Electrical Stimulation Enhanced Mesenchymal Stem Cell Gene Expression for Orthopaedic Tissue Repair
Electrical Stimulation Enhanced Mesenchymal Stem Cell Gene Expression for Orthopaedic Tissue Repair
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DOI：
10.1166/jbt.2013.1081
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发表时间：
2013-04-01
影响因子：
0.1
通讯作者：
Cartmell, Sarah
中科院分区：
文献类型：
--
作者：
Balint, Richard;Cassidy, Nigel J.;Cartmell, Sarah
骨不愈合和骨大小缺损性疾病影响着全球数百万患者的生活质量。然而，缺乏合适的供体组织限制了传统骨移植治疗的范围。组织工程植入物具有克服这些问题的潜力，而电刺激已被证明可以促进体外培养的骨细胞的增殖、基因表达和细胞外基质的沉积。将同样的方法应用于间充质干细胞，可以为骨不连的治疗和骨组织工程提供宝贵的工具。在定制的生物反应器中培养的原代人间充质干细胞受到不同电压脉冲的刺激。用Alamar Blue染色检测细胞数量/活力，用Live/Dead染色检测细胞活力、形态和排列。检测11种分化标志物和血管内皮生长因子在生长和成骨培养上清液中的表达。在成骨培养液中，除CNN1水平显著降低外，基因表达没有受到影响，而在成骨介质中，刺激显著改变了ALPL、Acan、CNN1和GFAP的表达。在所应用的大多数电制度中，增殖都被降低了。本研究表明，直流电刺激对人骨髓间充质干细胞的分化和增殖均有影响。
Non-union fractures and large-size defects in bone affect the quality of life of millions of patients around the world. However, the lack of suitable donor tissue limits the scope of traditional bone graft treatments. Tissue engineered implants have the potential to overcome these problems, whilst electrical stimulation has been shown to enhance the proliferation, gene expression and ECM deposition of bone cells in vitro. Applying the same modality to Mesenchymal Stem Cells could provide an invaluable tool for the treatment of non-unions and bone tissue engineering. Primary human Mesenchymal Stem Cells cultured in custom-made bioreactors were stimulated with voltage pulses of varying regimes. Cell numbers/viability were measured using Alamar Blue staining, while viability, morphology and alignment were assessed using Live/Dead staining. The expression of 11 markers of differentiation and Vascular Endothelial Growth Factor was assayed in growth and osteogenic medium. Gene expression was unaffected in the growth medium, with the exception of a significant decrease of CNN1 levels whilst in the osteogenic medium, the stimulation significantly altered ALPL, ACAN, CNN1 and GFAP expressions. Proliferation was lowered in the majority of electrical regimes applied. This study demonstrates that direct electrical stimulation can influence both the differentiation and proliferation of human Mesenchymal Stem Cells.