Abstract: Objective The purpose of this study was to explore the effects of pure titanium discs treated with hydrogen peroxide and heat on attachment, proliferation and differentiation of MC3T3-E1 cells of mice. Methods Sandblasted, dual acid-etched titanium discs were set as control group, whereas sandblasted, acid-etched and hydrogen peroxide and heat treated titanium discs were as test group. Scanning electron microscope (SEM) and X-ray diffraction (XRD) were used for surface characterization. Pre-osteoblasts MC3T3-E1 were seeded on these discs in 2 groups, and then early cell attachment, cell proliferation and cell differentiation were evaluated. Results SEM analysis showed that the titanium discs in the 2 groups both had roughened surfaces with porous network structure. However, XRD examination indicated that anatase only covered on the surfaces in the test group. Early attachment of osteoblasts was much stimulated by the titanium discs in the test group (P<0.05). Besides cell proliferation on the surfaces of titanium discs was faster in the test group (P<0.05). Meanwhile, alkaline phosphatase activity and osteocalcin production on surfaces of titanium discs were dramatically elevated in the test group, whether in protein level or in transcriptional level. Conclusions Sandblasted, acid-etched and hydrogen peroxide and heat treated pure titanium implants are prone to integrate with surrounding bone tissue.