The capacity to reossify a calvarial bone defect is very small in mature animals and in humans greater than 2 y of age. The clinical treatment of injured tissue sites of bones by low-intensity pulsed ultrasound is widespread, but little is known about the precise effects of ultrasound on the fundamental processes that promote repair and regeneration. In this study, we used real-time polymerase chain reaction (RT-PCR) to investigate the expression of osteogenesis-associated genes after stimulation by low-intensity ultrasound in adult mouse osteoblast from the parietal calvaria. The gene associated with the Runx2 pathway had notably higher levels after 1, 2 and 3 days of stimulation. Therefore, low-intensity ultrasound seems to have an effect on the transcriptional gene expression of the calvarial bone in vitro.