Table Of ContentAdvances in Regenerative Medicine: Role of
Nanotechnology, and Engineering Principles
NATO Science for Peace and Security Series
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Series A: Chemistry and Biology
Advances in Regenerative
Medicine: Role of
Nanotechnology,
and Engineering Principles
Edited by
Venkatram Prasad Shastri
Universität Freiburg
Institute of Makromolekulare Chemie
Germany
George Altankov
ICREA & Institute of Bioengineering Catalunya
Barcelona
Spain
Andreas Lendlein
Institute of Polymer Research and Berlin-Brandenburg Center for
Regenerative Therapies
GKSS Research Center Geesthacht GmbH
Teltow
Germany
Published in Cooperation with NATO Public Diplomacy Division
Proceedings of the NATO Advanced Research Workshop on Nanoengineered
Systems for Regenerative Medicine Varna, Bulgaria
21–24 September 2007
Library of Congress Control Number: 2010929479
ISBN 978-90-481-8789-8 (PB)
ISBN 978-90-481-8788-1 (HB)
ISBN 978-90-481-8790-4 (e-book)
Published by Springer,
P.O. Box 17, 3300 AA Dordrecht, The Netherlands.
www.springer.com
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© 2010 Springer Science + Business Media B.V.
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Preface
This book summarizes the NATO Advanced Research Workshop (ARW) on
“Nanoengineered Systems for Regenerative Medicine” that was organized under
the auspices of the NATO Security through Science Program. I would like to thank
NATO for supporting this workshop via a grant to the co-directors.
The objective of ARW was to explore the various facets of regenerative medi-
cine and to highlight role of the “the nano-length scale” and “nano-scale systems”
in defining and controlling cell and tissue environments. The development of novel
tissue regenerative strategies require the integration of new insights emerging from
studies of cell-matrix interactions, cellular signalling processes, developmental and
systems biology, into biomaterials design, via a systems approach. The chapters in
the book, written by the leading experts in their respective disciplines, cover a wide
spectrum of topics ranging from stem cell biology, developmental biology, cell-
matrix interactions, and matrix biology to surface science, materials processing and
drug delivery. We hope the contents of the book will provoke the readership into
developing regenerative medicine paradigms that combine these facets into clini-
cally translatable solutions.
This NATO meeting would not have been successful without the timely help of
Dr. Ulrike Shastri, Sanjeet Rangarajan and Ms. Sabine Benner, who assisted in the
organization and implementation of various elements of this meeting. Thanks are
also due Dr. Fausto Pedrazzini and Ms. Alison Trapp at NATO HQ (Brussels,
Belgium). The commitment and persistence of Ms. Wil Bruins at Springer in get-
ting this book published is very much appreciated. Last but not least, I would like
to thank Prof. George Altankov my co-Director and Prof. Andreas Lendlein for
their supportive role.
