regulation, and applications. Chem Rev 119(6):4357–4412 Huo M, Wang L, Wang Y, Chen Y, Shi J (2019) Nanocatalytic tumor therapy by single-atom catalysts. ACS Nano 13(2):2643–2653 Jiang D, Ni D, Rosenkrans ZT, Huang P, Yan X, Cai W (2019) Nanozyme: new horizons for responsive biomedical applications. Chem Soc Rev 48(14):3683–3704 Korsvik C, Patil S, Seal S, Self WT (2007) Superoxide dismutase mimetic properties exhibited by vacancy engineered ceria nanoparticles. Chem Commun (camb) 14:1056–1058 Lin Y, Liu X, Liu Z, Xu Y (2021) Visible-light-driven photocatalysis-enhanced nanozyme of Tio2 nanotubes@ MoS2 nanoflowers for efficient wound healing infected with multidrug-resistant bacteria. Small 17(39):2103348 Liu H, Hua Y, Cai Y, Feng L, Li S, Wang H (2019) Mineralizing gold-silver bimetals into heminmelamine matrix: a nanocomposite nanozyme for visual colorimetric analysis of H2 O2 and glucose. Anal Chim Acta 1092:57–65 Luo S, Liu F, Gu S, Chen K, Yang G, Gu Y, Qu LL (2022) Nanozyme-mediated signal amplification for ultrasensitive photoelectrochemical sensing of Staphylococcus aureus based on Cu–C3N4– TiO2 heterostructure. Biosens Bioelectron 216:114593 Mahmudunnabi RG, Farhana FZ, Kashaninejad N, Firoz SH, Shim YB, Shiddiky MJ (2020) Nanozyme-based electrochemical biosensors for disease biomarker detection. Analyst 145(13):4398–4420 Manea F, Houillon FB, Pasquato L, Scrimin P (2004) Nanozymes: gold-nanoparticle-based transphosphorylation catalysts. Angew Chem Int Ed 43:6165−6169 Meng X, Zare I, Yan X, Fan K (2020) Protein-protected metal nanoclusters: an emerging ultra-small nanozyme. Wiley Interdiscip Rev: Nanomedicine Nanobiotechnol 12(3):e1602 Munir S, Shah AA, Rahman H, Bilal M, Rajoka MSR, Khan AA, Khurshid M (2020) Nanozymes for medical biotechnology and its potential applications in biosensing and nanotherapeutics. Biotech Lett 42(3):357–373 Muz B, de la Puente P, Azab F, Azab AK (2015) The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia 3:83 Niu X, Shi Q, Zhu W, Liu D, Tian H, Fu S, Lin Y (2019) Unprecedented peroxidase-mimicking activity of single-atom nanozyme with atomically dispersed Fe–Nx moieties hosted by MOF derived porous carbon. Biosens Bioelectron 142:111495 Pathakoti K, Manubolu M, Hwang H-M (2018) Nanotechnology applications for environmental industry. Handbook of nanomaterials for industrial applications. Accessed 22 Sept 2022 Pei J, Zhao R, Mu X, Wang J, Liu C, Zhang XD (2020) Single-atom nanozymes for biological applications. Biomater Sci 8(23):6428–6441 Serewko MM, Popa C, Dahler AL, Smith L, Strutton GM, Coman W, Saunders NA (2002) Alterations in gene expression and activity during squamous cell carcinoma development. Can Res 62(13):3759–3765 Sindhu RK, Najda A, Kaur P, Shah M, Singh H, Kaur P, Rahman MH (2021) Potentiality of nanoenzymes for cancer treatment and other diseases: current status and future challenges. Materials 14(20):5965 Singh S (2019) Nanomaterials exhibiting enzyme-like properties (nanozymes): current advances and future perspectives. Front Chem 7:46 Tao X, Wang X, Liu B, Liu J (2020) Conjugation of antibodies and aptamers on nanozymes for developing biosensors. Biosens Bioelectron 168:112537

382

B. G. Prajapati et al.

Thangudu S, Su CH (2021) Peroxidase mimetic nanozymes in cancer phototherapy: progress and perspectives. Biomolecules 11(7):1015 Veroniaina H, Wu Z, Qi X (2021) Innate tumor-targeted nanozyme overcoming tumor hypoxia for cancer theranostic use. J Adv Res 33:201–213 Wang D, Wu H, Phua SZF, Yang G, Qi Lim W, Gu L, Zhao Y, et al (2020a). Self-assembled single-atom nanozyme for enhanced photodynamic therapy treatment of tumor. Nat Commun 11(1):1–13 Wang Q, Wei H, Zhang Z, Wang E, Dong S (2018) Nanozyme: an emerging alternative to natural enzyme for biosensing and immunoassay. TrAC Trends Anal Chem 105:218–224 Wang X, Fan L, Cheng L, Sun Y, Wang X, Zhong X, Zha Z (2020b). Biodegradable nickel disulfide nanozymes with GSH-depleting function for high-efficiency photothermal-catalytic antibacterial therapy. IScience 23(7):101281 Wang Z, Dong K, Liu Z, Zhang Y, Chen Z, Sun H, Qu X (2017) Activation of biologically relevant levels of reactive oxygen species by Au/g-C3N4 hybrid nanozyme for bacteria killing and wound disinfection. Biomaterials 113:145–157 Wei H, Wang E (2013) Nanomaterials with enzyme-like characteristics (nanozymes): nextgeneration artificial enzymes. Chem Soc Rev 42(14):6060–6093 Xu B, Wang H, Wang W, Gao L, Li S, Pan X, Liu H et al (2019) A single-atom nanozyme for wound disinfection applications. Angewandte Chemie 131(15):4965–4970 Xu M, Lu Q, Song Y, Yang L, Ren C, Li W, Li N (2020) NIR-II driven plasmon-enhanced cascade reaction for tumor microenvironment-regulated catalytic therapy based on bio-breakable Au–Ag nanozyme. Nano Res 13(8):2118–2129 Yi Z, Jiang L, Zhao L, Zhou M, Ni Y, Yang Y, Zhu Y (2019) Glutathione peroxidase 3 (GPX3) suppresses the growth of melanoma cells through reactive oxygen species (ROS)-dependent stabilization of hypoxia-inducible factor 1-α and 2-α. J Cell Biochem 120(11):19124–19136 Yim G, Kim CY, Kang S, Min D-H, Kang K, Jang H (2020) Intrinsic peroxidase-mimicking Ir nanoplates for nanozymatic anticancer and antibacterial treatment. ACS Appl Mater Interfaces 12:41062–41070. https://doi.org/10.1021/acsami.0c10981 Yu Z, Lou R, Pan W, Li N, Tang B (2020) Nanoenzymes in disease diagnosis and therapy. Chem Commun 56(99):15513–15524 Zhang X, Chen X, Zhao Y (2022) Nanozymes: versatile platforms for cancer diagnosis and therapy. Nano-Micro Lett 14(1):1–27 Zhang Y, Jin Y, Cui H, Yan X, Fan K (2020) Nanozyme-based catalytic theranostics. RSC Adv 10(1):10–20 Zhao Y, Shi C, Cao J (2021) Biomimetic phototherapy in cancer treatment: from synthesis to application. Drug Deliv 28(1):2085–2099 Zhu J, Peng X, Nie W, Wang Y, Gao J, Wen W, Wang S (2019) Hollow copper sulfide nanocubes as multifunctional nanozymes for colorimetric detection of dopamine and electrochemical detection of glucose. BiosensS Bioelectron 141:111450 Zou MZ, Liu WL, Chen HS, Bai XF, Gao F, Ye JJ, Zhang XZ, et al (2021) Advances in nanomaterials for treatment of hypoxic tumor. Natl Sci Rev 8(2):nwaa160

NMR-Based Pharmacometabonomics of Nanoparticles for Treating Hypoxia Isha Gupta, Sonia Gandhi , and Sameer Sapra

