Extracellular Vesicles: Applications to Regenerative Medicine, Therapeutics and Diagnostics

Cover
Extracellular Vesicles: Applications to Regenerative Medicine, Therapeutics and Diagnostics
Preface
Contents
Chapter 1 - Therapeutic Potential of Mesenchymal Stromal Cell- derived Small Extracellular Vesicles
1.1 Concepts in Regenerative Medicine
1.2 MSCs Act in a Paracrine Rather than Cellular Manner
1.3 MSC- EVs Mediate Their Therapeutic Effects via Extracellular Vesicles
1.4 MSC- sEVs Improve Disease Symptoms in Many Different Disease Models
1.5 MSC- sEVs Have Successfully Been Applied to Patients
1.6 Immunomodulatory Activities of MSC- EV Products
1.7 Other Activities Mediated by MSC- EV Products
1.8 Heterogeneity of MSC- sEV Preparations
1.9 Take- home Message
References
Chapter 2 - Separation and Purification of Extracellular and Bio- fluid Vesicles – State of the Art: Past, Present, and Future of Extracellular Vesicle Separation and Purification
2.1 Introduction
2.2 EV Separation Methods
2.2.1 Density- based Separation
2.2.1.1 Ultracentrifugation (UC) and Differential Ultracentrifugation (dUC)
2.2.1.2 Density Gradient Ultracentrifugation (DGUC)
2.2.2 Size- based Separation
2.2.2.1 Ultrafiltration (UF)
2.2.2.2 Tangential Flow Filtration (TFF)
2.2.2.3 Size Exclusion Chromatography (SEC)
2.2.3 Affinity (AFF)
2.2.4 Precipitation (PPT)
2.3 Comparative Evaluation of EV Separation Methods
2.4 Combination of Methods for EV Separation
2.5 Emerging Technologies
2.6 Conclusions and Future Perspectives
References
Chapter 3 - Physicochemical Characterisation of Extracellular Vesicles
3.1 Introduction
3.1.1 Perceived Significance of Single Particle Analysis
3.2 Characterisation Methods
3.2.1 Characterisation of the Size, Size Distribution and Concentration of EVs
3.2.1.1 Nano Flow Cytometry (nFCM)
3.2.1.1.1
How Does It Work.Flow cytometry (FCM) is commonly used for the characterisation of EVs. FCM can be used for both quantitative an...
3.2.1.1.2
What Information Can You Obtain.nFCM provides three key physicochemical properties of EVs: size, concentration and molecular com...
3.2.1.1.3
Advantages of the Technique.Three of the most common advantages of FCM include short analysis time (two minutes per sample), rep...
3.2.1.1.4 Disadvantages of the Technique.Although nFCM is designed for high- throughput analysis of EVs, the drawbacks of nFCM include coi...
3.2.1.2 Tunable Resistive Pulse Sensing (TRPS)
3.2.1.2.1
How Does It Work.Tunable resistive pulse sensing (TRPS) is used to measure size and concentration of submicron colloidal particl...
3.2.1.2.2
What Information Can You Obtain.Since TRPS measures changes of the current caused by individual EVs passing through the pore, it...
3.2.1.2.3 Advantages of the Technique.The main advantage of TRPS is that this method is an impedance- based method, which eliminates the l...
3.2.1.2.4 Disadvantages of the Technique.Although TRPS is one of the high- resolution techniques, which can accurately measure size and co...
3.2.1.3 Resonant Mass Measurement (RMM)
3.2.1.3.1
How Does It Work.Resonant mass measurement (RMM), also known as suspended microchannel resonator (SMR), is a technique that meas...
3.2.1.3.2
What Information Can You Obtain.RMM can measure buoyant mass and density of nanoparticles, which can be used to compute size dis...
3.2.1.3.3
Advantages of the Technique.The major advantage of RMM is the ability to measure the size, buoyant mass and dry mass of individu...
3.2.1.3.4 Disadvantages of the Technique.Although RMM was used to measure the size distribution and mass of submicron- sized particles, th...
3.2.1.4 Dynamic Light Scattering (DLS)
3.2.1.4.1
How Does It Work.Dynamic light scattering (DLS) is a commonly used technique for particle size measurement in the range of nano ...
3.2.1.4.2
What Information Can You Obtain.By identifying the diffusion coefficient of the particles in suspension, DLS can analyse their h...
3.2.1.4.3
Advantages of the Technique.One major advantage of DLS is that it measures particles ranging from 1 nm to 6 µm.33 Simple sample ...
3.2.1.4.4
Disadvantages of the Technique.Despite the multiple advantages, DLS has a few limitations. DLS has a high tendency to overestima...
