Fig. 2. Effects of BD on osteogenic differentiation and mineralization of osteogenic-induced MC3T3-E1 cells. The ALP assay (a) and matrix mineralization assay (b, c) were performed to examine ALP activity and mineralization in MC3T3-E1 cells after the addition of BD to the osteogenic medium and subsequent incubation for 14 and 21 d, respectively. (a) BD increased the level of ALP activity in osteogenic-induced MC3T3-E1 cells, especially at 10 and 14 d, with a pronounced effect at 0.10 μmol/L. (b, c) BD promoted the formation of extracellular mineralized nodules, especially at 0.10 μmol/L. Data are expressed as mean±SD (n=3). ^* P<0.05, ^** P<0.01, ^*** P<0.001, and ^**** P<0.0001 represent comparisons with the control group; ^# P<0.05, ^## P<0.01, and ^### P<0.001 represent comparisons between experimental groups. BD: benidipine; ALP: alkaline phosphatase.