Abstract Hypoxia is a prominent factor in cancer, chronic heart and kidney diseases, and reproductive diseases. This factor can trigger tumour metastasis, angiogenesis and is even responsible for chemotherapy resistance. A hypoxic environment compromises the effectiveness of conventional therapies. Hence, integrating multifunctional and intelligent nanomedicine in therapies and imaging can improve human health by overcoming limitations such as low drug distribution and perfusion, distribution in off-target-healthy tissues, no precision therapy and non-selective toxicity. These smart nanosystems can sense, respond and interact with the host environment. Currently, nanomedicine therapy is in its infancy. Variable patient responses to conventional drugs are a key issue for drug failure and increased mortality. At this stage, Pharmacometabonomics will help predict drug dosing effects such as safety, efficacy, pharmacokinetics and metabolism. This will ultimately pave the way for personalised medicine by using the local environment to optimise smart nanomedicine behaviour and improve the healthcare sector. Nuclear magnetic resonance (NMR) spectroscopy and Mass spectrometry (MS) are used for analysing metabolite profiles in biological fluids for Pharmacometabonomics. This chapter will provide an outline for evaluating the hypoxia related nanomedicine dosing effects through Pharmacometabonomics, helping in possible future developments. Keywords Hypoxia · Nanoparticles · Pharmacometabonomic · Pharmaceutical nanotechnology

I. Gupta · S. Gandhi (B) Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organisation (DRDO), Brig. S. K. Mazumdar Marg, Delhi 110054, India e-mail: [email protected] I. Gupta · S. Sapra Department of Chemistry, Indian Institute of Technology Delhi (IITD), Delhi, India © The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2023 S. Chawla et al. (eds.), Smart Nanomaterials Targeting Pathological Hypoxia, Smart Nanomaterials Technology, https://doi.org/10.1007/978-981-99-1718-1_20

383

384

I. Gupta et al.

1 Background Humans are aerobic eukaryotic organism which requires oxygen (O2 ) to meet metabolic demand (Ortiz-Prado et al. 2019). Oxygen act as a scavenger for removing harmful electrons or any critical substrate to maintain all enzymatic reactions (Chen et al. 2020). During aerobic respiration, oxygen is responsible for the breakdown of nutrients and energy production in the form of adenosine triphosphate (ATP) (Meletis and Wilkesa 2019; Ortiz-Prado et al. 2019). Oxygen also maintains homeostasis through the pressure gradient between membranes and tissues (Ortiz-Prado et al. 2019). The normal oxygen level (normoxia state) varies from 10 to 21% for healthy mammalian tissues (Zenewicz 2017). Among all tissues, the brain tissue consumes the maximum amount of oxygen pumped from the heart (Meletis and Wilkesa 2019). The condition of insufficient oxygen level (1–5%) can lead to a hypoxic state that can last for a short (acute hypoxia) or long (chronic hypoxia) period (Chen et al. 2020). Some tissues can tolerate some forms of hypoxia/ischaemia for longer, while the low oxygen levels can severely damage other tissues. Here, cells will perform adaptive physiological mechanisms for survival, such as anaerobic glycolysis (Chen et al. 2020) and hypoxia-inducible factor (HIF) signalling pathways (Zenewicz 2017), and autophagy (Luo et al. 2022). As soon as the oxygen levels are restored, these mechanisms are stopped. The low haemoglobin level, high altitude, poor tissue perfusion, decreased diffusion and impaired ventilation induces hypoxia in the mammalian cells. However, hypoxia is the most common pathological response involved in developing diseases such as anaemia, pneumonia, cancer, diabetes, depression, chronic heart, neuromuscular, kidney and reproductive diseases (Meletis and Wilkesa 2019). The hypoxic state is categorised into hypoxic, anaemic, stagnant and histotoxic. Hypoxic hypoxia occurs at high altitudes or due to respiratory diseases such as hypoventilation and pulmonary oedema. The military faces this condition during high-altitude operations (Shaw 2021). Stagnant hypoxia, also known as hypoperfusion, occurs when blood flow is abnormal due to shock, anaesthesia, or cardiac arrest. Anaemic hypoxia occurs when the O2 -carrying capacity of haemoglobin is reduced due to extreme blood loss, methemoglobinaemia and carbon monoxide poisoning. Histotoxic hypoxia occurs when cellular usage of oxygen is reduced due to reduced ATP production or cyanide poisoning (Manninen and Unger 2016). This chapter focuses on the role of hypoxia in different pathological conditions, the nanoparticles and their classifications, and recent nanotherapies for treating hypoxiarelated pathological conditions. The last section elaborately explains metabonomics, the integration of metabonomics in pharmacy leading to ‘pharmacometabonomics’ and also investigates the importance of pharmacometabonomics in understanding and designing better nanotherapeutic regimens against hypoxia-related diseases.

NMR-Based Pharmacometabonomics of Nanoparticles for Treating …

385

2 Role of Hypoxia in Different Pathological Conditions As discussed in the above section, hypoxia is associated with several diseases, which is illustrated in Fig. 1 (Luo et al. 2022). This section will briefly discuss a few diseases associated with hypoxia.

2.1 Cancer Cancer is the fifth leading cause of death in India. Cancer cases and death rates will rise to 29.5 million and 16.3 million, respectively, by 2040. Around 0.8 million new cancer cases are added yearly (Kulothungan et al. 2022). In this disease, uncontrolled cell division takes place due to genetic mutations. Tumour hypoxia occurs when the tumour reaches 1–2 mm in size, and it is responsible for cancer development, progression, immunosurveillance, glycolysis, angiogenesis, apoptotic resistance, genomic instability and metastasis (Ruan et al. 2009; Chen et al. 2018a, b). This condition also resists radiotherapy and chemotherapy (Chen et al. 2018a, b).

2.2 Infectious Diseases The novel coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which attacks the pulmonary tissues and damage gas exchange causing systemic hypoxia and acute respiratory distress

Fig. 1 Hypoxia-related pathological conditions

386

I. Gupta et al.

syndrome (ARDS) (Serebrovska et al. 2020). Pulmonary vasoconstriction leads to a hypoxic state in COVID-19, aiding viral replication, inflammation and progressive lung inflammation. Hypoxia also encourages viral replication in hepatitis C (Somers et al. 2020).

2.3 Diabetes Diabetes is a chronic, heterogeneous and metabolic disease. It is due to impaired functioning of the pancreas to produce enough insulin for maintaining glucose levels in the blood (Luo et al. 2022). Diabetes has become an epidemic in developing countries such as China and India due to obesity and unhealthy lifestyles of youth. In 2019, 77 million people had diabetes in India, which will rise to over 134 million by 2045. Around 57% of people remain undiagnosed (Pradeepa and Mohan 2021). It has been found that kidney, adipose, pancreatic islets, retina, skin and wound tissues are hypoxic in diabetes, ultimately complicating diabetes. HIF-1-mediated adaptive responses are compromised in diabetes, resulting in cellular dysfunction (Gunton 2020).

3 Nanoparticles Nanoscience and nanotechnology have revolutionised healthcare by introducing an interdisciplinary field, ‘nanomedicine’, which is in the infancy stage (Ventola 2012). European Union (EU) has recognised nanomedicine as a Key Enabling Technology which can provide a new and innovative medical solution to unresolved medical needs by developing better drugs for the early diagnosis, treatment or prevention of diseases with improved specificity, efficacy, bioavailability, personalisation, safety, dose–response and targeting (Soares et al. 2018; Ventola 2012). Here, the synergism of therapy and diagnosis in nanomedicine opens an area of nanotheranostics, which is even more sensitive and helps in the real-time monitoring of drug delivery (Ventola 2012; Muthu et al. 2014; Ladju et al. 2022). Nanomedicines have nanoparticles (NPs) as the main constituent, responsible for the improved properties and can mimic biological macromolecules. These medicines are usually engineered nanomaterials or nonbiological complex drugs (NBCDs) (Pelaz et al. 2017; Soares et al. 2018). The development in the past decade has resulted in the translation of ‘nanomedicine’ into the commercial world (Wagner et al. 2006). As a result, in 1995, the Food Drug and Administration (FDA) approved the first nanomedicine, Doxil, which marked the beginning of the nanotechnology era. Thus, this field has attracted the scientific community and the pharmaceutical sector. The nanomedicine market is anticipated to expand globally by 17.1% from 2017 to 2023 (Kad et al. 2022).

