Cartilage: From Biology to Biofabrication

Description

This book provides a comprehensive overview of cartilage structure, functions, and approaches for the regeneration of cartilage tissue. It reviews multiple signaling pathways that are involved in the growth and repair of cartilage tissue. The initial chapter of the book examines the etiology, diagnosis, and pathological features of various cartilage diseases. Subsequently, the book presents recent advances in biomaterial sciences, regenerative medicine, and fabrication technology for cartilage regeneration. It discusses hydrogels as a promising scaffold for cartilage tissue engineering, focusing on recapitulating microenvironments present during development or in adult tissue to induce the formation of cartilaginous constructs with biochemical and mechanical properties of native tissue. Lastly, it covers the applications of 3 D printing techniques for the fabrication of scaffolds for cartilage tissue regeneration for the production of biological implants capable of treating a range of conditions. Preface Contents Editors and Contributors 1: Introduction to Cartilage Tissue: Development, Structure, and Functions 1.1 Introduction 1.2 Cartilage Tissue in Mammalian Body 1.3 Articular Cartilage 1.3.1 Origin of Articular Cartilage: Embryology of Cartilage 1.3.2 Postnatal Articular Cartilage Growth and Expansion 1.4 Articular Cartilage Macrostructure 1.4.1 Cartilage Composition 1.4.2 The Mature Cartilage Matrix Structure and Function 1.5 Microstructure of Articular Cartilage: Chondrocyte Structure and Function 1.5.1 Chondrocyte Biology 1.6 Ultrastructure of Adult Articular Cartilage: Articular Cartilage Zone Organization 1.6.1 Radial Structure (The Chondron) 1.7 Biomechanical Functions and Mechanical Characteristics of Articular Cartilage 1.7.1 Compressive Properties of Articular Cartilage 1.7.2 Tensile and Shear Properties 1.7.3 Swelling Behavior of Articular Cartilage 1.8 Metabolism of Articular Cartilage 1.9 Maintaining a Healthy State: Articular Injuries and Disease 1.10 Conclusion References 2: Cartilage Defects and Diseases: Conventional Therapies and Its Limitations 2.1 Introduction 2.2 OA 2.2.1 Progress and Limitations of Drug Therapy for OA 2.2.2 Progress and Limitations of Non-drug Therapy for OA 2.2.3 Progress and Limitations of Exercise Therapy for OA 2.2.4 Advances and Limitations of Cell Preparations and Stem Cell Therapy for OA 2.2.5 Progress and Limitations of Surgical Treatment of OA 2.3 RA 2.3.1 Progress and Limitations of Drug Therapy for RA 2.3.2 Progress and Limitations of Biologic Therapy for RA 2.3.3 Progress and Limitations of Exercise Therapy in RA 2.4 GA 2.4.1 Progress and Limitations of Drug Therapy for GA 2.4.2 Progress and Limitations of Exercise Therapy in GA 2.5 Conclusion References 3: Strategies to Control Mesenchymal Stem Cell Differentiation for Regenerating Phenotypically Defined Articular Cartilage 3.1 Introduction 3.2 Spatial and Temporal