The materials (C-ODTi) with different topographical surfaces that possess interstitial oxygen atoms into the host titanium lattice and an upper nanometric surface layer of anatase-TiO(2) covered by a carbon thin layer were fabricated in this study. The carbon thin layer on the surface of C-ODTi was composed of amorphous carbon and nano-graphite crystals. In vitro tests, using human bone marrow-derived mesenchymal cells (hBMCs), were performed to check cytotoxicity, examining in particular cell morphology, cell proliferation, cell differentiation, and mineralization capability. After 10 days of culture a higher degree of cell viability was observed on the surface of C-ODTi with an abraded surface. We also observed that hBMCs cultured in direct contact with C-ODTi maintained their capability to express alkaline phosphatase activity (ALP) and formed mineralized nodules similar to the control cultures. Our results demonstrate that the carbon layer coating on the surface of C-ODTi possess better biological response than commercially pure titanium (cp Ti), which was evidenced by the higher proliferation rates of osteoblasts, higher osteo-differentiation and a higher mineralization capability.