Handbook of Intelligent Scaffolds for Tissue Engineering and Regenerative Medicine

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Handbook of Intelligent Scaffolds for Tissue Engineering andRegenerative Medicine

Copyright c© 2012 Pan Stanford Publishing Pte. Ltd.

All rights reserved. This book, or parts thereof, may not be reproduced in anyform or by any means, electronic or mechanical, including photocopying,recording or any information storage and retrieval system now known or tobe invented, without written permission from the publisher.

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ISBN 978-981-4267-85-4 (Hardcover)

ISBN 978-981-4267-86-1 (eBook)

Printed in the USA


I would like to dedicate this handbook to my wife,

Isabella Seong Hee Koh; my children, Jerome Taeuk

and Daniel Taehoon; and my mother.

—G.K.


Contents

Preface xxix

I. INTRODUCTION

1 Biomaterials and Manufacturing Methods for Scaffoldin Regenerative Medicine 3Gilson Khang1.1 Introduction 4

1.2 Biomaterials for Regenerative Medicine and Tissue

Engineering 6

1.2.1 Importance of Scaffold Matrices in

Regenerative Medicine and Tissue Engineering 6

1.2.2 Bioceramic Scaffolds 9

1.2.2.1 Calcium phosphate 10

1.2.2.2 Tricalcium phosphate 10

1.2.2.3 Hydroxyapatite 11

1.2.2.4 Bioglass 11

1.2.2.5 Demineralized bone particle 11

1.2.3 Synthetic Polymers 13

1.2.3.1 Poly(α-hydroxy ester)s 14

1.2.3.2 Polyanhydride 16

1.2.3.3 Poly(propylene fumarate) 16

1.2.3.4 PEO and its derivatives 17

1.2.3.5 Polyvinylalcohol 18

1.2.3.6 Oxalate-based polyesters

(polyoxalate) 19

1.2.3.7 Polyphosphazene 19

1.2.3.8 Biodegradable polyurethane 20

1.2.3.9 Other synthetic polymers 20


viii Contents

1.2.4 Natural Polymers 21

1.2.4.1 Fibrin 21

1.2.4.2 Collagen 22

1.2.4.3 Alginate 23

1.2.4.4 Small intestine submucosa 25

1.2.4.5 Silk 25

1.2.4.6 Hyaluronan 26

1.2.4.7 Chitosan 27

1.2.4.8 Agarose 28

1.2.4.9 Acellular dermis 28

1.2.4.10 Polyhydroxyalkanoates 29

1.2.4.11 Other natural polymers 29

1.2.5 Bioactive Molecules Release System for

the Regenerative Medicine and Tissue

Engineering 30

1.3 Scaffold Fabrication and Characterization 32

1.3.1 Fabrication Methods of Scaffolds 32

1.3.1.1 Electrospinning method 32

1.3.1.2 PGA nonwoven sheet 33

1.3.1.3 Porogen-leaching methods 33

1.3.1.4 Gas-foaming method 34

1.3.1.5 Phase separation method 34

1.3.1.6 Rapid prototyping 35

1.3.1.7 Injectable gel method 37

1.3.2 Physicochemical Characterization of Scaffolds 37

1.3.3 Sterilization Method for Scaffolds 39

1.4 Conclusions 40

II. CERAMIC AND METAL SCAFFOLD

2 Innovative Bioinspired SIC Ceramics from VegetableResources 51M. Lopez-Alvarez, P. Gonzalez, J. Serra, A. de Carlos,S. Chiussi, and B. Leon2.1 Introduction 52

2.2 Bioinspired SiC Ceramics 54

2.3 Biocompatibility Studies 60

2.4 Conclusions and Outlook 64


Contents ix

3 Production of Three-Dimensional Hierarchical NanoTi-Based Metals Scaffolds for Bone Tissue Grafts 69Shuilin Wu, Xiangmei Liu, Paul K. Chu, Tao Hu,Kelvin W. K. Yeung, Jonathan C. Y. Chung, and Zushun Xu3.1 Introduction 70

3.2 Fabrication and Characteristics of Macroporous

Ti-Based Alloys 73

3.3 Natural Growth and Characterization of

1D Nano Titanates 75


4 Bioceramic Scaffold—Bone Tissue Engineering 83Willi Paul and Chandra P. Sharma4.1 Introduction 84

4.2 Bioceramics 85

4.3 Bone Tissue Engineering 86

4.4 Research Perspective 87

4.5 Basic Questions in Bone Tissue Engineering 90

4.6 Conclusion 91

III. INTELLIGENT HYDROGEL

5 Induction of Soft-Tissue Regeneration Using HydrogelsOptimized for Inflammatory Response 99Nicholas P. Rhodes and John A. Hunt5.1 Introduction 100

