Fig 1: Ceramide 24:1 stimulates osteoclast differentiation. (A) Primary mouse BMMs were incubated with 30 ng/mL M-CSF and 100 ng/mL RANKL in the absence or presence of the indicated concentration of C24:1 for 4 days. After staining cells with TRAP, the number of TRAP-positive MNCs (= 3 nuclei/cell) was determined to assess osteoclast differentiation. (B) Mouse BMMs were cocultured with primary calvaria osteoblasts for ten days in a medium containing 10-8 M 1a,25-OH(2) D3 and 10-6 M prostaglandin E2 without or with 0.01 µM C24:1. (C) Mouse BMMs were cultured with 30 ng/mL M-CSF and 100 ng/mL RANKL on dentine discs in the absence or presence of 0.01 µM C24:1 for ten days. Resorption pits were visualized by staining with hematoxylin. (D) qRT-PCR expression analysis of osteoclast differentiation markers in mouse BMMs exposed to 30 ng/mL M-CSF and 100 ng/mL RANKL in the absence or presence of 0.01 µM C24:1 for 4 days. (E) qRT-PCR analysis to determine relative Rankl and Opg expression in mouse calvaria osteoblasts exposed to 50 µg/mL ascorbic acid and 10 mM ß-glycerophosphate in the absence or presence of 0.01 µM C24:1 for seven days. Scale bars: 500 µm for (A–C). Data are presented as mean ± SEM. *P < 0.05 vs. untreated control using the Mann-Whitney U test or ANOVA followed by Tukey’s posthoc analysis.
Fig 2: Ceramide 18:0 stimulates osteoclast differentiation. (A) Primary mouse BMMs were incubated with 30 ng/mL M-CSF and 100 ng/mL RANKL in the absence or presence of the indicated concentration of C18:0 for four days. After staining cells with TRAP, the number of TRAP-positive multinucleated cells (MNCs) (=3 nuclei/cell) was determined to assess osteoclast differentiation. (B) Mouse BMMs were cocultured with primary calvaria osteoblasts for 10 days in medium containing 10-8 M 1a,25-OH(2) D3 and 10-6 M prostaglandin E2 without or with 0.1 µM C18:0. (C) Mouse BMMs were cultured with 30 ng/mL M-CSF and 100 ng/mL RANKL on dentine discs in the absence or presence of 0.1 µM C18:0 for 10 days. Resorption pits were visualized by staining with hematoxylin. (D) qRT-PCR expression analysis of osteoclast differentiation markers in mouse BMMs exposed to 30 ng/mL M-CSF and 100 ng/mL RANKL in the absence or presence of 0.1 µM C18:0 for 4 days. (E) qRT-PCR analysis to determine relative Rankl and Opg expression in mouse calvaria osteoblasts exposed to 50 µg/mL ascorbic acid and 10 mM ß-glycerophosphate in the absence or presence of 0.1 µM C18:0 for 7 days. Scale bars: 500 µm for (A–C). Data are presented as mean ± SEM. *P < 0.05 vs. untreated control using the Mann-Whitney U test or ANOVA followed by Tukey’s posthoc analysis.
Fig 3: (A) VEGF and (B) OPG secreted form HUVEC in various Si-containing FGelMa hydrogels for different times. * Indicates a significant difference (p < 0.05) from Si0. # Indicates a significant difference (p < 0.05) from Si0.5. Data presented as mean ± SEM, n = 6 for each group.
Supplier Page from Abcam for Human Osteoprotegerin ELISA Kit