Influence of Bioactive Factors 3.3 Manipulation of Scaffold Microenvironment for Cartilage Tissue Engineering 3.4 Provision of Biomolecular Cues 3.5 Provision of Physical Cues 3.5.1 Substrate Elasticity 3.5.2 Surface Topography 3.6 Biomaterial Facilitation of Mesenchymal Condensation 3.7 3D Composite Multilayered Scaffolds 3.8 Manipulation of Cell Culture Conditions 3.8.1 Co-Culture Platform 3.8.2 Oxygen Tension 3.8.3 Dynamic Stimulation 3.9 Multifactorial Approach in Stimulating MSC Chondrogenesis 3.10 Future and Conclusion References 4: Single-Cell Analysis Approaches in Cartilage Diseases Diagnosis and Therapies 4.1 Introduction 4.2 Workflow in Single-Cell Technology 4.2.1 Single-Cell Isolation Technologies 4.2.1.1 Fluorescence-Activated Cell Sorting (FACS) 4.2.1.2 Laser Capture Microdissection (LCM) 4.2.1.3 Microfluidics 4.2.1.4 Magnetic-Activated Cell Sorting (MACS) 4.3 Various Type of Single-Cell Technologies 4.3.1 Single-Cell Genomics (SCG) 4.3.2 Single-Cell RNA Sequencing 4.3.2.1 Single-Cell Spatial Transcriptomics 4.3.3 Single-Cell Proteomics 4.3.3.1 Immunoassay-Based Techniques 4.3.3.2 Mass Spectrometry Tools 4.3.3.3 Microfluidics-Based Platforms 4.3.4 Single-Cell Epigenomics 4.3.4.1 Single-Cell DNA Methylation Profiling 4.3.4.2 Single-Cell Histone Modification Mapping 4.3.4.3 Single-Cell Chromatin Conformational Assessments 4.3.5 Single-Cell Multi-Omics 4.3.5.1 Single-Cell Genome and Transcriptome 4.3.5.2 Single-Cell Proteome and Transcriptome 4.3.5.3 Single-Cell Epigenome and Transcriptome 4.4 Single-Cell Analysis Applications 4.4.1 Biomarker Discovery 4.4.2 Single-Cell Genomics and Regenerative Medicine 4.4.2.1 Evaluation of Accuracy and Precision in Regenerative Medicine and Tissue Engineering 4.4.2.2 Single-Cell Atlases as a Reference for Tissue Regeneration 4.4.2.3 scRNA-seq-Related Methodologies to Guide Cell and Tissue Engineering Transcription Factor Combination Spatial Reconstruction CRISPR-Cas9 System 4.4.2.4 Importance of scRNA-Seq in Disease Modeling and Therapy 4.4.3 Single-Cell Sequencing and Personalized Medicine 4.4.3.1 Dissection of the Tumor Microenvironment 4.4.3.2 Tumor Heterogeneity Assessment 4.4.3.3 Study of Therapy Resistance 4.4.3.4 Drug Development 4.5 Concluding Remarks and Future Perspective References 5: The Importance of Mechanical Stimulation in Cartilage Formation: Applications of Bioreactors 5.1 Mechanical Functions and Properties of Articular Cartilage 5.2 Mechanical Milieu of Articular Cartilage 5.3 Development of Bioreactor Culture Systems 5.4 Cartilage-Specific Bioreactors Applying Mechanical Stimulation to Favor Neotissue Formation 5.4.1 Hydrostatic Pressure (HP) 5.4.2 Compression 5.4.3 Shear Stress 5.4.4 Multiaxial Loading 5.5 Mechanotransduction Mechanisms in Chondrogenic Cells: Evidence from Bioreactors 5.5.1 Mechanoreceptors on Chondrocyte Cytoplasmic Membranes 5.5.1.1 Ion Channels 5.5.1.2 Primary Cilia 5.5.1.3 Integrins 5.5.2 Downstream Signaling Cascades 5.6 New Perspectives