5.2 Hyaluronan as a Base Polymer for Regenerative

Therapies 101

5.2.1 Experience with Esterified Hyaluronan 102

5.2.2 Strategies for Controlling Inflammation 103

5.2.3 Amidated Hyaluronan Biomaterials 104

5.3 Results of Implantation of Amidated Gels 106


6 Enzymatically Triggered in situ Gel-FormingBiomaterials for Regenerative Medicine 111Yoon Ki Joung, Kyung Min Park, and Ki Dong Park6.1 Introduction 112


x Contents

6.2 In situ–Formed Hydrogels as Injectable Scaffolds 113

6.3 Enzyme-Triggered Hydrogels 114

6.3.1 HRP-Catalyzed Systems 115

6.3.2 TGase-Catalyzed Systems 120

6.3.3 Other Enzyme-Catalyzed Systems 121


7 Thermo-Sensitive Injectable Scaffolds forRegenerative Medicine 127Moon Suk Kim, Jae Ho Kim, Gilson Khang, and Hai Bang Lee7.1 Introduction 128

7.2 In situ–Forming Hydrogels Formed by Electrostatic

Interactions 129

7.2.1 In situ–Forming Chitosan Hydrogel Scaffolds 129

7.2.2 In situ–Forming Alginate Hydrogel Scaffolds 132

7.3 In situ–Forming Hydrogels Formed by Hydrophobic

Interactions 132

7.3.1 PEG–PPG Block Copolymers as in situ–

Forming Hydrogel Scaffolds 134

7.3.2 PEG–Other Degradable Polyesters as

in situ–Forming Hydrogel Scaffolds 135

7.3.3 Other Polymers as in situ–Forming Hydrogel

Scaffolds 136


8 Photocurable Hydrogel for Tissue Regeneration 143Min Soo Bae and Il Keun Kwon8.1 Introduction 143

8.2 Photopolymerization of Hydrogels 146

8.2.1 Photoinitiators of Photocurable Hydrogel 147

8.3 Photopolymerizable Materials 147

8.3.1 Photocurable Hydrogel from Natural Polymers 149

8.3.1.1 Photo-cross-linkable collagen and

gelatin 149

8.3.1.2 Photo-cross-linkable hyaluronic acid 151

8.3.1.3 Photo-cross-linkable chitosan 151

8.3.2 Photocurable Hydrogel from Synthetic

Polymers 152

8.3.2.1 Photo-cross-linkable poly(ethylene

glycol) 153


Contents xi

8.3.2.2 Transition in theoretical work 155

8.3.2.3 Photo-cross-linkable poly(hydroxyl

esters) 156

8.4 Summary and Outlook 157

9 Hyaluronan-Based Hydrogel Scaffolds 165Insup Noh9.1 Introduction 165

9.2 Characteristics of Hyaluronic Acid in Biomedical

Engineering 167

9.3 HA Derivatives 169

9.3.1 Ester Derivatives 169

9.3.2 Carbodiimide (R1N=C=NR2) 169

9.3.3 Sulfydrylation (HA–SH) 170

9.3.4 Sulfation 171

9.3.5 Acrylates 172

9.4 Fabrication of Hyaluronic Acid Hydrogels 172

9.4.1 Hydrogel Formation by Direct Cross-Linking

Methods 173

9.4.1.1 Diepoxy cross-linking method 173

9.4.1.2 Bifunctional amine cross-linkers 173

9.4.1.3 Divinyl sulfone 176

9.4.1.4 In situ HA hydrogels 177

9.4.1.5 HA–aldehyde hydrogels 184

9.4.1.6 Azaide 187

9.5 Hyaluronic Acid-Based Hybrid Hydrogels 188

9.5.1 HA–Collagen/Oligopeptide Hydrogels 188

9.5.2 HA–Natural Polymer Hybrid Hydrogels 190

9.5.3 HA–Synthetic Polymer Hybrid Hydrogels 191


IV. ELECTROSPINNING NANOFIBER

10 Guidance of Cell Adhesion, Alignment, Infiltration,and Differentiation on Electrospun NanofibrousScaffolds 201Sang Jin Lee and James J. Yoo10.1 Introduction 202

10.2 Electrospinning Technology 203


xii Contents

10.3 Cellular Interactions with Electrospun Fibrous

Scaffolds 204

10.3.1 Cell Adhesion 205

10.3.2 Cell Alignment 206

10.3.3 Cell Infiltration 210

10.3.4 Cellular Differentiation 211

10.4 Summary 212

11 Fabrication of Tissue Engineering Scaffolds byElectrospinning Techniques 219Jiang Chang, Wenguo Cui, Yue Zhou, and Lei Chen11.1 Introduction 220

11.2 Electrospun Nanofibers 222

11.3 One-Dimensional Electrospun Fibrous Bundle 224

11.4 Two-Dimensional Electrospun Fibrous

Membranes 225

11.5 Three-Dimensional Electrospun Fibrous

Scaffolds 227


12 Biodegradable Tunable Nanofibrous Matrix forRegenerative Medicine 233Shanta Raj Bhattarai, Madhab Prasad Bajgai,and Hak Yong Kim12.1 Introduction 234

