正弦交变电磁场促进大鼠骨髓间充质干细胞成骨性分化的研究  

Enhancement of Osteoblastic Differentiation of Bone Marrow Mesenchymal Stem Cells in Rats by Sinusoidal Electromagnetic Fields

作　　者：程国政[1] 陈克明[1] 李志锋[1] 周建[1] 韦哲[2] 白孟海[1] 赵宏斌[1] 

机构地区：[1]兰州军区兰州总医院骨科研究所,兰州730050 [2]兰州军区兰州总医院医学工程科,兰州730050

出　　处：《生物医学工程学杂志》2011年第4期683-688,共6页Journal of Biomedical Engineering

基　　金：甘肃省科技计划资助项目(092NKDA025)

摘　　要：研究50 Hz不同强度正弦交变电磁场（SEMFs）对体外培养大鼠骨髓间充质干细胞（rBMSCs）增殖与成骨性分化的影响,筛选出最佳磁场强度参数。采用贴壁筛选法培养原代rBMSCs,每天在频率为50 Hz,强度分别为0、1.4、1.6、1.8、2.0、2.2 mT的磁场环境中处理30 min。MTT法检测细胞增殖情况;于处理后的第3、6、9、12 d分别测定细胞碱性磷酸酶活性、钙盐沉积量、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6、12、24、48 h分别提取细胞总RNA,用RT Real-Time PCR法检测成骨性分化相关基因Osterix和IGF-1表达情况。实验结果显示电磁场干预组的细胞增殖率均低于对照组;但1.4~2.2 mT区间的磁场具有促成骨效应,以1.8 mT促进rBMSCs的成骨性分化更明显,表现在该组的碱性磷酸酶活性、钙盐沉积量、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）。由此可以认为频率为50 Hz、强度范围在1.4~2.2 mT内的SEMFs抑制rBMSCs的增殖;但促进其成骨性分化,以1.8 mT效果最为明显。The present research was aimed to investigate the effects of sinusoidal electromagnetic fields（SEMFs）on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells in rats（rBMSCs） and to find out the intensity with the best therapeutic efficacy.Primary rat bone marrow mesenchymal stem cells were obtained from Wistar rats and screened by the adhesive method.The rBMSCs were exposed to sinusoidal electromagnetic fields with 50Hz frequency and intensities of 0mT,1.4mT,1.6mT,1.8mT,2.0mT,and 2.2mT respectively,30min per day.The proliferation of the rBMSCs was analyzed by MTT reduction assay.The osteogenic differentiation markers including ALP activity,calcium deposition,mineralized bone modulus and collagenⅠexpression were compared between the rats in the exposed groups and those in the control group.The total cellular RNA was extracted after 6,12,24 and 48 hours,respectively.The gene expression of Osterix and IGF-1 was examined by quantitative reverse transcription-polymerase chain reaction（qRT-PCR）.The absorbance of exposed groups was suppressed significantly in comparison with that in the control group.The exposure to the rBMSCs with intensity of 1.8mT strongly enhances the osteogenic differentiation of rBMSCs,indicated by remarkably improved ALP activity,calcium deposition,collagenⅠexpression and the number of mineralized bone nodules compared to that in the control group and other groups.Osterix and IGF-1 were also significantly improved（P0.05）.The SEMFs with frequency and 50Hz and 1.4~2.2mT intensities enhanced the osteogenic differentiation of rBMSCs, but inhibited their proliferation in the presence of 0.1% serum culture.Among the rBMSCs used in the tests,the one with 1.8mT had the strongest activity,indicating that it could be the optimal intensity for the clinical application.

关 键 词：正弦交变电磁场 骨髓间充质干细胞 增殖 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]