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.

Includes supplementary material: sn.pub/extras

Klappentext
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.

Inhalt
Culture and Phenotyping of Chondrocytes in Primary Culture.- Culture of Chondrocytes in Alginate Beads.- Immortalization of Human Articular Chondrocytes for Generation of Stable, Differentiated Cell Lines.- Culture of Immortalized Chondrocytes and Their Use As Models of Chondrocyte Function.- Generation of Pluripotent Stem Cells and Their Differentiation to the Chondrocytic Phenotype.- Semiquantitative Analysis of Gene Expression in Cultured Chondrocytes by RT-PCR.- Quantification of mRNA Expression Levels in Articular Chondrocytes With PCR Technologies.- RNA Extraction From Cartilage.- Gene Expression Analysis in Cartilage by In Situ Hybridization.- Analysis of Differential Gene Expression in Healthy and Osteoarthritic Cartilage and Isolated Chondrocytes by Microarray Analysis.- High-Efficiency Nonviral Transfection of Primary Chondrocytes.- In Vitro Gene Transfer to Chondrocytes and Synovial Fibroblasts by Adenoviral Vectors.- Changes of Chondrocyte Metabolism In Vitro.- Analysis of Chondrocyte Functional Markers and Pericellular Matrix Components by Flow Cytometry.- A Simple and Reliable Assay of Proteoglycan Synthesis by Cultured Chondrocytes.- Assays of Proteoglycan and Collagen Degradation in Cultures of Rabbit Cartilage Explants.- Production of Antibodies Against Degradative Neoepitopes in Aggrecan.- Immunoassays for Collagens in Chondrocyte and Cartilage Explant Cultures.- Detection of Apoptosis in Cartilage In Situ and in Isolated Chondrocytes.- Expression, Activity, and Regulation of MAP Kinases in Cultured Chondrocytes.- Mechanical Loading of Chondrocytes Embedded in 3D Constructs.- In Vitro Physical Stimulation of Tissue-Engineered and Native Cartilage.