Synergistic effect of bone morphogenetic protein 2 and transforming growth factor beta2 on osteogenic differentiation of bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.1519

Abstract

Abstract:

BACKGROUND: It is revealed from the cell and gene level that bone morphogenetic protein 2 (BMP2) and transforming growth factor β2 (TGFβ2) synergistically promote the osteogenic differentiation of bone marrow mesenchymal stem cells, providing an experimental and theoretical basis for the bone regeneration.
OBJECTIVE: To investigate the overexpression of BMP2 and TGFβ2 in vitro to synergistically promote the osteogenic differentiation of bone marrow mesenchymal stem cells.
METHODS: Primary rat bone marrow mesenchymal stem cells were obtained by enzymatic digestion. Recombinant plasmids of BMP2 and TGFβ2 were packaged by lentivirus. The lentivirus carrying the recombinant plasmids of the target genes BMP2 and TGFβ2 was transfected into the third generation of bone marrow mesenchymal stem cells. MTS (cell proliferation assay) was used to detect the proliferation of bone marrow mesenchymal stem cells in different treatment groups at different time points. Quantitative PCR was used to detect the overexpression of BMP2 and TGFβ2 at mRNA levels. Immunohistochemistry of cell slides was performed to detect the expression level of osteogenic differentiation marker RUNX2 at the protein level under two-dimensional mixed culture conditions. The cell pellet obtained by three-dimensional culture in vitro was subjected to paraffin sectioning for Vonkossa staining (silver staining) to determine the positive region and expression intensity of osteogenic differentiation, and quantitative analysis was performed.
RESULTS AND CONCLUSION: (1) The proliferation of bone marrow mesenchymal stem cells transfected with BMP2 and TGFβ2 was not statistically different from that of untransfected cells (P > 0.05). (2) The BMP2 and TGFβ2 mRNA levels were significantly higher in the transfected cells than untransfected ones (P < 0.05). (3) RUNX2 immunohistochemistry under the in vitro two-dimensional culture conditions and Vonkossa staining under the in vitro three-dimensional culture showed that the BMP2 and TGFβ2 co-transfected bone marrow mesenchymal stem cells had stronger osteogenic ability than BMP2 or TGFβ2 alone transfected cells (P < 0.05). In conclusion, the overexpression of recombinant plasmids of BMP2 and TGFβ2 carried by lentivirus synergistically promotes the osteogenic differentiation of rat bone marrow mesenchymal stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cells, Bone Morphogenetic Proteins, Transforming Growth Factor beta2, Tissue Engineering

CLC Number:

Chang Xiaopeng, Chen Tao, Zhao Yin, Liang Ming. Synergistic effect of bone morphogenetic protein 2 and transforming growth factor beta2 on osteogenic differentiation of bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(1): 1-6.

Figures/Tables 5

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