(174au) Optimization of Chondrogenic Differentiation of Adipose-Derived Stem Cells through Co-Culture with Chondrocytes and Addition of Growth Factor and Nutraceutical Compounds

Osteoarthritis (OA), which occurs due to the degradation of native articular cartilage tissue, affects over 30 million adults in the United States. Cartilage tissue does not regenerate once injured, and there are currently no long term, effective treatment options for the disease that restore native joint function and structure. Thus, new therapies for the treatment of OA are necessary. The use of stem cell-based therapies to generate cartilage tissue is a promising technique as stem cells can be directed to differentiate and produce native-like cartilage tissue which, as the technology is perfected, can then be implanted into osteoarthritic joints. The addition of chondrogenic growth factors and the co-culture of mesenchymal stem cells with native chondrocyte cells has been shown to enhance differentiation of stem cells towards chondrogenic lineages and to promote the production of critical extracellular matrix (ECM) components such as glycosaminoglycans (GAG) and collagen II (Col-2). Further, in chondrocyte culture, nutraceutical compounds have been shown to reduce inflammation and promote the expression ECM components. This work investigates the combined roles of the growth factor dexamethasone, nutraceutical compounds, and the co-culture ratio of adipose derived stem cells and human chondrocytes for producing cartilage tissue rich in native cartilage markers and extracellular matrix components. A Box Behnken fractional factorial design will be used to investigate the significance of each variable at several levels, as well as identify significant interaction effects between dexamethasone, nutraceutical compounds, and the co-culture ratio on chondrogenesis. Chondrogenic mRNA markers and extracellular matrix components GAG and Col-2 will be quantified after 21 days of culture. We hypothesize that an optimum combination of growth factor and nutraceutical concentrations and co-culture ratio will be identified, leading to more efficient and cartilage-specific differentiation of co-cultured adipose-derived stem cells.