Effects of histatin-derived peptides immobilization by tresyl chloride-activation technique for MC3T3-E1 cellular responses on titanium (Ti) were evaluated. MC3T3-E1 were cultured on sandblasted and acid-etched Ti disks immobilized with histatin-derived peptides, including histatin-1, JH8194, and mixed histatin-1 with JH8194. Surface topography and cellular morphology were examined using a scanning electron microscope. Elemental composition and conformational peptides on Ti surface were examined using energy dispersive X-ray and fourier transform infrared spectroscopy, respectively. Cellular adhesion, proliferation, osteogenesisrelated genes, and alkaline phosphatase activity were evaluated. The results showed that peptides were successfully immobilized on Ti surface. Cell attachments on histatin-1 and mixed peptides coated groups are higher than control. Histatin-1 achieved the significantly highest cellular proliferation. Histatin-derived peptides improved the osteogenesis related-gene expression and alkaline phosphatase activity (p<0.05). This study suggested that histatin-1 immobilization by tresyl chloride-activation technique enhanced cellular responses and might be able to promote cellular activities around the dental implants.