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目的:研究不同治疗时间正弦电磁场(50 Hz,1.8 mT)对体外培养大鼠骨髓间充质干细胞成骨性分化的影响,筛选出最佳临床治疗时间。方法:采用贴壁筛选法培养原代大鼠骨髓间充质干细胞,每天在频率为50 Hz,强度为1.8 mT的磁场环境中处理0.5 h、1.0 h、1.5 h、2.0 h和2.5 h;同时设立未经电磁场处理的细胞作为对照组。于处理后的第3 d、6 d、9 d和12 d分别测定细胞碱性磷酸酶活性、骨钙素分泌量、钙化结节数以及Ⅰ型胶原表达量,并比较各组间差异;于处理后12 h提取细胞总RNA,用RT Real-TimePCR法检测成骨性分化基因Osterix表达情况。结果:正弦电磁场干预1.0 h能明显促进骨髓间充质干细胞的成骨性分化,表现在该组的碱性磷酸酶活性、骨钙素分泌量、钙化结节数、I型胶原表达量以及成骨性分化基因的表达量最高,亦显著高于对照组(P<0.05)。结论:50Hz、1.8mT强度的正弦电磁场能促进骨髓间充质干细胞的成骨性分化,以作用1.0 h成骨效果最为明显。
Objective: To study the effect of sinusoidal electromagnetic fields (50 Hz, 1.8 mT) at different treatment time on osteogenic differentiation of cultured rat bone marrow mesenchymal stem cells in vitro and to screen the best clinical treatment time. METHODS: Primary rat bone marrow-derived mesenchymal stem cells were cultured with adherent screening method and treated for 0.5 h, 1.0 h, 1.5 h, 2.0 h and 2.5 h every day in a magnetic field with a frequency of 50 Hz and an intensity of 1.8 mT. Simultaneously Cells without EMF were set as control group. Cell alkaline phosphatase activity, osteocalcin secretion, calcified nodules and collagen type I expression were measured on the 3 d, 6 d, 9 d and 12 d after treatment, and the differences among groups were compared. The total RNA was extracted 12 h after treatment and Osterix expression was detected by RT Real-Time PCR. Results: Sinusoidal magnetic field intervention 1.0 h can significantly promote the osteogenic differentiation of bone marrow mesenchymal stem cells in the group showed alkaline phosphatase activity, osteocalcin secretion, calcified nodules, collagen type I expression and Osteogenic differentiation gene expression was highest, also significantly higher than the control group (P <0.05). Conclusion: Sinusoidal magnetic field of 50Hz and 1.8mT intensity can promote the osteogenic differentiation of MSCs. The effect of osteoblast at 1.0 h is the most obvious.