12.2 Electrospun Nanofiber Matrices 237

12.3 Electrospun Nanofiber Matrices as Tissue

Regenerative Matrices 239

12.3.1 Skin Grafts 240

12.3.2 Blood Vessel (Vascular and Cardiac) Grafts 241

12.3.3 Ligament Grafts 244

12.3.4 Nerve Grafts 245

12.3.5 Skeletal Muscle Grafts 247

12.3.6 Bone Tissue Grafts 248

12.3.7 Articular Cartilage Tissue Grafts 249

12.3.8 Drug, DNA, Protein, and Enzyme

Delivery 250

12.4 Conclusions 252


Contents xiii

13 PHBV/Proteins Composite Nanofibrous Scaffolds forTissue Engineering 257K. M. Kamruzzaman Selim, Zhi-Cai Xing, and Inn-Kyu Kang13.1 Introduction 258

13.2 Electrospinning Technique 259

13.3 Nanocomposite Preparation 260

13.3.1 Principle of the Blending Method 260

13.4 Typical PHBV/Protein Nanocomposite Preparation 261

13.4.1 Electrospun PHBV/Collagen Composite

Nanofibrous Scaffolds 261

13.4.2 Electrospun PHBV/Gelatin Composite

Nanofibrous Scaffolds 263

13.5 Interaction of As-Prepared Nanocomposites with

Cells and Results Obtained Thereby 265

13.5.1 PHBV–Col Nanocomposites 265

13.5.2 PHBV/Gelatin Nanocomposites 266


14 Nanofibrous Scaffolding for Bone Tissue Engineering 273Hae-Won Kim14.1 Introduction 273

14.2 Nanofiber Production Tools and Electrospinning 274

14.2.1 Phase Separation 275

14.2.2 Self-Assembly 275

14.2.3 Electrospinning 276

14.3 Nanofibrous Materials for Bone Tissue Engineering 277

14.3.1 Biodegradable Polymers 277

14.3.2 Bioactive Inorganics 279

14.3.3 Composite Nanofibers 282

14.4 Functionalization of Nanofibers for Bone Tissue

Engineering 284

14.4.1 Surface Modifications 284

14.4.2 Incorporation of Biomolecules 286

14.5 Concluding Remarks 287

15 Strategies to Engineer Electrospun ScaffoldArchitecture and Function 291Aaron S. Goldstein, Christopher A. Bashur, and Joel Berry15.1 Overview 291


xiv Contents

15.2 Design of Fiber Topography to Affect Cell Function 292

15.2.1 Effect of Electrospun Fiber Alignment on

Cell Morphology 293

15.2.2 Effect of Fiber Diameter on Cell

Morphology 294

15.2.3 Effect of Fiber Roughness 295

15.3 Creation of Larger Pores to Facilitate Cell Entry

into Scaffolds 296

15.3.1 Co-Electrospinning of a Sacrificial Polymer 296

15.3.2 Incorporation of Extruded Fibers 296

15.3.3 Incorporation of Porogens into Electospun

Meshes 298

15.3.4 Chemotaxis and other Considerations

Regarding Cell Infiltration into Electrospun

Meshes 298

15.3.5 Electrospraying or Electrospinning of Cells 299

15.4 Creation of Three-Dimensional Architectures for

Tissue-Specific Applications 299

15.4.1 Processing Techniques for Tube- and

Cord-Shaped Structures 300

15.4.2 Variations on the Tube Structure for Blood

Vessel and Annulus Fibrosis Applications 300

15.5 Composite Scaffolds and the Spatial

Heterogeneity 301

15.6 Mechanics of Scaffold Deformation and Failure

Under Strain 302


V. NOVEL BIOMATERIALS FOR SCAFFOLD

16 Synthetic/Natural Hybrid Scaffold for TissueRegeneration 311Gilson Khang, Soon Hee Kim, Su Hyun Jung,and Yun Sun Yang16.1 Introduction 312