NMR-Based Pharmacometabonomics of Nanoparticles for Treating …

387

Nanomedicines are widely studied and being developed due to their biocompatibility, efficacy and possible potential (Kad et al. 2022). Nanomedicines are materials having at least one dimension of 100 nm or smaller (Makabenta et al. 2021). However, the European Commission and the International Organisation for Standardisation (ISO) Technical Committee 229 have defined nanomaterials as materials with dimensions ranging from 1 nm to 1 μm (Leon et al. 2020). Due to their size, shape and surface, these materials possess unique physicochemical characteristics compared to their bulk counterparts; therefore, they have gained prominence for different applications such as industries, healthcare, textiles, electronics and cosmetics. NPs are further classified into inorganic NPs (Metal and Metal oxides), organic NPs (dendrimers, micelles and liposomes), carbon-based NPs (graphene, fullerene, carbon nanotubes), silicate-based NPs (montmorillonite, hectorite and saponite) and ceramics NPs (Kad et al. 2022; Muthu et al. 2014). Figure 2 gives insight into the various NPs investigated for hypoxia-related pathological conditions (Pelaz et al. 2017). The limitations restricting the translation of nanoparticles into the commercial world are Pharmacokinetics, Pharmacodynamics, Toxicological assays and biocompatibility. The next section of the chapter will broadly explain Pharmacometabonomics, its emergence, application, workflow and recent developments in the NMR pharmacometabonomics of nanoparticles for treating hypoxia.

Fig. 2 Application of NPs for Hypoxia-related pathological conditions

388

I. Gupta et al.

4 Pharmacometabonomics The introduction of ‘omics’ revolutionised molecular biology. This term was derived from the Greek word ‘ome’, meaning whole or complete. Omics is the complete assessment of all biological molecules of the organism by integrating system biology and bioinformatics (Vailati-Riboni et al. 2017; Nalbantoglu and Karadag 2019). This branch of science is broadly categorised into four layers: Genomics, Transcriptomics, Proteomics and Metabolomics. Genomics was the first to emerge and deal with genomes related disease, prognosis and response to treatment. Transcriptomics investigates qualitatively and quantitatively the complete set of transcripts (RNA) produced by the genome (Hasin et al. 2017). These two omics have been explored in different fields, such as pharmaceuticals, theranostics, gene therapy and disease. The third layer, proteomics, examines the complete set of proteins, their structure and function. Metabonomics is the multiparametric comprehensive analysis of metabolome present in biological samples such as urine, serum, cerebrospinal fluid, amniotic fluid, cells and tissues. Here, the metabolome is the low molecular weight metabolites (5500 m) (Bakonyi and Radak 2004). Mountaineers have climbed Mount Everest, which has an elevation of 8,848 m above sea level and is the highest point on the lithosphere. Besides the Himalayas, several other mountain ranges such as Andes and Swiss Alps have the peaks reaching the heights of up to 6,096 m and provide just as many tourist attractions at elevations between 3000 and 5000 m. Higher altitudes result in thinner air because of reduced atmospheric pressure and a decline in oxygen partial pressure. Figure 3

402

A. Arya et al.

Fig. 3 High Altitudes and reference points with change in atmospheric pressure. Altitude above 1500 m is considered high altitude, 1500 to 3000 m as very high altitude and beyond 5500 m is considered as extreme altitude. [Photo-Sneha Singh-Greater Himalayas—Himachal Pradesh, India]

shows variations in barometric pressure in relation to height and a few reference locations (Peacock 1998). The decrease in oxygen partial pressure causes decrease in alveolar pO2 , consequently resulting in a decrease in the cellular oxygen concentration. When the partial pressure of oxygen in alveoli (pAO2 ) is normal, the red blood cells (RBCs) quickly diffuse oxygen molecules across the alveolar capillary membrane before combining them with haemoglobin. The passage of an RBC via a pulmonary capillary takes about 0.75 s while the body is at rest. At sea level, blood-oxygen tension in capillaries approaches alveolar oxygen tension in less than 0.25 s (Teppema and Dahan 2010). However, pAO2 is decreased at altitude, resulting in a slower increase in blood-oxygen tension in capillaries as compared to that of the sea level. At rest, the blood flow in capillary is still sufficient to bring oxygen tension closer to that of the alveoli, but when the transit-time is prolonged by activity, hypobaric hypoxia is significantly worsened. Above a certain point, these modifications trigger compensatory adaptive cardiopulmonary reactions, which include hyperventilation and the production of free radicals. This causes a state of oxidative distress and affects glucose metabolism, which eventually results in the onset of altitude sickness. Altitude sickness results from the partial pressure of oxygen (pO2 ) decreasing at low atmospheric pressure, which has a detrimental impact on human physiology (Rick 1995). The harshness of the sickness is largely affected by the rate of climb. Symptoms typically start to show up 48 h after arriving at a high altitude. The most typical signs and symptoms are a headache, shortness of breath, exhaustion, nausea and vomiting, as well as swollen hands, feet, and face (Hultgren 1996). The ability to combine coverage has been made possible by new networking and informatics techniques, despite the fact that high-altitude sickness is difficult to monitor internationally. In 2004, Hackett and colleagues reported on a global sample of high-altitude sickness, which was one such integration of altitude sickness data.

Nanoceria and Hypoxia: Promises and Challenges

403

Rapid climb to high altitudes is linked to physiological changes like shortness of breath, drowsiness, appetite loss (often accompanied by nausea), poor sleep, fatigue, insomnia and irritability in most common cases. In some exceptional and extreme cases, high-altitude pulmonary oedema (HAPE) and high-altitude cerebral oedema (HACE) may emerge as more challenging life-threatening situations. Although the exact causes of AMS are unknown, it is important to climb quickly and be exposed for a long time. The onset of AMS requires a few hours of exposure, but cases have been documented at exposure times as short as 12 h and at elevations as low as 2500 m (8000 ft). The emergence of inter-continental flights has also increased the prevalence of acute mountain sickness, which is yet another similar condition associated with air travel. It is generally suggested from the acclimatisation studies that the affected person should stay at the altitude reached for at least 24 h and rest if symptoms of altitude sickness start to occur. After the symptoms have fully subsided, the person can begin slowly ascending (Wilson et al. 2009). Among the most common symptoms of HAPE are elevated heart rate, rapid breathing, crackles in the lungs’ bases and a dry cough. A severe variant of AMS called HACE causes ataxia, hallucinations, comas and ultimately death. If HAPE or HACE symptoms appear, the victim should be supplemented with external oxygen supply and descend at lower altitudes right away. Ravenhill initially outlined HAPE in 1913. (West 1996), showing that within a couple of days of reaching high altitude, HAPE sets in and is characterised by symptoms like elevated breathing rate at rest, reduced tolerance to exercise and a feeling of tightness in the chest. Other symptoms of HAPE include tachypnea, central cyanosis, resting tachycardia and rales or wheezes in at least one lung fluid (Hackett et al. 1992). Haemoptysis and fever might also happen. Chest radiographic findings can vary, but typically reveal patchy alveolar intratations that could become diffuse as the disease worsens (Marticorena and Hultgren 1979; Vock et al. 1991). Although the precise cause of HAPE is not entirely understood, several theories suggest that it may be caused by abnormally enlarged pulmonary arteries and increased hypoxic pulmonary vasoconstriction (Penaloza and Sime 1969; Hultgren 1996; Allemann et al. 2000). Additionally, this causes localised hyperperfusion and increased capillary pressure, which finally result in trans microvascular fluid leakage (Schoene et al. 1988; Maggiorini et al. 2000; Zhao et al. 2001; Swenson and Maggiorini 2002). HACE is the final stage of AMS and is a potentially lethal brain-related illness that appears in patients over the course of a few hours to a couple of days, especially in people who are already affected by AMS or HAPE. HACE can manifest itself within a week after reaching an altitude of 2750 m (9022 ft) or above. It is most frequently observed in isolated regions considerably higher than aforementioned altitude. The primary characteristics of HACE, such as alterations in awareness and ataxia, are used to make the clinical diagnosis (Sutton 1992). Irrational behaviour that quickly develops into lethargy, fainting and coma are only a few examples of mental state changes. The presence of swelling of eye-disc or increased intracranial pressure (ICP), retinal haemorrhages and neurological impairments are additional physical symptoms of HACE that might aid in clinical diagnosis. Patients with HACE who experience these clinical circumstances in its exacerbated form develop cerebral

