Selvamurugan Nagarajan, Kwok Sukyee, Vasilov Anatoliy, Jefcoat Stephen C, Partridge Nicola C
Department of Physiology and Biophysics, UMDNJ--Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey 08854, USA.
J Orthop Res. 2007 Sep;25(9):1213-20. doi: 10.1002/jor.20409.
Bone morphogenetic proteins (BMPs) strongly promote osteoblast differentiation. Pulsed electromagnetic fields (PEMFs) promote fracture healing in non-union fractures. In this study, we hypothesized that a combined BMP-2 and PEMF stimulation would augment bone formation to a greater degree than treatment with either single stimulus. BMP-2 maximally increased the proliferative activity of rat primary osteoblastic cells at 25 ng/ml concentration. Real-time reverse transcription-polymerase chain reaction (RT-PCR) showed that BMP-2 stimulated mRNA levels of alkaline phosphatase (ALP), alpha(1) (I) procollagen, and osteocalcin (OC) in the differentiation phase and only OC mRNA expression in the mineralization phase after 24-h treatment. Both BMP-2 and PEMF (Spinal-Stim) increased cell proliferation, which was additive when both agents were combined. PEMF alone or together with BMP-2 increased only ALP mRNA expression and only during the differentiation phase 24 h after one 4-h treatment. This effect was additive when both agents were combined. Continuous daily 4-h treatment with PEMF alone or together with BMP-2 increased expression of all three osteoblast marker genes during the differentiation phase and increased the mineralized matrix. This effect was additive when both agents were combined, suggesting that the two interventions may be working on different cellular pathways. Thus, a combined effect of BMP-2 and PEMF in vitro could be considered as groundwork for in vivo bone development that may support skeletal therapy.
骨形态发生蛋白(BMPs)能强力促进成骨细胞分化。脉冲电磁场(PEMFs)可促进骨不连骨折的愈合。在本研究中,我们假设BMP - 2与PEMF联合刺激比单一刺激治疗能在更大程度上增强骨形成。BMP - 2在浓度为25 ng/ml时能最大程度提高大鼠原代成骨细胞的增殖活性。实时逆转录 - 聚合酶链反应(RT - PCR)显示,BMP - 2在24小时处理后的分化阶段刺激碱性磷酸酶(ALP)、α1(I)前胶原和骨钙素(OC)的mRNA水平,在矿化阶段仅刺激OC mRNA表达。BMP - 2和PEMF(Spinal - Stim)均能增加细胞增殖,两者联合时具有相加作用。单独的PEMF或与BMP - 2联合使用仅在一次4小时处理后24小时的分化阶段增加ALP mRNA表达。两者联合时此效应具有相加性。单独使用PEMF或与BMP - 2联合进行连续每日4小时的处理,在分化阶段增加了所有三种成骨细胞标志物基因的表达,并增加了矿化基质。两者联合时此效应具有相加性,表明这两种干预可能作用于不同的细胞途径。因此,BMP - 2和PEMF在体外的联合效应可被视为体内骨发育的基础,这可能支持骨骼治疗。
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