Figure 8

Effect of exogenous BMP4 on osteoblastic and adipocytic differentiation of hBMSC^−Bone cells. (a) Quantification of cell viability of hBMSC^−Bone cells in the presence or absence of recombinant BMP4. (b) qRT-PCR quantification for TAGLN, TPM1, and Col1A2 in hBMSC^−Bone cells in the presence or absence of recombinant BMP4. The expression of each target gene was normalized to GAPDH. Data are presented as mean ± SD from three independent experiments, n = 9; ***p < 0.0005. (c) OsteoImage™ staining (20× magnification) of hBMSC^−Bone cells which were induced into the osteoblast in the presence or absence of recombinant BMP4. The lower panel shows Alizarin Red S staining. The quantification of mineralized matrix formation for vehicle or recombinant BMP4-treated hBMSC^−Bone cells is shown (right panel). Data are presented as relative mean mineralization ± SD from three independent experiments, n = 9; *p < 0.0005. (d) qRT-PCR quantification of ALPL, OCN, ON, and and COL1A1 osteogenic markers in hBMSC^−Bone cells in the presence or absence of recombinant BMP4 under osteogenic induction conditions. The expression of each target gene was normalized to GAPDH. Data are presented as the means ± SD from three independent experiments, n = 9;, *p < 0.05**p < 0.005, ***p < 0.0005. (e) hBMSC^−Bone cells were differentiated into adipocytes for 7 days under the indicated experimental conditions. Upper panel shows fluorescence Nile red staining of mature oil filled adipocytes (20× magnification), whilst the lower panel shows Oil red O staining for adipocytes (20× magnification). The lower panel shows the relative quantification of Nile red staining of mature oil-filled adipocytes. (f) qRT-PCR quantification for LPL and CEBPA adipocytic markers. The expression of each target gene was normalized to GAPDH. Data are presented as mean ± SD from three independent experiments, n = 9; **p < 0.005, ***p < 0.0005. (g) Schematic model illustrating the convergence of BMP and TGFβ in regulating hBMSC differentiation.