Journal: Cytotechnology

Article Title: Apigenin inhibits osteoblastogenesis and osteoclastogenesis and prevents bone loss in ovariectomized mice

doi: 10.1007/s10616-014-9694-3

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

Journal: PLoS ONE

Article Title: Toddaculin, Isolated from of Toddalia asiatica (L.) Lam., Inhibited Osteoclastogenesis in RAW 264 Cells and Enhanced Osteoblastogenesis in MC3T3-E1 Cells

doi: 10.1371/journal.pone.0127158

Figure Lengend Snippet: Effect of toddaculin on osteoblast mineralization in ascorbic acid and β-glycerophosphate-treated MC3T3-E1 cells that were evaluated by Alizarin red S staining (microplate reader analysis (A) and light microscopic evaluation (B)). Values are expressed as means ± SEM (n = 3). *, P

Article Snippet: Ascorbic acid and β-glycerophosphate were purchased from Wako Pure Chemical (Osaka, Japan).

Techniques: Staining

Journal: Scientific Reports

Article Title: Magnesium prevents vascular calcification in vitro by inhibition of hydroxyapatite crystal formation

doi: 10.1038/s41598-018-20241-3

Figure Lengend Snippet: Crystal or particle clusters formed in BGP cell culture supernatant are morphologically and chemically distinct. Morphology of crystal clusters formed in BGP and in BGP supplemented with Mg 2+ cell culture supernatants, as overview (upper) and focused (lower) for EDX analysis ( a ). The averaged EDX-spectrum and resulting quantification of the crystal cluster isolated from Mg 2+ supplemented culture supernatant reveals reduced Ca (green) and P (purple), as visualized in the elemental map ( c ) compared to crystal clusters found in BGP treated cultures ( b ). In addition, Mg 2+ supplemented crystal clusters showed increased Na (blue) and Cl (orange) compared to BGP crystal clusters. Data are presented as mean of the analyses of ten individual crystal clusters. BGP, β -glycerophosphate; CPS, counts per second; EDX, energy dispersive spectroscopy; (k)eV, (kilo)-electron volt.

Article Snippet: Experimental design For calcification, medium was supplemented with 5% (v/v) FBS and 10 mM β -glycerophosphate (BGP, Merck Millipore, Massachusetts, USA).

Techniques: Cell Culture, Isolation, Spectroscopy

Journal: Scientific Reports

Article Title: Magnesium prevents vascular calcification in vitro by inhibition of hydroxyapatite crystal formation

doi: 10.1038/s41598-018-20241-3

Figure Lengend Snippet: Impact of Mg 2+ on BGP-induced Ca 2+ -apatite formation. X-ray powder diffraction analysis of cell culture supernatants show presence of NaCl (*) and Ca-apatite (arrows) in BGP treated cultures (black line), but not in BGP cultures supplemented with Mg 2+ (gray line). Of note: as the BGP culture supplemented with Mg 2+ contained less precipitates; one measurement represents 21 cultures (6-well format) compared to 7 in the BGP condition in order to reach the detection threshold. BGP, β -glycerophosphate.

Article Snippet: Experimental design For calcification, medium was supplemented with 5% (v/v) FBS and 10 mM β -glycerophosphate (BGP, Merck Millipore, Massachusetts, USA).

Techniques: Cell Culture