CYTOKINE STATUS OF THE BLOOD SERUM OF RABBITS WITH MECHANICAL NON-PENETRATING CORNEA INJURY AND CORRECTION WITH THE USE OF XENORNEAL STROMA OBTAINED BY THE DECELLULARIZATION METHOD

Abstract

Introduction. Researchers in the field of ophthalmology have proven the important role of cytokines in the immunopathogenesis of eye diseases as active bioregulators of inflammatory and reparative processes.

The aim of the study – to establish the peculiarities of changes in the cytokine profile of the blood of rabbits in the dynamics of surgical correction with a keratoxenoimplant obtained by the decellularization method, under the condition of experimental mechanical non-penetrating corneal injury.

Research Methods. Corneal damage was reproduced on Chinchilla rabbits. In both eyes, under local epibulbar anesthesia with a 0.5 % alkaine solution and retrobulbar anesthesia with a 2 % lidocaine solution, a concentric epithelial incision was made in the upper half of the cornea with a trephine, within which the epithelium was removed together with the anterior layer of the corneal stroma with a disposable ophthalmic scalpel. The transplant material is the stroma of the pig xenocornea, obtained by the decellularization method, which was carried out by treating it with a 0.5 % solution of sodium dodecyl sulfate and, subsequently, with ultrasound followed by incubation in the presence of a 0.1 % papain solution. Determination of the concentration of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-4 (IL-4) and interleukin-6 (IL-6) was carried out by the immunoenzymatic method.

Results and Discussion. Our research indicates differences in the dynamics of cytokine levels in groups of animals that were simulated mechanical non-penetrating corneal injury and those that underwent surgical correction. In particular, the concentration of pro-inflammatory cytokines – TNF-α – increases when simulating mechanical trauma to the cornea. IL-1β. IL-6 with a maximum on the 3rd day of observation. By the 28th day, their concentration decreases, but is significantly higher than in animals without simulated pathology. The concentration of the anti-inflammatory cytokine IL-4 increases on the 3rd day with a further decrease until the 28th day. Surgical correction with closure of the defect using porcine xenocorneal stroma obtained by the decellularization method was accompanied by less pronounced changes in pro-inflammatory and anti-inflammatory cytokines, and they completely normalized by the end of the experiment.

Conclusion. The obtained results indicate a difference in the type of immune response after mechanical corneal injury: in animals that were simulated mechanical corneal injury and no surgical correction was performed, it was hyperreactive, and under the conditions of surgical correction using the corneal stroma obtained by the decellularization method, it was compensatory.