Applications of Nanovesicular Drug

Delivery Amit Kumar Nayak

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Applications of Nanovesicular Drug Delivery
Applications of
Nanovesicular Drug
Delivery

Edited by
Amit Kumar Nayak
Department of Pharmaceutics, Seemanta Institute of Pharmaceutical Sciences,
Mayurbhanj, Odisha, India

Md Saquib Hasnain
Department of Pharmacy, Palamau Institute of Pharmacy,
Daltonganj, Jharkhand, India

Tejraj M. Aminabhavi
School of Advanced Sciences, KLE Technological University,
Hubballi, Karnataka, India

Vladimir P. Torchilin
Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University,
Boston, MA, United States
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Contents

List of contributors xiii 2.4.3 Self-assembling 27

Preface xvii 2.4.4 Solvent dispersion methods 27
2.4.5 Detergent removal method 27
1. Targeting cellular and molecular 2.4.6 Microfluidization method 27
mechanisms of nanovesicular systems 2.4.7 Handjani Vila method 28
for the treatment of different diseases 1 2.5 Nanodrug codelivery systems 28
2.5.1 Nanovesicles-hydrogels for
Natassa Pippa, Hector Katifelis, Maria Gazouli codelivery of drugs 28
and Stergios Pispas 2.5.2 Nanovesicles for codelivery of
1.1 Introduction 1 anticancer drugs 31
1.2 Lipid nanovesicular systems 2 2.5.3 Nanovesicles for codelivery of
1.2.1 Liposomes 2 cardiovascular drugs 33
1.2.2 Elastic liposomes: ethosomes and 2.5.4 Nanovesicles for codelivery of
transferosomes 2 antibacterial/antiinflammatory drugs 33
1.2.3 Niosomes 5 2.6 Conclusion 34
1.2.4 Ufasomes 5 References 34
1.3 Polymer nanovesicular systems 5
1.3.1 Polymersomes/polymer vesicles 5
3. Theranostic nanovesicles 39
1.3.2 Nanovesicular systems for targeting to Arnab De, Shilpa Das, Santanu Ghosh,
cellular mechanisms 7 Bhaskar Das, Sonalinandini Samanta,
1.3.3 Nanovesicular systems for targeting Bolay Bhattacharya and Amalesh Samanta
molecular mechanisms and the era of
3.1 Introduction 39
CRISPR/CAS9 10
3.2 Imaging strategies 40
1.3.4 Nanovesicular systems for the
3.2.1 Optical imaging 41
treatment of different diseases 11
3.2.2 Magnetic resonance imaging 41
1.4 Conclusions 13
3.2.3 Radionuclide-based imaging 41
References 14
3.2.4 Computed tomography 41
3.2.5 Ultrasound 42
2. Nanovesicles for drug codelivery 21 3.3 Different nanovesicles used as theranostic
system 42
Nafiu Aminu 3.3.1 Liposomes 42
2.1 Introduction 21 3.3.2 Ethosomes 44
2.2 Combination drug therapy 22 3.3.3 Transferosomes 45
2.3 General overview of nanovesicles 23 3.3.4 Niosomes 45
2.3.1 Liposomes 24 3.3.5 Polymersomes 47
2.3.2 Niosomes 25 3.4 Conclusion 48
2.3.3 Exosomes 26 References 48
2.3.4 Spanlastics 26
2.4 Design and preparation techniques of 4. Nanovesicles for ocular drug delivery 53
codelivery nanovesicles 26
Sophia G. Antimisiaris and Evangelos Natsaridis
2.4.1 Mechanical dispersion/film hydration
method 26 4.1 Introduction 53
2.4.2 Ultrasonication method 26

v
vi Contents

4.2 Physiology, routes of drug administration 5.12.3Central nervous system disorders 93

and ocular barriers for drug penetration 53 5.12.4Migraine 93
4.2.1 Physiology of the eye 53 5.12.5Hypertension 93
4.2.2 Routes of drug administration to 5.12.6Anxiety disorders 93
the eye and corresponding ocular 5.12.7Antinociceptive 94
barriers 54 5.12.8Oxytocin and insulin delivery 94
4.3 Ocular diseases 57 5.12.9Cancer 94
4.3.1 Anterior segment diseases 57 5.12.10Neurodegenerative/brain
4.3.2 Posterior segment diseases 58 inflammatory disease 94
4.4 Nanovesicles for ocular drug delivery 59 5.12.11 Cerebral arteriosclerosis,
4.4.1 Preclinical studies 61 thrombosis, and vertigo
4.4.2 Clinical studies and approved disorders 94
products 74 5.13 Conclusion 96
4.5 Conclusions and future perspectives 77 References 96
Acknowledgments and funding 77
References 77
6. Nanovesicles for transdermal drug
5. Nanovesicles for nasal drug delivery 81 delivery 103
Hongda Zhu, Chaobo Yang and Kai Ma
Gourav Paudwal, Nagma Banjare and
Prem N. Gupta 6.1 Introduction 103
6.1.1 The mechanisms of interactions
5.1 Introduction 81
between nanovesicle systems and
5.2 Intranasal drug delivery system 82
skin 104
5.3 Dosage forms and absorption
6.2 Lipid-based vesicular nanostructures for
enhancers 82
transdermal drug delivery 104
5.3.1 Nasal drops 82
6.2.1 Traditional liposomes as skin drug
5.3.2 Nasal spray 83
delivery systems 104
5.3.3 Nasal gel 83
6.2.2 Transfersomes 104
5.3.4 Nasal powders 83
6.2.3 Ethosomes 107
5.4 Benefits of intranasal drug delivery 83
6.2.4 Invasome 108
5.5 Barriers in nasal distribution 84
6.2.5 Glycerosomes 109
5.5.1 Poor bioavailability 84
6.2.6 Hyalurosomes 110
5.5.2 Biliary clearance 84
6.3 Nanovesicles formed by nonlipid building
5.5.3 Enzyme degradation 84
blocks 110
5.6 Need for intranasal drug delivery
6.3.1 Niosomes as transdermal drug
system 85
delivery systems 110
5.7 Anatomy and physiology of nasal
6.3.2 Polymersomes as transdermal
route 85
drug delivery systems 111
5.8 Mechanism of absorption of drugs via
6.4 Conclusion and future perspective 111
nasal route 86
References 112
5.8.1 Intracellular pathway 86
5.8.2 Transcellular transport 86
5.9 Nasal devices 87 7. Nanovesicles for intravenous drug
5.10 Role of nanotechnology intranasal drug delivery 115
delivery 87
Hazal Ezgi Gültekin, Ezgi Oner, Miray İlhan
5.11 Nanovesicles for intranasal drug delivery 88
and Merve Karpuz
5.11.1 Lipid based nanovesicles 88
5.11.2 Nonionic surfactant based 7.1 Introduction 115
nanovesicles 91 7.2 Intravenous drug administration 116
5.11.3 Biologically derived nanovesicles 92 7.3 Nanovesicular systems 121
5.12 Applications of nanovesicular intranasal 7.3.1 Liposomes 121
delivery system 92 7.3.2 Niosomes 124
5.12.1 Viral infection 92 7.3.3 Polymersomes 125
5.12.2 Osteoclastic bone resorption 93 7.3.4 Transfersomes 126
 Contents vii

7.3.5 Ethosomes and ethosomal 9. Blood brain barrier and nanovesicles

nanovesicles 127 for brain-targeting drug delivery 167
7.3.6 Phytosomes 127
7.3.7 Extracellular vesicles 128 Yadollah Omidi, Hossein Omidian,
7.4 Intravenous nanovesicles for imaging 128 Young Kwon and Ana Castejon
7.5 Intraveneous nanovesicles for therapy 133 9.1 Introduction 167
7.5.1 Tumor targeting and cancer therapy 133 9.2 Neurovascular unit 169
7.5.2 Fungal infections 134 9.2.1 Blood brain barrier and blood-
7.5.3 Pain management and inflammatory cerebrospinal fluid barrier roles 169
diseases 134 9.2.2 Immunosurveillance in
7.5.4 Others 134 neurovascular unit 170
7.6 Intravenous nanovesicles for gene therapy 135 9.2.3 Tight junctional molecular
7.6.1 Intravenous nanovesicular systems machinery 170
developed for gene augmentation 136 9.2.4 Blood brain barrier models 170
7.6.2 Intravenous nanovesicular systems 9.2.5 Blood brain barrier transport
developed for gene silencing machinery 172
(suppression) 136 9.2.6 Endocytosis, transcytosis, and
7.6.3 Intravenous nanovesicular systems vesicular trafficking 172
developed for genome editing 136 9.2.7 Nanovesicles delivery mechanisms 174
7.7 Intravenous nanovesicles for theranostic 136 9.3 Issues with the targeted therapy of brain
7.8 Conclusion 138 diseases 174
References 139 9.4 Nanoscale brain-targeting delivery
systems 176
8. Nanovesicles for target specific drug 9.5 Nanovesicles 176
delivery 149 9.5.1 Lipid-based nanovesicles for brain
Amna Zafar, Asim-ur-Rehman and targeting 177
Naveed Ahmed 9.5.2 Translation of brain-targeting
lipid-based nanovesicles 185
8.1 Introduction 149 9.5.3 Polymer-based nanovesicles 188
8.2 Liposomes as drug delivery vesicles 150 9.6 Concluding remarks 190
8.2.1 Types of liposomes 150 References 190
8.2.2 Applications 151
8.3 Polymeric micelles as drug delivery
vehicles 152 10. Nanovesicles for hepatic-targeted
8.3.1 Applications 153 drug delivery 201
8.4 Exosomes as drug delivery vesicles 153
Manish Kumar, Abhishek Jha, Kanchan Bharti
8.4.1 Applications 155
and Brahmeshwar Mishra
8.5 Niosomes—drug delivery vesicles 155
8.5.1 Applications 155 10.1 Introduction 201
8.6 New era of vesicular drug delivery systems 155 10.2 Nanovesicular systems for drug delivery
8.6.1 Transferosomes 155 to liver 201
8.6.2 Ethosomes 157 10.3 Mechanism of nanovesicles-targeted
8.6.3 Sphingosomes 157 delivery 202
8.6.4 Cubosomes 157 10.3.1 Passive targeting 202
8.6.5 Ufasomes 158 10.3.2 Active targeting 202
8.6.6 Colloidosomes 158 10.4 Role in improving the drug distribution
8.6.7 Aquasomes 159 and pharmacokinetic parameters 205
8.6.8 Polymerosomes 159 10.5 Applications 205
8.6.9 Emulsomes 159 10.5.1 Nanovesicles for hepatocellular
8.6.10 Virosomes 160 carcinoma 205
8.6.11 Enzymosomes 160 10.5.2 Nanovesicles for hepatic
8.6.12 Pharmacosomes 160 infections 211
8.7 Conclusions 163 10.5.3 Nanovesicles for hepatoprotective
References 163 effect 211
viii Contents

