Osteogenic Activity of Lupeol Isolated from Clinacanthus nutans Lindau: Activity and Mode of Action

Figure 4

Lupeol induced mineralization activity of osteoblast cells MC3T3-E1. Osteoblast cells were cultured in the MEMα medium containing vitamin C (50 μg/ml) and β-glycerolphosphate (10 mM) for 21 days with/without lupeol and then stained with 40 mM alizarin red-S (pH 4.2) for 1 h. The cells were destained for 15 minutes with 10% cetylpyridium chloride in 10 mM sodium phosphate buffer (pH 7.0); then, calcium formation was observed and determined spectrophotometrically at 562 nm. The results are calculated as percentage compared to the nontreated sample (control, DMSO). Each value is the average of triplicate cultures, and each bar indicates means ± SD;  ≤ 0.05;  ≤ 0.01.