Zhu Baoying, Zhou Jian, Gao Yuhai, Shi Wengui, Wei Zhenlong, Li Wenyuan, Wang Yuanyuan, Chen Keming
Institute of Orthopedic Research, Lanzhou General Hospital of PLA, Lanzhou 730050, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2017 May 25;46(6):585-592. doi: 10.3785/j.issn.1008-9292.2017.12.03.
To compare the effects of 50 Hz 1.8 mT sinusoidal magnetic field (SEMF) and 50 Hz 0.6 mT pulsed electromagnetic field(PEMF) on the maturation and mineralization of rat calvaria osteoblasts. Primary cultured rat calvarial osteoblasts were divided into 3 groups:blank control group, SEMF group and PEMF group. The rats in SEMT and PEMT groups were treated with 50 Hz 1.8 mT SEMF or 50 Hz 0.6 mT PEMF for 90 min/d, respectively. Western blotting and Real-time RT-PCR were used to detect the protein and mRNA expressions of Collagen-1, bone morphogenetic protein 2 (BMP-2), osterix (OSX) and Runt-associated transcription factor 2(Runx-2). The alkaline phosphatase(ALP) activity was detected by ALP test kits at d6 and d9 after treatment, and by ALP staining using azo coupling at d10 after treatment. The formation of calcium nodules was observed by alizarin red staining. Compared with blank control group, the protein and mRNA expressions of Collagen-1, BMP-2, OSX and Runx-2 in SEMT and PEMT groups were significantly increased ( <0.01 or <0.05); while the mRNA expressions of Collagen-1 and BMP-2 in PEMF group were significantly higher than those in SEMF group. After 6 days treatment, the activity of ALP in PEMF group was significantly higher than that in blank control group (<0.05), while such difference was not observed in SEMF group (>0.05); after 9 days treatment, the activities of ALP in both PEMF and SEMP groups were significantly higher than that in blank control group (all <0.05), but the difference between PEMF and SEMF groups was not significant (>0.05). After 10 days treatment, ALP staining was increased in both PEMF and SEMF groups compared with that in blank control group (all <0.01), and the stained area was bigger in PEMF group than that in SEMF group (<0.05). After 12 days treatment, calcium nodules were increased in PEMF and SEMF groups compared with that in blank control group (all <0.01), and more calcium nodules were observed in PEMF group than SEMF group (<0.05). Both 50 Hz 1.8 mT that in SEMF and 50 Hz 0.6 mT PEMF can promote the maturation and mineralization of osteoblasts, and the effect of PEMF is more marked.
比较50Hz 1.8mT正弦磁场（SEMF）和50Hz 0.6mT脉冲电磁场（PEMF）对大鼠颅骨成骨细胞成熟和矿化的影响。将原代培养的大鼠颅骨成骨细胞分为3组：空白对照组、SEMF组和PEMF组。SEMT组和PEMT组的大鼠分别用50Hz 1.8mT的SEMF或50Hz 0.6mT的PEMF处理，每天90分钟。采用蛋白质免疫印迹法（Western blotting）和实时荧光定量聚合酶链反应（Real-time RT-PCR）检测Ⅰ型胶原（Collagen-1）、骨形态发生蛋白2（BMP-2）、osterix（OSX）和Runt相关转录因子2（Runx-2）的蛋白和mRNA表达。处理后第6天和第9天，用碱性磷酸酶（ALP）检测试剂盒检测ALP活性，处理后第10天用偶氮偶联法进行ALP染色。用茜素红染色观察钙结节的形成。与空白对照组相比，SEMT组和PEMT组中Collagen-1、BMP-2、OSX和Runx-2的蛋白和mRNA表达均显著增加（P<0.01或P<0.05）；而PEMF组中Collagen-1和BMP-2的mRNA表达显著高于SEMF组。处理6天后，PEMF组的ALP活性显著高于空白对照组（P<0.05），而SEMF组未观察到这种差异（P>0.05）；处理9天后，PEMF组和SEMP组的ALP活性均显著高于空白对照组（均P<0.05），但PEMF组和SEMF组之间的差异不显著（P>0.05）。处理10天后，与空白对照组相比，PEMF组和SEMF组的ALP染色均增加（均P<0.01），且PEMF组的染色面积大于SEMF组（P<0.05）。处理12天后，与空白对照组相比，PEMF组和SEMF组的钙结节均增加（均P<0.01），且PEMF组观察到的钙结节比SEMF组更多（P<0.05）。50Hz 1.8mT的SEMF和50Hz 0.6mT的PEMF均可促进成骨细胞的成熟和矿化，且PEMF的作用更显著。
