Corneal endothelium functional activity in pigs
Heading: Ophtalmology Article type: Original article
Authors: Baturina G.S., Katkova L.E., Solenov E.I., Palchikova I.G., Iskakov I.A.
Organization: Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, Novosibirsk State University, S. Fyodorov Eye Microsurgery Federal State Institution, Technological Design Institute of Scientific Instrument Engineering of the Siberian branch of the Russian Academy of Sciences
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.
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