404

A. Arya et al.

herniation, which terminates in death (Hultgren 1996; Yarnell et al. 2000; Hackett 1999). Although research on intracranial pressure, the cerebrovascular system and magnetic resonance imaging (MRI) have improved our understanding of the overall effects of hypobaric hypoxia, there is still much to learn about the cellular and vascular mechanisms underlying these effects. Three processes can be used to categorise the beginning and development of high-altitude sickness: (a) the hypoxic trigger, (b) sensing/perception and (c) molecular alterations. Although symptoms of acute altitude sickness appear 6–7 h after exposure, they increase with increasing altitude and are relieved by returning to normal levels of inspired PO2 (Savonitto et al. 1992). Hypoxia has been the fundamental trigger for the disease since it began (Wu et al. 2007). The cellular oxygen sensing system detects these stimuli, which causes a compensating reaction. Higher mechanical pressure and increased permeability are the two main compensating events that are evoked (molecular). Nitric oxide (NO), a powerful vasodilator, can control enhanced permeability while the hyperventilatory reaction raises pressure. Due to the increased capillary pressure that results in HAPE as well as HACE, there is further a negative impact on the cardiopulmonary and cerebral systems. The development of oxidative stress-causing radicals, notably the reaction between superoxide and nitric oxide, which results in peroxynitrite, a powerful reactive molecule that causes localised histological damage, is another aspect that aggravates the illness.

2 Oxidant Generation is Central to High-Altitude Sickness One of the environmental stresses that affect organ function and consequent oxygenation of the organism is physiological hypoxia (Voelkel et al. 1981; Bartsch et al. 2011; Sylvester et al. 2012). The role of electron transport chain (mtETC), and several other associated enzymes to redox machinery such as nitric oxide synthase, nicotinamide adeninedinucleotide phosphate oxidase, xanthine oxidase reductase enzymes are more likely to produce reactive oxygen and nitrogen species (RONS) under these oxygen-limiting conditions, which also favour the development of an inflammatory process (Turrens 2003; Dosek et al. 2007; Winterbourn 2008). The production of RONS is also aided by the concurrent exhaustion of canonical antioxidant defence consisting of enzyme and non-enzyme entities (Singh et al. 2012). This redox imbalance brought on by hypoxia plays a significant role in pathogenic processes. Rapid variations in the oxygen environment and a decreased physiological adaptability make pathological progression more obvious. The organs most susceptible to variations in oxygen concentration are the brain and the lungs (Sharma et al. 2011). The primary duty of the highly specialised organ known as the lungs is to ensure that the body has an appropriate supply of oxygen. Lung functions and oxygen delivery to the body are compromised by pathological hypoxia and hypobaric hypoxia’s reduced pO2 as well as oxygen-limiting situations (Sylvester et al. 2012). By encouraging the production of RONS, these circumstances change the redox balance of the lung and cause severe structural damage to the key biomolecules of the cell (Dosek

Nanoceria and Hypoxia: Promises and Challenges

405

et al. 2007; Rosanna and Salvatore 2012). There is mounting evidence linking lifethreatening respiratory, inflammatory and vascular illnesses to oxidative stress in the lung (Bartsch et al. 2001; Faiss et al. 2013). The brain is another important organ that can be damaged by changes in oxygen concentration. Unlike the lungs, the brain has a high energy demand (and therefore a high oxygen consumption) and relies primarily on glucose as an energy source. The ratio of glucose to oxygen is crucial for the regulation of glucose metabolism. Low glucose transport to the brain or low oxygen levels have been linked to impaired homeostasis of glucose metabolism and associated with the generation of free radicals (Gadoth and Gobel 2011). These radicals further damage lipids and proteins through additive reactions, and high lipid content in the brain increases susceptibility to hypoxia. Altered synthesis, uptake and release of neurotransmitters formation of RONS and altered expression of a variety of genes have been clinically linked to pathophysiological conditions such as acute altitude sickness and HACE (Wilson et al. 2009). These conditions are generally characterised by insomania, anxiety and cognitive deficits. While some of these features are reversible upon acclimatisation, the irreversible cognitive deficit leads to mental illness and proves to be a potential socio-economic problem for people (Basnyat 2003). The hippocampus is the most important site for the formation of mammalian memory and cognitive functions (Turner 1969). Excessive formation of RONS and altered expression of various genes have been reported to cause hippocampal cell death (Choi 1996; Ruan et al. 2003). Since the role of RONS in neurodegeneration leading to cognitive deficits is now well established, the supplementation of antioxidants is considered one of the likely prophylaxis/interventions to improve the situation. The use of antioxidants in various studies has merely reached the clinical trials, as they are limited by the tight cross BBB (blood–brain-barrier) obstructions or rapid degradation causing repetitive doses. Therefore, better antioxidants are still being sought to improve the cognitive deficit caused by hypobaric hypoxia (Halliwell 2003).

3 Antioxidants as Potential Agents to Combat High-Altitude Sickness Exogenous/nutritional supplementation has been hailed as a significant modality for reducing oxidative dis-stress, despite the fact that all cells are endowed with a repertoire of antioxidant defence that rapidly combats any radical mediated harms to biological systems (Halliwell, et al. 2006). Recent epidemiological evidence shows that levels of recognised antioxidants and associated diseases are negatively correlated. cancer, cardiovascular disease and mortality from these diseases (Cameron and Pauling 1976; Willett and MacMahon 1984; Radimer et al. 2004). Antioxidant supplementation is therefore a growingly common technique to preserve healthy physiological function. New antioxidants with synthetic or natural origins are being developed. Consuming antioxidants has been suggested as one method of

406

A. Arya et al.

preventing the pathophysiology linked to oxidative stress. Several plant extracts such as those from Withania somnifera Seabuckthorn and metabolites such as huperzine A, quercetin, ascorbic acid, -tocopherol, melatonin, vitamin E, carotene and polyphenols are examples of antioxidant supplements with positive effects that can reduce hypoxia-induced oxidative stress (Barhwal et al. 2007; Wu et al. 2011; Patir et al. 2012; Shi et al. 2012; Baitharu et al. 2013; Gattererer et al. 2013). Alpha-ketoglutaric acid (α-KG) and 5-hydroxymethylfurfural (5-HMF) taken together before being exposed to hypoxia reduce carbonyl levels and oxidative stress, but had no effect on performance (Gattererer et al. 2013). Similar to this, giving arginine to hypoxic broiler chickens increased their pulmonary vascular function; the effect was further amplified by giving them antioxidant vitamins E and C. Nitric oxide (NO) bioavailability could be increased by arginine and antioxidant vitamins, reducing oxidative stress damage, and enhancing cardiac function. This study showed the beneficial effects of an antioxidant combination (Bautista-Ortega and and Ruiz-Feria 2010). Morris Water Maize performance, an indicator of cognitive functions was shown to be improved by oral administration of a standard antioxidant molecule, acetyl-L-carnitine. Furthermore, the production of free radicals, lactate dehydrogenase activity, lipid peroxidation and the apoptosis cascade in rat brain hippocampus could be reduced by the aforementioned molecule (Barhwal et al. 2007). The author’s lab has also demonstrated the use of artificial short peptides (NAP to enhance cognition and reduce oxidative stress in hypobaric hypoxia (Sharma et al. 2011). The quantity constraints, cos and the chemicals’ unclear mechanism of action, however, have restricted their utilisation. Therefore, novel antioxidants with regeneration capability are being investigated for improved and efficient activity.