10.6 Conclusion 213 13.2 Factors affecting colonic drug delivery 253
References 214 13.3 Advantages and limitations of
colon-targeted drug delivery systems 255
11. Nanovesicles for tumor-targeted 13.4 Application of nanocarriers other than
drug delivery 219 nanovesicles for colon-targeted drug
delivery 256
Merve Karpuz, Miray İlhan, 13.5 Applications of nanovesicles for the
Hazal Ezgi Gültekin, Emre Ozgenc, treatment of colonic disease 257
Zeynep Şenyiğit and Evren Atlihan-Gundogdu 13.5.1 Liposomes 257
11.1 Introduction 219 13.5.2 Niosomes 259
11.2 Nanovesicles 221 13.5.3 Phytosomes 260
11.2.1 Liposomes 222 13.5.4 Cubosomes 260
11.2.2 Niosomes 223 13.5.5 Emulsosome 260
11.2.3 Phytosomes 224 13.6 Applications of nanovesicles in the
11.2.4 Ethosomes 224 detection of colonic disease 262
11.2.5 Polymersomes 225 13.7 Conclusion and future prospects 263
11.2.6 Exosomes 225 References 263
11.2.7 Transfersomes 226
11.3 Targeting mechanisms of nanovesicles 14. Nanovesicles for delivery of
for tumor 226
anticancer drugs 267
11.3.1 Passive targeting 226
11.3.2 Active targeting 228 Jithu Joseph
11.4 Nanovesicles for tumor imaging 232
14.1 Introduction 267
11.4.1 Magnetic resonance imaging 232
14.2 Classification and development of the
11.4.2 Computed tomography imaging 232
nanovesicles 267
11.4.3 Nuclear medicine imaging 233
14.2.1 Classification of the
11.5 Nanovesicles for tumor treatment 233
nanovesicles 267
11.6 Nanovesicles for theranostic approach 234
14.3 Applications of the nanovesicles for the
11.7 Conclusion 235
delivery of anticancer drugs 269
References 236
14.4 Conclusion and future prospects 278
References 278
12. Tumor microenvironment-responsive
nanovesicular drug delivery systems 245 15. Nanovesicles for the treatment of
Monira Ghoniem, Kholoud K. Arafa and skin disorders 285
Ibrahim M. El-Sherbiny Ayesha Waheed, Abdul Ahad,
12.1 Introduction 245 Dipak Kumar Gupta, Mohd. Aqil,
12.1.1 Doxorubicin-based nanovesiculars 246 Fahad I. Al-Jenoobi and
12.1.2 Paclitaxel-based nanovesiculars 246 Abdullah M. Al-Mohizea
12.1.3 Physiological tumor 15.1 Introduction 285
microenvironment characteristics 247 15.1.1 Skin permeation pathways 285
12.1.4 Tumor biochemical characteristics 247 15.2 Types of nanovesicles 286
12.1.5 Immune microenvironment 15.2.1 Liposomes 288
characteristics 250 15.2.2 Ethosomes 288
12.2 Conclusion 251 15.2.3 Niosomes 289
References 251 15.2.4 Transfersomes 289
15.2.5 Cubosomes 290
13. Nanovesicles for colon-targeted 15.2.6 Solid lipid nanoparticles 290
drug delivery 253 15.2.7 Nanostructured lipid carriers 290
15.2.8 Nanoemulsion 290
Pooja Das Bidla, Pritish K. Panda, Amit Verma,
15.2.9 Polymeric nanoparticles 290
Sarjana Raikwar and Sanjay K. Jain
15.2.10 Nanofibers 290
13.1 Introduction 253 15.2.11 Dendrimers 291
 Contents ix

15.3 Skin disorders 291 17.3.5 Transferosomes 316

15.3.1 Skin cancer 291 17.3.6 Proteasomes 317
15.3.2 Psoriasis 292 17.3.7 Sphingosomes 317
15.3.3 Acne 293 17.3.8 Archaesomes 317
15.3.4 Alopecia 295 17.3.9 Ethosomes 317
15.3.5 Fungal infections 295 17.3.10 Polymersomes 317
15.3.6 Atopic dermatitis 296 17.4 Nanovesicles for central nervous
15.4 Conclusion 297 system disorders 317
References 297 17.4.1 Nanovesicles for Alzheimer’s
disease 317
16. Nanovesicles for the delivery of 17.4.2 Nanovesicles for Parkinson’s
nonsteroidal anti-inflammatory drugs 303 disease 320
17.4.3 Nanovesicles for migraine 321
Shohreh Fahimirad
17.4.4 Nanovesicles for epilepsy 322
16.1 Introduction of nonsteroidal 17.4.5 Nanovesicles for psychosis 324
anti-inflammatory drug 303 17.4.6 Nanovesicles for central nervous
16.2 Nonsteroidal anti-inflammatory agents 303 system infection 325
16.3 Nanotechnology and nonsteroidal 17.4.7 Nanovesicles for depression 326
anti-inflammatory drugs delivery 304 17.4.8 Nanovesicles for brain tumors 327
16.4 Liposomes 305 17.4.9 Nanovesicles for neuroprotection 328
16.5 Nonliposomal lipid-based nanovesicles 306 17.4.10 Nanovesicles for multiple sclerosis
16.5.1 Niosomes 306 and amyotrophic lateral sclerosis 330
16.5.2 Transfersomes 306 17.4.11 Nanovesicles for cerebral
16.5.3 Ethosomes 306 ischemia 331
16.5.4 Sphingosomes 306 17.5 Current challenges and future prospects 333
16.5.5 Ufasomes 307 17.6 Conclusion 334
16.5.6 Pharmacosomes 307 Conflicts of interest 334
16.5.7 Virosomes 307 References 335
16.5.8 Quatsomes 307
16.6 Methods of preparation 307 18. Nanovesicles for the delivery of
16.6.1 Conventional preparation cardiovascular drugs 341
methods 307
Domenico Marson, Suzana Aulic,
16.6.2 Thin lipid film hydration method 308
Alice Fermeglia, Erik Laurini and Sabrina Pricl
16.6.3 Solvent injection technique 311
16.7 Novel preparation methods 311 18.1 Introduction 341
16.7.1 Supercritical fluids methods 311 18.2 A primer of cardiovascular diseases 342
16.7.2 Recent application of 18.2.1 Atherosclerosis and
nanovesicles for delivery of hyperlipidemia 342
nonsteroidal anti-inflammatory 18.2.2 Venous thromboembolism 343
drugs 311 18.2.3 Acute myocardial infarction 343
16.8 Conclusion 311 18.2.4 Hypertension 344
References 311 18.2.5 Pulmonary hypertension 344
18.2.6 Stroke 345
17. Nanovesicles for delivery of central 18.3 Nanovesicles for the delivery of
nervous system drugs 315 cardiovascular drugs 345
18.3.1 Nanovesicles for the treatment of
Reshu Virmani, Tarun Virmani and Kamla Pathak
atherosclerosis and
17.1 Introduction 315 hyperlipidemia 345
17.2 Nanovesicles 315 18.3.2 Nanovesicles for the treatment of
17.3 Categories of nanovesicles 315 venous thromboembolism 349
17.3.1 Liposomes 315 18.3.3 Nanovesicles for the treatment of
17.3.2 Virosomes 316 hypertension 351
17.3.3 Niosomes 316 18.3.4 Nanovesicles for the treatment of
17.3.4 Proniosomes 316 pulmonary hypertension 353
x Contents

