压力对体外培养的牛角膜内皮细胞的影响

目的观察压力对体外培养的牛角膜内皮细胞形态结构及细胞活力的影响。方法对体外培养的牛眼角膜内皮细胞施以2.67kPa、5.33kPa及8.00kPa的压力各6、12、24、36、48h,以不加压组作为对照组。用倒置相差显微镜及电镜观察细胞形态结构,以台盼蓝染色观察比较48h时各组细胞活力的差别,用单因素方差分析比较不同压力下阳性细胞率。结果当作用于细胞的压力为2.67kPa(1kPa=7.5mmHg),作用时间为6、12、24、36、48h时,与对照组比较,细胞形态结构均无明显变化;48h时台盼蓝计数与对照组比较差异无统计学意义(P=0.776)。当压力为5.33kPa,作用时间为24h时,细胞形态结构出现轻度的损伤,在36、48h时加重;48h时台盼蓝计数与对照组比较,差异有统计学意义(P=0.00)。当压力为8.00kPa时,6h时即出现明显细胞形态结构损害,随着压力和作用时间的进一步增加,细胞的损伤程度也进一步加重;48h时台盼蓝计数与对照组比较差异有统计学意义(P=0.000)。结论牛角膜内皮细胞在加压48h时只能承受2.67kPa的压力,当压力为5.33kPa及8.00kPa,施压48h时,将造成细胞...

The effects of pressure on cultured bovine corneal endothelial cells in vitro

Objective To explore whether the exposure of bovine corneal endothelial cells to pressure stress in vitro causes changes in cellular morphology and cell viability.Methods Cultured bovine corneal endothelial cells were incubated in a pressurized chamber at 2.67 kPa,5.33 kPa and 8.00 kPa for 6,12,24,36 and 48 h.Control cells were incubated at atmospheric pressure.The morpholo-gy of the cells was investigated using an inverted phase contrast microscope and electron microscope.The viability of the cells was assessed by measuring with Trypan blue staining in response to el-evated pressure for 48 h.The differences in Trypan blue staining of positive cells at different pressure conditions when compared at 48 h were evaluated by a one-way ANOVA.Results Cell morphology did not change following exposure to 2.67 kPa(1 kPa=7.5 mmHg)pressure for 6,12,24,36 and 48 h.There were no statistical differences in Trypan blue staining of positive cells for different pressure conditions at 2.67 kPa for 48 h(P=0.776).Cells subjected to 5.33 kPa for 24 h demonstrated cell damage characterized by morphological changes and the number of damaged cells further in-creased at 36 h and 48 h.A significant morphological difference was observed under 8.00 kPa pressure at 6 h and further increased in a time and pressure-dependent manner.There were statistical differences in Trypan blue staining of positive cells when pressure conditions were compared at 5.33 kPa and 8.00 kPa for 48 h(P= 0.000).Conclusion These observations demonstrate that the exter-nal pressure limit for bovine corneal endothelial cells is 2.67 kPa in vitro.They exhibit morphological changes as well as decreased cell viability following exposure to 5.33 kPa and 8.00 kPa up to 48 hours' duration.