In Vitro Model of Osteoarthritis for Drug Discovery

The overall goal of this project is the development of a high throughput, in vitro drug screening semploys immortalized human chondrocytes as a model of human cartilage matrix degradation as occurs iThe specific aim of Phase I is to demonstrate that a clonal line of immortalized human chondrocytesin vitro model for cartilage matrix degradation. The in vitro responses of IHCs and normal, nonimmorchondrocytes will be compared under conditions that induce the extracellular release of collagen IIenzyme assay, immunocytochemistry, western blot analysis, amino acid sequence analysis of proteoglycfragments, and quantitation of collagen II and proteoglycan loss from the matrix by reversed-phase Hrelease, respectively. The Phase II goal is to develop a quantitative, high-volume-drug screening syIHCs to identify compounds that can inhibit cartilage matrix destruction. The need for identificationovel therapeutics for the treatment of osteoarthritis is overwhelming, as symptomatic relief with ninflammatory drugs, or total joint replacement are currently the only therapeutic options.National Cancer Institute (NCI)