3.2.1.5 Particle Tracking Analysis (PTA)
3.2.1.5.1
How Does It Work.Similar to DLS, PTA measurements are based the Brownian motion of the suspended particles.36 Both PTA and DLS d...
3.2.1.5.2
What Information Can You Obtain.PTA allows quantitative analysis of the sample at a single vesicle level. By calculating the dif...
3.2.1.5.3
Advantages of the Technique.A significant advantage of PTA over DLS is its ability to analyse polydisperse samples where there a...
3.2.1.5.4
Disadvantages of the Technique.Although PTA has similar principles of mechanism with DLS, PTA has narrower size measurement rang...
3.2.2 Morphological Descriptive Analysis
3.2.2.1 Cryo- transmission Electron Microscopy (Cryo- TEM)
3.2.2.1.1 How Does It Work.Cryo- transmission electron microscopy (cryo- TEM) is used for morphological study of biological samples such a...
3.2.2.1.2 What Information Can You Obtain.By image processing the vitrified sample, cryo- TEM generates high- resolution images that allow...
3.2.2.1.3 Advantages of the Technique.The main advantage of cryo- TEM is the preservation of EV morphology through vitrification. With pro...
3.2.2.1.4 Disadvantages of the Technique.Cryo- TEM involves vitrification that entails a complex sample preparation procedure. For example...
3.2.2.2 Atomic Force Microscopy
3.2.2.2.1
How Does It Work.Atomic Force Microscopy (AFM) is a versatile technique that enables quantitative and qualitative analysis of EV...
3.2.2.2.2
What Information Can You Obtain.Regardless of the mode of operation, AFM provides both qualitative and quantitative data for ind...
3.2.2.2.3
Advantages of AFM Imaging and Force Spectroscopy.The main advantage of AFM and force spectroscopy stems from the high sensitivit...
3.2.2.2.4
Disadvantages of AFM Imaging and Force Spectroscopy.Force spectroscopy uses different mathematical models to calculate the appar...
3.2.2.3 Vibrational Spectroscopy – Nanoscale Infrared Spectroscopy (AFM- IR, nanoIR) and Surface Enhanced Raman Spectroscopy (SER...
3.2.2.3.1
How Does It Work.Vibrational spectroscopy, including Infrared (IR) spectroscopy and Raman spectroscopy, measures molecular vibra...
3.2.2.3.2
What Information Can You Obtain.Both IR and Raman spectroscopy are methods of exploring the physical properties of materials in ...
3.2.2.3.3 Advantages of AFM- IR and SERS.AFM- IR and SERS enable the measurement of the adsorption of individual molecules to the EV surfa...
3.2.2.3.4
Disadvantages of AFM-IR and SERS.While both techniques provide complementary information about EV chemical and structural compos...
3.3 Summary
3.3.1 Advantages and Disadvantages of Each Technique
3.3.2 Types of Analysis Conducted With Each Technique
References
Chapter 4 - Biological Characterization of Extracellular Vesicles – Methodologies to Characterize Molecular Composition of Extracellular Vesicles – Advances Towards Single Vesicle Characterization
4.1 Introduction
4.2 The Demand for Single Vesicle Characterization Methods
4.3 Imaging Methods
4.4 Light Microscopy
4.4.1 Widefield Fluorescence Microscopy
4.4.2 Confocal Microscopy
4.4.3 Super- resolution Techniques
4.4.4 Fluorescence Correlation Spectroscopy (FCS)
4.5 Electron Microscopy
4.5.1 Transmission Electron Microscopy (TEM)
4.5.2 Cryo- electron Microscopy
4.5.3 Scanning Electron Microscopy (SEM)
4.5.4 Immunoelectron Microscopy (iEM)
4.5.5 Correlative Imaging Techniques
4.5.6 Flow Cytometry
4.5.7 Label- free Techniques
4.6 Detection of Specific Molecules in EVs by Labeling Methods for Light Microscopy
4.6.1 Labeling Methods for EVs
4.6.2 Lipid Stains in EV Detection
4.6.3 Proteins
