Fig 1: Effect of CM of MSCs+BMP-9 primed with hypoxia on cell proliferation, migration and osteoblast differentiation of MC3T3-E1 cells. Cell proliferation at 24, 48 and 72 h (A) and migration at 0, 6, 12 and 24 h (B) of MC3T3-E1 cells cultured in CM of MSCs+BMP-9 cultured under either control (CM control) or hypoxia condition (CM hypoxia). Gene expression of Runx2 (C), Sp7 (D), Alp (E) and Oc (F) and protein expression of RUNX2 (G) and ALP (H) of MC3T3-E1 cells cultured either in CM control or CM hypoxia, on day 7. The data are presented as mean ± standard deviation, and the * indicates statistically significant differences (p = 0.05).
Fig 2: Effect of hypoxia on the expression of components and targets of the BMP signaling pathway in MSCs+BMP-9. Protein expression of BMP-9 (A), and gene expression of Bmp-2 (B), Bmp-4 (C), Bmpr1a (D), Smad1 (E), Smad5 (F), Hey1 (G) and Runx2 (H) in MSCs+BMP-9 cultured under either control or hypoxia condition, on day 8. The data are presented as mean ± standard deviation, and the * indicates statistically significant differences (p = 0.05).
Fig 3: Effect of CM of MSCs+BMP-9 primed with hypoxia on cell proliferation, migration and osteoblast differentiation of MSCs. Cell proliferation at 24, 48 and 72 h (A) and migration at 0, 6, 12 and 24 h (B) of MSCs cultured in CM of MSCs+BMP-9 cultured under either control (CM control) or hypoxia condition (CM hypoxia). Gene expression of Runx2 (C), Sp7 (D), Alp (E) and Oc (F) and protein expression of RUNX2 (G) and ALP (H) of MSCs cultured either in CM control or CM hypoxia, on day 7. The data are presented as mean ± standard deviation, and the * indicates statistically significant differences (p = 0.05).
Fig 4: Effect of MSCs+BMP-9 primed with hypoxia on bone repair of rat calvarial defects. Histological analysis of the bone tissue. Micrographs of rat calvarial defects treated with either control MSCs+BMP-9 (Control, (A–C)) or MSCs+BMP-9 primed with hypoxia (Hypoxia, (D–F)) locally injected 2 weeks post-defect creation and evaluated at 28 days post-treatment. The square in (A) is presented in (B), and the square in (D) is presented in (E). Scale bar: (A,D) = 1.25 mm; (B,E) = 200 µm; (C,F) = 50 µm. bv: blood vessel; cl: cement line; ct: connective tissue; ib: immature bone; lb: lamellar bone; ob: osteoblast; ot: osteocyte.
Fig 5: Hypoxia induction and duration of its effect on MSCs+BMP-9. Viability of MSCs+BMP-9 cultured in either absence (control) or presence of CoCl2 at 100, 150 and 200 µM for the last 3 (A), 5 (B) and 7 days (C) of culture and at 10, 20, 40 and 50 µM in the last 3 (D), 5 (E) and 7 days (F) of culture, measured on day 8. Protein expression of HIF-1a in MSCs+BMP-9 cultured in either absence (control) or presence of CoCl2 at 40 and 50 µM for the last 3 and 5 days, detected on day 8 (G). Gene expression of Glut1 (H) and Vegfa (I) in MSCs+BMP-9 cultured for 8 days, with the last 3 days in either absence (control) or presence of CoCl2 at 40 µM, detected 0, 4, 8 and 12 h after CoCl2 removal. The data are presented as mean ± standard deviation, and the * indicates statistically significant differences (p = 0.05).
Supplier Page from Abcam for Mouse BMP9 ELISA Kit