Although it is difficult to directly correlate the differences in surface topography and chemistry to the cellular events, it can be speculated that the obtained outcomes were greatly influenced by the different surface modifications. In this study, we utilized a sequence of analysis to evaluate different titanium surfaces through parameters of cytotoxicity, osteoblast adhesion, proliferation, and differentiation. The initial cell toxicity evaluation showed that all the different surface treatments did not result in cytotoxic outcomes, and that biological effects should be more related to surface contact than to alteration on culture media content. The subsequent method evaluated osteoblast behavior on the different surfaces for up to 48 hours. This time point was selected due to the possibility of identifying the Lorazepam proteins involved in the cell cycle progression. In this case, it was found that textured titanium surfaces resulted in an increase in the osteoblast number compared to the machined and polystyrene surfaces, which was in accordance with previous investigations. Curiously, our results showed that textured titanium surfaces stimulated CDK6 expression, a signaling protein known to control cell cycle progression, and which has been associated to conditions of increased proliferation of mouse osteoblasts However, different effects for diverse NSC 624206 nanotopographies were not evident, suggesting that this may be a general feature of nanostructured surfaces. FAK and Src phosphorylation were evaluated in response to different surfaces and it was observed that both markers are main signaling proteins during osteoblast adhesion to the textured titanium surfaces. Furthermore, FAK phosphorylation at Y397 profile was more evident in response to the nanostructured MBAA relative to the other surfaces, while Src remained active in all groups. From these evidences, we can conclude that chemical/morphological properties of surfaces define cell behavior. The present results also provided a possible explanation for the mechanism in which nanotopography may affect such processes, as revealed by the proteomic analysis performed on the surfaces after exposure to serum. The profile of protein adsorption varied with each nano-surface on both total protein content and the type of preferentially adsorbed proteins. This result becomes relevant on the light of the well-known role of Extracellular Matrix proteins on osteoblast adhesion mediated by integrin on biomaterial surfaces including functionalized Ti.