To improve the bioactivity of titanium implants, a homogeneous layer of TiO2 nanotubes with a diameter of approximately 110nm was prepared by anodization. Gelatin was immobilized onto TiO2 nanotubes through an intermediate layer of polydopamine. The surface characteristics of different substrates were evaluated using X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), atomic force microscopy (AFM) and contact angle measurements, respectively. These results demonstrate that gelatin was successfully immobilized onto TiO2 nanotubes. In vitro cell culture experiments including immunofluorescence staining, cell viability, alkaline phosphatase (ALP), mineralization and the expression of osteogenic genes including runt-related transcription factor 2 (Runx2), ALP, collagen type I (Col I), and osteopontin (OPN) confirm that cell spreading, proliferation and osteoblastic differentiation were improved when cells were seeded onto gelatin-immobilized TiO2 nanotubes. This resulting material shows great promise as a future material in titanium implant applications.