Corneal endothelium functional activity in pigs

The purpose of the study is to determine the potential for diagnostics of human corneal endothelial cells functional activity based on a study of the transport of water and sodium ions by the endothelial cells of a pig's cornea. Material and Methods. The study was conducted on the disks of pig's cornea. Evaluation of cell volume dynamics was studied with a method based on fluorescence quenching effect of Calcein dye by cytoplasmic proteins. The intracellular sodium concentration was investigated by fluorescence microscopy method using sodium-specific dye Sodium Green. Micro-fluidic flow chamber was used to study the dynamics of corneal matrix deswelling. Results. Increase of temperature from 20 to 37° С activates Na/K-ATPase and leads to a decrease in the volume of corneal endothelium cells (characteristic time 73.5±12.6 s) (n=6). The relative concentration of intracellular sodium decreased with the characteristic time 130.0±26.3 s (n=5). Studies with temperature activation of endothelial cell pumping activity allowed us to estimate the initial rate ofthe matrix volume reduction: 0.36±0.006 um/s (n=5). Conclusion. The study of the dynamics of endothelial cells' volume and intracellular content of sodium in it during cold preservation allows to improve the quality of assessment of viability of a preserved cornea.