Abstract:

Objective To investigate the morphological and functional of regulations under different pressures culture on rabbit corneal endothelial cells by pressure bionic culture system in vitro. Design Experimental study. Participants Rabbit corneal endothelial cells. Methods The rabbit corneal endothelial cells were divided into five groups: Group A: without pressure culture group, as a blank control group; Group B: normal pressure bionic culture group (15 mmHg); Group C: fluctuated pressures were 15 mmHg, 25 mmHg, 20 mmHg, 10 mmHg, each pressure was for 6 hours; Group D: the cells were exposed to 30 mmHg pressure in vitro; Group E: 50 mmHg pressure bionic culture group. The five groups were cultured for 24 h respectively. Cells origin was identified with neuronal-specific enolase immunoassay. Cellular change in the structure was observed with HE staining and electron microscopy (SEM) test. Cell activity was detected with flow cytometry. Main Outcome Measures Cell morphology and the survival rate of cell. Results NSE antibody of the primary corneal endothelial cells was positive without corneal epithelial cells and corneal stromal cells. Cells were hexagon in the majority, arranged closely and were rich in microvilli and nuclear chromatin in the normal pressure bionic culture group. Survival rate of cell in normal pressure bionic culture group, fluctuated pressures group, 30 mmHg group, 50 mmHg group was (98.16±0.45)%, (78.83±1.65)%, (70.2±3.54)%, (41.33±0.25)%, respectively (P=0.016). However, in other high pressures groups, corneal endothelial intercellular space increased and the cell activity was lower than that of Group B by HE staining and flow cytometry. Conclusions The experiment confirmed that cultivative rabbit corneal endothelial cells morphology and function were positive adjusted with the normal pressure bionic cultivation, but high pressures would cause damage to corneal endothelial cells, and the damage would serious with extended over time. (Ophthalmol CHN, 2017, 26: 56-60)
 

Key words: pressure, micro-environment, bionic culture, corneal endothelium