for Mechanically Stimulated Cartilage Models: Joint-On-Chip References 6: Signaling Pathways Regulating Cartilage Formation 6.1 Introduction 6.2 Development and Organization of the Cartilage 6.3 Signaling Pathways Regulate Chondrogenesis During Cartilage Formation 6.4 Early-Stage Regulator in the Signaling Cascade 6.4.1 TGF-beta Signaling Pathway 6.4.2 BMP Signaling Pathway 6.4.3 SOX-9 Signaling Pathway 6.4.4 IGF Signaling Pathway 6.4.5 FGF Signaling Pathway 6.4.6 Wnt/beta-Catenin Signaling Pathway 6.4.7 Growth Differentiation Factor 5 (GDF5) Signaling Pathway 6.4.8 Hedgehog Signaling Pathway 6.5 Late-Stage Signaling Pathways Involved in Cartilage Development 6.5.1 Prg4 Signaling Pathway 6.5.2 Notch Signaling Pathway 6.6 Conclusion References 7: Role and Application of Biomolecules for Regeneration of Cartilage Tissue 7.1 Introduction 7.2 Biomolecules Affecting Cartilage Development and Maturation 7.2.1 Cartilage Components 7.2.1.1 Type II Collagen (COL2A1) 7.2.1.2 Aggrecan (ACAN) 7.2.1.3 Lubricin (PRG4) 7.2.1.4 Tenascin-C (TN-C) 7.2.2 Signaling-Related Proteins 7.2.2.1 Transforming Growth Factor-betas (TGF-betas) 7.2.2.2 EGFR Signaling 7.2.2.3 Fibroblast Growth Factor 18 (FGF18) 7.2.2.4 Parathyroid Hormone-Related Peptide (PTHrP) 7.2.2.5 CD44 7.3 Transcription Factors 7.3.1 SRY-Box9 (SOX9) 7.3.2 Forkhead Box Class O (FOXO) 7.3.3 Nuclear Factor of Activated T Cells (NFATc) 7.3.4 cAMP Response Element-Binding Protein (CREB) 7.3.5 Hypoxia-Inducible Factor (HIF) 1α and 2α 7.3.6 Y-Box Binding Protein 1 (YBX1) 7.4 Biomolecules Affecting Cartilage Homeostasis 7.4.1 RNA-Binding Proteins 7.4.1.1 Pre-transcriptional Regulation 54-kDa Nuclear RNA-Binding Protein (p54nrb) 7.4.1.2 Splicing Nucleolar GTP-Binding Protein 3 (GNL3) Fused in Sarcoma (FUS) 7.4.1.3 mRNA Stability Methyltransferase 3 (METLL3) Tristetraprolin (TTP) Human Antigen R (HuR) Staphylococcal Nuclease and Tudor Domain-Containing 1 (SND1) 7.4.1.4 Stress Granule Assembly T-Cell-Restricted Intracellular Antigen 1 (TIA-1) Transactive Response DNA-Binding Protein 43kDa (TDP-43) 7.4.1.5 Regulation of Translocation Cytoplasmic Polyadenylation Element-Binding Protein 1 (CPEB1) Pumilio RNA-Binding Family Member 1 (PUM1) 7.4.2 MicroRNAs (miRNAs) 7.4.2.1 miRNA140 7.4.2.2 miRNA17 7.4.2.3 miRNA101 7.4.2.4 miRNA379-5p 7.4.2.5 miRNA455-5p and -3p 7.4.2.6 miRNA93-5p 7.4.2.7 miRNA126-5p 7.4.2.8 miRNA146a 7.4.3 Circular RNAs (circRNAs) 7.4.4 Ubiquitination 7.4.4.1 AXIN 7.4.4.2 Ubiquitin-Conjugating Enzyme E2 M (UBE2M) 7.4.4.3 PARKIN 7.4.4.4 F-Box Protein 6 (FBXO6) 7.4.4.5 Aurora Kinase A (AURKA) 7.4.4.6 WW Domain-Containing Protein 2 (WWP2) 7.5 Reactive Oxygen Species 7.5.1 Superoxide Dismutase 2 (SOD2) 7.5.2 Nuclear Factor (Erythroid-Derived 2)-Like 2 (NRF2) 7.5.3 Peroxiredoxins (PRDX) 7.5.4 Mitofusion 2 (MFN2) 7.6 Conclusion References 8: Extracellular Matrix Biomimicry for Cartilage Tissue Formation 8.1 Introduction 8.2 Physicochemical and Biological Properties of Biomimetic Constructs 