Limitations associated with the use of autologous chondrocytes (CH) for cartilage tissue engineering beget the need for alternative cell sources. Mesenchymal stem cells (MSC) are clinically attractive due to their ability to undergo chondrogenesis in three-dimensional culture [1,2]; however, when compared to CH, MSC fail to develop functional equivalence [2,3]. We have previously shown a marked depth-dependence in local equilibrium modulus of MSC-laden gels, with the superficial zones (where maximal media exchange occurs) considerably stiffer than regions removed from nutrient supply (center and bottom of construct); less dramatic depth-dependence was observed in CH-laden gels [4]. Similarly, other studies have shown depth-dependent properties in CH-laden gels with the construct edge generally stiffer than the center [5]. Given this apparent influence of nutrient supply, the objective of the current study was to assess the impact of dynamic culture (via orbital shaking) on the development of depth-dependent mechanical properties in both MSC and CH-laden hydrogels. Furthermore, we assessed cell viability and matrix content throughout the construct depth to determine the mechanism by which this depth-dependency arises. We hypothesized that improved nutrient transport would reduce construct inhomogeneity (particularly for MSC-laden constructs) and improve bulk mechanical properties.

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