18.3.5 Nanovesicles for the treatment of 20.2 Vesicular delivery systems 384
acute myocardial infarction 355 20.2.1 Liposomes 384
18.3.6 Nanovesicles for the treatment of 20.2.2 Niosomes 386
stroke 358 20.2.3 Transfersomes 388
18.4 Future outlook 361 20.2.4 Ethosomes 390
Acknowledgments 362 20.2.5 Transethosomes 392
References 362 20.2.6 Cubosomes 392
20.3 Conclusion 392
19. Nanovesicles for the delivery of References 393
antibiotics 371
21. Nanovesicles in antiviral drug
Qurat ul ain, Nazim Hussain, delivery 399
Syed Awais Attique and Muhammad Bilal
Mehvish Mumtaz, Zulqarnain Baqar,
19.1 Introduction 371 Nazim Hussain and Muhammad Bilal
19.2 Nanovesicles as potential antibiotic drug
delivery and/or targeting systems 371 21.1 Introduction 399
19.3 Nanoparticle bacterial resistance 372 21.2 What are nanovesicles? 400
19.4 Antimicrobial resistance mechanisms 372 21.3 Composition of nanovesicles 400
19.5 The impact of nanoparticles on 21.4 Development of nanovesicles 401
microbial strength 373 21.4.1 Thin film hydration 401
19.5.1 Treatment techniques as an 21.4.2 Nonshaken method 402
effective defense against 21.4.3 Proliposomes 402
microbial resistance 373 21.4.4 Method of freeze-drying 402
19.5.2 Overcoming the current 21.4.5 Ethanol injection method 402
mechanisms of antibiotic 21.4.6 Ether injection method 402
resistance 373 21.4.7 Hot method 402
19.6 Using numerous ways to combat 21.4.8 Cold method 402
microorganisms at the same time 373 21.4.9 Reverse-phase evaporation
19.7 Assisting in the transport of antibiotics 373 technique 403
19.8 Negative side: as a drug resistance 21.4.10 Ultrasonication 403
promoter 374 21.5 Nanovesicles characterization 403
19.9 Nanoparticles antibacterial application 375 21.5.1 Efficiency of entrapment 403
19.10 Dressings of wound 375 21.5.2 Visual representation of
19.11 Bone fortification 375 morphology 403
19.12 Dental equipment 375 21.5.3 Zeta potential and vesicle size 403
19.13 The mechanism for drug delivery 376 21.5.4 Temperature of transition 404
19.14 Types of nanovesicles used for the 21.5.5 Evaluation of surface tension
drug delivery 376 behavior 404
19.15 Efficiency of different nanovesicles for 21.5.6 Sustainability of vesicles 404
drug delivery system 376 21.5.7 Deformability or elasticity
19.16 Role of nanovesicles in the delivery of research 404
antibiotics 378 21.5.8 Drug content 404
19.17 Summary and future perspectives 380 21.5.9 Drug release 404
Acknowledgment 380 21.5.10 Permeability and absorption
Disclosure statement 380 research 404
References 380 21.6 Application of nanovesicles 404
21.6.1 Challenges of nanovesicles 404
21.7 Antiviral drugs 405
20. Nanovesicles for delivery of 21.8 Medical applications of antiviral
antifungal drugs 383 drugs 405
21.9 Designing of antiviral drugs 406
Biswarup Das, Amit Kumar Nayak and
21.9.1 Targeting antivirals 406
Subrata Mallick
21.9.2 Methodologies based on the
20.1 Introduction 383 point of the virus’s life phase 406
 Contents xi

21.9.3 Prior to entering into a cell 406 22.2.3 Dendritic cells 423
21.9.4 Inhibitor of entry 407 22.2.4 Mesenchymal stem cells 423
21.9.5 Inhibitor of uncoating 407 22.2.5 Milk 423
21.9.6 As during the propagation of 22.2.6 Plant 424
viral 407 22.3 Biological functions 424
21.9.7 Reverse transcription 407 22.4 Immune system response to generic
21.9.8 Integrase 407 nanovesicles 424
21.9.9 Translation/antisense 408 22.4.1 T/B cells formation and
21.9.10 Translation/ribozymes 408 nanovesicles 425
21.9.11 Protein targeting and 22.5 Nanovesicle production, cellular
processing 408 communication, and autoimmunity 425
21.9.12 Inhibitors of proteases 408 22.6 Nanovesicles and autoimmune
21.9.13 Long dsRNA helix targeting 408 diseases 426
21.9.14 Structure 409 22.6.1 Systemic lupus erythematosus 426
21.9.15 Step of release 409 22.6.2 Diabetes 427
21.10 Approved antiviral drugs 409 22.6.3 Rheumatoid arthritis 428
21.10.1 Acyclovir 409 22.6.4 Vitiligo 428
21.10.2 Valacyclovir 410 22.6.5 Preeclampsia 429
21.10.3 Ganciclovir 410 22.6.6 Multiple sclerosis 429
21.10.4 Penciclovir 410 22.6.7 Sjogren’s syndrome 429
21.10.5 Famciclovir 411 22.6.8 Autoimmune thyroid disease 430
21.10.6 Foscarnet 411 22.7 Nanovesicle-facilitated autoimmune
21.10.7 Ribavirin 411 disease treatment therapies 430
21.10.8 Lamivudine 411 22.7.1 Autoimmune diseases treatment
21.10.9 Amantadine and for mesenchymal stem
Rimantadine 411 cell-derived nanovesicles 430
21.10.10 Interferon alfa 412 22.7.2 Autoimmune disease therapy for
21.11 Nanovesicles in antiviral drug delivery 412 dendritic cell-derived
21.11.1 Liposomes 412 nanovesicles 430
21.11.2 Solid lipid nanovesicles 412 22.8 Modifications for the targeted
21.11.3 Nanoemulsions 412 delivery of extracellular nanovesicles 431
21.11.4 Self-nanoemulsify drug delivery 22.8.1 Nanovesicle donor cells
systems 413 manipulation 431
21.11.5 Lipid based nanovesicles for 22.8.2 Extracellular nanovesicles direct
small interfering RNA delivery 413 surface modification 431
21.11.6 Polymer-based nanovesicles 413 22.9 Utilization of nanovesicles in
21.11.7 Nanovesicles made of autoimmune clinical trials 432
polymers 414 22.10 Conclusion and future outlook 433
21.12 Conclusion 414 References 433
References 414
Further reading 419
23. Nanovesicular systems for protein
and peptide delivery 441
22. Nanovesicles for targeting
Theodore Sentoukas, Athanasios Skandalis
autoimmune diseases 421
and Stergios Pispas
Rahat Andleeb, Muhammad Umar Ijaz,
23.1 Introduction 441
Asma Ashraf, Rida Rafi, Derya Karataş Yeni,
23.2 Liposomes 442
Shabana Naz, Tayyaba Ali and
23.3 Polymersomes 444
Muhammad Asad Sajid
23.4 Exosomes 447
22.1 Introduction 421 23.5 Nonionic vesicles (niosomes) 448
22.2 Sources of extracellular nanovesicles 423 23.6 Organic inorganic hybrid nanovesicles 449
22.2.1 Tumor cells 423 23.7 Conclusions 451
22.2.2 Red blood cells 423 References 451
xii Contents