16.2 Fibrin/PLGA Hybrid Scaffolds 313

16.2.1 Fibrin/PLGA Hybrid Scaffolds for Cartilage

Regeneration in vivo and in vitro 313


Contents xv

16.2.2 Fibrin/PLGA Hybrid Scaffolds for IVD

in vitro 318

16.3 The Effect of DBPs on the Reduction of

Inflammatory Reaction of the PLGA/DBP Hybrid

Scaffold 323

16.3.1 Cell Viability 324

16.3.2 Inflammatory Cytokine Expression 325

16.3.3 In vivo Tissue Response 326

16.4 The Effect of SIS on the Host Tissue Response to

PLGA/SIS Hybrid Scaffolds 330


17 Artificial Binding Growth Factors 337Takashi Kitajima and Yoshihiro Ito17.1 Introduction 337

17.2 Diffusible and Nondiffusible Actions of Growth

Factors 338

17.3 Gene-Engineered Binding Growth Factors 340

17.3.1 Collagen-Binding Growth Factors 341

17.3.2 Fibrin-Binding Growth Factors 344

17.3.3 Cell-Binding Fusion Proteins 345

17.3.4 Other Binding Growth Factors 346

17.4 Application of Engineered Binding Growth Factors 346

17.4.1 Skin Wound Repair 347

17.4.2 Repair of Cardiovascular Tissues 347

17.4.2.1 Materials that induce

angiogenesis 348

17.4.2.2 Artificial blood vessel and heart

valve 348

17.4.3 Nerve Regeneration 349

17.4.4 Bone Regeneration 349


18 Porous Poly(Lactic-Co-Glycolic Acid) Microsphere asCell Culture Substrate and Cell TransplantationVehicle 355Byung-Soo Kim18.1 Introduction 356


xvi Contents

18.2 Fabrication of Macroporous PLGA Microspheres 357

18.3 Macroporous PLGA Microsphere as an ASC Culture

Substrate 358

18.4 Macroporous PLGA Microsphere as an ASC

Transplantation Vehicle 361


19 Suppression of Inflammatory Reactions on MPCPolymer Surfaces 365Yasuhiko Iwasaki and Kazuhiko Ishihara19.1 Introduction 365

19.2 Molecular Design and Fundamental Property of

MPC Polymers 367

19.3 Secretion of HSP mRNA from Adherent Cells on

MPC Copolymers 369

19.4 Reduction of in vivo Host Responses to MPC

Polymer Hydrogels 373

19.5 Newly Extracellular Matrices Generated from

MPC Polymers 376

19.6 Conclusion 380

20 Extracellular Matrix–Based Scaffolds from Scratch 385Willeke F. Daamen, Kaeuis A. Faraj, Martin J. W. Koens,Gerwen Lammers, Katrien M. Brouwer, Peter J. E.Uijtdewilligen, Suzan T. M. Nillesen, Luc A. Roelofs,Jody E. Nuininga, Paul J. Geutjes, Wouter F. J. Feitz,and Toin H. van Kuppevelt20.1 Introduction 386

20.2 Scaffolds with a Specific Three-Dimensional

Structure 387

20.2.1 Flat Films 387

20.2.2 Porous Scaffolds 388

20.2.2.1 Unidirectional scaffolds 389

20.2.3 Tubular Porous Scaffolds 389

20.3 Scaffolds with Defined Molecular Composition 391

20.3.1 Purification of Scaffold Components 391

20.3.2 Cross-Linking and Covalent Attachment of

Glycosaminoglycans 392

20.3.3 Binding of Growth Factors 392


Contents xvii

20.4 Acellular Scaffolds for in vivo Tissue Regeneration 393

20.5 Future Outlook 395

21 Design of Biomimetic Scaffolds for Liver TissueEngineering 399Chong-Su Cho, Hu-Lin Jiang, Takashi Hoshiba,and Toshihiro Akaike21.1 Introduction 400

21.2 Specific Interaction between Galactose Residue

and ASGPR 401

21.3 Bulk Modification of Biomaterials 401

21.4 Surface Modification of Biomaterials 404

21.5 Criteria to Design Biomimetic Scaffolds for Liver

Tissue Engineering 404

21.5.1 Topology of the ECM 404

21.5.2 Coculture 407

21.5.3 Cell Sources 409

21.6 Summary 411

22 Hybrid Porous Scaffolds of Biodegradable SyntheticPolymers and Collagen for Tissue Engineering 417Guoping Chen, Naoki Kawazoe, and Tetsuya Tateishi22.1 Introduction 418

22.2 PLGA-Collagen Hybrid 419

22.3 PLGA-Collagen Hybrid Mesh 422

22.4 PLLA-Collagen Hybrid Braid 427

22.5 Biphasic Hybrid Porous Scaffold 428

22.6 Leak-Proof Hybrid Scaffolds 430


23 Chitin and Chitosan for Tissue EngineeringApplication 435Sang Jun Park and Chun-Ho Kim23.1 Introduction 435

23.2 Chitin and Chitosan 436

23.3 Chitosan Sponge Scaffolds 439

23.3.1 Preparation of Chitosan Sponge Scaffolds 439

23.3.2 Cell Culture on Chitosan Sponge Scaffolds 441

23.4 Chitosan Bead Scaffolds 442


xviii Contents

23.4.1 Preparation of Chitosan Bead Scaffolds 443

23.4.2 Cell Culture on Chitosan Bead Scaffolds 446

23.5 Chitosan Hydrogels 447

23.5.1 Preparation of Chitosan Hydrogels 448

23.5.2 Cell Culture on Chitosan Hydrogels 449


VI. NOVEL FABRICATION METHODS FOR SCAFFOLD

24 Controlling a Cellular Niche in Scaffold Designs forEpithelial Tissue Engineering 455Zhilian Yue, Yan-Ru Lou, Nur Aida Abdul Rahim,and Hanry Yu24.1 Introduction 456

