Cartilage Tissue Engineering: Methods and Protocols

303 Pages·2015·7.11 MB·English

by  Pauline M. Doran

Methods in
Molecular Biology 1340
Pauline M. Doran Editor
Cartilage
Tissue
Engineering
Methods and Protocols
M M B
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life and Medical Sciences
University of Hertfordshire
Hatfield, Hertfordshire , AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651

Cartilage Tissue Engineering
Methods and Protocols
Edited by
Pauline M. Doran
Faculty of Science, Engineering and Technology, Swinburne University
of Technology, Hawthorn, Melbourne, VIC, Australia
Editor
Pauline M . D oran
Faculty of Science, Engineering and Technology
Swinburne University of Technology
Hawthorn, Melbourne, V IC , A ustralia
ISSN 1064-3745 ISSN 1940-6029 (electronic)
Methods in Molecular Biology
ISBN 978-1-4939-2937-5 ISBN 978-1-4939-2938-2 (eBook)
DOI 10.1007/978-1-4939-2938-2
Library of Congress Control Number: 2015952786
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Prefa ce
Cartilage in articular joints is a relatively vulnerable tissue, being subject to common i njuries
and degenerative conditions such as arthritis. Motivated by the need to develop new treat-
ment strategies, some of the earliest attempts at tissue engineering targeted cartilage as a
feasible goal for in vitro synthesis. For more than 20 years, interdisciplinary teams of biolo-
gists, engineers, materials scientists, and clinicians have studied the culture and differentia-
tion of cartilage cells and tissues. Many cornerstone technologies that distinguish tissue
engineering from routine cell culture, such as three-dimensional culture systems and the
use of scaffolds and bioreactors, were developed, tested, and widely adopted within the
context of cartilage tissue engineering.
So far, the goal of producing laboratory-grown functional cartilage has eluded us but
remains an active ambition. Irrespective of whether chondrocytes or stem cells are used as
starting material, exerting adequate control over cellular differentiation is a major chal-
lenge. We do not yet know how to integrate engineered constructs with host cartilage
in vivo and this continues to restrict clinical translation of cartilage engineering technology.
Other important areas requiring further research include the response of chondrogenic
cells to physical and mechanical stimuli, the heterogeneity of cell populations, and the com-
plex molecular networks and regulatory cascades that direct cell lineage commitment and
tissue development.
To answer all the outstanding questions in cartilage tissue engineering, further sig-
nifi cant creative and intellectual input is required. This should come not only from
established contributors but also, perhaps more importantly, from new and/or cross-
disciplinary researchers in the area. How does a newcomer to cartilage tissue engineer-
ing become familiar with the techniques that underpin this fi eld? I hope this question
may be answered herein, as this book aims to describe clearly and in detail the key practi-
cal skills involved. Methods are outlined for isolation and expansion of chondrocytes and
stem cells, differentiation, synthesis and application of three-dimensional scaffolds,
design and operation of bioreactors, in vivo testing of engineered constructs, and molec-
ular and functional analysis of cartilage cells and tissues. Frequently used techniques are
covered, as well as more recent advances in inspirational areas such as “smart” biomaterial
development, novel bioreactor design, –omics analysis, and genetic manipulation of
matrix synthesis. The book does not attempt to be comprehensive; instead, it provides a
snapshot of selected practical technologies that are either responsible for the progress
already achieved in cartilage tissue engineering or indicative of the direction of future
related research.
The chapters have been written by 45 authors and coauthors who have personal practi-
cal experience in cartilage tissue engineering. In the interests of informing the scientifi c
community and expanding the engagement of researchers in this fi eld, the contributors
have provided careful and detailed protocols for experimental work covering a broad range
of objectives for cartilage synthesis and regeneration. I thank all of the experts who have
v
vi Preface
generously contributed their knowledge, insights, and valuable tips to prepare this volume.
We hope that readers will fi nd it a useful resource. I would also like to acknowledge the kind
guidance and encouragement of Professor John Walker, Series Editor of M ethods in
Molecular Biology , throughout the duration of this project.
Melbourne, VIC, Australia P auline M. D oran
Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i x
PART I OVERVIEW
1 Cartilage Tissue Engineering: What Have We Learned in Practice?.. . . . . . . . . 3
Pauline M. Doran
PART II CELL ISOLATION, EXPANSION, AND DIFFERENTIATION
2 Human Fetal and Adult Chondrocytes.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Kifah Shahin , N astaran M ahmoudifar , and Pauline M. Doran
3 Mesenchymal Stem Cells Derived from Human Bone Marrow . . . . . . . . . . . . 4 1
Oliver F . W. Gardner , Mauro Alini , and Martin J. Stoddart
4 M esenchymal Stem Cells Derived from Human Adipose Tissue. . . . . . . . . . . . 5 3
Nastaran M ahmoudifar and Pauline M . D oran
5 D erivation and Chondrogenic Commitment of Human Embryonic
Stem Cell-Derived Mesenchymal Progenitors . . . . . . . . . . . . . . . . . . . . . . . . . 65
