МОРФОЛОГІЧНІ ЗМІНИ РОГІВКИ ПРИ НЕПРОНИКНІЙ МЕХАНІЧНІЙ ТРАВМІ ТА ЗА УМОВ КОРЕКЦІЇ КОРНЕРЕГЕЛЕМ І СТОВБУРОВИМИ КЛІТИНАМИ

This study presents the results of research on morphological changes in the cornea under the condition of non-penetrating mechanical injury and against the backdrop of correcting traumatic damage with Corneregel and stem cells. A mechanical injury was modeled by creating a concentric epithelial notch with a 7.0 mm trephine on the upper half of the cornea. Within this area, an ophthalmic scalpel was used to remove the epithelium along with the anterior stromal layer of the cornea (flap thickness up to 0.2 mm). Control of erosion recovery was conducted by staining the cornea with a 0.5 % fluorescein solution. The experimental model of corneal damage was reproduced on both eyes of a rabbit under local episcleral anesthesia with a 0.5 % solution of Alcaine and retrobulbar anesthesia with 1.0 ml of a 2 % lidocaine solution. Stem cells were obtained from the umbilical cord. The mesenchymal stem cells (MSCs) were immunophenotyped using the BD Accuri™ C6 Plus flow cytometer (Becton Dickinson, USA) with murine monoclonal antibodies against CD73, CD90, CD105, CD34, and CD45 (Invitrogen, USA). MSCs were characterized as CD73+, CD90+, CD105+, CD34-, CD45-. For the experiment, MSCs at the 10th passage were used, preparing a suspension with 4.35 million MSCs per 1 ml of sodium chloride saline per gram of Corneregel. For histological studies, corneal tissue samples were collected, fixed in 10% neutral buffered formalin, processed, and embedded in paraffin blocks following standard methodology. Sections 4-5 µm thick were prepared using a rotary microtome AMR-400 and stained with hematoxylin-eosin. Histological specimens were examined using a light optical microscope MICROmed SEO SCAN, and photo documentation was performed with a Vision CCD Camera. The results revealed that stem cells are more effective than Corneregel alone in stimulating regeneration of the anterior corneal epithelium, anterior limiting membrane, and improving the structural organization of the corneal stroma.