8.2.1 Physicochemical Properties 8.2.1.1 Stiffness 8.2.1.2 Porosity 8.2.1.3 Surface Properties 8.2.1.4 Piezoelectric Properties 8.2.1.5 Electrical Conductivity 8.2.1.6 Fluid Absorption 8.2.1.7 Surface Wettability 8.2.2 Biocompatibility 8.2.2.1 Ability to Stimulate Chondrogenesis 8.2.2.2 Proliferation 8.2.2.3 Adherence 8.2.2.4 Maintenance of Cell-Cell Interactions 8.2.2.5 Degradation 8.3 Biomimetic Materials for Cartilage Tissue Restoration 8.3.1 Natural ECM-Based Biomimetic Constructs for Cartilage Repair 8.3.2 Natural Non-cartilage ECM-Based Biomimetic Materials for Cartilage Repair 8.3.3 Synthetic Materials for Cartilage Repair 8.3.4 Mixed Biomimetic Materials for Cartilage Repair 8.4 The Effects of External Physical and Chemical Stimuli on ECM-Based Scaffolds/Structures 8.5 Application of Scaffolds for In Vivo Studies 8.6 Conclusions References 9: Cartilage Tissue Engineering: Advances and Frontiers 9.1 Introduction 9.2 Scaffold-Based Techniques for Cartilage Regeneration 9.2.1 3D Bioprinting 9.2.2 In Situ Printing 9.2.2.1 Handheld 9.2.2.2 Robotic 9.2.3 Organ-on-a-Chip Platforms 9.2.4 Injectable Hydrogel-Based Drug Delivery System 9.3 Emerging Technologies for Cartilage Tissue Engineering 9.3.1 4D Printing 9.3.2 5D Printing 9.3.3 6D Printing 9.4 Scaffold-Free Techniques for Cartilage Regeneration 9.4.1 Kenzan Bioprinting 9.4.2 Organoids 9.4.3 Cell Sheet 9.4.4 Cell Imprinting 9.5 Conclusion References 10: Advances in Hydrogels for Cartilage Regeneration 10.1 Introduction 10.2 Cross-Linking Strategies for Hydrogel Formation in Cartilage Tissue Engineering 10.2.1 Physically Cross-Linked Hydrogels 10.2.2 Chemically Cross-Linked Hydrogels 10.2.3 Double-Network Hydrogels 10.3 Hydrogels as Vehicles for Delivery of Chondroinductive Factors 10.4 Bioprinting of Hydrogels for Cartilage Regeneration 10.5 Conclusion and Outlook References 11: Shape-Memory Polymers in Cartilage Tissue Engineering 11.1 Introduction 11.2 Shape-Memory Polymer (SMP) as a Biomaterial 11.3 Shape-Memory Polymers in Biomedical Applications 11.3.1 Polymers with Shape Memory for Delivery of Drugs 11.3.2 Shape-Memory Polymers for Cardiovascular 11.3.3 Polymers with Shape Memory and Antibacterial Properties 11.4 Biodegradable Shape-Memory Polymers (BSMP) 11.5 Tissue Engineering 11.6 Cartilage Tissue Engineering 11.7 Using Shape-Memory Polymers in Cartilage Tissue Engineering Scaffolds 11.7.1 Alginate as a Smart Natural Polymer in Cartilage Tissue Engineering 11.7.2 Alginate Scaffolds in Cartilage Tissue Engineering 11.7.3 Alginate as a Cell Carrier in Cartilage Tissue Engineering 11.7.4 Alginate in the Electrospinning Process 11.7.5 Alginate in a Multiresponsive System 11.8 More Studies and Reviews 11.9 Conclusion References 12: Widely Used Biomaterials in Cartilage Biofabrication 12.1 Introduction 12.2 Properties of Cartilage Tissues 12.3 Cartilage Diseases 12.4 Cartilage Tissue Regeneration Approaches 12.4.1 Clinically Used Approaches 12.4.2 Surgical Approaches 12.4.3 