Due to mechanical loading, cartilage experiences distortional change, volumetric change, and fluid flow. Research has shown cells to be responsive to unconfined compression, a load that produces all three conditions. To isolate the factor(s) responsible for chondrogenesis, the first goal of this research was to design and implement a device for the application of shear deformation to cells. Secondly, using this device, Stage 23/24 chick limb bud cells were suspended in 2% alginate and subjected to 20% shear deformation at 1 Hz. for two hours daily for three days. Gene expression, DNA content, sGAG content, and cartilage nodule formation were determined after eight days in culture and compared to results obtained for non-loaded cells. Results indicated that shear deformation at the applied level did not have a significant effect on chondrogenesis in Stage 23/24 chick limb bud cells, suggesting that this cell type is not extremely sensitive to distortional change.

The global objective of this research was to identify the proteins secreted by stem cells in response to mechanical stress. Since it has been shown in previous studies that conditioned medium from compressed chick limb bud cells cultured in alginate can initiate chondrogenesis in non-compressed cells, it was hypothesized that the conditioned medium contains valuable growth/differentiation factors. Due to cartilage?s limited capacity for repair, factors that stimulate stem-cell mediated regeneration are highly sought. To discern these proteins, conditioned medium was collected from cyclically compressed stage 23/24 chick limb buds suspended in alginate. The proteins were extracted, separated by 2-D gel electrophoresis, and evaluated by mass spectroscopy. While a few regulators of chondrogenesis were observed, such as FGF receptor, actin, and IP3 receptor, many potential peptides were not found in the database. However, this study showed that ascertaining proteins produced by chondrocytes in response to mechanical stimulation should be pursued.