Wang Yu, Cui Haitao, Wu Zhenxu, Wu Naipeng, Wang Zongliang, Chen Xuesi, Wei Yen, Zhang Peibiao
Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, People's Republic of China.
University of Chinese Academy of Sciences, Beijing, People's Republic of China.
PLoS One. 2016 May 5;11(5):e0154924. doi: 10.1371/journal.pone.0154924. eCollection 2016.
Electrical stimulation (ES) is therapeutic to many bone diseases, from promoting fracture regeneration to orthopedic intervention. The application of ES offers substantial therapeutic potential, while optimal ES parameters and the underlying mechanisms responsible for the positive clinical impact are poorly understood. In this study, we assembled an ES cell culture and monitoring device. Mc-3T3-E1 cells were subjected to different frequency to investigate the effect of osteogenesis. Cell proliferation, DNA synthesis, the mRNA levels of osteosis-related genes, the activity of alkaline phosphatase (ALP), and intracellular concentration of Ca2+ were thoroughly evaluated. We found that 100 Hz could up-regulate the mRNA levels of collagen I, collagen II and Runx2. On the contrary, ES could down-regulate the mRNA levels of osteopontin (OPN). ALP activity assay and Fast Blue RR salt stain showed that 100 Hz could accelerate cells differentiation. Compared to the control group, 100 Hz could promote cell proliferation. Furthermore, 1 Hz to 10 Hz could improve calcium deposition in the intracellular matrix. Overall, these results indicate that 100Hz ES exhibits superior potentialities in osteogenesis, which should be beneficial for the clinical applications of ES for the treatment of bone diseases.
电刺激(ES)对许多骨疾病具有治疗作用,从促进骨折再生到骨科干预。ES的应用具有巨大的治疗潜力,然而,最佳的ES参数以及产生积极临床影响的潜在机制仍知之甚少。在本研究中,我们组装了一种ES细胞培养和监测装置。将MC-3T3-E1细胞置于不同频率下,以研究其对成骨作用的影响。对细胞增殖、DNA合成、骨相关基因的mRNA水平、碱性磷酸酶(ALP)活性以及细胞内Ca2+浓度进行了全面评估。我们发现100Hz可上调I型胶原、II型胶原和Runx2的mRNA水平。相反,ES可下调骨桥蛋白(OPN)的mRNA水平。ALP活性测定和固蓝RR盐染色表明,100Hz可加速细胞分化。与对照组相比,100Hz可促进细胞增殖。此外,1Hz至10Hz可改善细胞内基质中的钙沉积。总体而言,这些结果表明100Hz的ES在成骨方面具有卓越的潜力,这将有利于ES在骨疾病治疗中的临床应用。