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. 2018 Dec 7;27(2):394–410. doi: 10.1016/j.ymthe.2018.11.019

Figure 6.

Figure 6

miR-296 Directs toward Osteoblastic Differentiation in Immunodeficient Mice and Bone Formation from BMSCs

(A and B) ARS (A) and ALP (B) analysis revealed that BMSCs treated with miR-296 mimic potentiated osteogenesis and miR-296 AMO weakened osteoblast differentiation compared with NC after osteogenic differentiation of 14 days. Scale bar, 100 μm. (C and D) Real-time qPCR confirmed that miR-296 mimic (C) increased the mRNA levels of osteoblast genes and miR-296 AMO (D) decreased the expression relative to NC, respectively. (E) Expression of osteogenic genes in BMSCs were diminished by miR-296 AMO and increased by miR-296 mimic. (F) Fluorescent images of Runx2 revealed significantly enhanced osteogenic ability caused by miR-296 mimic while this was decreased by miR-296 AMO. Runx2, red; DAPI, blue. Scale bar, 100 μm. (G) The ratio of new bone was analyzed by H&E staining from immunodeficient mice. The black arrows indicate bone formation, and the red arrows indicate hydroxyapatite. Scale bar, 50 μm. (H and I) ARS and ALP analysis verified miR-296 can restore the osteogenic ability of BMSCs from osteoporotic mice caused by OVX surgery (H) or overexpression of lncRNA-ORLNC1 (I) compared with control mice. Scale bar, 100 μm. *p < 0.05, **p < 0.01, and ***p < 0.001.