In this paper, a thin transparent titania (TiO2) film was coated on the surface of flexible poly(ethylene terephthalate) (PET) film using the sol–gel method. The surface properties of the obtained TiO2/PET film were further improved by RF glow discharge oxygen plasma as a function of exposure time and discharge power. The changes in hydrophilicity of TiO2/PET films were analyzed by contact angle measurements and surface energy. The influence of plasma on the surface of the TiO2/PET films was analyzed by atomic force microscopy (AFM) as well as the change in chemical state and composition that were investigated by X-ray photo electron spectroscopy (XPS). The cytotoxicity of the TiO2/PET films was analyzed using human osteoblast cells and the bacterial eradication behaviors of TiO2/PET films were also evaluated against Staphylococcus bacteria. It was found that the surface roughness and incorporation of oxygen containing polar functional groups of the plasma treated TiO2/PET films increased substantially as compared to the untreated one. Moreover the increased concentration of Ti3 + on the surface of plasma treated TiO2/PET films was due to the transformation of chemical states (Ti4 + → Ti3 +). These morphological and chemical changes are responsible for enhanced hydrophilicity of the TiO2/PET films. Furthermore, the plasma treated TiO2/PET film exhibited no citotoxicity against osteoblast cells and antibacterial activity against Staphylococcus bacteria which can find application in manufacturing of biomedical devices.