The Effect of Bioregulators Isolated from Blood Serum and Cornea of the Bovin’s Eye on the Condition of Tissue and Cells of the Rabbit Corneas during Cultivation and Storage

Abstract

Objective: to study the condition of the cornea, as well as its epithelial and endothelial cells, while maintaining in vitro at various temperature conditions, under the influence of a number of factors, including bioregulators isolated from blood serum and cornea of the bovine, and epidermal growth factor.Methods. The study was carried out on rabbit corneas stored at temperatures of +4, –86 °C, as well as the cultivation of endothelial and epithelial cells isolated from the cornea after storage at these temperatures, followed by histological and immunohistochemical studies.Results. Storage of the cornea at +4 °C for 10 days leads to corneal edema and significantly reduces their transparency, both bioregulators partially prevent a decrease in the transparency of the cornea, while the endothelial layer lyses in groups with the addition of epidermal growth factor and corneal bioregulator; but remains in the cornea with the addition of a serum bioregulator. All three factors contribute to the preservation of the Bowman membrane. In the corneas stored at –86 °C on the 30th day, a preserved endothelial layer was observed, and the epithelium retained its multilayering in all groups with the addition of factors other than the control group. In the control samples, the epithelial layer partially exfoliated, the endothelial layer was almost completely lysed. Both bioregulators stimulated the proliferation of cells isolated from the native cornea and enhanced the action of the epidermal growth factor. Similar results were obtained on cells isolated from stored corneas for 2 weeks at –86 °C. In the case of combined use of the epidermal growth factor and bioregulators on the 30th day, the endothelial layer was mainly preserved, the Descemet’s membrane was not broken. In the control samples, the epithelium was mainly single-layered, partially exfoliated, and the endothelial layer was completely lysed.Conclusion. Storage of cornea during hypothermia (+4 °С) does not provide corneal viability for longer than 10 days. Storage under conditions of cryopreservation (–86 °C) ensures the viability of the cornea for 60 days. Adding bioregulators and an epidermal growth factor to the basic preservation medium allows one to obtain a structurally safe and viable cornea, while all cellular layers of the cornea, including the endothelial layer, are preserved and viable.