24. Nanovesicles for the delivery of 24.3 Some applications of nanovesicles for
siRNA 457 the delivery of small interfering RNA
in target cells/drug delivery 462
Samuel Eshorame Sanni, Ifi Favour and 24.4 Conclusion 463
Adedayo Adeyanju References 464
24.1 Introduction 457
24.2 Preparation of nanovesicles and small 25. Clinical trials of nanovesicles for
interfering RNA-loaded nanovesicles 459 drug delivery applications 467
24.2.1 Nanovesicle preparation 459
Mourelatou Elena, Galatou Eleftheria,
24.2.2 Loading nanovesicles with
Sarigiannis Yiannis, Zacharia C. Lefteris,
small interfering RNAs 459
Plioukas Michael, Aislaitner Georgios and
24.2.3 Preparing a western blot 459
Petrou C. Christos
24.2.4 Small interfering RNA
quantification 460 25.1 Introduction 467
24.2.5 Size and zeta potential 25.2 The legal framework for clinical trials 467
measurement of nanovesicles 460 25.3 Regulatory challenges in clinical trials
24.2.6 Short harpin RNA transduction, in the field of nanovesicles 468
PKH67 labeling and nanovesicle- 25.4 Liposomes 469
uptake 460 25.5 Peptide-based nanovesicles 476
24.2.7 In vitro treatment of nanovesicles 25.6 Exosomes 480
in human umbilical vein 25.7 Phytosomes 482
endothelial cells and λ820 cells 460 25.8 Niosomes 483
24.2.8 The quantitative real time-PCR 25.9 Conclusions 484
kit for nanovesicle-small interfering References 484
RNAs gene detection/recognition 461
24.2.9 Cell count and proliferation Index 487
assays 461
List of contributors
Adedayo Adeyanju Department of Chemical
Engineering, Covenant University, Ota, Ogun State,
Nigeria
Abdul Ahad Department of Pharmaceutics, College of
Pharmacy, King Saud University, Riyadh, Saudi Arabia
Naveed Ahmed Department of Pharmacy, Faculty of
Biological Sciences, Quaid-i-Azam University,
Islamabad, Pakistan
Tayyaba Ali Department of Zoology, Government
College University Faisalabad, Faisalabad, Pakistan
Fahad I. Al-Jenoobi Department of Pharmaceutics,
College of Pharmacy, King Saud University, Riyadh,
Saudi Arabia
Abdullah M. Al-Mohizea Department of Pharmaceutics,
College of Pharmacy, King Saud University, Riyadh,
Saudi Arabia
Nafiu Aminu Department of Pharmaceutics and
Pharmaceutical Microbiology, Faculty of
Pharmaceutical Sciences, Usmanu Danfodiyo
University, Sokoto, Nigeria
Rahat Andleeb Department of Zoology, Government
College University Faisalabad, Faisalabad, Pakistan
Sophia G. Antimisiaris Laboratory of Pharmaceutical
Technology, Department of Pharmacy, School of
Health Sciences, University of Patras, Rio Patras,
Greece; Institute of Chemical Engineering Sciences,
Foundation for Research and Technology Hellas
(FORTH/ICE-HT), Rio Patras, Greece
Mohd. Aqil Department of Pharmaceutics, School of
Pharmaceutical Education and Research, Jamia
Hamdard (Deemed University), New Delhi, India
Kholoud K. Arafa Nanomedicine Research
Laboratories, Center for Materials Science, Zewail
City of Science and Technology, Giza, Egypt
Asma Ashraf Department of Zoology, Government
College University Faisalabad, Faisalabad, Pakistan
Asim-ur-Rehman Department of Pharmacy, Faculty of
Biological Sciences, Quaid-i-Azam University,
Islamabad, Pakistan
Evren Atlihan-Gundogdu Department of Radiopharmacy,
Faculty of Pharmacy, Ege University, İzmir, Turkey
Syed Awais Attique School of Interdisciplinary
Engineering & Science (SINES), NUST, Islamabad,
Pakistan
Suzana Aulic Molecular Biology and Nanotechnology
Laboratory (MolBNL@UniTS)—DEA, University of
Trieste, Trieste, Italy
Nagma Banjare PK-PD Tox & Formulation Division,
CSIR-Indian Institute of Integrative Medicine, Jammu,
Jammu and Kashmir, India; Academy of Scientific
and Innovative Research (AcSIR), Ghaziabad, Uttar
Pradesh, India
Zulqarnain Baqar Centre for Applied Molecular
Biology (CAMB), University of the Punjab, Lahore,
Pakistan
Kanchan Bharti Department of Pharmaceutical
Engineering & Technology, Indian Institute of
Technology (BHU), Varanasi, Uttar Pradesh, India
Bolay Bhattacharya School of Pharmacy, Sister
Nivedita University, Kolkata, West Bengal, India
Muhammad Bilal School of Life Science and Food
Engineering, Huaiyin Institute of Technology, Huaian,
China
Ana Castejon Department of Pharmaceutical Sciences,
College of Pharmacy, Nova Southeastern University,
Fort Lauderdale, FL, United States
Petrou C. Christos Department of Life and Health
Sciences, University of Nicosia, Nicosia, Cyprus
Bhaskar Das Directorate of Drugs Control, Department
of Health & Family Welfare, Government of West
Bengal, Kolkata, West Bengal, India
Biswarup Das Department of Pharmaceutics, Seemanta
Institute of Pharmaceutical Sciences, Mayurbhanj,
Odisha, India
Shilpa Das Department of Pharmaceutical Technology,
Jadavpur University, Kolkata, West Bengal, India
Pooja Das Bidla Pharmaceutics Research Projects
Laboratory, Department of Pharmaceutical Sciences,
xiii
xiv List of contributors
Dr. Harisingh Gour Vishwavidyalaya, Sagar, Madhya
Pradesh, India
Arnab De School of Pharmacy, Sister Nivedita
University, Kolkata, West Bengal, India
Galatou Eleftheria Department of Life and Health
Sciences, University of Nicosia, Nicosia, Cyprus
Mourelatou Elena Department of Life and Health
Sciences, University of Nicosia, Nicosia, Cyprus
Ibrahim M. El-Sherbiny Nanomedicine Research
Laboratories, Center for Materials Science, Zewail
City of Science and Technology, Giza, Egypt
Shohreh Fahimirad Molecular and Medicine Research
Center, Arak University of Medical Sciences, Arak,
Iran
Ifi Favour Department of Chemical Engineering,
Covenant University, Ota, Ogun State, Nigeria
Alice Fermeglia Molecular Biology and Nanotechnology
Laboratory (MolBNL@UniTS)—DEA, University of
Trieste, Trieste, Italy
Maria Gazouli School of Medicine, Laboratory of
Biology, Department of Basic Medical Science,
National and Kapodistrian University of Athens,
Athens, Greece; School of Medicine, 2nd Department
of Radiology, National and Kapodistrian University of
Athens, Athens, Greece
Aislaitner Georgios Department of Life and Health
Sciences, University of Nicosia, Nicosia, Cyprus;
Federal Institute for Drugs and Medical Devices
(BfArM), Bonn, Germany
Monira Ghoniem Department of Chemistry, College of
Science, Imam Mohammad Ibn Saud Islamic
University (IMSIU), Riyadh, Saudi Arabia
Santanu Ghosh Department of Pharmaceutical
Technology, JIS University, Kolkata, West Bengal,
India
Hazal Ezgi Gültekin Department of Pharmaceutical
Technology, Faculty of Pharmacy, İzmir Katip Çelebi
University, İzmir, Turkey
Dipak Kumar Gupta Department of Pharmaceutics,
School of Pharmaceutical Education and Research,
Jamia Hamdard (Deemed University), New Delhi,
India
Prem N. Gupta PK-PD Tox & Formulation Division,
CSIR-Indian Institute of Integrative Medicine, Jammu,
Jammu and Kashmir, India; Academy of Scientific
and Innovative Research (AcSIR), Ghaziabad, Uttar
Pradesh, India
Nazim Hussain Centre for Applied Molecular Biology
(CAMB), University of the Punjab, Lahore, Pakistan
Muhammad Umar Ijaz Department of Zoology,
Wildlife and Fisheries, University of Agriculture,
Faisalabad, Pakistan
Miray İlhan Department of Pharmaceutical Technology,
Faculty of Pharmacy, İzmir Katip Çelebi University,
İzmir, Turkey; Department of Pharmaceutical
Technology, Faculty of Pharmacy, Düzce University,
Düzce, Turkey
Sanjay K. Jain Pharmaceutics Research Projects
Laboratory, Department of Pharmaceutical Sciences,
Dr. Harisingh Gour Vishwavidyalaya, Sagar, Madhya
Pradesh, India
Abhishek Jha Department of Pharmaceutical
Engineering & Technology, Indian Institute of
Technology (BHU), Varanasi, Uttar Pradesh, India
Jithu Joseph Department of Applied Chemistry, Cochin
University of Science and Technology, Kochi, Kerala,
India
Merve Karpuz Department of Radiopharmacy, Faculty
of Pharmacy, İzmir Katip Çelebi University, İzmir,
Turkey
Hector Katifelis School of Medicine, Laboratory of
Biology, Department of Basic Medical Science,
National and Kapodistrian University of Athens,
Athens, Greece
Manish Kumar Department of Pharmaceutical
Engineering & Technology, Indian Institute of
Technology (BHU), Varanasi, Uttar Pradesh, India
Young Kwon Department of Pharmaceutical Sciences,
College of Pharmacy, Nova Southeastern University,
Fort Lauderdale, FL, United States
Erik Laurini Molecular Biology and Nanotechnology
Laboratory (MolBNL@UniTS)—DEA, University of
Trieste, Trieste, Italy
Zacharia C. Lefteris Department of Life and Health
Sciences, University of Nicosia, Nicosia, Cyprus
Kai Ma Department of Pharmaceutical Engineering,
Hubei University of Technology, Wuhan, China
Subrata Mallick Department of Pharmaceutics, School
of Pharmaceutical Sciences, Siksha “O” Anusandhan
(Deemed to be University), Bhubaneswar, Odisha,
India
Domenico Marson Molecular Biology and
Nanotechnology Laboratory (MolBNL@UniTS)—
DEA, University of Trieste, Trieste, Italy

Plioukas Michael Department of Life and Health
Sciences, University of Nicosia, Nicosia, Cyprus
Brahmeshwar Mishra Department of Pharmaceutical
Engineering & Technology, Indian Institute of
Technology (BHU), Varanasi, Uttar Pradesh, India
Mehvish Mumtaz Centre for Applied Molecular
Biology (CAMB), University of the Punjab, Lahore,
Pakistan
Evangelos Natsaridis Laboratory of Pharmaceutical
Technology, Department of Pharmacy, School of
Health Sciences, University of Patras, Rio Patras,
Greece; Institute of Chemical Engineering Sciences,
Foundation for Research and Technology Hellas
(FORTH/ICE-HT), Rio Patras, Greece
Amit Kumar Nayak Department of Pharmaceutics,
Seemanta Institute of Pharmaceutical Sciences,
Mayurbhanj, Odisha, India
Shabana Naz Department of Zoology, Government
College University Faisalabad, Faisalabad, Pakistan
Yadollah Omidi Department of Pharmaceutical
Sciences, College of Pharmacy, Nova Southeastern
University, Fort Lauderdale, FL, United States
Hossein Omidian Department of Pharmaceutical
Sciences, College of Pharmacy, Nova Southeastern
University, Fort Lauderdale, FL, United States
Ezgi Oner Department of Pharmaceutical Biotechnology,
Faculty of Pharmacy, İzmir Katip Çelebi University,
İzmir, Turkey
Emre Ozgenc Department of Radiopharmacy, Faculty of
Pharmacy, Ege University, İzmir, Turkey
Pritish K. Panda Pharmaceutics Research Projects
Laboratory, Department of Pharmaceutical Sciences,
Dr. Harisingh Gour Vishwavidyalaya, Sagar, Madhya
Pradesh, India
Kamla Pathak Faculty of Pharmacy, Uttar Pradesh
University of Medical Sciences, Etawah, Uttar
Pradesh, India
Gourav Paudwal PK-PD Tox & Formulation Division,
CSIR-Indian Institute of Integrative Medicine, Jammu,
Jammu and Kashmir, India; Academy of Scientific
and Innovative Research (AcSIR), Ghaziabad, Uttar
Pradesh, India
Natassa Pippa Faculty of Pharmacy, Department of
Pharmaceutical Technology, National and
Kapodistrian University of Athens, Athens, Greece;
Theoretical and Physical Chemistry Institute, National
Hellenic Research Foundation, Athens, Greece
List of contributors xv
Stergios Pispas Theoretical and Physical Chemistry
Institute, National Hellenic Research Foundation,
Athens, Greece
Sabrina Pricl Molecular Biology and Nanotechnology
Laboratory (MolBNL@UniTS)—DEA, University of
Trieste, Trieste, Italy; Department of General
Biophysics, Faculty of Biology and Environmental
Protection, University of Lodz, Lodz, Poland
Rida Rafi Department of Applied Chemistry,
Government College University Faisalabad,
Faisalabad, Pakistan
Sarjana Raikwar Pharmaceutics Research Projects
Laboratory, Department of Pharmaceutical Sciences,
Dr. Harisingh Gour Vishwavidyalaya, Sagar, Madhya
Pradesh, India
Muhammad Asad Sajid Department of Zoology,
Government College University Faisalabad,
Faisalabad, Pakistan
Amalesh Samanta Department of Pharmaceutical
Technology, Jadavpur University, Kolkata, West
Bengal, India
Sonalinandini Samanta ESI-PGIMSR, ESIC Medical
College and Hospital, Joka, Kolkata, West Bengal,
India
Samuel Eshorame Sanni Department of Chemical
Engineering, Covenant University, Ota, Ogun State,
Nigeria
Theodore Sentoukas Center of Polymer and Carbon
Materials, Polish Academy of Sciences, Zabrze,
Poland
Athanasios Skandalis Department of Materials, Imperial
College London, London, United Kingdom
Qurat ul ain Centre for Applied Molecular Biology
(CAMB), University of the Punjab, Lahore, Pakistan
Amit Verma Babulal Tarabai Institute of Pharmaceutical
Sciences, Sagar, Madhya Pradesh, India
Reshu Virmani School of Pharmaceutical Sciences,
MVN University, Palwal, Haryana, India
Tarun Virmani School of Pharmaceutical Sciences,
MVN University, Palwal, Haryana, India
Ayesha Waheed Department of Pharmaceutics, School
of Pharmaceutical Education and Research, Jamia
Hamdard (Deemed University), New Delhi, India
Chaobo Yang Department of Pharmaceutical
Engineering, Hubei University of Technology, Wuhan,
China
xvi List of contributors

