低频正弦交变磁场磁力线方向对体外培养成骨细胞增殖与分化的影响  

Effect of Sinusoidal Electricity Magnetic Fields on the Proliferation and Differentiation of Osteoblasts in vitro

作　　者：王嘉琪[1] 葛宝丰[1] 周建[1] 韦哲[2] 韩桂秋[1] 朱瑞清[1] 陈克明[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工程医疗器械科,兰州730050

出　　处：《四川大学学报（医学版）》2012年第3期367-372,共6页Journal of Sichuan University(Medical Sciences)

基　　金：甘肃省科技重大专项项目(09ZNKDA025)资助

摘　　要：目的研究磁场强度1.8mT、频率50Hz,正弦交变磁场磁力线方向对体外培养大鼠成骨细胞(ratsosteoblasts,ROB)的影响。方法原代培养大鼠颅骨成骨细胞,传代后随机分为3组,对照组、平行组和垂直组,平行组和垂直组分别暴露在磁场中,磁场强度1.8mT、频率50Hz、磁力线方向分别与培养皿平行和垂直,30min/次/d。倒置相差显微镜下观察细胞形态;MTT法测定细胞增殖;第3d、6d、9d、12d测定碱性磷酸酶(alkalinephosphatase,ALP)活性和钙盐沉积量;第10d进行ALP组织化学染色,第12d进行茜素红钙化结节染色;在正弦交变磁场(sinusoidal electromagnetic fields,SEMFs)处理0h、24h、48h和96h后,Real-time RT-PCR检测ALP、骨形态发生蛋白-2(bone morphogenetic protein-2,BMP-2)和Osterix(OSX)mRNA表达平行变化。结果成骨细胞于磁场暴露处理3～4d后呈漩涡样分布;与对照组相比,磁场组均抑制细胞增殖(P<0.01或P<0.05),其中与垂直组比较平行组显著抑制细胞增殖;与对照组比较,磁场组均促进细胞分化成熟,表现为提高细胞的ALP活性,促进钙盐沉积量,增加钙化结节数量,提高ALP、BMP-2和OSX mRNA表达水平,尤以平行组最为明显。结论 1.8mT、50Hz,磁力线方向与培养皿平行和垂直放置的正弦交变磁场均抑制体外培养成骨细胞的增殖,但能促进其分化成熟,此作用与磁力线方向有关,尤以平行组促分化效果最为明显。Objective To investigate the effect of sinusoidal electricity magnetic fields（SEMFs） on the proliferation and differentiation of osteoblasts in vitro.Methods Calvarial osteoblasts of newborn rats were isolated by enzyme digestion and randomly divided into 3 groups after subculture.Two groups of cells were exposed to 50 Hz 1.8 mT SEMFs for 30 min/d in parallel and vertical,respectively.Those without SEMFs exposure served as control.The cells were observed under the contrast phase microscope each day.After 48 h,cell proliferation was assayed by MTT method.The alkaline phasphatase（ALP） activities and calcium contents were measured after 3,6,9,and 12 days.The ALP positive colonies were histochemically stained after 10 days and the calcified nodules were stained by Alizarin Bordeaux after 12 days.Expressions of ALP,bone morphogenetic protein-2（BMP-2） and Osterix（OSX） mRNA were measured at 0 h,24 h,48 h and 96 h.Results The cells exposed to the SEMFs were arranged in spiral appearance after 3 days.Compared with control,SEMFs inhibited cell proliferation（P0.01 or P0.05）,but enhanced the maturation and mineralization of the osteoblasts.The results showed that SEMFs improved ALP activities,promoted calcium contents,increased calcified nodulues numbers,boosted expressions of ALP,BMP-2 and OSX mRNA.SEMFs with magnetic lines of force in parallel has stronger activities than those in vertical.Conclusion The SEMFs at 1.8 mT and 50 Hz inhibit the proliferation of osteoblasts,but enhance the maturation and mineralization of osteoblasts.

关 键 词：成骨细胞 磁场 磁力线方向 增殖 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]