Park Sang-Hyug, Sim Woo Young, Park Sin Wook, Yang Sang Sik, Choi Byung Hyune, Park So Ra, Park Kwideok, Min Byoung-Hyun
Department of Biomedical Engineering, Ajou University, Wonchon-dong, Youngtong-gu, Suwon, Korea.
Tissue Eng. 2006 Nov;12(11):3107-17. doi: 10.1089/ten.2006.12.3107.
In this study, we present a biological micro-electromechanical system and its application to the chondrogenic differentiation of rabbit bone marrow-derived mesenchymal stem cells (MSCs). Actuated by an electromagnetic force, the micro cell exciter was designed to deliver a cyclic compressive load (CCL) with various magnitudes. Two major parts in the system are an actuator and a cartridge-type chamber. The former has a permanent magnet and coil, and the latter is equipped with 7 sample dishes and 7 metal caps. Mixed with a 2.4% alginate solution, the alginate/MSC layers were positioned in the sample dishes; the caps contained chondrogenic defined medium without transforming growth factor-beta (TGF-beta). Once powered, the actuator coil-derived electromagnetic force pulled the metal caps down, compressing the samples. The cyclic load was given at 1-Hz frequency for 10 min twice a day. Samples in the dishes without a cap served as a control. The samples were analyzed at 3, 5, and 7 days after stimulation for cell viability, biochemical assays, histologic features, immunohistochemistry, and gene expression of the chondrogenic markers. Applied to the alginate/MSC layer, the CCL system enhanced the synthesis of cartilage-specific matrix proteins and the chondrogenic markers, such as aggrecan, type II collagen, and Sox9. We found that the micromechanically exerted CCL by the cell exciter was very effective in enhancing the chondrogenic differentiation of MSCs, even without using exogenous TGF-beta.
在本研究中,我们展示了一种生物微机电系统及其在兔骨髓间充质干细胞(MSCs)软骨分化中的应用。微细胞刺激器由电磁力驱动,设计用于施加不同大小的循环压缩载荷(CCL)。该系统的两个主要部分是一个致动器和一个盒式腔室。前者有一个永磁体和线圈,后者配备有7个样品盘和7个金属盖。将藻酸盐/间充质干细胞层与2.4%的藻酸盐溶液混合后置于样品盘中;盖子中含有不含转化生长因子-β(TGF-β)的软骨形成限定培养基。一旦通电,致动器线圈产生的电磁力会将金属盖向下拉,从而压缩样品。循环载荷以1赫兹的频率每天施加两次,每次10分钟。没有盖子的样品盘中的样品作为对照。在刺激后的第3、5和7天对样品进行细胞活力、生化分析、组织学特征、免疫组织化学以及软骨形成标志物基因表达的分析。应用于藻酸盐/间充质干细胞层的CCL系统增强了软骨特异性基质蛋白和软骨形成标志物的合成,如聚集蛋白聚糖、II型胶原蛋白和Sox9。我们发现,即使不使用外源性TGF-β,细胞刺激器施加的微机械CCL在增强间充质干细胞的软骨分化方面也非常有效。