24.2 Native Extracellular Microenvironment for

Epithelial Cells 458

24.2.1 Extracellular Matrix and Growth Factors 459

24.2.2 Epithelial Polarity, Differentiation,

and Function 461

24.2.3 Other Physical Factors: Biomechanics and

Microfluidics 462

24.3 Engineering an Extracellular Microenvironment

for Epithelial Cells 464

24.3.1 The State of Art 464

24.3.1.1 2D plastic substrata 464

24.3.1.2 3D polymeric scaffolds 466

24.3.2 Spatial and Temporal Presentation of

Extracellular Cues in Scaffolds for Liver


24.3.3 Biomechanical Issues 470

24.3.4 Fluid Dynamics and Mass Transfer 472

24.4 Applications and Outlook 473

25 Biological Implications of Polymeric Scaffolds forBone Tissue Engineering Developed via SolidFreeform Fabrication 483Andrew B. Yeatts and John P. Fisher25.1 Introduction 484

25.1.1 The Need for Bone Tissue Engineering 484


Contents xix

25.1.2 Benefits of Scaffolds Developed via Solid

Freeform Fabrication 485

25.2 Stereolithography 486

25.2.1 Scaffold-Manufacturing Process 486

25.2.2 Biological Implications 488

25.3 Three-Dimensional Printing 490



25.4 Selective Laser Sintering 494



25.5 Fused Deposition Modeling 498




26 Considerations on the Structure of Biomaterials forSoft- and Hard-Tissue Engineering 509Hideaki Kagami, Hideki Agata, Makoto Satake,and Yuji Narita26.1 Introduction 510

26.2 Material Design for Soft-Tissue Engineering:

Small-Caliber Vascular Grafts 512

26.2.1 Tissue Engineering for Cardiovascular

Surgery 512

26.2.2 Decellularized Tissue Scaffolds for

Tissue-Engineered Small-Caliber Vascular

Grafts: Methodology, Biocompatibility, and

Mechanical Properties 513

26.2.3 Biodegradable Synthetic Polymer Scaffolds

for Tissue-Engineered Small-Caliber

Vascular Grafts 515

26.2.4 How to Create Scaffolds for

Tissue-Engineered Small-Diameter

Vascular Grafts Using Electrospun

Nanofibers 516

26.2.5 Biocompatibility and Mechanical

Properties of Electrospun Synthetic

Scaffolds 518


xx Contents

26.2.6 Prospects of Designing a Scaffold for

Cardiovascular Tissue Engineering 521

26.3 Scaffold Design for Hard-Tissue Engineering:

Alveolar Bone 521

26.3.1 Bone Reconstruction/Regeneration in

Orthopedic and Oral Applications 521

26.3.2 Ideal Ceramic Scaffolds for Bone Tissue

Engineering from a Clinical Point of View 523

26.3.3 Fate of Transplanted Scaffolds in the

Human Body: A Clinical Study of Alveolar

Bone Tissue Engineering Using Bone

Marrow Stromal Cells and β-TCP 525

26.3.4 Considerations for Designing Scaffolds for

Clinical Bone Tissue Engineering 527

26.3.5 Prospective Novel Biomaterials for

Hard-Tissue Engineering 530

26.3.5.1 Composite and combined

materials 530

26.3.5.2 Growth factor incorporation into

scaffolds 531


27 Mechano-Active Scaffolds 537Sang-Heon Kim, Youngmee Jung, Young Ha Kim,and Soo Hyun Kim27.1 Introduction 538

27.2 Mechano-Active Scaffolds 539

27.2.1 Elastic Biodegradable PLCL Copolymer 539

27.2.2 Tubular PLCL Scaffold 541

27.2.3 Seamless Double-Layered Scaffold 542

27.2.4 Sheet-Form PLCL Scaffold 545

27.3 Mechano-Active Tissue Engineering 545

27.3.1 Vascular Tissue Engineering 545

27.3.2 Cartilage Tissue Engineering 550


28 Reinforced Scaffold for Tissue Engineering 561Young-Kwon Seo and Jung-Keug Park28.1 Introduction 562


Contents xxi

28.2 Biocompatibility of Reinforced Composite

Scaffolds 564

28.3 Reinforced Composite Scaffold for Bioartificial

Tissue 565

28.3.1 Bioartificial Ligament and Tendon 566

28.3.2 Bioartificial Bone 568

28.3.3 Bioartificial Vessel 569

28.3.4 Bioartificial Tracheae 570

28.3.5 Bioartificial Skin 571

28.4 Conclusion and Outlook 574

29 Three-Dimensional Shape Control of Implant Devices 579Ung-il Chung, Hideto Saijoh, Kazuyo Igawa, Yuki Kanno,Yoshiyuki Mori, and Tsuyoshi Takato29.1 Introduction 580