Hicham Drissi , Jason D . G ibson , R osa M . Guzzo , and Ren-He Xu
6 Differentiation of Human Induced Pluripotent Stem Cells
to Chondrocytes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
R osa M . Guzzo and H icham D rissi
7 Gene Transfer and Gene Silencing in Stem Cells
to Promote Chondrogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 7
Feng Zhang and Dong-An W ang
PART III BIOMATERIALS AND SCAFFOLDS
8 Hydrogels with Tunable Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
Peggy P.Y. C han
9 D ecellularized Extracellular Matrix Scaffolds for Cartilage Regeneration . . . . . 1 33
Shraddha Thakkar , H ugo Fernandes , and L orenzo M oroni
10 U se of Interim Scaffolding and Neotissue Development to Produce
a Scaffold-Free Living Hyaline Cartilage Graft. . . . . . . . . . . . . . . . . . . . . . . . . 153
Ting T ing L au , Wenyan L eong , Yvonne Peck , K ai Su ,
and Dong-An Wang
11 Bioprinted Scaffolds for Cartilage Tissue Engineering . . . . . . . . . . . . . . . . . . . 1 61
Hyun-Wook K ang , James J. Yoo , and Anthony Atala
vii
viii Contents
12 Scaffolds for Controlled Release of Cartilage Growth Factors.. . . . . . . . . . . . . 1 71
Marie M orille , Marie-Claire V enier-Julienne ,
and Claudia N. M ontero-Menei
13 N anostructured Capsules for Cartilage Tissue Engineering . . . . . . . . . . . . . . . 1 81
Clara R . C orreia , R ui L . Reis , and João F. Mano
14 Stratified Scaffolds for Osteochondral Tissue Engineering. . . . . . . . . . . . . . . . 1 91
Patcharakamon N ooeaid , G undula S chulze-Tanzil , and Aldo R . Boccaccini
PART IV BIOREACTORS
15 Mechanobioreactors for Cartilage Tissue Engineering. . . . . . . . . . . . . . . . . . . 2 03
Joanna F. Weber , R oman P erez , and Stephen D. W aldman
16 Shear and Compression Bioreactor for Cartilage Synthesis. . . . . . . . . . . . . . . . 2 21
Kifah Shahin and Pauline M . D oran
17 Microbioreactors for Cartilage Tissue Engineering.. . . . . . . . . . . . . . . . . . . . . 2 35
Yu-Han Chang and Min-Hsien W u
PART V IN VIVO APPLICATION
18 Transplantation of Tissue-Engineered Cartilage in an Animal Model
(Xenograft and Autograft): Construct Validation. . . . . . . . . . . . . . . . . . . . . . . 247
Hitoshi N emoto , Deborah Watson , and Koichi Masuda
PART VI EVALUATION
19 Proteomic Analysis of Engineered Cartilage.. . . . . . . . . . . . . . . . . . . . . . . . . . 2 63
Xinzhu P u and J ulia T hom O xford
20 M echanical Testing of Cartilage Constructs. . . . . . . . . . . . . . . . . . . . . . . . . . . 2 79
Dinorath Olvera , A ndrew D aly , and Daniel John Kelly
Index. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 289
Contributors
MAURO ALINI • Musculoskeletal Regeneration, AO Research Institute Davos , D avos Platz,
Switzerland
ANTHONY ATALA • Wake Forest Institute for Regenerative Medicine, W ake Forest School
of Medicine , Winston-Salem, NC, USA
ALDO R. BOCCACCINI • Department of Materials Science and Engineering,
Institute of Biomaterials, U niversity of Erlangen-Nuremberg , E rlangen, G ermany
PEGGY P . Y. CHAN • Faculty of Science, Engineering and Technology, S winburne University
of Technology , M elbourne, VIC, Australia
YU-HAN CHANG • Department of Orthopaedic Surgery , Chang Gung Memorial Hospital ,
Linko , T aiwan
CLARA R. C ORREIA • 3B’ s Research Group—Biomaterials, Biodegradables and Biomimetics ,
University of Minho, Headquarters of the European Institute of Excellence on Tissue
Engineering and Regenerative Medicine , Guimarães, P ortugal ; ICVS/3B’s –PT
Government Associate Laboratory , Guimarães, B raga, Portugal
ANDREW D ALY • Trinity Centre for Bioengineering, Trinity Biomedical Sciences Institute,
Trinity College Dublin , Dublin, Ireland ; Department of Mechanical and Manufacturing
Engineering, School of Engineering, T rinity College Dublin , D ublin, I reland ;
Advanced Materials and Bioengineering Research Centre (AMBER),
Trinity College Dublin , Dublin, Ireland
PAULINE M. D ORAN • Faculty of Science, Engineering and Technology,
Swinburne University of Technology , Melbourne, V IC, Australia
HICHAM DRISSI • Department of Orthopaedic Surgery , UConn Health , F armington,
CT, U SA ; D epartment of Genetics and Genome Sciences, UConn Health ,
Farmington, C T , U SA ; S tem Cell Institute, UConn Health , Farmington, C T , U SA
HUGO FERNANDES • University of Maastricht – MERLN Institute for Technology Inspired
Regenerative Medicine, Complex Tissue Regeneration Department, The Netherlands
OLIVER F. W. G ARDNER • Musculoskeletal Regeneration, AO Research Institute Davos ,
Davos Platz, S witzerland ; C ardiff University School of Biosciences , Cardiff, U K
JASON D. GIBSON • Department of Orthopaedic Surgery , U Conn Health , F armington, C T ,
USA
ROSA M. GUZZO • Department of Orthopaedic Surgery , UConn Health , F armington, C T ,
USA ; S tem Cell Institute, UConn Health , Farmington, CT , U SA
HYUN-WOOK KANG • Wake Forest Institute for Regenerative Medicine, W ake Forest School
of Medicine , Winston-Salem, NC, U SA
DANIEL JOHN KELLY • Trinity Centre for Bioengineering, Trinity Biomedical Sciences
Institute, T rinity College Dublin , D ublin, I reland ; D epartment of Mechanical and
Manufacturing Engineering, School of Engineering, T rinity College Dublin , D ublin,
Ireland ; A dvanced Materials and Bioengineering Research Center (AMBER), T rinity
College Dublin , D ublin, Ireland
TING TING LAU • Division of Bioengineering, School of Chemical and Biomedical
Engineering, N anyang Technological University , Singapore
ix