[Biological effect of WO-1 on human embryonic osteoblasts]

To investigate the effect of WO-1 on the proliferation and differentiation of human embryonic osteoblasts (HEO) and to provide research methods of bone tissue engineering. Methods HEO were isolated from periosteum and calvaria and then cultured in vitro. The dose-effect relationship between WO-1 concentration and biological effect of HEO was evaluated by growth curve and 3H-TdR count. The effect of WO-1 on cell activity and proliferation was investigated by cloning efficiency, cell cycle analysis was determined by flow cytometer and morphological was examined through transmission electron microscope. Moreover, the effect of WO-1 on osteoblastic function was evaluated at protein and mRNA levels by ALP activity, 3H-proline incorporation, osteocalcin secretion (RIA) and mRNA expression of type I collagen and osteocalcin (RT-PCR). Results The proliferation of HEO was inhibited in high concentration of WO-1, while it was promoted in low concentration of WO-1. The optimal dose was 8 microg/ml, and there was dose-effect relationship in the certain range of WO-1 concentration (0.25 microg/ml to 8 microg/ml). In 8 microg/ml of WO-1, the cloning efficiency and cloning volume of HEO were increased, population doubling time was decreased. All indexes of osteoblastic function including ALP activity, type I collagen synthesis and osteocalcin secretion were increased, the more sufficed cell organs were observed under transmission electron microscope than control group (P < 0.05).

WO-1 can promote the cell activity and proliferation of HEO cultured in vitro in low concentration, enhance the synthesis of extracellular matrix, such as type I collagen and osteocalcin, and accelerate the mineralization of osteoid. WO-1 can be used as a stimulant of proliferation and differentiation of HEO in the research of bone tissue engineering, which provide the theoretical basis in clinical application.