Crosslinking density influences the morphology of chondrocytes photoencapsulated in PEG hydrogels during the application of compressive strain.

Chondrocyte deformation, which occurs during mechanical loading, is thought to play an important role in the mechanotransduction pathway. In designing a scaffold that can be gelled in situ for cartilage tissue engineering, an important consideration is the influence of mechanical loading. This study tested the hypothesis that changes in the crosslinking density of a hydrogel scaffold influence the morphology of encapsulated chondrocytes in response to an applied load. Chondrocytes were entrapped in photo-crosslinkable hydrogel scaffolds based on poly(ethylene glycol) (PEG) with two crosslinking densities, 0.119 and 0.376 mol/l, with the higher density having a 11-fold higher compressive modulus. The cell-embedded hydrogels were subjected to static compressive strains between 0% and 20% after 1 and 6 days of culture. Using confocal laser scanning microscopy, chondrocytes in the highly crosslinked gel at day 1 deformed more than gels in the more loosely crosslinked gel. By day 6, this finding was reversed. When single cells within a region were followed, heterogeneities in cell deformation were observed on both a macroscopic and microscopic scale. These heterogeneities were greater in the highly crosslinked gel. These findings demonstrate that different levels of cell deformation and heterogeneity may be obtained by varying the crosslinking density in PEG hydrogels.