Hoxa9 silencing promotes tibial fracture healing by regulating osteogenic differentiation

doi:10.3969/j.issn.2095-4344.2919

Abstract

Abstract:

BACKGROUND: Studies have shown that Hoxa10 gene has a certain correlation with osteogenic differentiation after microRNA regulation. However, the expression and function of Hoxa9 in osteogenic differentiation and tibial fracture healing have not been reported.

OBJECTIVE: To preliminarily investigate the effects of Hoxa9 on osteogenic differentiation of bone marrow mesenchymal stem cells in vitro and on fracture healing in an in vivo model.

METHODS: In vitro experiment: In bone marrow mesenchymal stem cells cultured in osteogenic induction medium, Hoxa9 was inhibited by shHoxa9 or overexpressed by MSCV-Hoxa9. In vivo experiment: A tibial fracture model was constructed in rats, followed by intervention with Hoxa9 and shHoxa9.

RESULTS AND CONCLUSION: In the in vitro test, qRT-PCR and western blot results indicated that Hoxa9 overexpression could significantly inhibit the osteogenic differentiation of bone marrow mesenchymal stem cells (P < 0.05), downregulate osteogenesis-related gene markers osteocalcin, osteopontin, Runx2 and collagen type I at mRNA and protein levels (P < 0.05), and reduce the number of calcified nodules, osteogenic differentiation ability and alkaline phosphatase activity (P < 0.05). In contrast, transfection of BMSCs with silenced Hoxa9 gene could reverse the above indicators. In the in vivo test, histological staining results showed that Hoxa9 intervention significantly aggravated bone damage in rats, and no interstitial collagen fibers and cartilage in the callus were found. The rats intervened by shHoxa9 showed a significant increase in bone size and number, and inter-tissue collagen fibers and cartilage in callus were detected. Findings from qRT-PCR and western blot in the in vivo test showed that the mRNA and protein expression of osteocalcin, osteopontin, Runx2 and collagen type I were significantly down-regulated in the Hoxa9 group (P < 0.05), and up-regulated in the shHoxa9 group (P < 0.05). In summary, Hoxa9 silencing may promote osteogenic differentiation of bone marrow mesenchymal stem cells and thereby promote fracture healing.

Key words: Hoxa9, osteogenic differentiation, tibial fracture, fracture healing, cell proliferation, bone marrow mesenchymal stem cells, osteoblasts, osteogenic differentiation, animal model

CLC Number:

Zhang Chao, Li Xingyong, Ma Guifu, Pu Xingyu, Luo Wenyuan. Hoxa9 silencing promotes tibial fracture healing by regulating osteogenic differentiation[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(35): 5600-5606.

Figures/Tables 8

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