探讨电磁场影响骨髓间充质干细胞增殖分化的时间依赖性  

The research on the time-dependent effect of electromagnetic fields on BMSCs proliferation and osteogenic differentiation

作　　者：宋明宇[1] 李峰[1] 杨勇[1] 虞冀哲[1] 易智谦[1] 刘阳[1] 吴华[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《骨科》2012年第3期113-116,144,共5页ORTHOPAEDICS

基　　金：国家自然科学基金(No.51077065);湖北省自然科学基金[No.2011CDC032]

摘　　要：目的探究正弦波电磁场诱导大鼠骨髓间充质干细胞(BMSCs)成骨方向分化、促进其增殖的时间依赖性。方法用差速贴壁法分离培养(bone marrow-derived mesenchymal stem cells,BMSCs),取用生长状态良好的第三代细胞进行实验。将细胞按照1×105个/孔接种于3.5 cm细胞培养皿中。将细胞培养皿按照完全随机分组的原则分为实验组与对照组,每组分别设有1 d组、4 d组、7 d组、10 d组、14 d组共五个亚组。将实验组置于15 Hz 1 mT正弦波电磁场中每天暴磁1 h,对照组在同条件无磁场的细胞培养箱中培养。暴磁结束后提取各组细胞总RNA,用荧光定量PCR方法检测比较各组中成骨指标:RUNX2、碱性磷酸酶(ALP)、骨唾液酸蛋白(BSP)、骨桥蛋白(OPN)基因相对表达量的变化趋势。另有一批细胞按照1×103个/孔接种于96孔细胞培养板中,每块细胞培养板设7个副孔,共接种10块细胞培养板。按照完全随机分组原则,随机分组为实验组与对照组,每组分别设有1 d组、4 d组、7 d组、10 d组、14 d组共五个亚组。将实验组置于15 Hz 1mT正弦波电磁场中每天暴磁1 h,对照组在同条件无磁场的细胞培养箱中培养。用CCK8法检测比较各组细胞增殖情况。结果电磁场作用骨髓间充质干细胞的早期阶段(1 d、4 d)可以诱导其向成骨方向分化,而在其作用的晚期阶段则抑制其进一步成骨方向分化;相应的电磁场作用初期抑制大鼠骨髓间充质干细胞增殖,电磁场作用后期可促进其增殖,尤其在7 d、10 d时效应明显。结论电磁场可以影响大鼠骨髓间充质干细胞增殖和成骨方向分化,并且这种影响具有时间依赖效应。Objective To explore the time-dependent effect of electromagnetic fields in vitro on BMSCs proliferation and osteogenic differentiation. Methods BMSCs were isolated and cultured from 4-week-old Sprague-Dawley rats （both sexes, weighing 80-120 g）. BMSCs （from the passage 3） were randomly divided into experimental group and control group. The rats in the experimental group were put into 15 Hz sinusoidal electromagnetic fields for presetting stimulation durations （one hour every day at 1st, 4th, 7th, 10th, and 14th day）, and those in the control group were put into the same environment without electromagnetic fields. The gen relative expression levels of RUNX2, alkaline phosphatase （ALP）, bone sialoprotein （BSP） and osteopontin （OPN） were detected by using real-time PCR. The proliferation of BMSCs was determined by mieroplate reader using CCK8 kit. Results In the early stages （1 st and 4th day） electromagnetic fields could induce BMSCs osteogenic differentiation to a certain extent, but further differentiation was inhibited in the late stages. Correspondingly the proliferation of BMSCs was inhibited by electromagnetic fields at the beginning, however, electromagnetic fields could promote the proliferation of BM- SCs in the later. Conclusion Electromagnetic fields can affect the proliferation and osteogenic differentiation of BMSCs in a time-dependent manner.

关 键 词：电磁场 骨髓祖代干细胞 成骨 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]