4.6.4 Live Cell (EV) Imaging by Light Microscopy
4.6.5 Analysis of Carbohydrates
4.7 Future Challenges and Opportunities
References
Chapter 5 - Extracellular Vesicles in Bone Tissue Engineering
5.1 Introduction
5.1.1 A Vesicle- based Solution
5.1.2 Advantages of Using EVs
5.1.3 The Role of EVs in Bone Formation
5.2 EVs in Bone Tissue Engineering
5.2.1 Priming EVs for Bone Tissue Engineering
5.2.2 Scaffold- based Approaches
5.2.3 EV Modification Strategies
5.2.4 EVs for the Delivery of Biotherapeutic Molecules
5.3 Concluding Remarks
References
Chapter 6 - Mesenchymal Stromal Cell (MSC)- derived Small Extracellular Vesicles as Next- generation Therapeutics for Cartilage Regeneration
6.1 Introduction
6.2 Stem Cells
6.3 Mesenchymal Stromal/Stem Cells (MSCs)
6.4 MSC Therapies for Cartilage Injury and Osteoarthritis
6.5 Paracrine Mechanisms of MSCs in Cartilage Repair
6.6 Extracellular Vesicles
6.7 Mesenchymal Stem Cell- derived Small Extracellular Vesicles (MSC- sEVs)
6.8 MSC- sEV Therapies for Cartilage Injury and Osteoarthritis
6.9 MSC- sEVs as Next- generation Therapeutics for Cartilage Repair
6.10 Considerations and Challenges of MSC- sEV Therapies for Cartilage Regeneration
List of Abbreviations
Acknowledgements
References
Chapter 7 - The Role of Extracellular Vesicles in the Neuronal System: Application of Extracellular Vesicles in Alzheimer's Disease, Parkinson's Disease, Multiple Sclerosis, and Dementia
7.1 Introduction
7.2 Extracellular Vesicles in the Healthy Central Nervous System
7.2.1 Synaptic Plasticity
7.2.2 Neurovascular Integrity
7.2.3 Feedback and Homeostasis
7.2.4 Myelination
7.3 Extracellular Vesicles in the Diseased Nervous System
7.3.1 Alzheimer's Disease
7.3.2 Parkinson's Disease
7.3.3 Multiple Sclerosis
7.3.4 Dementia
7.4 Extracellular Vesicles as Biomarkers for Central Nervous System Diseases
7.4.1 Alzheimer's Disease
7.4.2 Parkinson's Disease
7.4.3 Dementia
7.4.4 Multiple Sclerosis
7.5 Extracellular Vesicles as Therapeutics in Central Nervous System Diseases
7.5.1 Intrinsic Therapeutic Properties of Extracellular Vesicles for Parkinson's Disease, Alzheimer's Disease, and Dementia
7.5.2 Therapeutics Stimulating or Blocking Extracellular Vesicle Release
7.5.3 Using Extracellular Vesicles as Vehicles for Therapeutics
7.5.4 Extracellular Vesicle Therapeutics for Multiple Sclerosis
7.5.4.1 Remyelination
7.5.4.2 Neuroinflammation
7.6 Conclusions
List of Abbreviations
References
Chapter 8 - The Role of Extracellular Vesicles in Aging of the Human Placenta
8.1 Introduction: The Human Placenta
8.1.1 Biological Aging of the Placenta
8.1.2 Placental Pathologies Associated With Senescence and Aging
8.2 Fetal EVs Derived From Chorionic Villi
8.2.1 The Roles of Chorion- derived EVs in Aging- associated Placental Pathologies
8.2.2 Human Umbilical Cord
8.2.2.1 HUVEC- EVs
8.2.2.2 HUVEC- EVs in Aging- associated Pregnancy Disorders
8.2.2.2.1 HUMSC- EVs and WJMSC- EVs in Aging.HUMSC- EVs have regenerative potential as reviewed by Yaghoubi et al.51 For example, HUMSC- E...
8.2.2.2.2 HUMSC- EVs and WJMSC- EVs in Pregnancy Disorders Associated With Aging.HUMSC- EVs reduce cellular apoptosis and enhance angiogen...
8.2.3 Human Umbilical Cord Blood and EVs
8.2.3.1 UCB- EVs and Pregnancy Disorders Associated With Aging
8.2.4 Fetal Membranes
8.2.4.1 Amnion EVs
8.2.4.1.1 AEC- EVs.AEC- EVs are primarily studied for their potential therapeutic use and particularly for studying age- related diseases ...
8.2.4.1.2 AMSC- EVs.AMSC- EVs are also primarily studied for their therapeutic potential in animal models where they have multiple benefic...
