Arthritis Research: Methods and Protocols

441 Pages·2007·6.22 MB·English

by  Andrew P. Cope

Arthritis Research
M E T H O D S I N M O L E C U L A R M E D I C I N E™
John M. Walker, SERIES EDITOR
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135.Arthritis Research: Methods and Protocols, 114.Microarrays in Clinical Diagnostics,edited by
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125.Myeloid Leukemia:Methods and Protocols, ed- 102.Autoimmunity: Methods and Protocols,
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2005 Johnson Y. N. Lau, 2004
M E T H O D S I N M O L E C U L A R M E D I C I N E™
Arthritis Research
Methods and Protocols
Volume 1
Edited by
Andrew P. Cope
The Kennedy Institute of Rheumatology,
Imperial College London, London UK
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Arthritis research : methods and protocols / edited by Andrew P. Cope.
p. ; cm. — (Methods in molecular biology ; v. 135-136)
Includes bibliographical references and index.
ISBN 1-58829-344-0 (v. 1 : alk. paper) — ISBN 1-58829-918-X (v. 2 : alk. paper)
1. Arthritis—Laboratory manuals. 2. Arthritis—Molecular aspects. I. Cope, Andrew P. II. Series:
Methods in molecular biology (Clifton, N.J.) ; v. 135-136.
[DNLM: 1. Arthritis—Laboratory Manuals. 2. Laboratory Techniques and Procedures—Laboratory
Manuals. W1 ME9616J v.135-136 2007 / WE 25 A787 2007]
RC933.A665245 2007
616.7’220072—dc22 2006019975
Preface
“……………… do not go where the path may lead,
go instead where there is no path and leave a trail”
Ralph Waldo Emerson
The postgenomic era is upon us and with it comes a growing need to under-
stand the function of every gene and its contribution to physiological and patho-
logical processes. Such advances will underpin our understanding of the
molecular basis of common chronic inflammatory and degenerative diseases
and inspire the development of targeted therapy. Any postgenomic approach
for exploring gene function must necessarily address gene expression and regu-
lation, localization of gene products in diseased tissue, manipulation of expres-
sion by transgenesis or knockdown technology, and combine these studies with
appropriate manipulations in relevant in vivo models. To validate potential
therapeutic targets in any depth requires a growing repertoire of assays and
disease models that underpin key pathogenic pathways. The same repertoire of
tools must be employed to rigorously evaluate process specific biomarkers,
which may be of diagnostic and prognostic value. Indeed, measuring the im-
pact of our interventions remains a major challenge for the future.
The rheumatic diseases encompass prototypic chronic inflammatory and
degenerative diseases. It would be true to say that experimental procedures
adapted for investigating the pathogenesis of diseases such as rheumatoid arthri-
tis have contributed greatly to recent advances in biological therapy. Arthritis
Research: Methods and Protocols seeks to crystallize methods and protocols
that have contributed to such advances in molecular medicine.These volumes are
timely because the tools are now accessible to most laboratories. Also included
are newer technologies, some of which are still evolving and whose impact are
yet to be realized. It is important to note that in these volumes there is some-
thing for everyone—basic scientists, clinician scientists, and clinicians alike—
with contributions from leaders in their field covering imaging and immuno-
histochemistry, analysis of cartilage, and bone catabolism, as well as leukocyte
trafficking and migration. Combine volumes 1 and 2 and the end product is a
concise set of protocols condensing decades of experience and expertise. From
the outset of this project it was always the intention that this compendium
should provide a unique resource at the bench that would be used in ways that
will facilitate the endeavors of clinicians at the bedside in the future.
v
Acknowledgments
I wish to thank many friends and colleagues for their enthusiasm, support
and invaluable contributions toward this project. I am also very grateful to
Mandy Wilcox for her dedicated secretarial assistance in compiling the fin-
ished product. The research carried out by the Editor’s laboratory at the
Kennedy Institute is supported by grants from the Wellcome Trust and the
Arthritis Research Campaign, UK.
Andrew P. Cope
vii
Contents
Preface ..............................................................................................................v
Ackowledgments............................................................................................ vii
Contributors .................................................................................................xviii
Color Plate ....................................................................................................xvii
