Figure 1
Overexpression of H19 promoted osteogenesis.
(A–F) The hMSCs isolated from two independent donors’ bone marrow were initiated by osteogenic medium and harvested at indicated time points. The expression levels of several candidate lncRNAs, including CUDR (A), Linc-ROR (B), TINCR (C), H19 (D), ALP (E) and RUNX2 (F) were monitored by qRT-PCR. (G) The osteogenesis of H19-overexpressing hMSCs was initiated by osteogenic medium. At day 10 post-induction, the ALP activity was measured and ectopic expression of H19 enhanced ALP activity. (H) The osteoblast differentiation of H19-overexpressing hMSCs was started by osteogenic medium. At day 20 post-induction, the calcified nodules were visualized by Alizarin Red stainning and it was found that overexpression of H19 promoted hMSC osteogenesis. Scale bars: 1cm. (I) The RNA levels of osteogenesis-related marker genes were evaluated by qRT-PCR and ectopic expression of H19 upregulated the expression serial osteogenic maker genes. (J) H19-overexpressing hMSCs and corresponding control cells were implanted subcutaneously into the dorsal surfaces of nude mice. At 8 weeks post-implantation, the transplants were collected for histological analysis. Representative images of H&E staining and immunohistochemical staining of OCN were captured. The new bone matrix was indicated by black arrow and OCN proteins were highlighted by white arrow. Scale bars: 100 μm. The results of in vivo bone formation assay showed that H19 significnatly enhanced osteogenesis. (n = 3; *P < 0.05; **P < 0.01; ***P < 0.001).