4 Nanoceria as Potential Next-Gen Antioxidant Against Hypoxic-Perturbations Cerium, the second element in the lanthanide series and the most prevalent member of the rare earth family, is getting close to reaching industrial metal levels of other useful metals such as tin and zinc. Cerium’s strong redox abilities, especially, its alternating ionic forms, the cerous ion, Ce3+ and the ceric ion, Ce4+ are of considerable interest in redox biology, perhaps due to its unique electron configuration Xe- 4f1 5d1 6s2 in its ground state. The ionic radius of the more oxidised state is smaller, at 97 pm as opposed to 114 pm, as would be expected. Unexpectedly, even the Ce3+ ionic radius is substantially smaller than di-anion of oxygen in the crystal lattice of ceria, which has an ionic radius of 135 pm. This characterises the change between two electronic states that differ only by a single 4f electron as the Nernstian behaviour of cerium ions in liquids (Traovarelli 2002). Each cerium atom was found to be eightfold coordinated (connected to eight oxygen atoms) in the crystalline state, as opposed to each oxygen atom being fourfold coordinated. The crystal structure of Ce4 O8 is typically a fluorite structure which is a face-centred cubic (Traovarelli 2002).

Nanoceria and Hypoxia: Promises and Challenges

407

Fig. 4 Typical electron micrographs of nanoceria particles synthesised using solvothermal route. These nanoparticles of size 5–10 nm have shown enormous promise as an antioxidant specially to combat environmentally induced oxidative stress

Owing to oxygen vacancies in nanoceria crystals, a large proportion of unique biological activities of nanoceria mediated catalysis is reported. The idea of an oxygen vacancy is that one or more of the eight octants in a ceria unit cell have one or more oxygen atoms (or atoms for a di- or tri-vacancy) absent from them (Campbell and Peden 2005). Atomic models make this theoretically simple to represent. Esch and colleagues (Esch et al. 2005) using scanning tunnelling microscopy (STM) along with calculations of density functions demonstrated that surface and subsurface oxygen vacancies generated at 900 °C are perhaps, the most visually convincing crystal defects of nanoceria. There is a general consensus that the proportion of cerium atoms in the Ce3+ rises on decreasing the size of particles, while the issue of the charge of the cerium atoms and interaction with the oxygen vacancies is still up for debate. Seal and associates (Deshpande et al. 2005), using the high-resolution TEM, calculated a concurrent increase in Ce3+ proportion with reduction in particle size. Because of this, biological models showed that the reduction of cerium oxide to nanoscale was advantageous. Another notable characteristic of cerium oxide at the nanoscale is that as the particles get smaller, the lattice expands, which results in less oxygen release and recapture and more reduced cerium atoms (Reed et al. 2014). A few electron micrographs of typical nanoceria particles are shown in Fig. 4.

5 Promises of Future Use of Nanoceria Cerium oxide was employed as an abrasive catalyst in chemical mechanical planarization and polishing long before its biological activity was understood. It is also used as a catalyst in fuel cell and catalytic converter power generation as well as in fuel borne additives. Over the past ten years, studies showing the effective therapeutic or preventive effects of nanoceria have multiplied. For instance, the major event responsible for autocatalytic free radical scavenging has been recognised as the

408

A. Arya et al.

changeover between Ce3+ and Ce4+ oxidation states, making nanoceria a more effective antioxidant than current ones. Nanoceria can scavenge superoxide, hydrogen peroxide, hydroxyl and nitric oxide radicals because of oxygen defects in the crystal lattice. The nanoceria has shown to mimic several antioxidant enzyme activities, in particular, superoxide dismutase (SOD) (Heckert et al. 2008), oxidase (Asati et al. 2009), catalase (Pirmohamed et al. 2010), alkaline phosphatase (Hayat et al. 2013) and peroxynitrite scavenging activities (Dowding et al. 2013), which further expand the horizons of its benefits and also, its tuneable redox chemistry. Numerous in vitro studies show that CNPs have cytoprotective and antioxidant properties in endothelial cells, pancreatic cells, monocytes, (Niu et al. 2001; Hussain et al. 2012; Chen et al. 2013; Watson and Zhao 2013). According to studies, CNPs inhibit apoptosis through direct radial scavenging and modulation of apoptotic signalling pathways. Numerous studies in animal models have also shown the function of nanoceria in the prevention of light-induced damage to the retina (Chen et al. 2006), the reduction of liver inflammation (Suzanne and Steller 2009), the protection of the heart (Niu et al. 2007) and the protection of the nervous system (Niu et al. 2009; Estevez et al. 2011). Additionally, nanoceria has been demonstrated to be helpful in regenerative medicine and cancer prevention (Figueroa 2014; Das et al. 2014). Table 1 summarises biological studies that show nanoceria has positive benefits.

6 Challenges in Future Use of Nanoceria Traditional pharmacokinetics investigation and toxicity evaluation must be conducted in order to move nanoceria from basic research to clinical application level. Evidence of the ecotoxicity and biodistribution of nanoceria was described in a recent review study (Hirst et al. 2013; Yokel et al. 2012, 2014; Molina et al. 2014). From ecotoxicological studies, different modes of nanoceria absorption, such as oral, cutaneous, ophthalmic, pulmonary and intravenous, have been researched. Intravenous route was found to be of particular significance among these modes because it provides a benchmark for the pharmacokinetic fate of nanoceria once they are injected into blood. To push the bio-persistence of nanoceria and the majority of other inorganic nanomaterials into clinical trials, researchers are still up against challenges. Organs in the mononuclear phagocytic system receive the majority of nanoceria that enter the bloodstream. The body of research demonstrates that the dispersion of nanoceria is unaffected by dose, shape or dosing regimen. Less than one percent of administered dose of nanoceria is distributed to the rest of the body from lungs, and much less is distributed via the GI tract. Given that nanoceria within the cells and organ distribution can last for several months, the clearance rate appears to be exceedingly sluggish. Acute toxicity from nanoceria is also quite low. Granulomas and lung and liver fibrosis are the results of high/accumulative doses, though. Generally, toxicity increases with exposure time and effects wear off gradually. This may be because nanoceria are bio-persistent (Yokel et al. 2012).

