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电刺激对常氧和低氧条件下的人软骨细胞和间充质干细胞的影响。

Effect of electric stimulation on human chondrocytes and mesenchymal stem cells under normoxia and hypoxia.

机构信息

Department of Orthopaedics, Biomechanics and Implant Technology Research Laboratory, Rostock University Medical Centre, D‑18057 Rostock, Germany.

出版信息

Mol Med Rep. 2018 Aug;18(2):2133-2141. doi: 10.3892/mmr.2018.9174. Epub 2018 Jun 15.

Abstract

During joint movement and mechanical loading, electric potentials occur within cartilage tissue guiding cell development and regeneration. Exposure of cartilage exogenous electric stimulation (ES) may imitate these endogenous electric fields and promote healing processes. Therefore, the present study investigated the influence of electric fields on human chondrocytes, mesenchymal stem cells and the co‑culture of the two. Human chondrocytes isolated from articular cartilage obtained post‑mortally and human mesenchymal stem cells derived from bone marrow (BM‑MSCs) were seeded onto a collagen‑based scaffold separately or as co‑culture. Following incubation with the growth factors over 3 days, ES was performed using titanium electrodes applying an alternating electric field (700 mV, 1 kHz). Cells were exposed to an electric field over 7 days under either hypoxic or normoxic culture conditions. Following this, metabolic activity was investigated and synthesis rates of extracellular matrix proteins were analyzed. ES did not influence metabolic activity of chondrocytes or BM‑MSCs. Gene expression analyses demonstrated that ES increased the expression of collagen type II mRNA and aggrecan mRNA in human chondrocytes under hypoxic culture conditions. Likewise, collagen type II synthesis was significantly increased following exposure to electric fields under hypoxia. BM‑MSCs and the co‑culture of chondrocytes and BM‑MSCs revealed a similar though weaker response regarding the expression of cartilage matrix proteins. The electrode setup may be a valuable tool to investigate the influence of ES on human chondrocytes and BM‑MSCs contributing to fundamental knowledge including future applications of ES in cartilage repair.

摘要

在关节运动和机械加载过程中,软骨组织内会产生引导细胞发育和再生的电势。外源性电刺激(ES)的暴露可能模仿这些内源性电场并促进愈合过程。因此,本研究调查了电场对人软骨细胞、间充质干细胞及其共培养物的影响。从死后获得的关节软骨中分离出人软骨细胞,从骨髓(BM-MSCs)中分离出人间充质干细胞,分别或共培养到胶原基支架上。在 3 天的时间里用生长因子孵育后,使用钛电极施加交变电场(700 mV,1 kHz)进行 ES。在缺氧或常氧培养条件下,细胞在电场中暴露 7 天。之后,研究了细胞的代谢活性并分析了细胞外基质蛋白的合成率。ES 并未影响软骨细胞或 BM-MSCs 的代谢活性。基因表达分析表明,在缺氧培养条件下,ES 增加了人软骨细胞中 II 型胶原 mRNA 和聚集蛋白聚糖 mRNA 的表达。同样,在缺氧条件下暴露于电场后,胶原 II 合成显著增加。BM-MSCs 及其软骨细胞和 BM-MSCs 的共培养物在软骨基质蛋白的表达方面表现出类似但较弱的反应。电极设置可能是一种有价值的工具,可用于研究 ES 对人软骨细胞和 BM-MSCs 的影响,有助于包括 ES 在软骨修复中的未来应用在内的基础知识。

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