低频脉冲电磁场在三维支架下对人脂肪源干细胞增殖和成骨分化的影响
发布时间：2018-11-08 11:49
【摘要】：背景:目前已有较多实验研究证实低频脉冲电磁场在二维支架下促进多种干细胞向成骨细胞分化,但鲜有研究磁场在三维支架下对干细胞的增殖及成骨分化的影响。目的:探讨在聚已内酯3D培养支架环境下,外加低频脉冲电磁场对人脂肪源干细胞增殖及成骨分化的影响。方法:将第3代人脂肪源干细胞接种于聚已内酯3D培养支架上,分2组培养,实验组给予50 Hz、1 m T的低频脉冲电磁场干预,2 h/d,连续干预14 d;对照组不给于低频脉冲电磁场干预。培养7 d后,Live/Dead染色观察细胞存活情况;培养1,3,5,7 d后,MTT法检测细胞增殖;培养7,14 d,碱性磷酸酶染色及qR T-PCR检测细胞成骨分化水平。结果与结论:(1)Live/Dead染色:人脂肪源干细胞已长入到支架材料的空隙结构内部,在材料中可保持较高的活性;(2)细胞增殖:随着培养时间的延长,两组细胞A值均逐渐升高,实验组培养1,3 d的细胞A值高于对照组(P0.05);(3)碱性磷酸酶染色:实验组培养7,14 d的碱性磷酸酶阳性染色强于对照组;(4)qR T-PCR检测:培养7 d时,实验组碱性磷酸酶、Ⅰ型胶原基因表达量高于对照组(P0.01);培养14 d时,实验组骨桥蛋白、Runt相关转录因子2、碱性磷酸酶及Ⅰ型胶原的表达量均明显高于对照组(P0.05);(5)结果表明:在聚已内酯3D培养支架环境下,外加低频脉冲电磁场可促进人脂肪源干细胞的增殖及成骨分化。
[Abstract]:Background: at present, many experimental studies have confirmed that low-frequency pulsed electromagnetic fields can promote the differentiation of stem cells into osteoblasts under two-dimensional scaffolds, but little has been done on the effects of magnetic field on the proliferation and osteogenic differentiation of stem cells under three-dimensional scaffolds. Aim: to investigate the effects of low frequency pulsed electromagnetic field on proliferation and osteogenic differentiation of human adipose-derived stem cells in polycaprolactone 3D culture scaffold. Methods: the third generation of human adipose derived stem cells were inoculated on polycaprolactone 3D culture scaffold and cultured in two groups. The experimental group was treated with 50 Hz,1 Mt low frequency pulsed electromagnetic field for 2 h / d for 14 days. The control group was not treated with low frequency pulsed electromagnetic field. After 7 days of culture, the survival of cells was observed by Live/Dead staining, the proliferation of cells was detected by MTT assay, and the osteogenic differentiation level of cells was detected by alkaline phosphatase staining and qR T-PCR at 714 days after culture. Results and conclusions: (1) Live/Dead staining: human adipose-derived stem cells (HSCs) had grown into the interstitial structure of the scaffold and could maintain high activity in the scaffold. (2) Cell proliferation: with the extension of culture time, the A value of the two groups increased gradually, and the A value of the experimental group was higher than that of the control group (P0.05). (3) Alkaline phosphatase staining: the alkaline phosphatase positive staining in the experimental group was stronger than that in the control group for 714 days. (4) qR T-PCR detection: after 7 days of culture, the expression of alkaline phosphatase and collagen 鈪，
本文编号：2318418
[Abstract]:Background: at present, many experimental studies have confirmed that low-frequency pulsed electromagnetic fields can promote the differentiation of stem cells into osteoblasts under two-dimensional scaffolds, but little has been done on the effects of magnetic field on the proliferation and osteogenic differentiation of stem cells under three-dimensional scaffolds. Aim: to investigate the effects of low frequency pulsed electromagnetic field on proliferation and osteogenic differentiation of human adipose-derived stem cells in polycaprolactone 3D culture scaffold. Methods: the third generation of human adipose derived stem cells were inoculated on polycaprolactone 3D culture scaffold and cultured in two groups. The experimental group was treated with 50 Hz,1 Mt low frequency pulsed electromagnetic field for 2 h / d for 14 days. The control group was not treated with low frequency pulsed electromagnetic field. After 7 days of culture, the survival of cells was observed by Live/Dead staining, the proliferation of cells was detected by MTT assay, and the osteogenic differentiation level of cells was detected by alkaline phosphatase staining and qR T-PCR at 714 days after culture. Results and conclusions: (1) Live/Dead staining: human adipose-derived stem cells (HSCs) had grown into the interstitial structure of the scaffold and could maintain high activity in the scaffold. (2) Cell proliferation: with the extension of culture time, the A value of the two groups increased gradually, and the A value of the experimental group was higher than that of the control group (P0.05). (3) Alkaline phosphatase staining: the alkaline phosphatase positive staining in the experimental group was stronger than that in the control group for 714 days. (4) qR T-PCR detection: after 7 days of culture, the expression of alkaline phosphatase and collagen 鈪，
本文编号：2318418
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