Regenerative Medicine and Cell-Based Approaches 12.4.4 Tissue Engineering Approaches 12.5 Biomaterials for Damaged Cartilage Tissue Regeneration 12.5.1 Natural Materials 12.5.1.1 Collagen 12.5.1.2 HA 12.5.1.3 Fibrin 12.5.1.4 CS 12.5.1.5 Agarose 12.5.1.6 Alginate 12.5.1.7 Bacterial Cellulose (BC) 12.5.1.8 Chondroitin Sulfate (ChS) 12.5.2 Synthetic Materials 12.5.3 Bioactive Molecules Used for Cartilage Tissue Engineering 12.5.3.1 Kartogenin (KGN) 12.5.3.2 Simvastatin 12.5.4 Smart Biomaterials for Cartilage Tissue Regeneration 12.6 Conclusion and Perspective References 13: Importance of 3D Printing Techniques in Cartilage Tissue Engineering 13.1 Introduction 13.2 3D Printing Essentials in Cartilage Tissue Regeneration 13.3 Overview of 3D Printing Approaches in Cartilage Tissue Engineering 13.3.1 Scaffold-Based 3D Printing Approaches 13.3.1.1 Inkjet-Based 3D Printers 13.3.1.2 Extrusion-Based 3D Printers 13.3.1.3 Laser-Based 3D Printers 13.3.1.4 Stereolithography 3D Printers 13.3.1.5 In Situ 3D Printer 13.3.2 Scaffold-Free 3D Printing 13.4 Conclusion and Future Aspects References 14: Cell Therapy as a Novel Therapeutic Approach for Cartilage Diseases 14.1 Introduction 14.2 Mechanism of Cartilage Regeneration 14.3 Cellular Sources 14.3.1 Bone Marrow-Derived Mesenchymal Stem Cells (BD-MSCs) 14.3.2 Adipose-Derived Mesenchymal Stem Cells (AD-MSCs) 14.3.3 Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells (hUCB-MSCs) 14.3.4 Synovial-Derived Mesenchymal Stem Cells (SD-MSCs) 14.4 Cellular Delivery Techniques 14.4.1 Direct Implantation 14.4.2 Injection 14.5 Rehabilitation Following Cartilage Repair 14.6 Conclusion References 15: Extracellular Vesicles: A Potent Therapeutic Tool for Cartilage Regeneration 15.1 Introduction 15.2 Cartilage 15.3 Cartilage Injuries 15.4 Current Treatments and Challenges in Management of Cartilage Injuries 15.5 EV Biology 15.5.1 Biogenesis 15.5.2 Characteristics 15.6 Involvement of EVs in the Pathophysiology of Cartilage Diseases 15.7 Chondrogenic Role of EVs 15.8 Role of EVs in Cartilage Repair 15.9 EV-Based Cell-Free Therapy for Cartilage Repair 15.10 Future Perspectives and Conclusions References 16: Osteochondral Unit Approach for Articular Cartilage Regeneration 16.1 Introduction 16.2 Clinical Treatment Methods 16.3 Osteochondral Tissue Engineering Approaches for Articular Cartilage Regeneration 16.3.1 Requirement of Osteochondral Scaffolds 16.3.1.1 Biocompatibility, Bioactivity and Surface Topography 16.3.1.2 Biodegradability 16.3.1.3 Mechanical Strengths 16.3.1.4 Architecture and Porosity 16.3.2 Biomaterials for Osteochondral Regeneration 16.3.2.1 Biomaterials for Cartilage Regeneration 16.3.2.2 Biomaterials for Subchondral Bone Regeneration 16.3.3 Traditional TE Approaches 16.3.3.1 Cell-Free and Cell-Seeding TE Approaches 16.3.3.2 Cell-Based TE Approaches 16.3.3.3 Scaffold-Free TE Approaches 16.3.4 3D-Printed Osteochondral Scaffolds for Articular Cartilage Regeneration 16.3.4.1 Monophasic