共培养条件下电磁场干预对大鼠成骨细胞及骨髓间充质干细胞成骨分化的影响  

The effects of electromagnetic fields on osteogenesis during co-culture of bone marrow mesenchymal stem cells with osteoblasts

作　　者：虞冀哲[1] 杨勇[1] 刘朝旭[1] 宋明宇[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：《中华物理医学与康复杂志》2013年第4期250-255,共6页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：国家自然科学基金面上项目（51077065）

摘　　要：目的观察共培养条件下电磁场干预对大鼠成骨细胞及骨髓间充质干细胞（MSCs）成骨定向分化的影响，并探讨电磁场促进成骨分化的相关机制。方法体外分离培养SD大鼠BMSCs及成骨细胞，将第三代成骨细胞与BMSCs通过transwell培养小室建立共培养系统。采用随机数字表法将共培养细胞分为共培养组及共培养暴磁组，另随机选择单细胞培养的BMSCs及成骨细胞纳入单细胞培养组。共培养暴磁组细胞每日给予电磁场刺激4h。于实验进行14d后随机提取各组细胞总RNA，采用荧光定量PCR技术检测各组细胞Runx2、Sp7、碱性磷酸酶、I型胶原、骨形态形成蛋白-2及骨钙素基因表达情况，并选用茜素红染色法检测各组细胞矿化钙结节形成情况。结果单细胞培养模式下BMSCs及成骨细胞其各项成骨相关基因表达水平均较低，而共培养模式下BMSCs及成骨细胞其各项成骨相关基因表达均呈现不同程度增强，并且以共培养暴磁组成骨细胞上述指标增强幅度较显著。通过茜素红染色发现，与单纯共培养组比较，共培养暴磁组细胞矿化钙结节数量明显增多。结论低频电磁场干预可显著促进共培养条件下BMSCs及成骨细胞成骨定向分化，其可能机制为电磁场刺激能促进骨形态形成蛋白-2表达，使其通过自分泌或旁分泌方式介导细胞间信号转导，从而诱导BMSCs向成骨细胞分化及成骨细胞进一步成熟分化。Objective To explore the effects of electromagnetic fields （EMFs） on osteogenesis during co-culture of bone mesenchymal stem cells （BMSCs） with osteoblasts in rats. Methods BMSCs and osteoblasts were isolated from Sprague-Dawley rats and cultured. Sub-cuhured osteoblasts and BMSCs were seeded in transwell cell-cuhure-ehamber polyester inserts to establish the co-culture system. The co-cultures were then ran- domly divided into a normal co-culture group and a group exposed to an EMF. Single-cultured BMSCs and osteo- blasts were set as a single Culture group. The EMF group was exposed to an EMF for 4 hours per day. On the 14th day, cell culture plates or inserts were randomly selected for total RNA extraction and measurement of the mRNA expression levels of Runt-related transcription factor 2, transcription factor 7, alkaline phosphatase, col- lagen type I, bone morphogenetic protein 2 （BMP-2） and bone gamma-carboxyglutamate protein （ Osteocalcin gene, OC gene） using real-time PCR assays. Cell culture dishes or inserts were also randomly chosen for Alizarin red staining to detect mineralized nodules. Results The level of osteogenic gene expression in single-cul tured BMSCs and osteoblasts was low, while it was much higher in the co-culture group. The level of gene expression in the EMF-exposed and co-cultured group was even higher. Alizarin red staining also showed that calcium mineralized modules had increased in the stimulated, co-cultured system compared with the unstimulated, co-cultured cells. Conclusion EMF exposure can promote osteogenic differentiation of BMSCs and osteoblasts when they are co-cultured. BMP2-mediated cellular interaction might play an important role in osteogenic differentiation induced by EMF exposure.

关 键 词：电磁场 成骨细胞 骨髓间充质干细胞 共培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]