High density titania nanofiber (TiO2 NFs) arrays were fabricated in situ on a Ti–6Al–4V substrate by a simple oxidation process, and the in vitro cellular response of chondrocytes on the resulting surfaces was evaluated. Results show that the TiO2 nanofibrous substrate triggers enhanced chondrocytes adhesion, proliferation, and production of extracellular matrix (ECM) fibrils compared to untreated substrate. These results suggest that chondrocytes have an affinity to the surface structure produced by the oxidation process and therefore has potential use in implants designed for cartilaginous applications.