Effect of Chinese herbal recipe extract for replenishing kidney and activating blood circulation (CRKRABC) on osteoblast

AIM: To investigate the effect for Chinese herbal recipe exctract for replenishing kidney and activating blood circulation (CRERKABC) on cell proliferation, differentiation and mineralization in cultured rat osteoblasts. METHODS: Osteoblasts from craniums of newly born SD rats were cultured in vitro , and the third-passage osteoblasts were exposed to CRERKABC(20 mg/L, 40 mg/L, 80 mg/L) and vehicle, respectively. MMT was used to assess ability of cell proliferation. Activity of akaline phosphatase was assayed by p-nitrophenyl picolinate, and content of osteocalcin in suppernatants was examined by radioimmunoassay. In addition, alizarin bordeaux staining method was used to count mineralzation nodes under optical microscope. RESULTS: ①CRERKABC increased the proliferation of osteoblasts in a dose and time-dependant manner. ②Akaline phosphatase(ALP) activity and osteocalcin content were also increased in CRERKABC-treated groups at 48 h , 72 h, 96 h, except ALP in low concentration CRERKABC group at 48 h( P0.01 ). ③Amount of mineralized nodes in middle and high concentration CRERKABC-treated groups were obviously higher than that in control group ( P0.01 ); There was no difference in amount of mineralized nodes between low concentration CRERKABC-treated group and control ( P0.05 ). CONCLUSION: CRERKABC can promote proliferation, differentiation, maturation and mineralization of the osteoblasts cultured in vitro.