29.2 Current Status of Artificial Bones 581

29.3 3D Fabrication Technologies and Their

Comparison 583

29.4 Inkjet Printing Technology 584


30 Novel Fabrication and Characterization of Pore-Size-Gradient Scaffolds by a Centrifugation Technique 589Se Heang Oh and Jin Ho Lee30.1 Introduction 590

30.2 Fabrication of Pore-Size-Gradient Scaffolds 592

30.2.1 Materials 592

30.2.2 Pore-Size-Gradient Alginate Scaffolds 593

30.2.3 Pore-Size-Gradient Chitosan Scaffolds 594

30.2.4 Pore-Size-Gradient PCL Scaffolds 594

30.3 Characterization of Pore-Size-Gradient

Scaffolds 595

30.3.1 Measurements of Pore Sizes and Porosity 595

30.3.2 Measurements of Mechanical Properties 597

30.3.3 Evaluation of Wettability 600

30.3.4 In vitro Cell Interactions 600

30.3.5 In vivo Tissue Interactions 603



xxii Contents

31 Solid Freeform Fabrication Method Applied toTissue Scaffolds 609Dong-Woo Cho, Jin Woo Lee, Jong Young Kim,and Tae-Yun Kang31.1 Introduction 610

31.2 SFF Methods Applied to Scaffolds 610

31.2.1 Stereolithography 610

31.2.1.1 Photopolymer scaffold 611

31.2.1.2 Biopolymer scaffold 615

31.2.2 Fused Deposition Modeling 616

31.2.3 3D Printing 621

31.2.4 Selective Laser Sintering 623

31.3 Summary 628

32 Novel Microspheres for Prolonged Cell Survival 633Sing Muk Ng, Jeung Soo Huh, Syed Izhar Haider Abdi,and Jeong Ok Lim32.1 Introduction 634

32.2 Current Status and Development in Supplying

Oxygen for Tissue Engineering 636

32.2.1 The Use of Artificial Oxygen Carriers 637

32.2.2 Induction and Enhancement of

Vascularization 638

32.2.3 The Utilization of Oxygen-Generating

Biomaterials 639

32.3 Oxygen-Releasing Microspheres (ORMs) 640

32.3.1 The State of the Art 640

32.3.2 Materials as Building Blocks of

Microspheres 643

32.3.3 Techniques for Producing ORMs 647

32.3.3.1 Double-emulsion and solvent

evaporation technique 647

32.3.3.2 Functionalization of matrices

selected as building blocks 650

32.3.3.3 Instrumentations for the

preparation of microspheres 652

32.3.4 Evaluation of the Oxygen-Releasing Profile 653

32.3.4.1 Direct observation 653


Contents xxiii

32.3.4.2 Quantitative analytical approach 654

32.3.4.3 Biologically related study 656

32.4 ORMs in Applications for Efficient Cell Survival 657

32.4.1 Direct Integration in Scaffolds 657

32.4.2 Oxygen-Generating Reservoir 659

32.4.3 Medicine to Oxygenate Tissues 659

32.5 Conclusion 660

33 Emulsion Templating 665Elizabeth Cosgriff-Hernandez33.1 Introduction 665

33.2 Bone Tissue Engineering 667

33.3 Scaffold Fabrication 668

33.4 PolyHIPEs as Bone Scaffolds 671

33.4.1 Nondegradable: Styrene-Based PolyHIPEs 671

33.4.2 Semidegradable: Polyester-Based

PolyHIPEs 672

33.4.3 Fully Degradable: Fumarate-Based

PolyHIPEs 674

33.5 New Synthesis Routes 675


VII. SCAFFOLD FOR TARGET ORGAN

34 PGA Fiber for Soft Tissue Engineering 681Wei Liu and Yilin Cao34.1 Introduction 681

34.2 PGA Fibers for Tendon Engineering 683

34.3 PGA Scaffold for Cartilage Engineering 689

34.4 PGA Fibers for Skin Engineering 694

34.5 PGA Fibers for Corneal Stroma Engineering 698

34.6 PGA Fibers for Blood Vessel Engineering 701

34.7 PGA Fibers for Engineering Peripheral Nerve

Tissue 705

34.8 Conclusion 707

35 Tissue Engineering and Anti-Aging 711Minoru Ueda35.1 Introduction 711


xxiv Contents

35.2 Materials and Method 713

35.2.1 Tissue Preparation 714

35.2.2 Cell Culture 714

35.2.3 Medium and Autologous Serum

Preparation 716

35.2.4 Preparation of Cell Suspension and HA

Admixture 716

35.2.5 Safety Tests 717

35.2.6 Clinical Assessment of Aesthetic

Improvement 718

35.2.7 Skin Replica and Analysis 718

35.3 Result 719

35.3.1 Single-Case Report 724

35.4 Discussion 726

35.4.1 Fibroblast and HA for Skin Rejuvenation 726

35.4.2 Wrinkle Treatment in Dentistry 728

35.5 Conclusion 730

36 Matrices for Zonal Cartilage Tissue Engineering 733Daisy Irawan, Dietmar Hutmacher, and Travis Klein36.1 Introduction 734