8.2.4.2 Chorion and EVs
8.2.5 Amniotic Fluid and EVs
8.3 The Roles of EVs in Maternal Uterine Aging
8.3.1 EVs, Aging, and the Endometrium
8.3.2 EVs, Aging, and the Decidua
8.3.3 EVs, Aging, and the Myometrium
8.4 Conclusion
List of Abbreviations
References
Chapter 9 - Extracellular Vesicles in Lung Diseases
9.1 Introduction
9.2 Therapeutic Applications of Extracellular Vesicles (EVs) in Acute Lung Injury and Acute Respiratory Distress Syndrome
9.2.1 EVs in Preclinical Models of ARDS and ALI
9.2.2 EVs in Clinical Studies of ARDS
9.3 Therapeutic Applications of EVs in Bronchopulmonary Dysplasia
9.3.1 EVs in Preclinical Models of BPD
9.3.2 EVs in Clinical Studies of BPD
9.4 Therapeutic Applications of EVs in Pulmonary Hypertension
9.4.1 EVs in Preclinical Models of PH
9.5 Therapeutic Applications of EVs in Idiopathic Pulmonary Fibrosis
9.5.1 EVs in Preclinical Models of IPF
9.6 Therapeutic Applications of EVs in Other Lung Diseases
9.6.1 Therapeutic Applications of EVs in COPD
9.6.2 Therapeutic Applications of EVs in Asthma
9.7 Conclusion
References
Chapter 10 - Immunomodulatory Function of Extracellular Vesicles in Cancer and Tissue Repair/Regeneration
10.1 Introduction
10.2 EV- mediated Immunomodulation in Cancer
10.3 EVs in Immunomodulation for Promoting Tissue Repair/Regeneration
10.3.1 EVs in Musculoskeletal Disorders
10.3.2 EVs in Cardiovascular Diseases and Repair/Regeneration
10.3.3 EVs in Kidney and Lung Repair and Fibrosis
10.4 EV Immunomodulatory Role in ECM Remodelling in Cancer and Tissue Regeneration
10.5 Conclusion and Future Challenges
Acknowledgements
References
Chapter 11 - Diagnostic Applications of Extracellular and Bio- fluid Vesicles
11.1 Introduction
11.2 Molecular Biomarkers of EVs
11.2.1 Biogenesis of EVs
11.2.2 Biomarkers in EVs for Disease Detection
11.2.2.1 Classical Biomarkers—Nucleic Acids and Proteins
11.2.2.1.1
DNA.DNA encodes all genetic information, and is inherited during DNA replication and chromosome segregation, where parent cell D...
11.2.2.1.2
RNA.The flow of information from DNA (i.e., the genome) to produce RNA, or transcription, is regulated by dynamic factors known ...
11.2.2.1.3
Proteins.Proteins are the most commonly studied EV biomarkers. They are a class of macromolecules that carry out diverse functio...
11.2.2.2 Surface Modifications
11.2.2.3 Organisational State
11.3 Detection of EV Biomarkers
11.3.1 Challenges for Detection of EV Biomarkers
11.3.2 New Technologies for the Detection of EV Biomarkers
11.3.2.1 Nucleic Acids
11.3.2.2 Proteins
11.3.2.3 Surface Modifications
11.3.2.4 Organisational State
11.4 Conclusion and Outlook
Competing interests
Acknowledgements
References
Chapter 12 - Standardization and Commercialization of Extracellular Vesicles
12.1 Extracellular Vesicles: Hypes and Hopes
12.1.1 Biomarker Carriers and Analytical Substrates in Diagnosis and Bioprocess Optimization
12.1.2 Active Natural Effectors and Delivery Vectors in Healthcare and Beyond
12.1.3 Drivers and Indicators for Commercial Activities
12.1.3.1 Huge Market and Unmet (or Growing) Needs
12.1.3.2 Unique and Competitive EV Advantages Over Available Products
12.1.3.3 Large Patent Landscape
12.1.3.4 Investments
12.1.3.5 Intensive Research
12.1.3.6 Quality Stamps and Regulatory Initiatives
12.2 Bumps On the Road: Are We Skipping the Count- down Prior to Launch
12.2.1 Regulatory and Safety Concerns in Diagnostics
12.2.2 Regulatory and Safety Concerns in Therapeutics and Consumer Care
12.2.3 How Ready is the EV Technology Suite Today
12.2.3.1 Scalable Commercialization: Quality, Quantity and Costs
12.2.3.2 Biofluid EV Harvesting
12.2.3.3 Multimodal and Real- time Analytics
12.2.3.4 Sensitivity and Throughput – The EV Diagnostics Challenge
12.3 Where Do We Go Next
12.3.1 Best Positioned EV Providers
12.3.2 Best Positioned EV Tool Providers
12.3.3 Roads Less Travelled
12.3.3.1 Agri- food
12.3.3.2 Cosmetics
12.3.3.3 Microbes, Parasites and Animals
12.3.3.4 Building a Multilayered Innovation Ecosystem
Acknowledgements
References
Subject Index