Dynamic cell-adhesive microenvironments and their effect on myogenic differentiation

Integrin-mediated cell adhesion plays a central role in cell behavior on biomaterial surfaces and influences various cell functions. Photoactivatable RGD adhesive peptides were used to investigate the effect of the density and time point of bioadhesive ligand presentation on cell adhesion, proliferation and differentiation. PEGylated self-assembled monolayers were functionalized with RGD and caged RGD ligands and seeded with C2C12 myoblasts. The cultures were irradiated at various time points between 1 and 48 h after cell seeding in order to increase RGD surface concentration at defined time points. Attachment, spreading and myogenic differentiation of C2C12 myoblasts strongly varied with the density of RGD at the surface. Proliferation and myogenesis were further regulated by the time point at which RGD was presented to the cell, reaching highest levels when RGD exposure occurred ≤6 h after cell seeding. These results provide fundamental insights in cell–biomaterial interactions of C2C12 myoblasts in terms of temporal integrin-mediated cell responses.