Derya Karataş Yeni Veterinary Control Central Hongda Zhu Department of Pharmaceutical
Research Institute, Bacterial Disease Laboratory, Engineering, Hubei University of Technology, Wuhan,
Ankara, Turkey China
Sarigiannis Yiannis Department of Life and Health Zeynep Şenyiğit Department of Pharmaceutical
Sciences, University of Nicosia, Nicosia, Cyprus Technology, Faculty of Pharmacy, İzmir Katip Çelebi
Amna Zafar Department of Pharmacy, Faculty of University, İzmir, Turkey
Biological Sciences, Quaid-i-Azam University,
Islamabad, Pakistan
Preface

Nanotechnology has recently proven its importance and usefulness for guiding the revolutionary changes in the fields
of nanomedicines, where nanovesicles have extensively been studied and well recognized for the diagnosis and therapy.
Nanovesicles are the nanoscale colloidal carrier systems composed of an aqueous core with lipid coating. Over the past
few decades, different nanovesicular carrier forms (such as nanoemulsions, self-nanoemulsifying systems, nanolipo-
somes, transferosomes, proniosomes and niosomes, exosomes, polymerosomes, aquasomes, ethosomes, cubosomes,
phytosomes, hyalurosomes, glycerosomes, and nanobubbles) have been reported for the delivery of various drugs (such
as nonsteroidal antiinflammatory drugs, central nervous system drugs, cardiovascular drugs, antibiotics, anticancer
drugs, antiviral drugs, and protein and peptide drugs) through different routes of administrations (such as oral, nasal,
ocular, transdermal, and intravenous). These have been employed for the target-specific drug delivery applications
(such as brain targeting, colon targeting, liver targeting, and cancer targeting). In addition, some advanced trigger-
assisted systems (such as iontophoresis and ultrasound triggering) have been used for improved drug delivery by the
various nanovesicles. In this context the current book “Applications of Nanovesicular Drug Delivery” aims to present a
thorough insight into the complete and up-to-date discussions about the various multifunctional applications of nanove-
sicular drug delivery with a collection of 25 authoritative chapters by the leading academicians and researchers across
the world. A concise account on the contents of each chapter has been described to provide a glimpse of the book to
the readers.
Chapter 1 presents an insight into the role of targeting cellular and molecular mechanisms of nanovesicular systems
to treat different diseases. In the first part, an overview of the main categories of nanovesicles, formulation processes,
and characterization techniques is described. In the second part, the nanovesicular systems for targeting to cellular and
molecular mechanisms have been analyzed in depth. A comprehensive update about their application in different dis-
eases, marketed products, and potential challenges has also been discussed.
Chapter 2 highlights the applications of various nanovesicles for drug delivery, specifically codelivery of multiple
drugs. The chapter also provides an overview of drug combination therapy and different techniques used in preparing
nanovesicles for the codelivery of multiple drugs. A comprehensive recent update of the applications of various drug
codelivery nanovesicles in cancers, infectious, cardiovascular diseases, and inflammatory diseases has also been
discussed.
Chapter 3 provides a comprehensive review on theranostic applications of various nanovesicles (such as liposomes,
ethosomes, transferosomes, niosomes, and polymersomes).
Chapter 4 focuses on various strategies developed in the field of nanovesicle-assisted ocular drug delivery over the
past decade with an aim to overcome the limitations of ocular drug delivery.
Chapter 5 discusses about the potential of nanovesicular systems and their uses in nasal drug delivery. In addition,
dosage forms and absorption enhancers, benefits of intranasal drug delivery, barriers in nasal distribution, anatomy and
physiology of the nasal route, need for intranasal drug delivery systems, mechanism of absorption of drugs via the nasal
route, nasal devices and role of nanotechnology in intranasal drug delivery have been addressed.
Chapter 6 presents a comprehensive review on the various nanovesicular systems as excellent transdermal drug
delivery carriers to overcome the stratum corneum barrier in intact skin.
Chapter 7 describes the recent developments in the intravenous administration of nanovesicles, their administrations,
and commercially available products. In addition, this chapter covers the potential use of intravenous nanovesicles in
the treatment of different disease groups, gene therapy, imaging, and theranostic delivery by investigating their advan-
tages/disadvantages and the use of the mentioned areas.
Chapter 8 provides a detailed review of various natural and synthetic nanovesicular systems with their therapeutic
applications for the target-oriented drug delivery.
Chapter 9 imparts deep insights into various types of multifunctional nanovesicles capable of crossing the
blood brain barrier for active targeting and imaging in treating the unhealthy cells present in the brain.

xvii
xviii Preface

Chapter 10 offers a comprehensive insight of nanonovesicles for targeted drug delivery to the liver and their applica-
tions in the management of various liver disorders. In addition, mechanisms of nanovesicles for passive and active
hepatic targeting, the role of nanovesicles in improving drug distribution and pharmacokinetic parameters have been
well addressed.
Chapter 11 reviews different types of nanovesicles and their targeting mechanism for cancer imaging and treatment.
In addition, some studies performed to develop nanovesicles as imaging, treatment, or theranostic systems for cancers
have also been summarized.
Chapter 12 spotlights on the recent advances in the design of tumor microenvironment-responsive nanovesicles for
the effective delivery of chemotherapeutic drugs and tackling the challenges hindering their wide scale clinical
translation.
Chapter 13 covers the recent advancements in nanovesicles to design effective colon-targeted drug delivery carrier
systems for the treatment of colonic diseases.
Chapter 14 overviews the uses of various nanovesicles for the delivery of anticancer drugs in a facile manner and
ultimately helps to treat various types of cancers.
Chapter 15 deals with the uses of nanovesicles for the effective treatment/management of some important skin disor-
ders such as skin cancer, psoriasis, acne, alopecia, fungal infections, and atopic dermatitis. Different nanovesicles as
excellent alternatives to conventional dosage forms for efficient dermal delivery have also been addressed.
Chapter 16 describes the potential applications of different nanovesicles for efficient delivery of nonsteroidal antiin-
flammatory drugs.
Chapter 17 provides the current progress in applications of nanovesicles for effective drug delivery against central
nervous system diseases such as Alzheimer disease, Parkinson disease, psychosis, migraine, depression, epilepsy, and
brain tumor.
Chapter 18 focuses on the most recent advancements in nanovesicles as nanodelivery systems for cardiovascular
drugs for the treatment of various cardiovascular diseases such as atherosclerosis, hyperlipidemia, thromboembolism,
hypertension, acute myocardial infarction, and ischemic stroke.
Chapter 19 presents the potential applications of different nanovesicles for the efficient delivery of antibiotics. In
addition, mechanisms of antibiotic resistance and overcoming strategies by antibiotics-loaded nanovesicles have also
been discussed.
Chapter 20 discusses the uses of vesicular nanocarriers (nanovesicles) in the context of delivery of antifungal agents
through systemic and topical routes to treat fungal infections.
Chapter 21 overviews the potential applications of different nanovesicles for efficient delivery of antiviral drugs for
the treatment of various viral infections. The composition, preparation, and characterization of nanovesicles are dis-
cussed in this chapter. In addition, the designing of antiviral drugs and various approved antiviral drugs have also been
presented.
Chapter 22 summarizes the existing literature regarding the production, efficacy, and potential therapeutic uses of
nanovesicles in the contexts of targeting autoimmune diseases, from the disease pathology to diagnosis as well as
treatment.
Chapter 23 describes the basic characteristics of each group of nanovesicular structures, providing some of the latest
studies that show potential applications of these nanovesicular carrier systems for the delivery of therapeutic proteins
and peptides for the treatment/management of several diseases and conditions such as Parkinson’s disease, wound heal-
ing, diabetes, inflammation, cystic fibrosis, ischemic strokes, and several types of cancers.
Chapter 24 focuses on the role of nanovesicles for siRNA delivery. Other medical applications that bother on
approaches for siRNA delivery in relation to nanoparticles in chemically modified siRNAs have also been discussed.
Chapter 25 gives an insight into the regulatory framework governing the clinical trials of various nanovesicles that
are currently under clinical development based on bibliographic sources and clinical trial databases.
As editors, we would like to convey our special thanks to all the contributing authors for delivering their invaluable
chapter contributions in a timely manner, allowing us to publish this book in time. We would also like to express our
heartfelt gratitude to the Elsevier Inc., Andre Gerhard Wolff, Emma Hayes, and Patricia Gonzalez (Senior Editorial
Project Manager) for their invaluable assistance and kind support during the editing process of this book. We would
like to express our sincere thanks to Praveen Anand S. (Copyright Coordinator) for his outstanding support in obtaining
the copyright permissions and Punithavathy Govindaradjane (Production Manager) for the development and production
of the final book. All the copyright contents and reprinting licenses from different copyright sources have duly been
gratefully acknowledged.
 Preface xix