Scaffolds 16.3.4.2 Biphasic and Multiphasic Scaffolds 16.3.4.3 Gradient Scaffolds 16.4 Concluding Remarks References 17: Stem Cells Therapy for Cartilage Regeneration in Clinic: Challenges and Opportunities 17.1 Introduction 17.2 Stem Cells for Cartilage Regeneration 17.3 Stem Cell Delivery Strategies in Cartilage Regeneration 17.4 Mechanism of Stem Cells´ Function in Cartilage Regeneration 17.5 The Effect of Stem Cells on Cartilage Regeneration by Differentiating and Affecting Biological Processes 17.6 Regeneration Mechanisms of Mesenchymal Stem Cells in Damaged Cartilage 17.7 Embryonic and Induced Pluripotent Stem Cells in Cartilage Regeneration 17.7.1 Embryonic Stem Cells (ESCs) 17.7.1.1 Growth Factors 17.7.1.2 Coculture, Conditioned Medium, and Morphogenetic Factors 17.7.1.3 Small Molecules 17.7.1.4 Genetic Manipulation 17.7.1.5 Biomaterial-Assisted Chondrogenic Differentiation of ESCs and Cartilage Tissue Engineering 17.7.2 Induced Pluripotent Stem Cells 17.8 Mesenchymal Stem Cells in Cartilage Regeneration 17.8.1 MSCs as a Therapeutic Tool or Target 17.8.1.1 Endogenous MSCs 17.8.1.2 Exogenous MSCs 17.8.2 Combination Therapies with MSCs 17.8.2.1 Small Molecular Drugs 17.8.2.2 Growth Factors 17.8.2.3 Biomechanical Factors 17.9 Clinical Trial 17.10 Horizons and Challenges Ahead References 18: Validation of Tissue-Engineered Constructs: Preclinical and Clinical Studies 18.1 Introduction 18.2 Present Status for Articular Cartilage Repair 18.2.1 Traditional Surgical Regeneration Techniques 18.2.2 Osteochondral Transplantation (OT) 18.2.3 Autologous Chondrocyte Implantation (ACI) 18.2.4 Allogeneic and Autologous Stem Cells Implantation 18.3 Progress Toward Tissue-Engineered Cartilage 18.3.1 Tissue-Engineered Constructs 18.3.2 Cell-Free Approaches for Cartilage Regeneration 18.3.3 Scaffold-Free Constructs 18.3.4 Extracellular Vesicles: A Promising Cell-Free Therapy for Cartilage Repair 18.3.5 In Vivo Characterization for Bioconstructs Before Initiation of Clinical Studies (Preclinical Studies) 18.3.6 Mechanical Testing of Cartilage Implants 18.3.7 Evaluating Biomaterial-Host Interactions with Histological Scoring 18.3.8 Evaluating Biomaterial Interaction with Imaging 18.3.8.1 Magnetic Resonance Imaging (MRI) 18.3.8.2 Microcomputed Tomography (Micro-CT) 18.3.8.3 Noninvasive Tracking and Monitoring 18.3.8.4 Fluorescent Labeling 18.3.8.5 Bioluminescent Imaging 18.3.8.6 Radiolabeling 18.3.8.7 Magnetic Particle Labeling 18.4 Clinical Performance of Tissue-Engineered Constructs and Templates 18.4.1 Commercial or FDA-Approved Constructs 18.4.2 Clinical Trials Founded on Validated Constructs 18.4.2.1 VAS [Time Frame: x Days/Months] 18.4.2.2 The score for Lysholm [Time Frame: x Days/Months] 18.4.2.3 The Score According to KOOS [Time Frame: x Days/Months] 18.4.2.4 IKDC Score [Duration: x Days/Months] 18.4.3 Inclusion Criteria 18.4.3.1 Carticel 18.4.3.2 Chondron 18.4.3.3 Cartistem 18.4.4 MACI 18.5 Concluding Remarks and Future Trends References