Nanoceria and Hypoxia: Promises and Challenges

409

Table 1 Summary of some recent- studies on beneficial effects of nanoceria in vitro and in vivo S. No

Study model

Effect

References

In vitro studies 1

TM cell culture

Alleviation of ocular hypertension

Luo et al. (2021)

2

MC3T3-E1

Osteoporesis treatment

Pinna et al. (2021)

3

PEK cell culture

Gastroentritis virus inhibition

Rybalko (2021)

4

G-SIS (intestinal submucosa)

Tissue engineering

Singh et al. (2021)

5

Cancer cells

pH responsive drug release

Chen et al. (2021)

6

3T3 cells

Treatment of diabetic wound

Hussein et al. (2021)

7

MC3T3-E1

Promoting osteogenesis

Shao et al. (2021)

8

Osteoblasts

Matrix for bone therapeutics

Kurian et al. (2022)

9

HepG2

Phytotherapy of hepatocellular carcinoma

Ghorani-Azam et al. (2022)

10

Tumour cells

Target killing of cancer cells

Ma et al. (2022)

In vivo studies 1

Rats (CNS model)

Recovery of central nervous system injury

Yang et al. (2022)

2

Rat (ocular model)

Recovery of light-induced damage

Cui et al. (2022)

3

Rats (pulmonary model)

Alleviation of pulmonary toxicity

Quian et al. (2021)

4

Guinea Pig

Ophthalmic applications

Lyu et al. (2021)

5

Mice (Cancer model)

Anticancer roles

Thakur et al. (2021)

The investigations for determining the biodistribution in CD-1 mice were carried out using intravenous injections of nanoceria, at very low doses showed 15–20 nm agglomerate particles as depositions (Karakoti et al. 2008; Hirst et al. 2013). The findings showed that the spleen, liver, lung and kidney all had higher concentrations of cerium than the brain, which had none. Cerium excretion in the urine was also undetectable. The outcome suggests that there was very little organ damage during the one-month exposure suggesting non-saturation of organs with nanoceria during this period. For the 15, 30 and 55 nm nanoceria, the 2% proportion of the nanoparticles still in the bloodstream ten minutes later to an intravenous injection indicated highly fast translocation. While just 35% of 5 nm ceria could prevent opsonin adsorption or binding to RBCs, possibly to avoid being quickly targeted by macrophages and

410

A. Arya et al.

remained in the circulation for 10 min after 1 h infusion (Hirst et al. 2011). Similar to this, ~3 nm ceria coated in citrate-EDTA was eliminated from blood with a 3.7-h plasma half-life. Two to four hours after the injection of the 15- and 30-nm ceria, but not the 5- or 55-nm ceria or the cerium ion, blood ceria increased (Heckman et al. 2013). When IP injected 3–5 nm primary particles with fluorescent-tagged nanoceria into CD-1 mice at the same dose (0.5 mg kg1) and injected on the same schedule (2–5 weekly injections) as IV (see above), Yokel et al. provided exhaustive reports on organ-distribution of nanoceria across the body of experimental animals one week later than the dose intervention and suggested the possible routes of uptake of nanoceria (Yokel et al. 2014). Although nanoceria has been proposed as potential antioxidant with tuneable redox chemistry, as an irony, the toxicity of nanoceria on the other hand might be caused by oxidative stress. Displacement of cerium oxide nanoparticles may be the cause of adverse effects seen at the distant site of exposure. In biological contexts, nanoceria can alter its nature and impact biological components. Greater Ce3+ surface area has been linked to increased toxicity; this relationship becomes more important when particle size is reduced and surface-to-volume ratios rise. Table 2 provides a summary of the hazardous values of nanoceria at various concentrations and methods. There are speculations that prolonged nanoceria-contact or bio-persistence could result in negative health impacts and resultant toxicity that grows over time, yet we still have scantly evidences in humans and need further studies to prove the hypothesis. The current understanding of nanoceria, however, could reveal that the threshold of 50 mg per Kg body weight dose of nanoceria is substantially below dangerous concentrations and can thus be efficiently employed to evaluate the positive impacts of nanoceria in biological systems. This understanding is based on in vivo and in vitro research. Nanoceria has to be further studied at the sub-clinical and clinical levels in order to be evaluated as a viable therapeutic and preventative agent for Table 2 Summary of some recent toxicological studies on nanoceria in vitro and in vivo In vitro studies on toxic effects of nanoceria 1

Human cancer cells

Cytotoxicity

Jyothi and Tharayil (2022)

2

Human RBCs

Cytotoxicity (size dependent

Prokopiuk et al. (2022)

In vivo studies on toxic effects of nanoceria 1

Earthworm

Oxidative injury

290–2325 μmol/kg mg/m3

Li et al. (2022)

2

Several organisms

Ecotoxicity

641

Yi et al. (2021)

3

Microbes

Antimicrobial effects

290–4350 μmol/kg

Baker et al. (2022)

4

Rats

Male sterility

495 μmol/kg

Maciejewski et al. (2022)

5

Plants

Toxicity

495 μmol/kg

Fattahi et al. (2022)

Nanoceria and Hypoxia: Promises and Challenges

411

many diseases that are directly related to oxidative stress based on the considerable scientific knowledge acquired in the last 10 years.

7 Conclusion In light of the research finding for over a decade, nanoceria has emerged as an interesting redox-regulating molecule which has its own advantages such as redox cycling and low-dose requirements. Although nanoceria presents enormous opportunities and prospective prophylactic molecule, the concerns about the longterm toxicity and ill-effects on environment are yet to be understood completely. Despite this, a number of patents well-recognised scientific studies are evident to demonstrate the promising future of this novel nanomaterial for combating various environment-induced stresses. Acknowledgements Authors would like to acknowledge Prof. Mainak Das, Dr. Niroj Sethy, Dr. Kalpana Bhargava for being part of the DIP-254 project and mentorship while some of the studies were conducted by the Author (AA).

References Allemann Y, Sartori C, Lepori M, Pierre S, Melot C, Naeije R, Scherrer U, Maggiorini M (2000) Echocardiographic and invasive measurements of pulmonary artery pressure correlate closely at high altitude. Am J Physiol Hear Circ Physiol 279(4):H2013–6 Asati A, Santra S, Kaittanis C, Nath S, Perez JM (2009) Oxidase-like activity of polymer-coated cerium oxide nanoparticles. Angew Chem Int Ed Engl 48(13):2308–2312 Baitharu I, Jain V, Deep SN, Hota KB, Hota SK, Prasad D, Ilavazhagan G (2013) Withania somnifera root extract ameliorates hypobaric hypoxia induced memory impairment in rats. J Ethnopharmacol 145(2):431–441 Bakonyi T, Radak Z (2004) High altitude and free radicals. J Sports Sci Med 3(2):64–69 Barker E, Shepherd J, Asencio IO (2022) The use of cerium compounds as antimicrobials for biomedical applications. Molecules 27(9):2678 Bartsch P, Straub PW, Haeberli A (2001) Hypobaric hypoxia. Lancet 357(9260):955–956 Basnyat B (2003) Steadfast to the summit: living and climbing at altitude. Natl Med J India 16(5):276–277 Bautista-Ortega J, Ruiz-Feria CA (2010) L-arginine and antioxidant vitamins E and C improve the cardiovascular performance of broiler chickens grown under chronic hypobaric hypoxia. Poult Sci 89(10):2141–2146 Cameron E, Pauling L (1976) Supplemental ascorbate in the supportive treatment of cancer: prolongation of survival times in terminal human cancer. Proc Natl Acad Sci USA 73(10):3685–3689 Campbell CT & Peden CH (2005) Chemistry. Oxygen vacancies and catalysis on ceria surfaces. Science 309(5735):713–4. Chen M, et al (2021) Construction of nanoceria-capped mesoporous silica carriers for redox/pHresponsive drug release. J Drug Deliv Sci Technol 66:102763 Cui W, et al (2022) Nanoceria for ocular diseases: recent advances and future prospects. Mater Today Nano 100218

