Article Title: Apigenin inhibits osteoblastogenesis and osteoclastogenesis and prevents bone loss in ovariectomized mice
Figure Lengend Snippet: Effects of apigenin on collagen production, alkaline phosphatase activity, and mineralization of osteoblasts. MC3T3-E1 cells were cultured in 12-well plates (3.8 cm 2 /well) or 24-well plates (2.0 cm 2 /well) with α-MEM containing 10 % fetal bovine serum, 5 mM β-glycerophosphate, and 50 μg mL −1 ascorbic acid. After the cells reached confluence (day 3), the olive polyphenol apigenin was added at various concentrations (1–10 μM) to the culture medium. Fresh medium with test compound was supplied at 3-day intervals. a Collagen production was measured at day 7 as described in the method. Cell layers in 24-well plates (2.0 cm 2 /well) were fixed with Bouin’s solution and stained with Sirius Red dye reagent. Stained material was dissolved in 0.1 N sodium hydroxide and the absorbance was measured at 550 nm. b Alkaline phosphatase activity was measured at day 14 as described in the method. c Deposition of Ca 2+ ions was measured at days 21 and 24. Quantitative analysis of Ca 2+ ions was performed as described in the method. All values represent the mean ± SD of the results from three wells. Data are representative of results from three separate experiments. * P
Article Snippet: After reaching 70 % confluence, cells were detached by treatment with 0.05 % trypsin, replated in either 55-cm2 dishes or 12-well plates (area of each well, 3.8 cm2 ) at a density of 1 × 104 cells cm−2 , and grown in α-MEM supplemented with 10 % fetal bovine serum, 50 units mL−1 penicillin, 50 μg mL−1 streptomycin, 5 mM β-glycerophosphate (Sigma-Aldrich, Tokyo, Japan), and 50 μg mL−1 ascorbic acid (Sigma-Aldrich).
Techniques: Activity Assay, Cell Culture, Staining