In the present study, a nanoporous, biocompatible titanium dioxide (TiO 2 ) film was formed on the surface of titanium, using oxygen plasma immersion ion implantation (OPIII), and the influence of this film on the bio-functionalization, including the proliferation and differentiation properties of MG-63 osteoblast-like cells, was analyzed and investigated. The OPIII-treated surface was characterized by X-ray photoelectron, which showed that a TiO x layer was formed on Ti substrates. This TiO x surface exhibited nanoscale surface roughness in the form of nanoporous structures. The results also revealed that MG-63 cells expressed increased proliferation on the OPIII-treated surface as compared with the untreated Ti substrate. The Ti specimens treated with plasma energy of 1 kW revealed better expression of alkaline phosphatase (ALP) activity and showed higher average surface roughness than untreated specimens. Thus, it can be concluded that bioactivity of Ti implants can potentially be improved by OPIII.