412

A. Arya et al.

Chen S, Hou Y, Cheng G, Zhang C, Wang S, Zhang J (2013) Cerium oxide nanoparticles protect endothelial cells from apoptosis induced by oxidative stress. Biol Trace Elem Res 154(1):156– 166 Chen J, Patil S, Seal S, McGinnis JF (2006) Rare earth nanoparticles prevent retinal degeneration induced by intracellular peroxides. Nat Nanotechnol 1(2):142–150 Deshpande S, Patil S, Kuchibhatla S, Seal S (2005) Size dependency variation in lattice parameter and valency states in nanocrystalline cerium oxide. Appl Phys Lett 87(13) Dosek A, Ohno H, Acs Z, Taylor AW, Radak Z (2007) High altitude and oxidative stress. Respir Physiol Neurobiol 158(2–3):128–131 Dowding JM, Seal S, Self WT (2013) Cerium oxide nanoparticles accelerate the decay of peroxynitrite (ONOO). Drug Deliv Transl Res 3(4):375–379 Duong TT, Antao S, Ellis NA, Myers SJ, Witting PK (2008) Supplementation with a synthetic polyphenol limits oxidative stress and enhances neuronal cell viability in response to hypoxiare-oxygenation injury. Brain Res 1219:8–18 Esch F, Fabris S, Zhou L, Montini T, Africh C, Fornasiero P, Comelli G, Rosei R (2005) Electron localization determines defect formation on ceria substrates. Science 309(5735):752–755 Estevez AY, Pritchard S, Harper K, Aston JW, Lynch A, Lucky JJ, Ludington JS, Chatani P, Mosenthal WP, Leiter JC, Andreescu S, Erlichman JS (2011) Neuroprotective mechanisms of cerium oxide nanoparticles in a mouse hippocampal brain slice model of ischemia. Free Radical Biol Med 51(6):1155–1163 Faiss R, Pialoux V, Sartori C, Faes C, Deriaz O, Millet GP (2013) Ventilation, oxidative stress, and nitric oxide in hypobaric versus normobaric hypoxia. Med Sci Sports Exerc 45(2):253–260 Fattahi N, Tabrizi BH, Rani S, Sadeghi Z, Dehghanian Z, Lajayer BA, van Gadoth N, Gobel HH (2011) Oxidative stress and free radical damage in neurology. Springer, London Gatterer H, Greilberger J, Philippe M, Faulhaber M, Djukic R, Burtscher M (2013) Short-term supplementation with alpha-ketoglutaric acid and 5-hydroxymethylfurfural does not prevent the hypoxia induced decrease of exercise performance despite attenuation of oxidative stress. Int J Sports Med 34(1):1–7 Ghorani-Azam A, et al (2022) Resveratrol-mediated gold-nanoceria synthesis as green nanomedicine for phytotherapy of hepatocellular carcinoma. Front Biosci-Landmark 27.8:227 Hackett PH, Roach RC, Schoene RB, Harrison GL, Mills WJ Jr (1985) (1988) Abnormal control of ventilation in high-altitude pulmonary edema. J Appl Physiol 64(3):1268–1272 Hackett PH (1999) The cerebral etiology of high-altitude cerebral edema and acute mountain sickness. Wilderness Environ Med 10(2):97–109 Halliwell B (2003) Plasma antioxidants: health benefits of eating chocolate? Nature 426(6968):787 Halliwell B (2006) Reactive species and antioxidants. Redox biology is a fundamental theme of aerobic life. Plant Physiol 141(2):312–322 Heckert EG, Karakoti AS, Seal S, Self WT (2008) The role of cerium redox state in the SOD mimetic activity of nanoceria. Biomaterials 29(18):2705–2709 Heckman KL, DeCoteau W, Estevez A, Reed KJ, Costanzo W, Sanford D, Leiter JC, Clauss J, Knapp K, Gomez C, Mullen P, Rathbun E, Prime K, Marini J, Patchefsky J, Patchefsky AS, Hailstone RK, Erlichman JS (2013) Custom cerium oxide nanoparticles protect against a free radical mediated autoimmune degenerative disease in the brain. ACS Nano 7(12):10582–10596 Hirst SM, Karakoti A, Singh S, Self W, Tyler R, Seal S, Reilly CM (2013) Bio-distribution and in vivo antioxidant effects of cerium oxide nanoparticles in mice. Environ Toxicol 28(2):107–118 Hullebusch ED (2022) Toxicity of nanoparticles onto plants: Overview of the biochemical and molecular mechanisms. Toxic Nanoparticles Plants 69–94 Hultgren HN (1996) High-altitude pulmonary edema: current concepts. Annu Rev Med 47:267–284 Hussein MAM, et al (2021) Development and in vitro evaluation of biocompatible pla-based trilayer nanofibrous membranes for the delivery of nanoceria: a novel approach for diabetic wound healing. Polymers 13.21:3630

Nanoceria and Hypoxia: Promises and Challenges

413

Jyothi PP, Tharayil NJ (2022) Cyto-toxic Effect of Nano Ceria bio fabricated with DNA. IOP Conf Ser Mater Sci Eng 1263(1):012032. (IOP Publishing) Kapos V, Rhind J, Edwards M, Price MF, Ravilious C (2000) Developing a map of the world’s mountain forests. In: Price MF, Butt N (eds) Forests in sustainable mountain development: a state-of-knowledge report for 2000. Wallingford, UK, pp 4–9 Karakoti AS, Monteiro-Riviere NA, Aggarwal R, Davis JP, Narayan RJ, Self WT, McGinnis J, Seal S (1989, 2008) Nanoceria as antioxidant: synthesis and biomedical applications. JOM 60(3):33–37 Kurian AG, et al (2022) Surface-engineered hybrid gelatin methacryloyl with nanoceria as reactive oxygen species responsive matrixes for bone therapeutics. ACS Appl Bio Mater 5.3:1130–1138 Li W, He E, Zhang P, Li Y, Qiu H (2022) Multiomics analyses uncover nanoceria triggered oxidative injury and nutrient imbalance in earthworm Eisenia fetida. J Hazard Mater 437:129354 Luo L-J, Nguyen DD, Lai J-Y (2021) Harnessing the tunable cavity of nanoceria for enhancing Y-27632-mediated alleviation of ocular hypertension. Theranostics 11.11:5447 Lyu Q, Peng L, Hong X, Fan T, Li J, Cui Y, Zhao J (2021) Smart nano-micro platforms for ophthalmological applications: The state-of-the-art and future perspectives. Biomaterials 270:120682 Maciejewski R, Radzikowska-Büchner E, Flieger W, Kulczycka K, Baj J, Forma A, Flieger J (2022) An overview of essential microelements and common metallic nanoparticles and their effects on male fertility. Int J Environ Res Public Health 19(17):11066 Maggiorini M, Melot C, Pierre S, Pfeiffer F, Greve I, Sartori C, Lepori M, Hauser M, Scherrer U, Naeije R (2001) High-altitude pulmonary edema is initially caused by an increase in capillary pressure. Circulation 103(16):2078–2083 Marticorena E, Hultgren HN (1979) Evaluation of therapeutic methods in high altitude pulmonary edema. Am J Cardiol 43(2):307–312 Meybeck M, Green P, Vörösmarty C (2001) A new typology for mountains and other relief classes: an application to global continental water resources and population distribution. Mt Res Dev 21(1):11 Molina RM, Konduru NV, Jimenez RJ, Pyrgiotakis G, Demokritou P, Wohlleben W & Brain JD (2014) Bioavailability, distribution and clearance of tracheally instilled, gavaged or injected cerium dioxide nanoparticles and ionic cerium. Environ Sci Nano 1:13 Niu J, Azfer A, Rogers LM, Wang X, Kolattukudy PE (2007) Cardioprotective effects of cerium oxide nanoparticles in a transgenic murine model of cardiomyopathy. Cardiovasc Res 73(3):549– 559 Patir H, Sarada SK, Singh S, Mathew T, Singh B, Bansal A (2012) Quercetin as a prophylactic measure against high altitude cerebral edema. Free Radical Biol Med 53(4):659–668 Peacock AJ (1998) ABC of oxygen: oxygen at high altitude. BMJ 317(7165):1063–1066 Penaloza D, Sime F (1969) Circulatory dynamics during high altitude pulmonary edema. Am J Cardiol 23(3):369–378 Pinna A, et al (2021) Nanoceria provides antioxidant and osteogenic properties to mesoporous silica nanoparticles for osteoporosis treatment. Acta Biomaterialia 122:365–376 Pirmohamed T, Dowding JM, Singh S, Wasserman B, Heckert E, Karakoti AS, King JE, Seal S, Self WT (2010) Nanoceria exhibit redox state-dependent catalase mimetic activity. Chem Commun (camb) 46(16):2736–2738 Prokopiuk V, Onishchenko A, Yefimova S, Maksimchuk P, Seminko V, Nakonechna O, Tkachenko A et al (2022) Size-dependent effect of CeO2 nanoparticles on ROS generation in red blood cells. In: 2022 IEEE 12th international conference nanomaterials: applications and properties (NAP). IEEE, pp NRA07-1 Qian Q, Zhang Y, Chen Y, Ye C, Feng Q, Tu J et al (2021) Assessment of pulmonary toxicity of potential antioxidant drug PEGylated nanoceria after intratracheal instillation in rats. J Appl Toxicol 41(6):941–952

