Titanium has been widely used in the orthopedic and dental fields, however, the inert nature of Ti makes it unsuitable for application in promoting bone cell growth, osteogenic differentiation and antibacterial ability. The aims of the current study were to investigate the antimicrobial activity and biofunction of the polypeptide antibiotic bacitracin, and obtain a multi-biofunctional titanium implant by covalently-immobilizing titanium with the bacitracin. The results showed that the bacitracin possessed low minimum inhibitory concentration (MIC) to both Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA), with the non-cytotoxicity concentration up to 500 mu g/mL to human bone marrow mesenchymal stem cells (hBMSCs), furthermore, the bacitracin could improve the osteogenic differentiation of hBMSCs. The results of Scanning electron microscope (SEM) and X-ray photoelectron spectroscopy (XPS) indicated that bacitracin had been covalently immobilized on the surface of titanium. Immobilized bacitracin could improve the hydrophilic of immobilized titanium. The results of antimicrobial assay demonstrated that the covalently-immobilized bacitracin also had excellent antimicrobial property, and the bacitracin immobilized titanium could inhibit bacterial adhesion and colonization. The results of cell biology experiments proved that the bacitracin immobilized titanium could improve hBMSCs' adhesion, proliferation and osteogenic differentiation. We also found that the macrophages were difficult to spread or activate on the surface of bacitracin immobilized titanium, and the secretion of inflammatory factors had been inhibited. In conclusion, the novel bacitracin immobilized titanium has multi-biofunctions including outstanding antibacterial properties, excellent cell biology performance, and restraining inflammation, which has exciting application prospect. (C) 2016 Elsevier B.V. All rights reserved.