Finally, we appreciate our family members, all the respected teachers, friends, colleagues, and students for their con-
tinuous encouragement, inspiration, and support during the preparation of this voluminous book. We hope, along with
our contributing authors and publishers, that our efforts meet the demands of students, academicians, researchers, drug
delivery formulators, pharmaceutical specialists, polymer engineers, and biomedical experts.

Amit Kumar Nayak

Md Saquib Hasnain
Tejraj M. Aminabhavi
Vladimir P. Torchilin
Chapter 1

Targeting cellular and molecular

mechanisms of nanovesicular systems for
the treatment of different diseases
Natassa Pippa1,2, Hector Katifelis3, Maria Gazouli3,4 and Stergios Pispas2
1
Faculty of Pharmacy, Department of Pharmaceutical Technology, National and Kapodistrian University of Athens, Athens, Greece, 2Theoretical and
Physical Chemistry Institute, National Hellenic Research Foundation, Athens, Greece, 3School of Medicine, Laboratory of Biology, Department of
Basic Medical Science, National and Kapodistrian University of Athens, Athens, Greece, 4School of Medicine, 2nd Department of Radiology, National
and Kapodistrian University of Athens, Athens, Greece

1.1 Introduction
Nowadays, one of the key challenges in pharmaceutical nanotechnology is how to formulate platforms that selectively
deliver incorporated active substances to specific cells or tissues, when passive targeting is not possible, and the reduc-
tion of adverse reactions is required for patient compliance.1 10 Nanovesicles, such as lipid-based (liposomes, nio-
somes, transferosomes, ethosomes, etc.) and polymer-based (polymersomes, polymer nanovesicular structures, etc.), are
now well recognized as potential candidates for low-molecular-weight drug and antigen delivery and diagnostic
applications.1 25 Some of the nanovesicular systems have been already used in clinical practice as nanomedicines with
added value for the patients in comparison to the classical formulations.1,9
These consist of biocompatible and biodegradable materials (i.e., lipids, surfactants, and amphiphilic polymers), and
they can self-assemble in aqueous media into vesicular structures with various physicochemical characteristics and bio-
logical behaviors. Their advantage over other more conventional nanostructures is the property to encapsulate hydro-
philic active pharmaceutical ingredients (APIs) into their cavity and incorporate the lipophilic ones in their bilayers.
They can be also characterized as bio-inspired systems because they mimic the structural properties and the morphology
of cellular membranes and subcellular organelles.6 The recent trend in this direction of research is the fabrication of
nanovesicular systems composed of different materials (i.e., lipids/polymers and polymers/surfactants).20,21 Another
recent trend is the plant-derived nanovesicles that are a class of nanovesicles isolated from dietary vegetables and fruits
and exhibited several advantages and new properties.16 All the above nanovesicular systems can control the release of
the encapsulated APIs at the specific target tissue. The design and the development of stimuli-responsive nanovesicles
for spatially and temporally controlled release of APIs in response to intracellular stimuli, such as pH, redox potential,
reactive oxygen species, enzymes and temperature is very useful for the decrease of adverse reactions of the encapsu-
lated API.22 25
Nanovesicles can be used for the encapsulation of different categories of APIs, and they can be administered
through different routes. For example, exosomes (versatile, cell-derived nanovesicles naturally endowed with exquisite
target-homing specificity and the ability to surmount in vivo biological barriers) hold substantial promise for developing
exciting approaches in drug delivery, cancer immunotherapy, and as nanoscale cancer vaccines.12,13 Furthermore, nano-
vesicles are used as vaccine delivery platforms and as adjuvant systems. The adjuvant properties of nanovesicles are
very important because they can enhance the immune response in different molecular pathways and in parallel, they
can control the release of the antigen.17,18
This chapter provides an insight into the role of targeting cellular and molecular mechanisms of nanovesicular sys-
tems for the treatment of different diseases. In the first part, we present an overview of the main categories of nanovesi-
cles, the formulation processes and characterization techniques that are used in the recent literature. In the second part,
Applications of Nanovesicular Drug Delivery. DOI: https://doi.org/10.1016/B978-0-323-91865-7.00006-7
© 2022 Elsevier Inc. All rights reserved. 1
2 Applications of Nanovesicular Drug Delivery

the nanovesicular systems for targeting to cellular and molecular mechanisms are analyzed in depth and examples from
recently published studies are cited. A comprehensive update about their application in different diseases and the mar-
keted products, as well as potential challenges will be also discussed. Systematic search and review of papers regarding
the targeting cellular and molecular mechanisms of nanovesicular systems for the treatment of different diseases were
performed via MedLine, Scopus and Web of Science platforms.

1.2 Lipid nanovesicular systems

1.2.1 Liposomes
Liposomes are vesicular systems composed of phospholipid bilayers, and their size ranges from a few tens of nan-
ometers up to 1000 nm.26 36 They are biocompatible and biodegradable delivery nanosystems.26 36 Liposomal drugs
are marketed medicines with great potential in the field of cancer therapy as well as other diseases.30 31,34 According
to recent literature, patent publications on liposomal formulations have expanded innovative areas in pharmaceutical
nanotechnology.30 31,34 The composition of the liposomal dispersion and the formulation processes are the focus points
of most of the published patents. Special attention is given to formulate liposomal suspensions with great stability over
time and with controlled release properties of the encapsulated API.30,34 The Quality-by-Design and the scale-up of
liposomal formulations gain the interest of both the academic and industrial community.30,34 Liposomes are adminis-
tered through different routes including the oral route. They can improve the oral bioavailability of a variety of APIs
including peptides and proteins.29,33 Sometimes, the surface modification is mandatory for the oral administration of
these delivery platforms.33 Additionally, the design of effective liposomal forms for inhalation is also an important
research subject and depends highly on the composition of the vesicles, with the aim of reducing the detrimental effect
of shearing on liposome stability and maximizing their deposition in the lung tree.32 Finally, stealth liposomes are a
subcategory of liposomes with biocompatible and low protein binding polymer coatings exhibiting higher circulation
time in the human blood avoiding the decomposition from the plasma proteins.36 Stealth liposomes are also marketed
medicines for cancer therapy.31,36

