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The successful application of a nanostructured biomaterial as an implant is strongly determined by the nanotopography size triggering the ideal cell response. Here, nanoporous topography on 304L stainless steel substrates was engineered to identify the nanotopography size causing a transition in the cellular characteristics, and accordingly, the design of nanostructured stainless steel surface as orthopedic implants is proposed. A variety of nanopore diameters ranging from 100 to 220 nm were fabricated by one-step electrolysis process and collectively referred to as artificial microenvironments. Control over the nanopore diameter was achieved by varying bias voltage. MG63 osteoblasts were cultured on the nanoporous surfaces for different days. Immunofluorescence (IF) and scanning electron microscopy (SEM) were performed to compare the modulation in cell morphologies and characteristics. Osteoblasts displayed differential growth parameters and distinct transition in cell behavior after nanopore reached a certain diameter. Nanopores with 100-nm diameter promoted cell growth, focal adhesions, cell area, viability, vinculin-stained area, calcium mineralization, and alkaline phosphatase activity. The ability of these nanoporous substrates to differentially modulate the cell behavior and assist in identifying the transition step will be beneficial to biomedical engineers to develop superior implant geometries, triggering an ideal cell response at the cell-nanotopography interface.

The Spatiotemporal Control of Osteoblast Cell Growth, Behavior, and Function Dictated by Nanostructured Stainless Steel Artificial Microenvironments