A titanium surface nitrided by plasma contains nitrogen ions that guarantee resistance to corrosion and biocompatibility. Despite this, no descriptions concerning the influence of the expression of cell adhesion proteins and their influence on osteogenic cell differentiation are available. Thus, the present study aimed to assess the response of murine pre‐osteoblastic cells (MC3T3‐E1) cultured on nitrided titanium surfaces. Pre‐osteoblastic cells were grown on polished titanium discs, used as controls, and on previously characterized plasma‐nitrided titanium discs. Cells from both groups were submitted to the MTT cell viability test. The expressions of α5, α2, and β1 integrin were assessed by flow cytometry and immunofluorescence, while osteocalcin expression was assessed by flow cytometry. The nitrided surface presented higher α2 and β1 integrin expressions, as well as osteocalcin expression, when compared to the polished surface, with no alterations in cell viability. These findings seem to suggest that the plasma nitriding treatment produces a titanium surface with the potential for effective in vitro osseointegration.