Decoupling the effects of nanopore size and surface roughness on the attachment, spreading and differentiation of bone marrow-derived stem cells

The nanopore size and roughness of nanoporous surface are two critical variables in determining stem cell fate, but little is known about the contribution from each cue individually. To address this gap, we use two-dimensional nanoporous membranes with controlled nanopore size and roughness to culture bone marrow-derived mesenchymal stem cells (BMSCs), and study their behaviors such as attachment, spreading and differentiation. We find that increasing the roughness of nanoporous surface has no noticeable effect on cell attachment, and only slightly decreases cell spreading areas and inhibits osteogenic differentiation. However, BMSCs cultured on membranes with larger nanopores have significantly fewer attached cells and larger spreading areas. Moreover, these cells cultured on larger nanopores undergo enhanced osteogenic differentiation by expressing more alkaline phosphatase, osteocalcin, osteopontin, and secreting more collagen type I. These results suggest that although both nanopore size and roughness can affect BMSCs, nanopore size plays a more significant role than roughness in controlling BMSC behavior.