Experimental arthritis : in vitro and in vivo models

218 Pages·2017·6.93 MB·English

by  Hok Kei Tam

Experimental arthritis : in vitro and in vivo models
Citation for published version (APA):
Wang, X. (2008). Experimental arthritis : in vitro and in vivo models. [Phd Thesis 2 (Research NOT TU/e /
Graduation TU/e), Biomedical Engineering]. Technische Universiteit Eindhoven.
https://doi.org/10.6100/IR634844
DOI:
10.6100/IR634844
Document status and date:
Published: 01/01/2008
Document Version:
Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers)
Please check the document version of this publication:
• A submitted manuscript is the version of the article upon submission and before peer-review. There can be
important differences between the submitted version and the official published version of record. People
interested in the research are advised to contact the author for the final version of the publication, or visit the
DOI to the publisher's website.
• The final author version and the galley proof are versions of the publication after peer review.
• The final published version features the final layout of the paper including the volume, issue and page
numbers.
Link to publication
General rights
Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners
and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.
• Users may download and print one copy of any publication from the public portal for the purpose of private study or research.
• You may not further distribute the material or use it for any profit-making activity or commercial gain
• You may freely distribute the URL identifying the publication in the public portal.
If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license above, please
follow below link for the End User Agreement:
www.tue.nl/taverne
Take down policy
If you believe that this document breaches copyright please contact us at:
[email protected]
providing details and we will investigate your claim.
Download date: 20. Jan. 2023
Experimental Arthritis:
in vitro and in vivo Models
A catalogue record is available from the Eindhoven University of Technology Library 
ISBN: 978‐90‐386‐1270‐6 

Printed in the United States of America 
Wang, Susanne X. 
Experimental Arthritis: in vitro and in vivo Models/ Susanne X. Wang 
Cover design: Ryan H. Tam and Susanne X. Wang 

Copyright ©2008 by Susanne Xuanhui Wang 
All rights reserved. 
No part of this publication maybe reproduced or transmitted in any form or by any 
means, electronic or mechanical, including photocopy, recording, or any information 

storage and retrieval system, without permission in writing from the publisher.
Experimental Arthritis: in vitro and in vivo Models

PROEFSCHRIFT

ter verkrijging van de graad van doctor aan de
Technische Universiteit Eindhoven, op gezag van de
Rector Magnificus, prof.dr.ir. C.J. van Duijn, voor een
commissie aangewezen door het College voor
Promoties in het openbaar te verdedigen
op donderdag 3 juni 2008 om 16.00 uur

door

Xuanhui Wang

geboren te Liaoning, China
Dit proefschrift is goedgekeurd door de promotor:

prof.dr.ir. H.W.J. Huiskes

Copromotor:
dr. C.C. van Donkelaar
To my daughters Lauren and Martina
Thank you for being the most wonderful
children in the world.
For since the creation of the world God's invisible 
qualities—his eternal power and divine nature—have 
been clearly seen, being understood from what has been 
made, so that men are without excuse.
Table of Contents

