The response of osteoblast-like cells cultured on blasted and/or acid etching surfaces and the influence of surface texture or microtopography on cell attachment, cell proliferation, and the gene expression of the osteoblastic phenotype using ROS 17/2.8 cell lines were evaluated. The blasted and/or acid etching surfaces were significantly rougher in comparison to machined and etched surfaces (p < 0.05). On X-ray diffraction analysis, titanium hydride (TiH2) was observed on the surface etched with a mixture of HCl-H2SO4 solution, whereas TiH2 was not observed on machined and blasted surfaces. After 24 h incubation, most of the cells of all the groups had a flattened, polygonal shape and were fully spread, exhibiting the onset of proliferation. The MTT assay showed a significant decrease on the blasted surface compared to the machined surface at 7 day culture (p < 0.05). The expression of osteopontin mRNA, α1 (I) collagen mRNA, and alkaline phosphatase mRNA on rough surfaces was higher than on the machined surfaces, and was highest on the blasted surface at day 7.