低频电磁场对骨髓间充质干细胞增殖与分化成骨的影响研究

发布时间：2018-01-28 04:31

本文关键词： 电磁场骨髓间充质干细胞成骨　出处：《浙江大学》2012年硕士论文　论文类型：学位论文

【摘要】：实验目的：为了研究低频电磁场(EMF)对大鼠骨髓间充质干细胞(BMSCs)分化、增殖、细胞周期和体内作用的效果。实验方法：将原代的骨髓间充质干细胞培养至第三代,分成四个组别。辐照组细胞被置于0.5mT,50HZ的电磁场中；实验组细胞被置于含成骨诱导液的培养基中；联合组细胞被置于含成骨诱导液的培养基同时接受0.5mT,50HZ的电磁场的辐照。MTT法被用于估计细胞的增殖情况,碱性磷酸酶活性检测用于估计细胞的分化情况。分别在实验的第3天、第7天和第10天检测这两个指标。在第7天检测细胞周期的改变情况,在第12天检测骨结节的形成情况。此外,在第10天用RT-PCR检测Ⅰ型胶原的基因表达。辐照组和对照组的细胞分别被植入相同的PLGA支架后再分别植入到不同的大鼠股骨皮质中。分别在第4、第8周分别取材行病理切片并HE染色后观察成骨结果。实验结果：结果显示低频电磁场干预后加快了干细胞的增殖(p0.05),提高了干细胞的分化(p0.05),使G2/M+S期的细胞比例显著升高。与空白组相比,EMF辐照组有更显著的Ⅰ型胶原相关mRNA得以表达(p0.05)。EMF联合成骨诱导液组形成了明显的骨结节。在辐照组体内实验中,HE染色后观察到了明显的骨组织形成。结论：低频电磁场(0.5mT,50HZ)能显著促进骨髓间充质干细胞的增殖和分化。
[Abstract]:Aim: to study the effects of low frequency electromagnetic field (EMF) on differentiation, proliferation, cell cycle and in vivo action of rat bone marrow mesenchymal stem cells (BMSCs). Methods: primary bone marrow mesenchymal stem cells were cultured to the third generation and divided into four groups. The cells of the experimental group were placed in the medium containing osteogenic medium. The combined group of cells were placed in the medium containing osteoblast and exposed to the electromagnetic field of 0.5mTX 50HZ. MTT method was used to estimate the proliferation of the cells. Alkaline phosphatase activity was used to estimate cell differentiation. These two indexes were detected on day 3, day 7 and day 10, respectively. Changes in cell cycle were detected on day 7. The formation of bone nodules was detected on the 12th day. On the 10th day, the gene expression of type I collagen was detected by RT-PCR. The cells of irradiation group and control group were implanted with the same PLGA scaffold, and then implanted into different femur cortex of rats. 4. At the 8th week, the osteogenesis was observed by pathological sections and HE staining. The results showed that the low frequency electromagnetic field intervention accelerated the proliferation of stem cells and increased the differentiation of stem cells. The proportion of cells in G2 / MS phase was significantly increased compared with the blank group. The expression of type I collagen-associated mRNA in the EMF irradiation group was more significant, and the bone nodules were formed in the combined osteogenic inducer group. In vivo experiments were carried out in the irradiated group. The formation of bone tissue was observed after HE staining. Conclusion: low frequency electromagnetic field can significantly promote the proliferation and differentiation of bone marrow mesenchymal stem cells.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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