Contents for Volume 2...................................................................................xix
PART I SYNOVIAL JOINT MORPHOLOGY, HISTOPATHOLOGY, AND
IMMUNOHISTOCHEMISTRY
1 Imaging Inflamed Synovial Joints
Ai Lyn Tan, Helen I. Keen, Paul Emery, and Dennis McGonagle.........3
2 Arthroscopy as a Research Tool: A Review
Richard J. Reece..................................................................................27
3 Immunohistochemistry of the Inflamed Synovium
Martina Gogarty and Oliver FitzGerald.............................................47
4 In Situ Hybridization of Synovial Tissue
Stefan Kuchen, Christian A. Seemayer, Michel Neidhart,
Renate E. Gay, and Steffen Gay......................................................65
5 Subtractive Hybridization
Jörg H. W. Distler, Oliver Distler, Michel Neidhart,
and Steffen Gay..............................................................................77
6 Laser Capture as a Tool for Analysis of Gene Expression
in Inflamed Synovium
Ulf Müller-Ladner, Martin Judex, Elena Neumann,
and Steffen Gay..............................................................................91
7 Preparation of Mononuclear Cells from Synovial Tissue
Jonathan T. Beech and Fionula M. Brennan.....................................105
8 Quantitative Image Analysis of Synovial Tissue
Pascal O. van der Hall, Maarten C. Kraan, and Paul Peter Tak.......121
PART II CARTILAGE MATRIX AND BONE BIOLOGY
9 Cartilage Histomorphometry
Ernst B. Hunziker..............................................................................147
10 Image Analysis of Aggrecan Degradation in Articular
Cartilage with Formalin-Fixed Samples
Barbara Osborn, Yun Bai, Anna H. K. Plaas,
and John D. Sandy........................................................................167
ix
x Contents
11 In Situ Detection of Cell Death in Articular Cartilage
Samantha N. Redman, Ilyas M. Khan, Simon R. Tew,
and Charles W. Archer.................................................................183
12 Measurement of Glycosaminoglycan Release
from Cartilage Explants
John S. Mort and Peter J. Roughley..................................................201
13 Assessment of Collagenase Activity in Cartilage
Tim E. Cawston and Tanya G. Morgan.............................................211
14 Assessment of Gelatinase Expression and Activity
in Articular Cartilage
Rosalind M. Hembry, Susan J. Atkinson, and Gillian Murphy.........227
15 Analysis of MT1-MMP Activity in Cells
Richard D. Evans and Yoshifumi Itoh...............................................239
16 Analysis of TIMP Expression and Activity
Linda Troeberg and Hideaki Nagase.................................................251
17 Bone Histomorphology in Arthritis Models
Georg Schett and Birgit Tuerk..........................................................269
18 Generation of Osteoclasts In Vitro, and Assay of Osteoclast Activity
Naoyuki Takahashi, Nobuyuki Udagawa, Yasuhiro Kobayashi,
and Tatsuo Suda............................................................................285
PART III CELL TRAFFICKING, MIGRATION, AND INVASION
19 Isolation and Analysis of Large and Small Vessel Endothelial Cells
Justin C. Mason, Elaine A. Lidington, and Helen Yarwood..............305
20 Analysis of Flow-Based Adhesion In Vitro
Oliver Florey and Dorian O. Haskard..............................................323
21 Analysis of Leukocyte Recruitment in Synovial Microcirculation
by Intravital Microscopy
Gabriela Constantin..........................................................................333
22 Angiogenesis in Arthritis: Methodological and Analytical Details
Ursula Fearon and Douglas J. Veale.................................................343
23 Analysis of Inflammatory Leukocyte and Endothelial
Chemotactic Activity
Zoltán Szekanecz and Alisa Koch.....................................................359
24 Acquisition, Culture, and Phenotyping of Synovial Fibroblasts
Sanna Rosengren, David L. Boyle, and Gary S. Firestein.................365
25 Genotyping of Synovial Fibroblasts: cDNA Array in Combination
with RAP-PCR in Arthritis
Elena Neumann, Martin Judex, Steffen Gay,
and Ulf Müller-Ladner..................................................................377
Contents xi
26 Gene Transfer to Synovial Fibroblast: Methods and Evaluation
in the SCID Mouse Model
Ingmar Meinecke, Edita Rutkauskaite, Antje Cinski,
Ulf Müller-Ladner, Steffen Gay, and Thomas Pap.......................393
27 In Vitro Matrigel Fibroblast Invasion Assay
Tanja C.A. Tolboom and Tom W.J. Huizinga...................................413
28 Culture and Analysis of Circulating Fibrocytes
Timothy E. Quan and Richard Bucala...............................................423
Index ............................................................................................................435