The aim of this study was to evaluate the proliferation and mineralization ability of mouse osteoblastic MC3T3-E1 cells on F-containing TiO2 films with different morphology and nanostructure that previously confirmed antibacterial properties. F-containing TiO2 films were fabricated by anodizing Ti-6Al-4V alloy ELI -grade 23. By using a mixture of H2SO4/HF acid at 20 V for 5 and 60 min, a TiO2 film grows with nanoporous –NP– and nanotubular –NT–features, characterized with a pore diameter of 20 and 100 nm, respectively. Fluoride-TiO2 barrier films -FBL- were produced in 1 M NH4H2PO4/0.15M NH4F solution at constant voltage controlled at 20 V for 120 min. The amount of F incorporated in the nanostructured oxide films was 6 at.% and of 4 at.%, for the NP and NT respectively, while for the FBL film was 12 at.%. MC3T3-E1 cells exhibited different behaviour when seeded and grown onto these surfaces. Thus, F-doped TiO2 films with NP structures increased proliferation as well as osteogenic gene expression and the mineralization capacity of these osteoblastic cells. These results confirm that anodizing process is suitable to fabricate multifunctional surfaces on Ti-6Al-4V alloy with improved not only antibacterial but also osteogenic properties useful for bone fixation of prosthetic devices