Co-Director, Freiburg V. Prasad Shastri
v
Contents
1 Cell Adhesions and Signaling: A Tool for Biocompatibility
Assessment ................................................................................................. 1
1.1 Introduction ........................................................................................ 2
1.2 Cell-Matrix Adhesions ....................................................................... 3
1.2.1 Integrin Receptors and Integrin
Cytoplasmic Complexes ........................................................ 3
1.2.2 Focal Adhesions ..................................................................... 5
1.2.3 Fibrillar Adhesions ................................................................. 9
1.2.4 Three-Dimensional Matrix Adhesions ................................... 10
1.2.5 Dynamics of Cell Adhesions ................................................. 12
1.3 Concluding Remarks .......................................................................... 14
References ................................................................................................... 15
2 Development of Provisional Extracellular Matrix
on Biomaterials Interface: Lessons from In Vitro
Cell Culture ............................................................................................... 19
2.1 Introduction ........................................................................................ 20
2.2 Initial Cell–Materials Interaction ....................................................... 20
2.2.1 Development of Focal Adhesion Complex ............................ 21
2.2.2 Substratum Properties Affect Focal
Adhesions Formation ............................................................. 22
2.3 Development of Provisional Extracellular Matrix
at the Biomaterials Interface .............................................................. 23
2.3.1 Development of Early Matrix ................................................ 23
2.3.2 Development of Late Fibronectin Matrix .............................. 25
2.4 Integrin Receptors Dynamics and Provisional
Extracellular Matrix Formation ......................................................... 26
2.4.1 Studies on Integrin Dynamics ................................................ 26
2.4.2 Integrin Dynamics Depend on Substratum Properties ........... 29
2.5 Development of Extracellular Matrix at “Real”
Biomaterials Interface ........................................................................ 30
2.5.1 Effects of Substratum Chemistry on Matrix Formation:
A View of Biosensors Application ......................................... 30
vii
viii Contents
2.5.2 Development of Extracellular Matrix on Biomimetic
Hydroxyaptite Cements Surfaces ........................................... 33
2.5.3 Development of Extracellular Matrix
on Different Rough Titanium Surfaces .................................. 36
References ................................................................................................... 41
3 Endothelial Progenitor Cells for Tissue Engineering
and Tissue Regeneration .......................................................................... 45
3.1 Introduction ........................................................................................ 46
3.2 Vascular Networks for TE Constructs ................................................ 46
3.3 Sources of Human Endothelial Cells for TE
Vascular Networks ............................................................................. 47
3.3.1 Mature Vessel-Derived Endothelial Cells .............................. 47
3.3.2 Human Embryonic Stem Cells ............................................... 47
3.3.3 Blood-Derived EPCs .............................................................. 48
3.3.4 Bone Marrow-Derived Cells
for Tissue Vascularization ...................................................... 49
3.4 Blood-Derived EPCs for Creating Vascular Networks ...................... 50
3.4.1 Isolation and Culture of Human EPCs
and HSVSMCs ....................................................................... 50
3.4.2 In Vivo Assay for Vascularization .......................................... 51
3.4.3 Summary: Vasculogenic Potential of Human EPCs .............. 51
References ................................................................................................... 52
4 Dermal Precursors and the Origins of the Wound Fibroblast .............. 55
4.1 Introduction ........................................................................................ 55
4.2 Mesenchymal Stem Cells ................................................................... 56
4.2.1 Marrow Derived Fibroblast Populations ................................ 57
4.2.2 Fibrocytes ............................................................................... 58
4.2.3 Factors for Mobilization and Recruitment
of Marrow Populations ........................................................... 61
4.3 Defining the Dermal Fibroblast ......................................................... 62
4.3.1 Dermal Progenitors ................................................................ 62
4.3.2 Evidence for Progenitor Populations ..................................... 63
4.4 Clinical Applications ......................................................................... 65
4.5 Novel Healing Properties of MSC ..................................................... 66
4.6 Summary ............................................................................................ 66
References ................................................................................................... 67
5 Cell Based Therapies: What Do We Learn
from Periosteal Osteochondrogenesis? ................................................... 71
5.1 General Introduction .......................................................................... 72
5.1.1 Cell Based Therapies ............................................................. 72
5.2 Periosteum .......................................................................................... 73
Contents ix
5.2.1 Embryological Development of Bone, Cartilage,
and Periosteum ....................................................................... 74
5.2.2 Periosteum in Bone and Cartilage Repair .............................. 75
5.3 Differential Survival of Periosteal Progenitor Cells
Versus Chondrocytes .......................................................................... 77
5.3.1 Introduction ............................................................................ 77
