Cartilage and Osteoarthritis: Volume 1: Cellular and Molecular Tools

ISBN-13: 9781588292476 / Angielski / Twarda / 2004 / 358 str.

Massimo Sabatini; Frederic de Ceuninck; Philippe Pastoureau

Opis

Osteoarthritis (OA), the most common form of arthritis, is generally characterized by a slowly progressive degeneration of articular cartilage, particularly in the weight-bearing joints. It has a stronger prevalence in women, and its incidence increases with age. OA is a major and growing health concern in developed countries, owing to steadily increasing life expectancy and the demand for better quality of life. Because of its chronic nature and nonfatal outcome, OA affects the growing population of the elderly over an increasing time span. Moreover, despite its relatively benign character, OA is one of the most disabling diseases; it is responsible for increasing financial and social burdens in terms of medical treatments, forced inactivity, loss of mobility, and dependence. Despite a growing awareness of OA as a medical problem that has yet to reach its maximum impact on society, there is a surprising absence of effective medical treatments beyond pain control and surgery. So far, only symptom-modifying drugs are available, while there remains a major demand for disease-modifying treatments of proven clinical efficacy. This demand will hopefully be met in the future by some of the drugs that have been pressed into development and are now at different stages of clinical investigation. Nevertheless, the current lack of effective treatments reflects a still insufficient knowledge of cartilage with respect to its metabolism, interactions with other joint tissues, and causes and mechanisms (possibly of very different nature) leading to failure of its turnover.

Spis treści

Culture and Phenotyping of Chondrocytes in Primary Culture Sylvie Thirion and Francis Berenbaum Culture of Chondrocytes in Alginate Beads Frédéric De Ceuninck, Christophe Lesur, Philippe Pastoureau, Audrey Caliez, and Massimo Sabatini Immortalization of Human Articular Chondrocytes for Generation of Stable, Differentiated Cell Lines Mary B. Goldring Culture of Immortalized Chondrocytes and Their Use As Models of Chondrocyte Function Mary B. Goldring Generation of Pluripotent Stem Cells and Their Differentiation to the Chondrocytic Phenotype Luis A. Solchaga, Jean F. Welter, Donald P. Lennon, and Arnold I. Caplan Semiquantitative Analysis of Gene Expression in Cultured Chondrocytes by RT-PCR Gaëlle Rolland-Valognes Quantification of mRNA Expression Levels in Articular Chondrocytes With PCR Technologies Audrey McAlinden, Jochen Haag, Brigitte Bau, Pia M. Gebhard, and Thomas Aigner RNA Extraction From Cartilage Frédéric Mallein-Gerin and Jérôme Gouttenoire Gene Expression Analysis in Cartilage by In Situ Hybridization Frédéric Mallein-Gerin and Jérôme Gouttenoire Analysis of Differential Gene Expression in Healthy and Osteoarthritic Cartilage and Isolated Chondrocytes by Microarray Analysis Thomas Aigner, Joachim Saas, Alexander Zien, Ralf Zimmer, Pia M. Gebhard, and Thomas Knorr High-Efficiency Nonviral Transfection of Primary Chondrocytes Jean F. Welter, Luis A. Solchaga, and Matthew C. Stewart In Vitro Gene Transfer to Chondrocytes and Synovial Fibroblasts by Adenoviral Vectors Jean-Noel Gouze, Martin J. Stoddart, Elvire Gouze, Glyn D. Palmer, Steven C. Ghivizzani, Alan J. Grodzinsky, and Christopher H. Evans Changes of Chondrocyte Metabolism In Vitro: An Approach by Proteomic Analysis Anne-Marie Freyria and Michel Becchi Analysis of ChondrocyteFunctional Markers and Pericellular Matrix Components by Flow Cytometry Gust Verbruggen, Jun Wang, Lai Wang, Dirk Elewaut, and Eric M. Veys A Simple and Reliable Assay of Proteoglycan Synthesis by Cultured Chondrocytes Frédéric De Ceuninck and Audrey Caliez Assays of Proteoglycan and Collagen Degradation in Cultures of Rabbit Cartilage Explants Christophe Lesur and Massimo Sabatini Production of Antibodies Against Degradative Neoepitopes in Aggrecan John S. Mort and Peter J. Roughley Immunoassays for Collagens in Chondrocyte and Cartilage Explant Cultures R. Clark Billinghurst, Fackson Mwale, Anthony Hollander, Mirela Ionescu, and A. Robin Poole Detection of Apoptosis in Cartilage In Situ and in Isolated Chondrocytes Darryl D. D'Lima, Klaus Kuhn, and Martin K. Lotz Expression, Activity, and Regulation of MAP Kinases in Cultured Chondrocytes Jang-Soo Chun Mechanical Loading of Chondrocytes Embedded in 3D Constructs: In Vitro Methods for Assessment of Morphological and Metabolic Response to Compressive Strain David A. Lee and Martin M. Knight In Vitro Physical Stimulation of Tissue-Engineered and Native Cartilage Kelvin W. Li, Travis J. Klein, Kanika Chawla, Gayle E. Nugent, Won C. Bae, and Robert L. Sah Index

Blurb

Osteoarthritis, the most common form of arthritis, imposes a significant burden of suffering on a growing population of the elderly. Even today, its poorly understood pathophysiology limits the discovery of molecular targets for pharmacological intervention and there are few effective medical treatments beyond pain control and surgery. In Cartilage and Osteoarthritis a distinguished panel of researchers, physicians, and opinion leaders in this challenging field describe their updated classical, but still evolving, techniques, as well as many emerging methods that promise to add significantly to our understanding of cartilage metabolism in health and disease. Volume 1: Cellular and Molecular Tools describes proven molecular and cellular techniques for the in vitro study of normal and osteoarthritic cartilage through biochemical, biomolecular, immunological, and physical approaches, with emphasis on the genetic manipulation of cells. The protocols follow the successful Methods in Molecular Medicine™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. A companion volume, Volume 2: Structure and In Vivo Analysis, offers cutting-edge procedures for studies-at the tissue level-of turnover, structure, and functioning in normal and diseased cartilage by invasive and noninvasive means.
Comprehensive and up-to-date, the two volumes of Cartilage and Osteoarthritis provide researchers and bench scientists alike with an indispensable collection of readily reproducible protocols for new experiments-from the cellular to the animal level-designed to more clearly describe the pathophysiology of cartilage, as well as to discover novel molecular targets for pharmacological intervention.