BACKGROUND: At present, there is evidence that domestic porous tantalum has good biocompatibility and osteogenic properties, but the specific osteogenic mechanism and its effect on osteogenic factors are still unclear. OBJECTIVE: To observe the effects of domestic porous tantalum materials on the expression of collagen type I, tissue transglutaminase-2 and calcium-binding protein A4 in MG63 cells. METHODS: MG63 cells in logarithmic growth phase were inoculated onto 24-well plates and cultured in three groups: in blank group, conventional medium was added; in tantalum extract group, porous tantalum material extract was added; and in tantalum scaffold group, porous tantalum material and conventional medium were added. On 1, 3, 5, 7 and 9 days of culture, the cell proliferation of each group was detected by cell counting kit-8 method. On 5 days of culture, the levels of collagen type I, tissue transglutaminase-2 and calcium-binding protein A4 secreted by MG63 cells in each group were detected by ELISA. Western blot assay was used to detect the expression of three proteins in each group. RESULTS AND CONCLUSION: (1) With the prolongation of culture time, the number of cells in each group increased gradually. There was no difference in cell proliferation among the three groups at different time points (P> 0.05). (2) The secretory levels of collagen type I and tissue transglutaminase-2 in the tantalum scaffold group were significantly higher than those in the blank group and tantalum extract group (P < 0.05), while the secretion of collagen type I and tissue transglutaminase-2 in the tantalum extract group was significantly higher than that in the blank group (P < 0.05). The secretion of calcium-binding protein A4 in the tantalum scaffold group was significantly lower than that in the other two groups (P < 0.05). (3) The expression of collagen type I and tissue transglutaminase-2 protein in the tantalum scaffold group was significantly higher than that in the blank group and tantalum extract group (P < 0.05), while the expression of collagen type I and tissue transglutaminase-2 protein in the tantalum extract group was significantly higher than that in the blank group (P < 0.05). The expression of calcium-binding protein A4 in the tantalum scaffold group was significantly lower than that in the blank group and tantalum extract group (P < 0.05). To conclude, domestic porous tantalum materials could promote the secretion of collagen type I and tissue transglutaminase-2 by MG63 cells, and inhibit the secretion of calcium-binding protein A4.%背景:目前有证据已证实国产多孔钽具有良好的生物相容性及促成骨性能, 但具体的成骨机制及对成骨因子的影响尚不明确.目的:观察国产多孔钽材料对MG63细胞Ⅰ型胶原、谷氨酰转氨酶2及钙结合蛋白A4表达的影响.方法:将对数生长期的MG63细胞接种于24孔板, 分3组培养:空白组加入常规培养基, 钽浸提液组加入多孔钽材料浸提液, 钽支架组加入多孔钽材料与常规培养基.培养第1, 3, 5, 7, 9天, CCK-8法检测各组细胞增殖;培养第5天, 采用ELISA酶联免疫吸附法检测各组细胞分泌Ⅰ型胶原、谷氨酰转氨酶2和钙结合蛋白A4的水平, Western-blot法检测各组细胞中钙结合蛋白A4、Ⅰ型胶原、谷氨酰转氨酶2蛋白的表达.结果与结论: (1) 随着培养时间的延长, 各组细胞数量呈逐渐增加趋势, 3组不同时间点的细胞增殖比较无差异 (P> 0.05); (2) 钽支架组Ⅰ型胶原、谷氨酰转氨酶2分泌高于空白组、钽浸提液组 (P <0.05), 钽浸提液组Ⅰ型胶原、谷氨酰转氨酶2分泌高于空白组 (P <0.05);钽支架组钙结合蛋白A4分泌低于其他两组 (P <0.05); (3) 钽支架组中Ⅰ型胶原、谷氨酰转氨酶2蛋白表达均高于空白组、钽浸提液组 (P <0.05), 钽浸提液组中Ⅰ型胶原、谷氨酰转氨酶2蛋白表达高于空白组 (P <0.05);钽支架组钙结合蛋白A4蛋白表达均低于空白组、钽浸提液组 (P <0.05); (4) 结果表明, 国产多孔钽材料可促进MG63细胞分泌Ⅰ型胶原、谷氨酰转氨酶2, 抑制其分泌钙结合蛋白A4.
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