By Sylvie Thirion, Francis Berenbaum (auth.), Massimo Sabatini, Philippe Pastoureau, Frédéric De Ceuninck (eds.)
Osteoarthritis, the most typical type of arthritis, imposes an important burden of anguish on a becoming inhabitants of the aged. Even at the present time, its poorly understood pathophysiology limits the invention of molecular objectives for pharmacological intervention and there are few potent clinical remedies past discomfort keep watch over and surgical procedure. In Cartilage and Osteoarthritis a unique panel of researchers, physicians, and opinion leaders during this hard box describe their up to date classical, yet nonetheless evolving, strategies, in addition to many rising tools that promise so as to add considerably to our realizing of cartilage metabolism in well-being and disorder. quantity 1: mobile and Molecular instruments describes confirmed molecular and mobile ideas for the in vitro examine of standard and osteoarthritic cartilage via biochemical, biomolecular, immunological, and actual methods, with emphasis at the genetic manipulation of cells. The protocols stick with the winning equipment in Molecular drugs™ sequence layout, every one providing step by step laboratory directions, an creation outlining the primary in the back of the strategy, lists of the mandatory apparatus and reagents, and pointers on troubleshooting and warding off identified pitfalls. A spouse quantity, quantity 2: constitution and In Vivo research, bargains state-of-the-art tactics for studies-at the tissue level-of turnover, constitution, and functioning in general and diseased cartilage by way of invasive and noninvasive means.
entire and up to date, the 2 volumes of Cartilage and Osteoarthritis supply researchers and bench scientists alike with an fundamental number of without difficulty reproducible protocols for brand spanking new experiments-from the mobile to the animal level-designed to extra in actual fact describe the pathophysiology of cartilage, in addition to to find novel molecular goals for pharmacological intervention.
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Extra info for Cartilage and Osteoarthritis: Volume 1: Cellular and Molecular Tools
B. (1996) Human chondrocyte cultures as models of cartilage-specific gene regulation, in Methods in Molecular Biology: Human Cell Culture Protocols (Jones, G. ), Humana, Totowa, NJ, pp. 217–231. Robbins, J. R. and Goldring, M. B. (1998) Methods for preparation of immortalized human chondrocyte cell lines, in Methods in Molecular Medicine: Tissue Engineering Methods and Protocols (Morgan, J. R. and Yarmush, M. ), Humana, Totowa, NJ, pp. 173–192. , Robbins, J. , and Goldring, M. B. (2000) Proteoglycan production by immortalized human chondrocyte cell lines cultured under conditions that promote expression of the differentiated phenotype.
M. (1998) Matrix degradation by chondrocytes cultured in alginate: IL-1` induces proteoglycan degradation and proMMP synthesis but does not result in collagen degradation. Osteoarthritis Cartilage 6, 330–340. 22 De Ceuninck et al. 11. 11 Loeser, R. , Todd, M. , and Seely, B. L. (2003) Prolonged treatment of human osteoarthritic chondrocytes with insulin-like growth factor-I stimulates proteoglycan synthesis but not proteoglycan matrix accumulation in alginate cultures. J. Rheumatol. 30, 1565–1570.
Hyaluronidase may be added during these medium changes to facilitate the infection efficiency (see Note 6). 3. Begin infection by removing the medium and adding 5 mL of freshly thawed virus-containing medium supplemented with polybrene at 8 μg/mL to promote the binding of virus to the cells. Replace the dishes in the 37°C incubator and incubate for 4–6 h or overnight (see Note 7). 4. Remove the polybrene-containing medium and replace with fresh growth medium. Incubate for at least 24 h, until the cultures become confluent, or for at least one cell division cycle.