To clarify the effects of micron/submicron hybrid topography on cell morphology and functionalization, we investigated the adhesion and differentiation of human mesenchymal stem cells (hMSCs) to titanium (Ti) surfaces with three different topographies: micron, submicron, and hybrid grooves created using a femtosecond laser. hMSCs cultured on Ti specimens showed high alignment on micron and hybrid surfaces after 6 h of incubation, whereas cells attached to submicron and hybrid surfaces were elongated. An examination of vinculin-positive adhesion plaques indicated that micron grooves affected cellular alignment by modifying the initial cell polarization, whereas submicron grooves affected cellular extension. A superposition effect of topography was evidenced by the highly aligned and elongated morphology of hMSCs grown on the hybrid surface, which promoted osteogenic and chondrogenic differentiation. These findings provide a basis for the design of novel biomaterial surfaces that can control specific cellular functions.
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