Hirano Yuji, Ishiguro Naoki, Sokabe Masahiro, Takigawa Masaharu, Naruse Keiji
Department of Orthopaedic Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan.
J Biotechnol. 2008 Jan 20;133(2):245-52. doi: 10.1016/j.jbiotec.2007.07.955. Epub 2007 Aug 11.
Tensile and compressive strains are commonly used in mechanobiological models. Here we report on the development of a novel three-dimensional cell-culture method, which allows both tensile and compressive loads to be applied. Preliminary results were obtained using HCS2/8 chondrocytic cells embedded in type I collagen gel. This construct was subjected to either 16% tension or 14% compression. Confocal laser scanning microscopy showed that both tension and compression caused significant cell deformation. The collagen gel-embedded HCS2/8 cells were subjected to static tension, dynamic tension, static compression or dynamic compression for 24h. Dynamic compression led to significantly decreased 5-bromo-2'-deoxyuridine incorporation compared with the control group. PCR analysis revealed upregulation of type II collagen caused by dynamic tension, upregulation of aggrecan caused by static compression, and downregulation of type II collagen and aggrecan caused by dynamic compression. Nitric oxide production was significantly increased by static tension and static compression compared with the control group. Our experimental system effectively applied several types of strain to HCS2/8 cells embedded in collagen gel. Our results suggest that the mode of mechanical strain affects the response of HCS2/8 cells.
拉伸应变和压缩应变常用于力学生物学模型。在此,我们报告一种新型三维细胞培养方法的开发,该方法能够施加拉伸和压缩载荷。使用包埋于I型胶原凝胶中的HCS2/8软骨细胞获得了初步结果。该构建体分别承受16%的拉伸或14%的压缩。共聚焦激光扫描显微镜显示,拉伸和压缩均导致细胞显著变形。将包埋于胶原凝胶中的HCS2/8细胞进行静态拉伸、动态拉伸、静态压缩或动态压缩处理24小时。与对照组相比,动态压缩导致5-溴-2'-脱氧尿苷掺入显著减少。PCR分析显示,动态拉伸导致II型胶原上调,静态压缩导致聚集蛋白聚糖上调,动态压缩导致II型胶原和聚集蛋白聚糖下调。与对照组相比,静态拉伸和静态压缩使一氧化氮生成显著增加。我们的实验系统有效地对包埋于胶原凝胶中的HCS2/8细胞施加了几种类型的应变。我们的结果表明,机械应变模式会影响HCS2/8细胞的反应。