414

A. Arya et al.

Radimer KL, Ballard-Barbash R, Miller JS, Fay MP, Schatzkin A, Troiano R, Kreger BE, Splansky GL (2004) Weight change and the risk of late-onset breast cancer in the original Framingham cohort. Nutr Cancer 49(1):7–13 Reed K, Cormack CM, kulkarni A, Mayton M, Sayle DC, Klaessig F, Stadler B (2014) Exploring the properties and applications of nancoeria: is there plenty of room at teh bottom? Environ Sci. Nano 1(1):14 Rick C (1995) Outdoor action guide to high altitude: acclimatization and illnesses. Princeton University, United States Rosanna DP, Salvatore C (2012) Reactive oxygen species, inflammation, and lung diseases. Curr Pharm Des 18(26):3889–3900 Ruan YW, Ling GY, Zhang JL, Xu ZC (2003) Apoptosis in the adult striatum after transient forebrain ischemia and the effects of ischemic severity. Brain Res 982(2):228–240 Rybalko S, et al (2021) Nanoceria can inhibit the reproduction of transmissible gastroenteritis virus: Consideration for use to prevent and treat coronavirus disease. Mikrobiolohichnyi Zhurnal 67–75 Savonitto S, Cardellino G, Doveri G, Pernpruner S, Bronzini R, Milloz N, Colombo MD, Sardina M, Nassi G, Marraccini P (1992) Effects of acute exposure to altitude (3,460 m) on blood pressure response to dynamic and isometric exercise in men with systemic hypertension. Am J Cardiol 70(18):1493–1497 Schoene RB, Swenson ER, Pizzo CJ, Hackett PH, Roach RC, Mills WJ Jr, Henderson WR Jr, Martin TR (1985) (1988) The lung at high altitude: bronchoalveolar lavage in acute mountain sickness and pulmonary edema. J Appl Physiol 64(6):2605–2613 Singh M, Arya A, Kumar R, Bhargava K, Sethy NK (2012) Dietary nitrite attenuates oxidative stress and activates antioxidant genes in rat heart during hypobaric hypoxia. Nitric Oxide Biol Chem/Off J Nitric Oxide Soc 26(1):61–73 Singh H, et al (2021) Biomatrix from goat-waste in sponge/gel/powder form for tissue engineering and synergistic effect of nanoceria. Biomed Mater 16.2:025008. Shao D, et al Titania nanotube array supported nanoceria with redox cycling stability ameliorates oxidative stress-inhibited osteogenesis. Chem Eng J 415(2021):128913 Sharma NK, Sethy NK, Meena RN, Ilavazhagan G, Das M, Bhargava K (2011) Activity-dependent neuroprotective protein (ADNP)-derived peptide (NAP) ameliorates hypobaric hypoxia induced oxidative stress in rat brain. Peptides 32(6):1217–1224 Shi Q, Fu J, Ge D, He Y, Ran J, Liu Z, Wei J, Diao T, Lu Y (2012) Huperzine A ameliorates cognitive deficits and oxidative stress in the hippocampus of rats exposed to acute hypobaric hypoxia. Neurochem Res 37(9):2042–2052 Suzanne M, Steller H (2009) Letting go: modification of cell adhesion during apoptosis. J Biol 8(5):49 Sutton JR (1992) Mountain sickness. Neurol Clin 10(4):1015–1030 Swenson ER, Maggiorini M (2002) Salmeterol for the prevention of high-altitude pulmonary edema. N Engl J Med 347(16):1282–1285; Author reply 1282–1285 Sylvester JT, Shimoda LA, Aaronson PI, Ward JP (2012) Hypoxic pulmonary vasoconstriction. Physiol Rev 92(1):367–520 Teppema LJ, Dahan A (2010) The ventilatory response to hypoxia in mammals: mechanisms, measurement, and analysis. Physiol Rev 90(2):675–754 Turner E (1969) Hippocampus and memory. Lancet 2(7630):1123–1126 Turrens JF (2003) Mitochondrial formation of reactive oxygen species. J Physiol 552(Pt 2):335–344. https://doi.org/10.1113/jphysiol.2003.049478 Vock P, Brutsche MH, Nanzer A, Bartsch P (1991) Variable radiomorphologic data of high altitude pulmonary edema. Features from 60 patients. Chest 100(5):1306–1311 Voelkel NF, McMurtry IF, Reeves JT (1981) Hypoxia impairs vasodilation in the lung. J Clin Investig 67(1):238–246 Wason MS, Zhao J (2013) Cerium oxide nanoparticles: potential applications for cancer and other diseases. Am J Transl Res 5(2):126–131

Nanoceria and Hypoxia: Promises and Challenges

415

West JB (1985, 1996) T.H. Ravenhill and his contributions to mountain sickness. J Appl Physiol 80(3):715–724 Willett WC, MacMahon B (1984) Diet and cancer–an overview (second of two parts). N Engl J Med 310(11):697–703 Wilson MH, Newman S, Imray CH (2009) The cerebral effects of ascent to high altitudes. Lancet Neurol 8(2):175–191 Winterbourn CC (2008) Reconciling the chemistry and biology of reactive oxygen species. Nat Chem Biol 4(5):278–286 Wu TY, Ding SQ, Liu JL, Jia JH, Dai RC, Zhu DC, Liang BZ, Qi DT, Sun YF (2007) High-altitude gastrointestinal bleeding: an observation in Qinghai-Tibetan railroad construction workers on Mountain Tanggula. World J Gastroenterol WJG 13(5):774–780 Wu YC, Wang YJ, Tseng GF (2011) Ascorbic acid and alpha-tocopherol supplement starting prenatally enhances the resistance of nucleus tractus solitarius neurons to hypobaric hypoxic challenge. Brain Struct Funct 216(2):105–122 Yarnell PR, Heit J, Hackett PH (2000) High-altitude cerebral edema (HACE): the denver/front range experience. Semin Neurol 20(2):209–217. https://doi.org/10.1055/s-2000-9830 Wang Y, et al (2022) Central nervous system injury meets nanoceria: opportunities and challenges. Regen Biomater Yi X, Shihong Y, Guoxiang Y, Jun H (2021) Review of the potential ecological risks and toxicity mechanisms of nanoceria. Asian J Ecotoxicol 1:43–55 Yokel RA, Au TC, MacPhail R, Hardas SS, Butterfield DA, Sultana R, Goodman M, Tseng MT, Dan M, Haghnazar H, Unrine JM, Graham UM, Wu P, Grulke EA (2012) Distribution, elimination, and biopersistence to 90 days of a systemically introduced 30 nm ceria-engineered nanomaterial in rats. Toxicol Sci: off J Soc Toxicol 127(1):256–268 Yokel RA, Hussain S, Garantziotis S, Demokritou P, Castranova V, Cassee FR (2014) The Yin: an adverse health perspective of nanoceria: uptake, distribution, accumulation, and mechanisms of its toxicity. Environ Sci Nano 1(5):406–428 Zhao L, Mason NA, Morrell NW, Kojonazarov B, Sadykov A, Maripov A, Mirrakhimov MM, Aldashev A, Wilkins MR (2001) Sildenafil inhibits hypoxia-induced pulmonary hypertension. Circulation 104(4):424–428