Quantitative analysis of cell proliferation and orientation on substrata with uniform parallel surface micro-grooves.

In order to quantify the effect of the substrata surface topography on cellular behaviour, planar and micro-textured silicon substrata were produced and made suitable for cell culture by radio frequency glow discharge treatment. These substrata possessed parallel surface grooves with a groove and ridge width of 2.0 (SilD02), 5.0 (SilD05) and 10 microns (SilD10). Groove depth was approximately 0.5 micron. Rat dermal fibroblasts (RDFs) were cultured on these substrata and a tissue culture polystyrene control surface for 1, 2, 3, 5 and 7 days. After incubation the cell proliferation was quantified with a Coulter Counter, and RDF size, shape and orientation with digital image analysis. Cell counts proved that neither the presence of the surface grooves nor the dimension of these grooves had an effect on the cell proliferation. However, RDFs on SilD02, and to a lesser extent on SilD05 substrata, were elongated and aligned parallel to the surface grooves. Orientation of the RDFs on SilD10 substrata proved to be almost comparable to the SilD00 substrata. Finally, it was observed that the cells on the micro-textured substrata were capable of spanning the surface grooves.

Effect of parallel surface microgrooves and surface energy on cell growth.

To evaluate the effect of surface treatment and surface microtexture on cellular behavior, smooth and microtextured silicone substrata were produced. The microtextured substrata possessed parallel surface grooves with a width and spacing of 2.0 (SilD02), 5.0 (SilD05), and 10 microns (SilD10). The groove depth was approximately 0.5 microns. Subsequently, these substrata were either left untreated (NT) or treated by ultraviolet irradiation (UV), radiofrequency glow discharge treatment (RFGD), or both (UVRFGD). After characterization of the substrata, rat dermal fibroblasts (RDF) were cultured on the UV, RFGD, and UVRFGD treated surfaces for 1, 3, 5, and 7 days. Comparison between the NT and UV substrata revealed that UV treatment did not influence the contact angles and surface energies of surfaces with a similar surface topography. However, the contact angles of the RFGD and UVRFGD substrata were significantly smaller than those of the UV and NT substrata. The dimension of the surface microevents did not influence the wettability characteristics. Cell culture experiments revealed that RDF cell growth on UV-treated surfaces was lower than on the RFGD and UVRFGD substrata. SEM examination demonstrated that the parallel surface grooves on the SilD02 and SilD05 substrata were able to induce stronger cell orientation and alignment than the events on SilD10 surfaces. By combining all of our findings, the most important conclusion was that physicochemical parameters such as wettability and surface free energy influence cell growth but play no measurable role in the shape and orientation of cells on microtextured surfaces.