1.2.2 Elastic liposomes: ethosomes and transferosomes

Elastic liposomes are a category of liposomes for cutaneous administration, including dermal and transdermal delivery
of the APIs.37 In this class of vesicular nanosystem, ethosomes and transferosomes are categorized.15,38 55 Ethosomes
consist of phospholipids, ethanol at high percentages (up to 40% 50%) and aqueous medium.15,42 46 The fluidity of
ethosomal membrane is quite fluid due to the presence of ethanol. Additionally, the ethanol high percentage in ethoso-
mal formulations allows the vesicles to penetrate deeper into the skin.15,42 46 According to the recent literature, gly-
cerosomes and trans-ethosomes are phospholipid vesicles containing glycerol or a mixture of ethanol and edge
activators. The edge activators also enhance the skin penetration.49 These vesicular systems have been already used for
the treatment of skin bacterial and fungal infections, acne, skin cancers and inflammation.15,42 55 For example, fisetin
loaded binary ethosomes formulation is a potential dermal delivery system for the management of skin cancer.47
Ethosomes have potential applications in the development of phytomedicines, for the treatment of challenging dis-
eases.54 For example, fluorouracil ethosomes have been developed for skin deposition and melanoma treatment in mice
models. Stabilization and effective application in skin of antioxidants and nanocosmeceutical compounds, like caffeic
acid and rosmarinic acid, have been achieved by their encapsulation into ethosomes.53 55 Transfersomes offer a versa-
tile delivery concept for facing the instability limitations as well as the potential to be used with a wide range of
APIs.56 61 Comparative study of liposomes, ethosomes and transfersomes as carriers for enhancing the transdermal
delivery of different APIs in vitro and in vivo have appeared in the literature, showing the added value of transfersomes
in dermal and ethosomes in transdermal delivery. They are (quasi)metastable, which makes the membrane ultra-flexible
and are characterized as “soft vesicles.” For this reason, they can easily squeeze through pores in the stratum corneum.
Surfactants/edge activators (i.e., sodium cholates; Tween-80 and Span-80) are also components of transferosomes as
flexibility agents. The percentage of surfactants /edge activators is between 10% and 25%, while the percentage of etha-
nol ranges between 3% and 10%. This is the main difference between ethosomes and transferosomes (the percentage of
ethanol in the formulation).49 59 Lyophilized transfersomal gel containing oleic acid was considered as a promising
nanosystem for hydrophilic APIs like buspirone hydrochloride.58 Transferosomes are promising delivery platforms for
anticancer agent for the treatment of skin cancers as well.59 61 In Table 1.1, examples of research reports on using
transfersomes as carriers for the delivery of anticancer agents are presented.
Another random document with
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machine. Fortunately I had presence of mind enough to keep my
watch going, as well as the captain’s chronometer, for otherwise I
would have had no knowledge of the passage of time. Once or twice
the scarlet women visited the ship, but seemed nervous and wary,
and made no effort to approach or molest me, merely gazed about
as if searching for something—perhaps for me—and then retiring.
Several times, too, I ventured on deck, and peered over the ship’s
side, but saw none of the giantesses, although with the glasses I
could see crowds of the beings about the city in the distance.
“Also, I noticed among them, several individuals who were much
smaller than the rest, and who appeared to be men, although I could
not be sure. I also discovered, and almost lost my life in the
discovery, that the atmosphere of this place is unfit for human beings
to breathe, and is thick with sulphurous fumes. Close to the ground
these fumes are so dense that a person would succumb in a few
moments, but at the height of the Chiriqui’s decks, nearly seventy
feet above the rocky bed on which she rests, the air is breathable,
although it causes one to choke and cough after a few minutes. And
I am sure that the houses of these giant beings have been built on
the summits of the basalt columns in order to avoid the suffocating
fumes of the lower levels. Later, too, I learned that the membrane-
like frills upon these creatures are a sort of gills, or as I might say,
natural gas-masks, which by some means enable the beings to
breathe the sulphur-laden air. But even with these, they avoid the
lower areas where the fumes are the worst, and only visit them when
necessity arises, which accounts for my being left in peace, with
none of the horrible women near the ship, for days at a time. I
discovered the presence of the sulphur gas on the first day when,
attempting to eat, I removed my gas-mask. Suffocating as I found
the fumes, I was compelled to endure them, and gradually I became
slightly accustomed to them, so that now I have little trouble in
breathing during the short time it takes me to eat my meals. At all
other times I must wear the apparatus, and I thank God that this is
so, for I know now that it is the gas-mask which so far has preserved
my life.
“On the tenth day after my arrival I noticed a number of the
giantesses gathering about the huge, spherical airship which still
rested on its cradle near the Chiriqui, but which, I have forgotten to
state, ceased to emit its green or red lights after it had landed. Lying
there it resembled nothing so much as a gigantic can-buoy or a
floating mine, if one can imagine a buoy two hundred yards in
diameter.
“On the day I mentioned, all interests seemed to be centered on
the thing, and cautiously peering from the shelter of the deck-house,
I watched the proceedings. Presently several of the women entered
the sphere through an opening in its middle band; the aperture
closed behind them, and immediately there was a low, humming
sound as of machinery. As the sounds issued from the sphere, the
cables to which were attached the smaller spheres (which glowed
red when carrying the Chiriqui through the air) were drawn in until
the two smaller spheres were resting in recesses at the axes of the
large sphere, and where they appeared merely as hemi-spherical
projections. Then, slowly at first, but with ever increasing speed, the
slender rods about the large sphere began to move back and forth,
or rather in an oscillating manner, until they were vibrating with such
rapidity that they appeared merely rays of light. Slowly, majestically,
the immense globe rose from its cradle, and gathering headway,
leaped upward to an immense height. Then, tilting at an angle, it
passed over the city and headed for an immense pinnacle of rock,
which, fully seven miles from where I stood, reminded me of a
gigantic chimney or funnel.
“Although it was barely visible to the naked eye, I could see it
distinctly through the glasses, and I watched it with the most intense
and concentrated interest. For a few moments it remained, poised a
hundred feet or so above the pinnacle. Then, from the towering,
tapering rock, a terrific jet of steam roared forth, and striking the
great spherical machine above it, hurled it upward and beyond my
vision. Give close heed to these words, whoever may, by God’s
grace, be listening to what I say, for upon them may hinge the fate of
the human race. Only by this means, by being shot upward by this
titanic jet of steam, can the airship leave this subterranean land and
emerge through the crater by which it entered bearing the Chiriqui.
Within this place it can sail at will; once above the crater opening it
can travel anywhere, although it cannot land; but by some unknown
force or magnetic attraction or freak of gravitation the machine
cannot ascend through the crater, although, when over it, it will drop
like a plummet through the opening. And herein—for the sake of
humanity, listen to this and remember my words—lies a means of
destroying the machine, for by surrounding the crater with powerful
guns the sphere can be shelled as it emerges and utterly destroyed.
To attempt to do so as it returns to the crater would be suicidal, for
once in the outer air, it emanates vast quantities of most poisonous
gas, and all living things within a radius of several miles would be
struck down unconscious, as were my companions on the Chiriqui.
Even if gas-masks were worn, it would be most difficult to destroy
the machine as it descended, for it travels with incredible speed in its
descent and, moreover, the terrible creatures who man the thing
would see that enemies lurked near and would find some means of
destroying them, or by the mysterious magnet force they control,
would draw even the heaviest cannon to the machine as an ordinary
magnet draws needles or iron filings. So if the thing is to be
destroyed, it must be done as the machine emerges from the crater.
Would to God that I could tell where the crater is, but beyond feeling
sure it is at the summit of an Andean peak, I have no means of
locating it.
“But I was telling of what occurred on that tenth day when the
spherical airship was projected from my sight by the blast of steam.
As the machine vanished, the women who had watched its
departure, returned to their city, and I swept the landscape with my
glasses, wondering at the bleak, terrible scenery and bizarre colors.

“As I focussed the binoculars upon a level plateau, perhaps a mile

from where the Chiriqui rested, I gasped in surprise. Clearly defined,
lay the remnants of what had once been a steamship! Had I given
the matter thought, I might have known that the Chiriqui was not the
first vessel to have fallen a victim to these awful beings; but the sight
of another ship’s skeleton came to me as a terrific shock. As nearly
as I could judge, the vessel had been dismantled, for only the great
steel frame remained, with the mighty boilers and other portions of
the ship scattered about, and gruesomely like some mammoth
creature lying disemboweled upon the earth.
 “I was consumed with a mad desire to visit that pathetic wreck, but
I knew not to what dangers I would be exposed, once I left the
security of my ship. Not a being was in sight, however, and carefully I
studied the land, visually measuring the relative distances between
myself and the wreck, and between the city and the route I must
traverse. Having already observed that the giantesses moved slowly
and cumbrously on foot, I at last decided that even if they attempted
to intercept me I could regain the Chiriqui before I was overtaken, so
I threw caution to the winds and prepared to undertake my
hazardous journey. Slinging the loaded rifle on my back, with the
revolver at my belt, and still further arming myself with a keen-edged
fireaxe, I hunted up the pilot’s ladder, lowered it over the lowest side
of the ship,—which was also the side farthest from the city,—and
clambering down the Chiriqui’s lofty sides, leaped down upon the
ground. To my amazement, I landed in a dense jungle of dry, tough
vegetation which rose to my shoulders. From the deck, looking
directly downwards, I had thought this dull-green growth a short, wiry
grass, and, of course, in its relative proportion to the gigantic women,
it was no higher than ordinary grass to a normal human being. It was
a wonderful example of the theory of relativity, but my mind was not
interested in scientific matters at the time, and I merely gave thanks
that the miniature jungle,—which I saw was composed of giant
lichens—would afford me cover through which I might sneak in
safety, and with little chance of detection.
“Without much difficulty I made my way to the other vessel, and
found her even more dissected than I had supposed. Why the
denizens of the place had torn her to bits I did not then know, but
certain portions of her machinery and fittings had been left intact,
and, as I examined these, I made another and most astounding
discovery. Deeply engraved upon a brass plate was the ship’s name
‘U. S. S. Cyclops!’ For a space I stood staring, scarcely able to
believe my eyes. Here then was the solution to that mystery of the
sea, the disappearance of the collier, as laden with manganese, she
vanished without word or trace when off the Barbados during the
World War. No doubt, I thought, many a mystery of the sea had been
caused by the damnable work of these beings with their infernal
machine. But why, for what reason, did they capture ships? Why did
they carry off the unconscious persons upon the vessels? And why
did they tear the vessels apart? It was all a mystery which, in all its
horrible, gruesome, ghoulish details I was soon to solve.
“There was nothing more to be learned from the remains of the
Cyclops, and in safety I returned to the Chiriqui to find, to my
surprise and terror, that a gang of the monstrous females had
boarded the ship in my absence and were stripping her of
everything. But as they caught sight of me, all threw down whatever
they had and fled precipitately, leaving me once more in undisputed
possession of the ship. I was relieved at this, for it was obvious that I
had no need to fear the creatures. By now, too, I had formulated a
theory to account for this strange dread of a being who was a puny,
miserable thing compared to them. Unquestionably my gas-mask
rendered me a most grotesque and unknown creature in their eyes.
My remaining alive and active while all others upon the ship had
succumbed to the noxious gas had probably caused them to think
that I was a supernatural being. The fact that I could go about and
breathe the sulphur-laden air would cause them to regard me with
even greater wonder and superstition, and, as I found later, the fact
that I was never seen to eat, confirmed their belief that I was some
mysterious being against whom their gases and their deviltries were
of no avail.
“I had not much time to devote to such matters, however. Soon
after regaining the Chiriqui I heard excited cries from the land, and
looking over the ship’s rails, I found an immense crowd had gathered
near the empty cradle of the airship, and that all were gazing
upward. Following their example, I stared into the greenish void and
instantly understood. Descending rapidly towards the plain, came the
great sphere, and, suspended below it, was the hull of another
captive ship. And as I focussed my glasses upon this, I rubbed my
eyes and gaped. The dull gray color, the lines, the raking funnels, the
barbettes and gun muzzles left no room for doubt. Incredible as it
seemed, the captive vessel was a warship! What hope then had my
fellow men upon earth? What chance was there if these giant
creatures could send forth their flaming machine, and by it, capture
the fastest, most powerful war-vessels—all within the space of a few
hours?
 “Rapidly the machine and its burden approached, and presently
descended gently dropping the war vessel close to the Chiriqui. My
worst fears were confirmed. The vessel was an American destroyer,
the McCracken, and I knew that scores of my countrymen must lie
unconscious upon her, and in a few moments would be carried off to
some unknown horrible fate. What that fate was I had already
surmised. That first demonstration of the ferocious cannibalism of
the giantesses upon the Chiriqui’s deck had been enough to make
my blood run cold.
“But I had not yet guessed even a fraction of the true horror of it.
Scarcely had the McCracken been dropped upon the earth, when
the women swarmed upon her, and once more I saw the creatures
gathering the inert forms of men and carrying them to the city. And
rapidly, too, they commenced dismantling and tearing the destroyer
into bits. How they had accomplished this with the Cyclops had
puzzled me, but now I witnessed the process close at hand. From
the vicinity of the waterfall, lines or pipes were led to the vessel’s
side; presently there was the roaring sound of steam; dense clouds
of vapor arose from the cataract; the water ceased to flow, and from
the extremities of the lines or tubes twenty-foot jets of blinding flame
shot out. As easily as though made of wax, the steel sides, the
massive beams, the armored barbettes of the warship melted and
were cut by these jets, and as the pieces fell apart, the spherical
airship took a position above the vessel, and by its magnetic power,
lifted tons of the fragments, then sailing off, deposited them in some
spot beyond the city. It was then, as I saw the ship rapidly dissolving
before my eyes, that the inspiration came to me which may make it
possible for me to communicate with the outside world and may, if
God wills, serve to warn my fellow men of the fate which will
overtake them if these terrible creatures are allowed to follow out
their plans. As the jets of flame cut through the McCracken’s
superstructure, and the radio antennae fell in a tangled mass across
the deck, I forgot all else and rushed to the wireless room of the
Chiriqui. Here was my chance. If the ship’s radio transmitter was still
in working order; if the auxiliary battery was still charged, I might
send out messages which, small as the chances were, might reach
the ears of some of the countless thousands of persons who listened
each night at their receiving sets. I trembled with fear that I would
find the transmitter injured or dismantled. I shook with dread that the
battery might be dead. I felt faint with apprehension that the
message, if sent, might never penetrate the sulphur-laden
atmosphere or might never reach the outer world. And I realized,
with a sickening sinking of my heart, that even if heard my
communication might be regarded as a hoax, and no attention would
be given it. But I would do my best. The radio set had not been
molested. Everything was in working order, and I set myself the task
of transmitting my story each night at the same hour, repeating it
over and over again, until the storage batteries are exhausted, for to
get up steam and start the dynamos is beyond my powers. Had I
knowledge of Morse I would send my story by that code, but I have
not, and so—I must cease. For the love of your race and of your
dear ones listen, I beseech you, until I can resume.”

