Mechanism of development and progression of chronic lung injury --- An approach from mechanical property of cultured lung fibroblasts -

我们研究了在纤维连接蛋白包被的硅橡胶膜上培养的肺成纤维细胞的压力-容积曲线。在培养皿的底部制造一个大孔，并在培养皿的外部粘贴一层硅树脂膜（厚度为20微米）。皿的底部用小牛纤连蛋白包被。使人胚肺成纤维细胞粘附于RBMI-1640培养基中的纤连蛋白包被的硅酮膜（细胞片）。通过注入和抽出培养基使细胞片层膨胀和收缩，同时保持圆顶。检测了圆顶的体积变化和圆顶内的压力。将获得的压力和体积值绘制在X-Y坐标上。在本研究中施加的应力范围内，硅胶膜本身在同一部位扩张和收缩。纤维连接蛋白包被的硅橡胶膜和细胞片的压力-容积曲线均呈滞后回线。显微镜检查显示在压力-体积偏移之前和之后没有细胞从薄片上脱离。用含EDTA的胰蛋白酶处理分离成纤维细胞的细胞片层的滞后回线似乎比胰蛋白酶处理前的细胞片层的滞后回线小。我们的结论是，肺成纤维细胞的机械性能可以用这种方法来测量。

We studied the pressure-volume curve of a cell-sheet consisted of lung fibroblasts cultured on fibronectin-coated silicone membrane. A large hole was made in the bottom of a culture dish and a silicone membrane(20micron in thickness)was pasted up on the outside of the dish. The bottom of the dish was coated with calf fibronectin. Human embryo lung fibroblasts were allowed to adhere to the fibronectin-coated silicone membrane in RBMI-1640 medium(cell-sheet). Cell-sheet was inflated and deflated with keeping a dome by injecting and withdrawing medium. Volume change of the dome and the pressure inside the dome were detected-. Obtained pressure and volume values were plotted on a X-Y coordinate. Silicone membrane itself expanded and contracted on the same locus within the stress applied in this study. Pressure-volume curve of fibronectin-coated silicone membrane and cell-sheet showed hysteresis loop. microscopical check showed no detachment of the cells from the sheet before and after pressure-volume excursion. The hysteresis loop of cell-sheet from which fibroblasts were detached by EDTA-containing trypsin treatment seemed to become smaller than that of cell-sheet before trypsin treatment. We conclude that mechanical property of lung fibroblasts may be able to be measured in this way.