[1]王志平,柳峰,赵建华.脉冲电磁场对脱钙骨基质诱导人骨髓干细胞成骨分化的影响[J].陆军军医大学学报(原第三军医大学学报),2010,32(09):947-950.
 Wang Zhiping,Liu Feng,Zhao Jianhua.Effects of pulsed electromagnetic fields on osteogenic differentiation in demineralized bone matrix-induced human marrow stromal cells in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(09):947-950.
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脉冲电磁场对脱钙骨基质诱导人骨髓干细胞成骨分化的影响(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
32卷
期数:
2010年第09期
页码:
947-950
栏目:
论著
出版日期:
2010-05-15

文章信息/Info

Title:
Effects of pulsed electromagnetic fields on osteogenic differentiation in demineralized bone matrix-induced human marrow stromal cells in vitro
作者:
王志平柳峰赵建华
第三军医大学大坪医院野战外科研究所骨科
Author(s):
Wang Zhiping Liu Feng Zhao Jianhua
Department of Orthopaedics, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042, China
关键词:
脉冲电磁场脱钙骨基质人骨髓干细胞成骨分化
Keywords:
pulsed electromagnetic fields demineralized bone matrix human marrow stromal cells osteoblastic differentiation
分类号:
R322.71;R329.21;R339.57
文献标志码:
A
摘要:
目的   观察脉冲电磁场(pulsed electromagnetic fields, PEMF)对脱钙骨基质 (demineralized bone matrix, DBM)诱导人骨髓间充质干细胞(human mesenchymal stem cells, hMSCs)成骨分化的影响,探讨可能的发生机制。   方法   提取1例健康14周岁男性骨髓间充质干细胞,培养至第3代,以1×104/孔接种至8块24孔板,以1×105/孔接种至2块6孔板,各分成4组:细胞对照组(C组)、细胞+材料组(CD组)、细胞+脉冲电磁场组(CP组)和细胞+材料+脉冲电磁场组(CDP组)。CD组和CDP组加入一块5 mm×5 mm×3 mm大小的DBM,CP组和CDP组给予PEMF(频率15 Hz,场强5 Gs)照射;各组分别于种植后第1、7、14、21天进行碱性磷酸酶(ALP)活性、骨钙素(OC) 浓度等指标检测,在第21天进行钙结节茜素红染色,光镜观察。   结果   CD组、CP组和CDP组ALP活性及OC浓度在种植7 d后均明显升高(P<0.01);在14 d时CD组和CDP组ALP活性达到最高值,第7、14天时CDP组ALP活性较CD组、CP组均显著升高(P<0.01),仅在第14天时CD组ALP活性较CP组显著升高(P<0.01);CDP组OC值在7、14、21 d时均较CD组、CP组显著升高(P<0.01),仅在第21天时CD组OC浓度较CP组显著升高(P<0.01);21 d时形态学观察显示钙结节数量CDP组明显多于CD组和CP组(P<0.01),CD组明显多于CP组(P<0.01)。   结论   PEMF与DBM联合对hMSCs的成骨诱导要强于PEMF或DBM的单独作用,且随着磁场应用时间的延长更加明显。另外PEMF对DBM诱导hMSCs成骨分化具有明显的协同效应,可能是通过PEMF增强hMSCs对BMPs等成骨活性因子的反应性来实现的。
Abstract:
Objective   To investigate the effects of pulsed electromagnetic fields (PEMF) on osteogenic differentiation in demineralized bone matrix (DBM)induced human marrow stromal cells (hMSCs).    Methods   hMSCs were obtained from iliac crest marrow aspirates of a healthy boy aged 14. Then 1×104 hMSCs per well were plated in 8 24-well tissue culture plates, and 1×105 hMSCs per well in 2 6-well plates after they were expanded until passage 2. Those 24-well plates and 6-well plates were divided into 4 groups, cell control group (C group), cell-material group (CD group), cell-PEMF group (CP group) and cell-material-PEMF group (CDP group). CD group and CDP group were added with one DBM (size of 5 mm×5 mm×3 mm), CP and CDP groups were exposed to the PEMF (frequency of 15 Hz, intensity of 5 Gs), and hMSCs were cultivated in subculture medium with full media exchange in every 3 d. Alkaline phosphatase (ALP) activity and osteocalcin (OC) level were performed at 1, 7, 14 and 21 d, and alizarin red staining and counting of calcium nodules were performed at 21 d.    Results   Significant differences in ALP activity and OC level were observed between cell control group and the other 3 groups in 7 d after treatment (P<0.01). ALP activity of CD group was increased significantly than that of CP group only at 14 d (P<0.01), so did the OC level and calcium nodules only at 21 d (P<0.01). Compared with CD group and CP group, ALP activity in CDP group was increased significantly at 7 and 14 d (P<0.01), OC level in CDP group was increased significantly at 14 and 21 d (P<0.01), and the number of calcium nodules in CDP group was increased significantly at 21 d (P<0.01).    Conclusion   PEMF alone or together with DBM increases the expression of all osteoblast markers and the amount of mineralized matrix during the differentiation phase, but a combined treatment of DBM and PEMF is more effective than themselves alone. Also this effect was synergic on osteoblastic differentiation, suggesting that PEMF enhances the responsiveness of hMSCs to oteogenic factors such as BMPs.

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更新日期/Last Update: 2010-05-07