2022, Vol. 26 ›› Issue (30): 4757-4761

Adenovirus-mediated bone morphogenetic protein 2 induces osteogenic differentiation of rabbit bone marrow mesenchymal stem cells

Wu Chengcong1, Wang Fang2, Wan Jianshan1, Wu Zheng1, Sun Rong1, Huang Hefei1, Qian Xuankun1, Ou Hua1, Ren Jing1   

Abstract: BACKGROUND: Natural bone morphogenetic protein 2 cannot achieve sustained release in vivo and continuously induce bone marrow mesenchymal stem cells to differentiate into osteoblasts. How to obtain endogenous bone morphogenetic protein 2 protein is the focus of current research.
OBJECTIVE: To investigate osteogenic differentiation of rabbit bone marrow mesenchymal stem cells transfected by adenovirus with human bone morphogenetic protein 2 and its possible mechanism.
METHODS: The third passage of rabbit bone marrow mesenchymal stem cells were transfected by adenovirus with human bone morphogenetic protein 2 gene. At 48 hours after transfection, expression levels of bone morphogenetic protein 2 mRNA and protein in bone marrow mesenchymal stem cells were detected by qRT-PCR and western blot assay, respectively. The expression levels of proteins related to the Wnt/β-catenin signaling pathway were determined by western blot assay. After an induction with osteogenic medium, the alkaline phosphatase activity at 7 days, the type I collagen at 14 days, and the calcium nodules at 21 days were performed by alkaline phosphatase activity kit, immunohistochemical staining, and alizarin red S staining, respectively.
RESULTS AND CONCLUSION: (1) The expression of bone morphogenetic protein 2 mRNA and protein was significantly upregulated in Ad-BMP-2/EGFP transfected bone marrow mesenchymal stem cells. (2) Ad-BMP-2/EGFP induced alkaline phosphatase activity, promoted production of type I collagen and calcium nodules formation in rabbit bone marrow mesenchymal stem cells. The levels of β-catenin, cyclin D1, Runx2 and c-myc were upregulated in Ad-hBMP-2/EGFP transfected bone marrow mesenchymal stem cells, while the level of GSK3β significantly decreased. (3) Results indicated that Ad-BMP2-EGFP transfection promoted the differentiation of bone marrow mesenchymal stem cells into osteoblasts and its mechanism may be achieved by up-regulating the expression level of bone morphogenetic protein 2 protein to activate the Wnt/β-catenin signaling pathway.
Key words: adenovirus, bone marrow mesenchymal stem cells, bone morphogenetic protein 2, osteogenesis, Wnt signaling pathway