Effect of BMP-2 combined with strontium ranelate on osteogenic differentiation of rat bone marrow mesenchymal stem cells

doi:10.19438/j.cjoms.2024.05.003

Abstract

Abstract: PURPOSE: To study the effect of bone morphogenetic protein-2(BMP-2) combined with strontium ranelate (Sr) on osteogenic differentiation of rat bone mesenchymal stem cells(BMSCs), and explore the related mechanisms. METHODS: Log-phase BMSCs were taken and randomly divided into control group (without any induction solution treatment), BMP-2 group (0.1 mg/L BMP treatment), Sr group (5 mmol/L Sr treatment), BMP-2+Sr group (after treatment with 0.1 mg/L BMP for 6 days, changed the solution and added 5 mmol/L Sr for treatment). Cell proliferation was detected by MTT experiment. Alkaline phosphatase(ALP) activity was detected by nitrophenyl phosphate method. Mineralization was observed by alizarin red staining. Protein expression of transforming growth factor β1(TGF-β1), Smad2, p-Smad2, and Runt-related transcription factor 2 (Runx2) were detected by Western blot. SPSS 24.0 software package was used for data analysis. RESULTS: Compared with the control group, the ALP activity of BMP-2 group, Sr group, and BMP-2+Sr group were enhanced, of which MTT experimental absorbance(A) value, the percentage of mineralized area and the expression of TGF-β1 and Runx2 protein, p-Smad2/Smad2 were significntly increased(P<0.05). Compared with BMP-2 group and Sr group, the ALP activity of the BMP-2+Sr group were enhanced, of which MTT experimental A value, the percentage of mineralized area and the expression of TGF-β1 and Runx2 protein, p-Smad2/Smad2 were significantly increased(P<0.05). CONCLUSIONS: BMP-2 combined with Sr can promote the proliferation and osteogenic differentiation of BMSCs in rats, the mechanism of which may be related to the activation of TGF-β1/Smad signaling pathway.

Key words: Rat, Bone mesenchymal stem cells, Osteogenic differentiation, Bone morphogenetic protein-2, Strontium ranelate

CLC Number:

R336

YUAN Run-lin, LI Ya-ming. Effect of BMP-2 combined with strontium ranelate on osteogenic differentiation of rat bone marrow mesenchymal stem cells[J]. China Journal of Oral and Maxillofacial Surgery, 2024, 22(5): 430-434.

References

[1] Nakamura K, Koide M, Kobayashi Y, et al.Sclerostin deficiency effectively promotes bone morphogenetic protein-2-induced ectopic bone formation[J]. J Periodontal Res, 2023, 58(4): 769-779.
[2] Uijlenbroek HJJ, Lin X, Liu T, et al.Bone morphogenetic protein-2 incorporated calcium phosphate graft promotes peri-implant bone defect healing in dogs: a pilot study[J]. Clin Exp Dent Res, 2022, 8(5): 1092-1102.
[3] Hu T, Liu L, Lam RWM, et al.Bone marrow mesenchymal stem cells with low dose bone morphogenetic protein 2 enhances scaffold-based spinal fusion in a porcine model[J]. J Tissue Eng Regen Med, 2022, 16(1): 63-75.
[4] 俞灏, 刘燕, 杨翔文, 等. 雷奈酸锶对大鼠骨髓间充质干细胞成软骨分化的影响[J]. 上海口腔医学, 2022, 31(1):24-28.
Yu H, Liu Y, Yang XW, et al.Effect of strontium ranelate on chondrogenic differentiation of rat bone mesenchymal stem cells[J]. Shanghai Journal of Stomatology, 2022, 31(1): 24-28.
[5] Zhang H, Xu R, Li B, et al.LncRNA NEAT1 controls the lineage fates of BMSCs during skeletal aging by impairing mitochondrial function and pluripotency maintenance[J]. Cell Death Differ, 2022, 29(2): 351-365.
[6] Tang B, Chen Y, Zhao P, et al.MiR-601-induced BMSCs senescence accelerates steroid-induced osteonecrosis of the femoral head progression by targeting SIRT1[J]. Cell Mol Life Sci, 2023, 80(9): 261.
[7] Yu D, Li Z, Cao J, et al.microRNA-25-3p suppresses osteogenic differentiation of BMSCs in patients with osteoporosis by targeting ITGB3[J]. Acta Histochem, 2022, 124(6): 151926.
[8] Tong X, Chen J, Wang R, et al.The paracrine effect of hyaluronic acid-treated endothelial cells promotes BMP-2-mediated osteogenesis[J]. Bioengineering (Basel), 2023, 10(10): 1227.
[9] Huang K, Wu G, Zou J, et al.Combination therapy with BMP-2 and psoralen enhances fracture healing in ovariectomized mice[J]. Exp Ther Med, 2018, 16(3): 1655-1662.
[10] Miranda TS, Napimoga MH, De Franco L, et al.Strontium ranelate improves alveolar bone healing in estrogen-deficient rats[J]. J Periodontol, 2020, 91(11): 1465-1474.
[11] Wang D, Yan C, Zhou L, et al.Changes in BMP-2 expression and mechanical properties during treatment of rats with osteoporotic hindlimb fracture with strontium ranelate[J]. J Musculoskelet Neuronal Interact, 2020, 20(1): 136-141.
[12] Zhang J, Shi H, Zhang N, et al.Interleukin-4-loaded hydrogel scaffold regulates macrophages polarization to promote bone mesenchymal stem cells osteogenic differentiation via TGF-β1/Smad pathway for repair of bone defect[J]. Cell Prolif, 2020, 53(10): e12907.
[13] Ma L, Zhang Y, Wang C.Coaction of TGF-β1 and CDMP1 in BMSCs-induced laryngeal cartilage repair in rabbits[J]. J Mater Sci Mater Med, 2020, 31(12): 130.
[14] 黄晓丹, 吕辉珍, 靳思思, 等. 雷奈酸锶通过TGF-β1/Smad通路促进骨髓间充质干细胞向成骨细胞分化[J]. 中国病理生理杂志, 2013, 29(2): 302-302.
Huang XD, Lyu HZ, Jin SS, et al.Strontium ranelate promotes osteogenic differentiation of rat bone mesenchymal stem cells through TGF-β1/Smad signaling pathway[J]. Chinese Journal of Pathophysiology, 2013, 29(2): 302-302.
[15] 吴钰坤, 韩杰, 温帅波. 骨折愈合过程中Runx2基因的作用机制[J]. 中国组织工程研究, 2021, 25(14):148-153.
Wu YK, Han J, Wen SB.Mechanism of Runx2 gene in fracture healing[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(14): 148-153.
[16] Kim KM, Son HE, Min HY, et al.Vitexin enhances osteoblast differentiation through phosphorylation of Smad and expression of Runx2 at in vitro and ex vivo[J]. Mol Biol Rep, 2020, 47(11): 8809-8817.