Here the message broke off abruptly, and Frank and I sat staring at
each other, fearing to speak lest we might interrupt or miss the words
which might come, and listening with straining ears at the head-sets.
For an hour we sat there and then, once more the voice spoke.
“The doom that I feared is approaching. I have been here for three
months and this will, I know, be my final message. Oh that I could
only be sure that someone has heard my words, that my fate has not
been in vain but has served to warn my fellow beings. But I must
hurry on. I have learned everything of importance. I have watched,
studied and have even learned to understand much of the language
of these beings. I found that there were men. They are puny beings
compared to the women, though ten-foot giants compared to normal
men, and they are cowed, abject, mere slaves of the females. Only
enough male children are permitted to survive to propagate the race.
All others are killed.
“As they reach manhood only those males of super-intelligence,
strength and virility are permitted to live. The others are destroyed
and—yes, horrible as it sounds, their bodies, like those of the
murdered infants and of the aged, sick or infirm, are devoured. And
as fast as the males attain middle age their lives are forfeited. Long
ago these beings subsisted upon the few wild creatures which
roamed their land; but long ago all these were exhausted and human
flesh became the only meat. There is no vegetable food, and for a
time the sacrificed surplus males, and the aged, provided food for
the race. But gradually the male births decreased, female children
preponderated, and with the increased population resulting, the
males were too few to nourish the others. Then, through what
damnable accident or design I do not know, the creatures went forth
in their airship and discovered the teeming millions of human beings
on earth.
“But the bulk of humanity was and still is safe from them, at least
until new means of attacking mankind are devised, for the globular
airship cannot approach the land. The very power it uses to lift the
greatest steamships and carry them off, draws the machine to the
earth and holds it fast. But above water, which acts as an insulator
apparently, the apparatus can operate at will. And they have a two-
fold purpose in capturing ships. All the available metal in this land
was exhausted in constructing two of the spherical machines. One of
these never returned from its first trip, and only the one remains. To
construct more, these giant women plan to use the metal salvaged
from captured ships, until a vast fleet of the infernal things is ready to
go forth and wipe the seas clean of ships and human beings. And
the bodies of the men and women, struck down by the gas, are to
serve as food for these demons in human form.
“This is the most horrible, blood-curdling thing of all. Rendered
unconscious by the gas, the victims remain in a state of suspended
animation indefinitely, exactly as do grubs, spiders and insects when
stung by certain species of wasps and placed in their nests to
provide food for their young. Stacked in great storage vaults these
breathing, living, but paralyzed human beings are kept, and as
needed, are taken out.
“Already they have a supply on hand sufficient to last them for
over a year. Some of the Cyclops company are still preserved; there
are over three hundred from the Chiriqui, hundreds from other ships,
and the entire crew of the McCracken.
“All these things I learned little by little, and mainly through a
friend, for marvelous as it may seem, I have a friend—if friend he
can be called, a miserable, trembling, terrified male, who, doomed to
death, sought to escape his fate and sought refuge with me,
dreading my presence less than his doom, and hoping that such a
feared and almost reverenced being as myself might protect him. For
two months he has been my companion, but he cannot eat anything
but meat and the supply of meat upon the ship is getting low, and
sooner or later he must succumb. And the women, maddened at his
escape from their clutches, though not yet daring to approach too
closely to me, are getting bolder. Some time, at some unguarded
moment, they will find the poor fellow alone and will fall upon him.
And in his terror, in an effort to buy his life, he will, I know, reveal to
them that I am but an ordinary mortal, a man who eats and drinks
and who survived the gas by mechanical and not supernatural
means. But I will not be taken alive by these fearful female
cannibals. When the time comes, as I know it will, I will blow my
brains out, and though they may devour my body they will not rend
me alive. No more ships have been brought in here since the
McCracken was captured. But this I know is due to the fact that all
the energies of these creatures are being devoted to building
additional air machines. This work goes on in a vast cavern beyond
the city where tremendous forces, furnaces with heat beyond human
conception and machines of which we know nothing, are controlled
by the internal steam, the radiant energy and the magnetic powers of
the earth’s core.
“And now, again let me implore any and all who may hear my
words to give close attention to what I say, for here again is a means
by which humanity may combat and destroy these ghastly, gigantic
cannibals. The spherical air-machines are helpless from above.
Their magnetic or electrical forces extend only downwards. The
gasses they throw out are heavier than air and descend but cannot
ascend, and by means of swift planes, huge bombs and machine
guns, the things can be easily destroyed. And they cannot travel
without throwing off the dazzling green light. Only when motionless
are they dark. And so they will offer easy marks and can be readily
detected. So, I beseech you who may hear, that the governments
are notified and warned and that a fleet or many fleets of airplanes
properly equipped patrol the seas, and at first sight of one of the
green meteors rise above it and utterly destroy it without mercy.
 “Wait! I hear a terrified scream.... I am back again at the
transmitter. It was the fellow who has been with me. Poor devil! He
has met his fate, but after all it was the custom of his people, and,
moreover, he would have starved to death in a few days. For that
matter I, too, face starvation. The ship’s stock is running low; all the
food upon the McCracken was destroyed in cutting up that vessel,
and unless another ship is captured I will have no food after two
weeks more. What a strange thought! How terrible an idea! That the
awful fate of hundreds of my fellows would be my salvation! But I will
never live to die from hunger. I can hear the terrible screams of my
late companion on the deck outside. God! It is the end! The fellow
must have told the enraged females. His body has been torn to
shreds. With bloody hands and reeking lips they are rushing towards
the upper deck where I sit. They are here! This is my last word! God
grant that I have been heard! I am about to⸺”
Crashing in our ears came the report of a pistol.
The End

1 The message as it came in, was halting, and interrupted, with many unintelligible
words and repetitions, as if the sender were laboring under an intense strain or was an
amateur. For the sake of clarity and continuity, the communication has been edited and
filled in, but not altered in any detail.
2 The metropolitan papers reported the meteor on the eighteenth and stated it was
observed by those on the Chiriqui on the evening of the seventeenth, but it must be
remembered that the Chiriqui was in the western Pacific and hence had gained a day
in time.

Transcriber’s Note: This story appeared in the July 1927 issue of

Amazing Stories Magazine.
*** END OF THE PROJECT GUTENBERG EBOOK A VOICE FROM
THE INNER WORLD ***

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