5.3.2 Cell Labeling and Scaffolds ................................................... 77
5.3.3 Chondrocytes Survive Transplantation
Better than Periosteal Progenitor ........................................... 78
5.3.4 Hyaluronan Increases the Number of Viable
Periosteal Progenitors ............................................................ 78
5.3.5 Discussion .............................................................................. 79
5.4 A Model to Study Periosteal Osteochondrogenesis In Vivo .............. 79
5.4.1 Introduction ............................................................................ 79
5.4.2 Periosteal Callus Recapitulates the Sequential
Steps of Endochondral Bone Formation ................................ 80
5.4.3 HIF-1a Activation and BMP Expression ............................... 81
5.4.4 Periostin Activation During Periosteal
Callus Formation .................................................................... 83
5.5 Repair of Osteochondral Defects with Cartilage
from Periosteum ................................................................................. 84
5.5.1 Introduction ............................................................................ 84
5.5.2 Improved Repair of Osteochondral Defects ........................... 84
5.5.3 Discussion .............................................................................. 86
5.6 Evidence That Endochondral Bone Formation
Can Be Manipulated in the IVB ......................................................... 87
5.6.1 Introduction ............................................................................ 87
5.6.2 Growth Factors ....................................................................... 87
5.6.3 Hypoxia .................................................................................. 87
5.6.4 Calcium .................................................................................. 88
5.7 Discussion .......................................................................................... 88
References ................................................................................................... 89
6 Bioreactor Systems in Regenerative Medicine ....................................... 95
6.1 Introduction ........................................................................................ 95
6.2 Bioreactors in Regenerative Medicine: Key Features ........................ 96
6.2.1 Cell Seeding on Three-Dimensional Matrices ....................... 96
6.2.2 Maintenance of a Controlled Culture Environment ............... 97
6.2.3 Physical Conditioning of Developing Tissues ....................... 99
6.2.4 Predicting Mechanical Functionality
of Engineered Tissues ............................................................ 100
6.3 Bioreactor-Based Manufacturing of Tissue
Engineering Products ......................................................................... 101
6.3.1 Automating Conventional Cell Culture Techniques .............. 103
x Contents
6.3.2 Automating Tissue Culture Processes .................................... 104
6.3.3 Streamlining Tissue Engineering Processes........................... 106
6.3.4 Different ‘Manufacturing’ Concepts ...................................... 109
6.4 Conclusions and Future Perspectives ................................................. 110
References ................................................................................................... 111
7 Biomimetic Approaches to Design of Tissue
Engineering Bioreactors ........................................................................... 115
7.1 Introduction ........................................................................................ 116
7.2 Cardiac Tissue Engineering ............................................................... 116
7.2.1 Myocardium (Cardiac Muscle) .............................................. 116
7.2.2 Tissue Engineering ................................................................. 117
7.3 Cartilage Tissue Engineering ............................................................. 120
7.3.1 Articular Cartilage ................................................................. 120
7.3.2 Tissue Engineering ................................................................. 121
7.4 Conclusions ........................................................................................ 125
References ................................................................................................... 126
8 The Nature of the Thermal Transition Influences
the Shape-Memory Behavior of Polymer Networks .............................. 131
8.1 Introduction ........................................................................................ 131
8.2 Architectures of Different Polymer Networks ................................... 136
8.2.1 Synthesis ................................................................................ 136
8.2.2 Thermomechanical Properties ............................................... 139
8.3 Shape-Memory Capability ................................................................. 143
8.4 Degradation ........................................................................................ 151
8.5 Summary ............................................................................................ 153
8.6 Outlook .............................................................................................. 154
References ................................................................................................... 155
9 Nanoengineered Systems for Regenerative Medicine
Surface Engineered Polymeric Biomaterials
with Improved Bio-Contact Properties ................................................... 157
9.1 Introduction ........................................................................................ 157
9.2 Polymeric Materials with Improved
Bio-Contact Properties ....................................................................... 158
9.2.1 Strong Hydrophilic “Water-Like” Protein
Repellent Surfaces.................................................................. 158
9.2.2 Protein Repelent Plasma Films .............................................. 164
9.2.3 Polydimetylesiloxane (PDMS) with Improved
Interactions with Living Cells ................................................ 164
9.3 Conclusions ........................................................................................ 172
References ................................................................................................... 172
