Simvastatin promotes osteogenic differentiation of bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.1769

Abstract

Abstract:

BACKGROUND: In vitro studies have shown that simvastatin can stimulate osteogenic differentiation of bone marrow mesenchymal stem cells, but the mechanism is unclear. Recent studies have shown that the Hedgehog signaling pathway is crucial for osteogenic differentiation of bone marrow mesenchymal stem cells.

OBJECTIVE: In combination with Hedgehog pathway blocker (cyclopamine), to observe the effect of simvastatin on osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro.

METHODS: Eight 4-week-old Sprague-Dawley female rats, SPF grade, were used in this study. The rats were killed by cervical dislocation and removed of bilateral femur and tibia under sterilization conditions. The second generation of bone marrow mesenchymal stem cells was randomly divided into four groups: control group cultured with osteogenic induction medium; simvastatin group cultured in the induction medium containing 10^-7 mol/L simvastatin; simvastatin+cyclopamine group (combination group) cultured in complete culture medium containing 5 μmol/L cyclopamine for 2 hours and then cultured in the induction medium containing 10^-7 mol/L simvastatin; cyclopamine group cultured in the osteogenic induction medium containing 5 μmol/L cyclopamine. After 7 days of culture, alkaline phosphatase staining was used. The levels of type I collagen and osteocalcin were evaluated by immunofluorescence and western blot assay. Alizarin red S staining was performed at 28 days of culture.

RESULTS AND CONCLUSION: Compared with the control group, the simvastatin group had more alkaline phosphatase positive cells and calcium nodules, and higher type I collagen and osteocalcin fluorescence intensities (P < 0.05). All the measurement indexes except for osteocalcin fluorescence intensity and protein expression were significantly lower in the combination group than the simvastatin group (P < 0.05). All the measurement indexes in the cyclopamine group were significantly lower than those in the control group (P < 0.05). To conclude, simvastatin can promote osteogenic differentiation of bone marrow mesenchymal stem cells via Hedgehog signaling pathway that cannot be completely blocked by cyclopamine.

Key words: osteoporosis, bone marrow mesenchymal stem cells, simvastatin, cyclopamine, type I collagen, osteocalcin, osteogenic differentiation, Natural Science Foundation of Hebei Province

CLC Number:

Xing Lei, Gao Jingyuan, Wang Xiaoxu, Chen Yujie, Chi Bojing, Tian Faming.

Simvastatin promotes osteogenic differentiation of bone marrow mesenchymal stem cells [J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 3961-.

Figures/Tables 5

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