EXPERIMENTAL STUDY OF THE INFLUENCE OF NEURAL CREST-DERIVED MULTIPOTENT STEM CELLS ON THE STATE OF THE VISUAL ANALYZER IN GLAUCOMA

Abstract

Development of new effective treatments for glaucomatous optic neuropathy is one of the most acute aspects of a modern ophthalmology. The aim of our study was to design adrenal-induced glaucoma (AIG) condition in rats, followed by evaluation of morphological changes in retina and effects of different cell administration modes using cultured adult neural crest-derived multipotent stem cells (NCMSCs) on retina restoration. Materials and methods: Modeling of AIG was performed on Wistar rats (males, 10-12 months) via intraperitoneal administration of 0.18% solution of adrenaline hydrotartrate in a dose ranging from 10-15 µg per 100 g (20 injections over 6 weeks). Measurement of intraocular pressure (IOP) was carried with use of applanation tonometer. Adult NC-MSCs were isolated from the bulge region of whisker follicle by explant methods. Adult NC-MSCs were expanded in vitro and then characterized by immunocytochemistry and directed differentiation assay into NC-derived cell types. NC-MSCs administration modes: i/v 5×106 cells per animal; retro- and parabulbar injections of 0.5×106 cells per animal. Retina and optic nerve histomorphometric analysis was done on H&E stained sections. Results: NC-MSCs have a phenotype of nestin+p75+Sox10+cytokeratin-. IOP in rats before modeling was 7-10 mmHg. During the injection IOP was increased in all experimental animals from 15 to 22 mmHg and remained increased after modeling. AIG modeling led to development of stable through 3 months glaucoma-characteristic dystrophic and degenerative changes. I/v administration reduced the swelling of the optic nerve, and all the retina layers, but didn’t restore retina structure. Parabulbar administration of NC-MSCs resulted in a reduction of swelling and restoration of layers’ structure, most pronounced in the ganglionic layer and inner plexiform layer. In the outer plexiform layer, outer nuclear layer and layer of rods and cones significant swelling remained. Retrobulbar administration of NC-MSCs decreased swelling and layers’ structure restoration was most pronounced. Compared with a model, the number of nuclei in the inner and outer nuclear layers was statistically increased, although it was lower than in intact animals. NC-MSCs administration induced positive changes for all cell delivery methods. Conclusion: Positive effect of the NC-MSCs administration was observed in terms of swelling regression, retina and optic nerve restoration. Retrobulbar mode had the most prominent effect on the recovery of ganglionic layer, restoration of the structure of inner and outer nuclear layers and layer of rods and cones. Further studies of mechanisms involved in retina recovery should be done.