Effect of a novel compound from Lycopodium obscurum L. on osteogenic activity of osteoblasts in vitro

Abstract

We investigated the potential of an extract of Lycopodium obscurum L.; stigmastane-3-oxo- 21-oic acid (SA), to enhance osteogensis of mouse osteoblastic MC3T3-E1 cells. SA at a concentration of 16 µM was found to have no significant effect upon the viability of the cells, thus concentrations of 8 µM and 16 µM of SA were used in all further experiments. Both con- centrations of SA had an inhibitory affect upon alkaline phosphatase activity (ALP) after 8 days incubation, however, after 16 days activity was restored to control levels. However Alizarin red S staining showed increased levels of miner- alization for both concentrations after 16 days culture. Real time PCR showed inhibition of genes Runx2 and Osterix genes responsible for the up-regulation of ALP. However early time point (8 days) up-regulation of bone matrix min- eralization genes OPN and OCN, and late time point (16 days) up-regulation of both Jun-D and Fra-2 mRNA expression was significantly en- hanced. These results suggest a potential me- chanism of SA in enhancing bone fracture heal- ing is through the up-regulating bone matrix mineralization.