Effect of microenvironment on adhesion and differentiation of murine C3H10T1/2 cells cultured on multilayers containing collagen I and glycosaminoglycans

Polyelectrolyte multilayer coating is a promising tool to control cellular behavior. Murine C3H10T1/2 embryonic fibroblasts share many features with mesenchymal stem cells, which are good candidates for use in regenerative medicine. However, the interactions of C3H10T1/2 cells with polyelectrolyte multilayers have not been studied yet. Hence, the effect of molecular composition of biomimetic multilayers, by pairing collagen I (Col I) with either hyaluronic acid or chondroitin sulfate, based primarily on ion pairing and on additional intrinsic cross-linking was studied regarding the adhesion and differentiation of C3H10T1/2 cells. It was found that the adhesion and osteogenic differentiation of C3H10T1/2 cells were more pronounced on chondroitin sulfate-based multilayers when cultured in the absence of osteogenic supplements, which corresponded to the significant larger amounts of Col I fibrils in these multilayers. By contrast, the staining of cartilage-specific matrixes was more intensive when cells were cultured on hyaluronic acid-based multilayers. Moreover, it is of note that a limited osteogenic and chondrogenic differentiation were detected when cells were cultured in osteogenic or chondrogenic medium. Specifically, cells were largely differentiated into an adipogenic lineage when cultured in osteogenic medium or 100 ng mL^-1 bone morphogenic protein 2, and it was more evident on the oxidized glycosaminoglycans-based multilayers, which corresponded also to the higher stiffness of cross-linked multilayers. Overall, polyelectrolyte multilayer composition and stiffness can be used to direct cell-matrix interactions, and hence the fate of C3H10T1/2 cells. However, these cells have a higher adipogenic potential than osteogenic or chondrogenic potential.