Objective: To determine the effect of simvastatin (Sim) and saponified simvastatin (Sim+) on bone formation in vitro by rat bone marrow (RBM) cells. Methods: RBM cells harvested from 7- week old inbred LEWIS rats were either not stimulated or stimulated with dexamethasone (Dex, 10^-8 M), 0.075% or 0.0075% ethanol (EtOH) with Dex (negative controls 1-4), 10^-6 M or 10^-7 M Sim with Dex, or 10^-6 M or 10^-7 M Sim+ with Dex (experimental groups 1-4). Alkaline phosphatase (ALP) assays were taken at 4 and 7 days, and von Kossa assays were performed at 7 and 14 days following stimulation. Results: At day 4, ALP levels of the 10^-6 M Sim and Sim+ groups were greater than all groups (p < 0.05) and more than the 0.075% EtOH group at p=0.076 and 0.051 levels, respectively. At day 7, ALP levels of all groups were greater than the unstimulated group (p<0.05), the 10^-7 M Sim+ group was greater than all groups except the 10^-7 M Sim group (p<0.05), the 10^-7 M Sim group was greater than both EtOH groups and the 10^-6 M Sim group (p<0.05) and the Dex and the 10^-6 M Sim+ group at p=0.088 and 0.059 levels, respectively. Mineralized nodules were seen only at day 14 in the Dex, 10^-6 M Sim and 10^-6 M Sim+ groups, but were not statistically different from the negative controls due to one or two wells not mineralizing. However, mineralized nodules were virtually nonexistent in the negative controls. Conclusion: Simvastatin and saponified simvastatin both induce increased in vitro ALP production in RBM cells in a concentration dependent manner. Mineralized nodules are also formed by day 14 but were not greater than the Dex control. Currently this experiment is being repeated with more time points. Funded by the NIDCR Grant # 1 R03 DE14653-01.

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