In the work reported here, titanium oxide film were synthesized using magnetron sputtering from a high-purity Ti metal target while sustaining a differential oxygen partial pressure of from 0.02 to 0.30 Pa. The biological behavior of endothelial cells grown on the film surface was studied by in vitro human umbilical vein endothelial cell (HUVEC) culture investigation. Our studies revealed that the adherence, growth, and proliferation of endothelial cells on different Ti-O film surfaces were strongly affected by the oxygen partial pressure. The biocompatibility mechanisms of ECs on Ti-O film surfaces were explored using X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), a-step profilometry, scanning electron microscopy (SEM), and atomic force microscopy (AFM). We conclude that the various film characteristics such as structure, composition and surface morphology due to the different oxygen partial pressures significantly influence the biological behavior of EC.