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2022, Vol. 26 ›› Issue (16): 2498-2503Effect of polymethyl methacrylate/mineralized collagen/Mg-Ca composite material on osteogenic differentiation of mouse preosteoblastsZhang Hongmei1, 2, Sun Xirao1, Wang Chengyue1
Abstract: BACKGROUND: The structure of mineralized collagen/Mg-Ca composite prepared by freeze-drying method is loose, and the structural stability of the composite can be enhanced by using polymethyl methacrylate bone cement as the outer strengthening layer. OBJECTIVE: To investigate the effects of polymethyl methacrylate/mineralized collagen/Mg-Ca composite on osteogenic differentiation of MC3T3-E1 cells. METHODS: The polymethyl methacrylate/mineralized collagen/Mg-Ca and polymethyl methacrylate/mineralized collagen composites were prepared respectively. The MC3T3-E1 cells were directly inoculated on the surface of polymethyl methacrylate/mineralized collagen/Mg-Ca composite (group A), polymethyl methacrylate/mineralized collagen composite (group B) and Mg-Ca alloy (group C). Cells cultured alone were blank controls (group D). The cell adhesion morphology on the material surface was observed under the scanning electron microscope. CCK-8, alkaline phosphatase activity, enzyme linked immunosorbent assay, RT-PCR, and western blot assay were used to analyze the effects of three materials on MC3T3-E1 cell proliferation and osteogenic differentiation. RESULTS AND CONCLUSION: (1) Scanning electron microscope showed that the largest adhesion number of MC3T3-E1 cells on the surface of polymethyl methacrylate/mineralized collagen/Mg-Ca composite, grew well, stretched obviously, pseudopodia extended more, and connected with each other to form a network structure. (2) CCK-8 assay showed that compared with the group D, the three materials could promote the proliferation of MC3T3-E1 cells (P < 0.05), and the composite had the most significant effect in group A (P < 0.05). (3) Compared with the group D, the three materials could promote the alkaline phosphatase activity in MC3T3-E1 cells (P < 0.05), and the group A was the most significant (P < 0.05). (4) Enzyme linked immunosorbent assay showed that the three materials could promote the secretion of osteocalcin and type I collagen in MC3T3-E1 cells, and the promotion effect of group A was the most significant (P < 0.05). (5) RT-PCR showed that the three materials could increase the mRNA expression of RUNX2, OSX, alkaline phosphatase and osteocalcin, especially in group A (P < 0.05). (6) Western blot assay showed that the three materials could increase the protein expression of RUNX2, OSX, alkaline phosphatase and osteocalcin, and the group A had the most significant increase (P < 0.05). (7) The results showed that polymethyl methacrylate/mineralized collagen/Mg-Ca composite could promote the expression of osteogenic differentiation related proteins and genes in MC3T3-E1 cells, and had a positive effect on the osteogenic differentiation. Key words:polymethyl methacrylate bone cement, mineralized collagen, osteogenic differentiation, magnesium calcium alloy, mouse preosteoblasts, transcription factor, osteocalcin, alkaline phosphatase |