Table of Contents ..............................................................................................................1 
Summary ............................................................................................................................9 
Chapter 1 ‐ Introduction ................................................................................................. 13 
1. Introduction to Arthritis ......................................................................................... 14 
1.1. Overview of OA ................................................................................................ 14 
1.2. Overview of RA ................................................................................................ 14 
2. Introduction of Articular Joints ............................................................................... 15 
3. Introduction to Articular Cartilage and Function ................................................... 16 
3.1. Chondrocytes in Articular Cartilage ................................................................. 16 
4. Problems of Articular Cartilage Repair ................................................................... 18 
5. Current OA Treatments .......................................................................................... 19 
5.1. Medications ..................................................................................................... 19 
5.2. Surgery ............................................................................................................. 19 
5.3. Alternatives ..................................................................................................... 20 
6. Development and Progression of OA ..................................................................... 21 
6.1. Typical OA Progression .................................................................................... 21 
6.2. OA as a Whole Joint Disease ........................................................................... 22 
6.3. Relationship between Articular Cartilage and Subchondral Bone .................. 23 
6.4. Bone Structure and Metabolism ..................................................................... 23 
6.5. Bone Structure, Material, and Mechanical Properties .................................... 24 
6.6. Bone Remodeling ............................................................................................ 25 
6.7. Possible Initiation of OA as a Bone Disease ..................................................... 25 
6.8. Anti‐resorptive Drugs as a Potential Treatment for OA .................................. 26 
6.9 Development of Osteophytes .......................................................................... 27 
6.10. Complexity of OA Development and Progression Pathways ......................... 27 
7. Need for Animal Models ......................................................................................... 28 
Page | 1
8. Objectives and outlines .......................................................................................... 29 
References .................................................................................................................. 30 
Chapter 2 ‐ Subchondral Bone Microarchitecture Changes in Animal Models of Arthritis 
Quantified by Micro‐CT .................................................................................................. 35 
1. Introduction ............................................................................................................ 37 
2. Bone changes in human arthritis ............................................................................ 38 
3. Bone changes in OA animal models ....................................................................... 38 
4.  Micro‐CT imaging in arthritis ................................................................................. 39 
5. Methodology of using micro‐CT in arthritis research ............................................. 40 
5.1 Sample preparation: ......................................................................................... 40 
5.2 Scanning procedures: ....................................................................................... 41 
5.3 Reconstruction: ................................................................................................ 41 
5.4 Three‐dimensional analysis .............................................................................. 42 
5.5 Three‐dimensional isosurfaces ......................................................................... 42 
6. Cartilage imaging using micro‐CT and micro MRI .................................................. 46 
7. The future of using micro‐CT in arthritis research ................................................. 46 
References .................................................................................................................. 48 
Chapter 3 ‐ Tissue Engineering of Biphasic Cartilage Constructs Using Various 
Biodegradable Scaffolds: an in vitro Study ..................................................................... 51 
1. Introduction ............................................................................................................ 53 
2. Materials and methods .......................................................................................... 54 
2.1. Articular chondrocyte isolation ....................................................................... 54 
2.2. Chamber seeding and construct formation .................................................... 54 
2.3. Histological evaluation .................................................................................... 56 
2.4. Confocal laser scanning microscopy (CLSM) cell viability (live/dead assay) ... 56 
2.5. Immunohistochemical staining for collagen types I, II and X .......................... 56 
2.6. Biochemical analyses (SDS‐PAGE) ................................................................... 56 
2.7. Transmission electron microscopy (TEM) ....................................................... 57 
2.8. Scanning electron microscopy (SEM) .............................................................. 57 
3. Results .................................................................................................................... 57 
Page | 2
3.1. Macroscopic and histological observations..................................................... 57 
3.2. Cell viability ...................................................................................................... 59 
3.3. Biochemistry and immunohistochemistry ...................................................... 60 
3.4. Transmission electron microscopy .................................................................. 60 
3.5. Scanning electron microscopy (SEM) .............................................................. 61 
4. Discussion ............................................................................................................... 63 
5. Conclusions ............................................................................................................. 65 
Acknowledgements .................................................................................................... 65 
References .................................................................................................................. 66 
Chapter 4 ‐ Disease‐Modifying Effects of N‐butyryl Glucosamine in a Streptococcal Cell 
Wall (SCW) Induced Arthritis Model in Rats ................................................................... 69 
1. Introduction ............................................................................................................ 71 
2. Materials and Methods .......................................................................................... 73 
2.1 Animal model and experiment design. ............................................................ 73 
2.2 Bone mineral density (BMD) measurement. .................................................... 74 
2.3 Micro‐CT imaging protocol. .............................................................................. 74 
2.4 Three‐dimensional trabecular bone measurements. ....................................... 74 
2.5 Three‐dimensional isosurface. ......................................................................... 75 
2.6 Two‐dimensional subchondral bone analysis. ................................................. 75 
2.7 Statistical analysis. ............................................................................................ 76 
3. Results .................................................................................................................... 76 
3.1 Effects of GlcNBu on the SCW‐induced inflammation. .................................... 76 
3.2 Decline of BMD by the arthritis and reversal by GlcNBu.................................. 77 
3.3 Micro‐architecture of the trabecular bone. ..................................................... 80 
3.4 Cancellous bone connectivity. .......................................................................... 82 
3.5 Site‐specific structure change. ......................................................................... 82 
3.6 Joint integrity. ................................................................................................... 86 
3.7 Structure model index (SMI). ........................................................................... 87 
4. Discussion ............................................................................................................... 87 
Page | 3