36.2 Carbohydrate-Based Matrices 736

36.2.1 Alginate 737

36.2.2 Agarose 739

36.3 Protein-Based Matrices 741

36.3.1 Collagen or Gelatin 741

36.3.2 Fibrin 742

36.4 Synthetic and Semisynthetic Matrices 744

36.4.1 Poly(Ethylene Glycol) 744

36.4.2 Extracel 747


37 Collagen-Based Scaffold for Bone TissueRegeneration 757Fu-Zhai Cui, Zong-Gang Chen, and Xue Xia37.1 Introduction 758

37.2 Compositional and Structural Characteristics of

Natural Bone 759


Contents xxv

37.2.1 Composition of the Natural Bone Matrix 759

37.2.2 Hierarchical Structure of the Natural Bone

Matrix by Self-Assembly 759

37.3 Biomimetic Fabrication with Self-Assembled

Collagen Mineralization 760

37.3.1 Mineralization Mechanism of

Hydroxyapatite Crystals on Collagen Fibers 761

37.3.2 Assembly of the Nano-Fibril of Mineralized

Collagen 763

37.4 Synthesis and Application of Collagen-Based

Scaffolds in Bone Tissue Regeneration 767

37.4.1 Synthesis of Nano-HA/Collagen-Based

Scaffolds 767

37.4.2 Applications and Development of

Nano-HA/Collagen Scaffolds for Bone



38 Scaffold Considerations for Osteochondral TissueEngineering 779Eric Farrell, Fergal J. O’Brien, and Gerjo J. V. M. van Osch38.1 Introduction 780

38.1.1 Tissue Engineering of the Bone

Cartilage Interface 780

38.2 Joint Homeostasis 780

38.3 Current and Recent Approaches to the Field of

Osteochondral Tissue Engineering 782

38.4 Functional Properties of Bone and Cartilage and

the Important Differences Between Them 783

38.5 Vascularization and its Absence in Cartilage 786

38.6 Scaffold Considerations for Osteochondral


38.7 Endochondral Ossification, a More Logical

Approach for Osteochondral Tissue Engineering 793

39 Application of Scaffolds for Artificial Skin inRegenerative Medicine 803Hyun Ju Lim and Ho Yun Chung39.1 Introduction 804


xxvi Contents

39.2 Scaffolds: Biomaterials as One of Important Factors

for Regeneration of Skin Tissue 806

39.3 Clinical Applications of Tissue-Engineered Skin

Products 808


40 Biodegradable Scaffolds for Bone Regeneration 817Yoichi Yamada40.1 Introduction 818

40.2 Biodegradable Scaffolds 819

40.2.1 Biodegradable Ceramic Composite (β-TCP) 819

40.2.2 Nanofibers Hydrogel Peptide 821

40.2.3 Injectable Tissue-Engineered Bone 824

40.3 Clinical Application 827

40.3.1 Preparation and Clinical Application of

MSCs, PRP, and Injectable TEB 827


41 An Efficient ex vivo Expansion of Adult MesenchymalStem Cells in Scaffolds 833Eui Kyun Park, Hong-In Shin, and Shin-Yoon Kim41.1 Introduction 834

41.2 The Use of Growth Factors and Glucocorticoids for

the Propagation of Adult MSCs 836

41.2.1 Growth Factors 836

41.2.1.1 Fibroblast growth factors 837

41.2.1.2 Epidermal growth factor 837

41.2.1.3 Platelet-derived growth factor 839

41.2.1.4 Other growth factors 840

41.2.2 Glucocorticoids 840

41.2.3 Combination of Growth Factors and

Steroids 841

41.3 Growth of MSCs in Scaffolds 843


42 Nanoparticles for Bioimaging in RegenerativeMedicine 855Dongwon Lee, John M. Rhee, and Gilson Khang42.1 Introduction 856


Contents xxvii

42.2 Fluorescent Probes for Imaging of Hydrogen

Peroxide 858

42.3 Luminescent Probes for Imaging of Hydrogen

Peroxide 861

42.4 Fluorescent Probes for Other ROS 866


43 Effect of Scaffolds with Bone Growth Factors onNew Bone Formation 871Hae-Ryong Song, Swee-Hin Teoh, Jun-Ho Wang,Hak-Jun Kim, Ji-Hoon Bae, Sung Eun Kim, Jerry Chan,Zhi-Yong Zhang, and Chang-Wug Oh43.1 Introduction 872

