Офтальмохирургия (Jan 2020)
Clinical results and corneal endothelial cells migration assusment after Hemi-DMEK in rabbit eyes
Abstract
Purpose. To assess clinical results of partial Descemet’s membrane endothelial keratoplasty (hemi-DMEK) performed on the rabbit eyes with the focus on migration vector of transplanted corneal endothelial cells.Material and methods. The study was performed on 10 female Chinchilla rabbits (10 eyes). Corneo-scleral discs of enucleated eyes of male rabbits were used for preparing donor transplants of the Descemet’s membrane. Half of the dissected 8,0 mm Descemet’s membrane disc was transplanted. In order to determine the sex of the endothelial cells and to determine the cell migration vector, cytological (staining with azureeosin and akrikhin solution) and molecular genetic (polymerase chain reaction) methods were used.Results. Early postoperative period was characterized by diffuse corneal oedema and andmixed injection of the eye. Two rabbits died from natural causes during the experiment (from 7 to 30 days). 1 month after the procedure in 3 rabbits (3 eyes), complete resorption of corneal oedema was achieved. In these eyescentralcornea regained transparencyand was surrounded by a circular whitish cloudy area. In all of these 3 corneas, periphery (1,0-1,5 mm in width adjacent to the limbus) was also transparent. In one rabbit, the cornea was having the same pattern of circular cloudiness, but the central zone was graded as semi-transparent. In 4 rabbits (4 eyes) intensive, thick central opacity of the cornea was formed, corresponding to the zone of descemetorhexis. Molecular genetic analysis was performed on 3 eyes with transparent grafts. Analysis showed that amplification using primers SRYfor and SRYrev to gSRY, localized on the Y-chromosome in the region determining sex, did not give positive responses for any of the studied areas of the cornea (centre, periphery).Conclusion. For the first time, technical feasibility of performing hemi-DMEK in on the rabbit eyes was demonstrated. Full transparency of the central cornea was achieved in 37,5% (3 out of 8 eyes). Regenerative response of the cornea to the surgical procedure included the formation of circular opacity of the stroma surrounding the transparent central zone. We were not being able to identify endothelial cells of the donor (male rabbit) on the middle periphery of the recipient’s cornea (female rabbit) which allows us to make a reasonable assumption about the absence of their migration to the iatrogenically defected zone. Transparent status of a transplanted half of the Descemet’s membrane with endothelial monolayer was due to endothelial cell migration from the periphery to the centre of the cornea.
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