Fig. 2. Individual supplementation of glutamine and magnesium enhanced osteogenesis.
(A) ALP activity on day 7 of osteogenic differentiation with various glutamine concentrations. Data are average SD. One-way ANOVA with Tukey’s multiple comparisons test, *P < 0.05, **P < 0.005, ***P < 0.0005, and ****P < 0.0001. n = 4. (B) mRNA expression of Runx2 on day 4 of hBM-MSCs osteogenic differentiation with various glutamine concentrations. Data are average SD. One-way ANOVA with Tukey’s multiple comparisons test, *P < 0.05, **P < 0.005, ***P < 0.0005, and ****P < 0.0001. n = 3. (C to F) mRNA expression of Alpl, Sp7, Spp1, and Ibsp on day 7 of hBM-MSCs osteogenic differentiation with various glutamine concentrations. Data are average SD. One-way ANOVA with Tukey’s multiple comparisons test, *P < 0.05, **P < 0.005, ***P < 0.0005, and ****P < 0.0001. n = 3. (G and H) Representative images and quantification of calcium nodules deposition (red) with various glutamine concentrations on day 14 of hBM-MSCs osteogenic differentiation. Scale bar, 500 μm. Data are average SD. One-way ANOVA with Tukey’s multiple comparisons test, *P < 0.05, **P < 0.005, ***P < 0.0005, and ****P < 0.0001. n = 4. (I) ALP activity on day 7 of osteogenic differentiation with various magnesium concentrations. Data are average SD. One-way ANOVA with Tukey’s multiple comparisons test, *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001. n = 4. (J) mRNA expression of Runx2 on day 4 of hBM-MSCs osteogenic differentiation with various magnesium concentrations. (K to M) mRNA expression of Alpl, Spp1, and Col1a1 on day 7 of hBM-MSCs osteogenic differentiation with various magnesium concentrations. Data are average SD. One-way ANOVA with Tukey’s multiple comparisons test, *P < 0.05, **P < 0.005, ***P < 0.0005, and ****P < 0.0001. n = 3. (N and O) Representative images and quantification of calcium nodules deposition (red) with various magnesium concentrations on day 14 of hBM-MSCs osteogenic differentiation. Scale bar, 500 μm. Data are average SD. One-way ANOVA with Tukey’s multiple comparisons test, *P < 0.05, **P < 0.005, ***P < 0.0005, and ****P < 0.0001. n = 3. OD, optical density.
