The aim of the present study is to investigate the effectiveness of low-intensity pulsed ultrasound(LIPUS) on cell proliferation, differentiation and mineralizationusing of MC3T3-E1 cell line. The MC3T3-E1 cells were exposed to LIPUS with different parameters(frenquency and intensity). The cell proliferations, differentiation, and mineralization were determined by CCK-8 method, alkaline phosphatase detection kit, and alizarin red staining, respectively. The ultrasound intensity of 30mW/cm² and 40mW/cm² could promote cell proliferation, while 50mW/cm² inhibit it. The lower intensity group under 1.5MHz and 1.7MHz present better alkaline phosphatase activity. The 1.5MHz and 40 mW/cm² groups showed better mineralization effect than others. LIPUS can promote MC3T3-E1 on cell proliferation, differentiation and mineralization. LIPUS may act as a new method forprevention and treatment of osteoporosis.