Surface Topographical Cues for Regulating Differentiation of Human Neural Stem Cells

Surface topographical cues has been highlighted to control the fate of neural stem cells (NSCs). Herein we developed a hierarchically patterned substrate (HPS) platform for regulating NSC differentiation. The HPS induced cytoskeleton alignment and highly activated focal adhesion in hNSCs as indicated by enhanced expression of focal adhesion proteins such as focal adhesion kinase (FAK) and vinculin. hNSCs cultured on HPS exhibited enhanced neuronal differentiation compared to flat group. We also developed a graphene oxide (GO)-based hierarchically patterned substrates (GPS) that promote focal adhesion formation and neuronal differentiation of hNSCs. Enhanced focal adhesion and differentiation of hNSCs on the HPS was reversed by blocking the ${\beta}1$ integrin binding and mechanotransduction-associated signals including Rho-associated protein kinase (ROCK) and extracellular-regulated kinase (ERK) pathway, which may suggest a potential mechanism of beneficial effects of HPS. In addition, hNSCs on the HPS differentiated into functional neurons exhibiting sodium currents and action potentials as confirmed by whole cell patch-clamping analysis. The hierarchical topography can direct differentiation of NSCs towards functional neurons, and therefore would be an important element for the design of functional biomaterials for neural tissue regeneration applications.