Implant-related infection is one of the most common and serious complications associated with biomedical implantation. To prevent bacterial adhesion, a series of porous TiO2 coatings with different concentrations of silver (designated as M0, M1, M2 and M3) were prepared on pure titanium substrates by a duplex-treatment technique combining magnetron sputtering with micro-arc oxidation. All coatings are porous with pore size less than 5 μm and the concentrations of silver in the M0, M1, M2 and M3 are 0, 0.95, 1.36 and 1.93 wt.%, respectively. Silver is found to be distributed throughout the thickness of the coatings by scanning electron microscopy. The release of silver from the TiO2 coatings was confirmed by an inductively-coupled plasma mass spectroscopy. The antibacterial effects of these coatings were tested against Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Escherichia coli (E. coli), and the cytotoxicity was evaluated using the mouse pre-osteoblast cells. The results indicate that the antibacterial activities of TiO2 coatings are greatly improved due to the incorporation of silver. No cytotoxic effect is found for the M1 surfaces from the observation of pre-osteoblast cell by MTT assay and fluorescence microscopy. Although the M2 and M3 coatings appeared to be toxic for pre-osteoblast cells after 1 day in culture, the cell viability on M2 and M3 surfaces was greatly raised after culturing for 2 days. Our results suggested that the TiO2 coatings incorporated with an optimum amount of silver can possess excellent antibacterial activities without cytotoxic effect, which has promising applications in biomedical devices.