Cranial sutures produce new bone at the sutural edges of the bone fronts in response to external stimuli. Little is known regarding the mechanism of osteogenesis in cranial sutures. Ets1 and Cbfa1 are two important osteogenic transcription factors regulating the differentiation and maturation of osteoblasts. But their function in cranial sutures is not still elucidated. We have investigated the gene expression of Ets1 and Cbfa1 in rat’s calvarial sutural osteoblast-like cells under a single period of mechanical strain. The cells were isolated from the cranial suture of SD rats and cultured in vitro, and subjected to a single 40 minutes mechanical strain using a four-point bending apparatus. The gene expression patterns of Ets1 and Cbfa1 were examined by RT-PCR. Both mRNA levels of Ets1 and Cbfa1 have increased significantly within 6 and 12 hours respectively after mechanical strain were applied, and the increase returned to control level thereafter. However, Ets1 and Cbfa1 exhibited different temporal expression patterns: Ets1 expressed immediately after the mechanical loading and reached the maximum transcription at 0.5h; whereas Cbfa1 experienced a latency period first, then increased slowly within 2 hours, and reached the maximum transcription at 6 h. The maximum transcription of Cbfa1 was about 2.58 fold of that of Ets1. Ets1and Cbfa1 may play different roles in regulating bone matrix protein expressions in osteoblast-like cells during suture distraction and their function is time-dependent. High frequency distraction (>2times/24h) is favourable to the maximal expression of the two genes.