43.2 Bone Lengthening in Preclinical Animal Studies 873

43.2.1 Calcium Sulfate in Tibial Lengthening 873

43.2.2 Cord Blood Stem Cells and rhBMP-2 in

Tibial Lengthening 877

43.3 Growth Factor–/Stem Cells–Mediated Scaffolds for

Bone Tissue Engineering 881

43.3.1 Use of Fibrin and Stem Cells for Bone

Defect Healing in Rabbits 881

43.3.2 Use of Bioreactors, Human Fetal Stem

Cells, and 3D Scaffolds for Bone Tissue

Engineering 888

43.3.3 The Use of Scaffolds with or without

Growth Factors and Cells for Clinical Trials 896

44 Temperature-Responsive Culture Surfaces forRegenerative Medicine 903Yoshikazu Kumashiro, Yoshikatsu Akiyama,Masayuki Yamato, and Teruo Okano44.1 Introduction 903

44.2 The Basic Mechanism of Cell Attachment to

and Detachment 905

44.3 Temperature-Responsive Cell Culture Surfaces

that Enable Affinity Control 908

44.4 Applications of Cell Sheet Technology 912

44.5 Corneal Surface Reconstruction 913


xxviii Contents

44.6 Periodontal Ligament Cell Sheets 916

44.7 Endoscopic Esophageal Epithelial Transplantation 918

44.8 Sealing of Lung air Leaks 920

45 Customized Nanocomposite Scaffolds Fabricated viaSelective Laser Sintering for Bone Tissue Engineering 925Bin Duan and Min Wang45.1 Introduction 926

45.2 Application of Rapid Prototyping Technologies to

Scaffold Fabrication 930

45.3 Design of Scaffolds and the Nanocomposite

Strategy 934

45.4 Fabrication of Nanocomposite Scaffolds via SLS

and Characteristics of the Scaffolds 938

45.5 Nanocomposite Scaffolds as Delivery Vehicles

for Biomolecules 941


Index 955


Preface

It has been recognized that regenerative medicine and tissue

engineering offer an alternative technique to whole-organ and

tissue transplantation for diseased, failed, or malfunctioning organs.

Millions of patients suffer from end-stage organ failure or tissue

loss annually. The only way to solve this problem might be organ

transplantation and biomaterials transplantation. However, in order

to avoid the shortage of donor organs and other problems caused

by poor biocompatibility of biomaterials, a new hybridized method

combined with cells and biomaterials had been introduced as

regenerative medicine and tissue engineering around 20 years ago.

The specialty of regenerative medicine and tissue engineering

continues to grow and change rapidly. This area saw major advances

in the past few years. This field for academic research and com-

mercialization is needed in multidisciplinary areas such as adult,

embryoinic, and induced pluripotent cells, genetic programming,

nuclear transfer, cloning, genomics, proteomics, nanotechnology,

biomaterials, etc. Thanks to the latest 20 years’ endeavor, several

tissue-engineered products (TEMPS) and regenerative medicinal

products (RMP) are on the boundary of the translation of benchside

discoveries to clinical therapies. For the reconstruction of a neo-

tissue by regenerative medicine and tissue engineering, triad

components such as (i) cells that are harvested and dissociated from

the donor tissue, including nerve, liver, pancreas, cartilage, and bone,

as well as embryonic stem cells, adult stem cells, induced pluripotent

cells (iPS), or precursor cells; (ii) biomaterials as scaffold substrateswhose cells are attached and cultured, resulting in the implantation

at the desired site of the functioning tissue; and (iii) growth factorsthat are promoting and/or preventing cell adhesion, proliferation,

migration, and differentiation by up-regulating or down-regulating


xxx Preface

the synthesis of protein; growth factors; and receptors must be

needed.

This handbook has concentrated on all the things for scaffolds

among triad components, especially “intelligent” scaffolds from

basic science, industries, to clinical applications. This textbook is

organized into seven major areas. Part I, Introduction, reveals some

of fundamentals of the biomaterials, scaffolds, and manufacturing

methods. Part II covers ceramic and metal scaffolds. Part III,

Intelligent Hydrogel, deals with various types of hydrogels for tissue

regenerations. In Part IV, topics of scaffolds from electrospinning

nanofibers have been covered. In Part V, novel biomaterials for

scaffolds have been introduced, especially to mimic Mother Nature.

The sixth part covers the recent novel fabrication methods for

smart scaffolds. The last part, Part VII, of this handbook deals with

the recent clinical trial of specific target organs using intelligent

scaffolds. The authors have tried to dedicate the 45 chapters to the

whole area of the recent topic of smart scaffolds for regenerative

medicine and tissue engineering. I am indebted to the authors for

their willing acceptance, devotion, and contribution to each recent

topic.

I express my thanks to my students Mrs. Yong Ki Kim, Jung Bo

Shim, and Young Un Kim for editing all manuscripts. Finally, I really

appreciate our publisher, Mr. Stanford Chong. Without his trust and

guidance, this huge work could not have been accomplished. Also, I

would like to give special appreciation to Mr. Sarabjeet Garcha and

Ms. Archana Ziradkar for the hard work.

Gilson Khang, PhD

IntelligentScaffolds.indd 1 1/4/12 12:42:39 AM

Handbook of Intelligent Scaffolds for Tissue Engineering and Regenerative Medicine

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