New approaches to the study of the functional activity of corneal endothelial cells

    Abstract

    New approaches to the study of the functional activity of corneal endothelial cells

    Baturina G.S., Katkova L.E., Palchikova I.G., Solenov E.I., Iskakov I.A.

    The Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences
Technological Design Institute of Scientific Instrument Engineering of the Siberian Branch of the Russian Academy of Sciences
The academician S.N. Fyodorov Federal State Institution National Medical Research Center «Intersectoral Research and Technology Complex «Eye microsurgery» Ministry of Health of the Russian Federation», Novosibirsk Branch

    The safety of the functional activity of corneal endothelial cells is the key to good outcomes of corneal transplantation. Existing invasive and non-invasive methods for assessing the viability of the donor cornea are indirect. The development and implementation of direct methods for studying the transport functions of endothelial cells can improve the quality of the assessment of corneal safety preservation.

    Purpose. To determine the potential for diagnostics of corneal endothelial cells 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 the pig's cornea. Evaluation of the dynamics of cell volume was studied by a method based on fluorescence quenching effect of Calcein dye by cytoplasmic proteins. The content of intracellular sodium was investigated by fluorescence microscopy using sodium-specific dye.

    Results. Increasing of temperature from 25 to 37 ° С activates Na / K-ATPase and leads to a decrease in the volume of corneal endothelium cells (characteristic time 34.8±1.3 s , the rate of decrease -3×10-3±10- 5 s-1. The rate of decrease in the relative concentration of intracellular sodium in this case was -7×10-4±10-6 s-1, (n=5). The cold preservation of the cornea (6 days or more) significantly reduces the intensity of excretion of sodium from the cell when the temperature rises to 37° C.

    Conclusion. The study of the dynamics of the volume of endothelial cells and the content of sodium in endothelial cells during cold preservation allows improving the quality of assessment of the viability of a preserved cornea.

    Key words: corneal endothelium, cold preservation, intracellular sodium, cell volume.