На сегодня известно, что биодеградируемые конструкции на основе фиброина шелка могут использоваться в качестве безопасного носителя биологически активных веществ и лекарственных препаратов. При этом последние исследования показали, что глиальный нейротрофический фактор является одним из необходимых факторов роста, участвующих в восстановлении роговичной ткани. Однако не изучен способ восстановления роговичных повреждений при помощи биодеградируемых конструкций на основе фиброина шелка, содержащих глиальный нейротрофический фактор, что объяснило цель данного исследования. Цель: изучить особенности восстановления роговичных повреждений при помощи биодеградируемых конструкций на основе фиброина шелка, содержащих глиальный нейротрофический фактор. Материалы и методы: Исследование проводилось на половозрелых самцах мышей линии C57BL/6. У животных формировали модель повреждения роговицы. В ходе эксперимента у мышей на 1-е, 3-и и 5-е сутки после повреждения проводили анализ площади эпителиального дефекта роговицы для оценки интенсивности регенеративного процесса. Результаты: установлено, что интенсивность регенеративного процесса была выше в группах мышей с применением биодеградируемых конструкций на основе фиброина шелка, содержащих глиальный нейротрофический фактор, по сравнению с контрольной группой. Выводы: Применение биодеградируемых конструкций на основе фиброина шелка, содержащих глиальный нейротрофический фактор, стимулирует процесс восстановления роговичной ткани после моделированного повреждения. Background: Today it is well known that biodegradable materials based on silk fibroin can be used as safe carriers of biologically active substances and drugs. At the same time, recent studies have shown that glial neurotrophic factor is one of the essential growth factors involved in corneal tissue regeneration. However, the method of restoring corneal damage with biodegradable materials based on silk fibroin containing glial neurotrophic factor has not been developed. Aim: To study pathogenetic features of corneal damage repair with biodegradable structures based on silk fibroin containing glial neurotrophic factor. Materials and Methods: The study was conducted on mature male C57BL/6 mice, in which corneal damage was modeled. During the experiment, the area of the epithelial corneal defect was assessed for the intensity of the regenerative process on days 1, 3, and 5 after injury. Results: In the groups of mice where biodegradable structures based on silk fibroin containing glial neurotrophic factor were used, the intensity of the regenerative process was higher compared to the control group (p < 0.05). Conclusion: The use of biodegradable structures based on silk fibroin containing glial neurotrophic factor stimulates the process of corneal tissue regeneration after corneal damage.

Corneal injury due to ocular trauma or infection is one of the most challenging vision impairing pathologies. The aim of the work was to study the effect of biodegradable silk fibroin-based scaffolds containing GDNF on the corneal regeneration process. During cultivate the highest keratocytes proliferative activity was registered with scaffolds containing 250 ng/ml and 500 ng/ml GDNF. In mice with an experimental model of epithelial-stromal damage to the cornea, silk fibroin-based scaffolds containing GDNF in various concentrations were used (in groups 1, 2 and 3 silk fibroin-based scaffolds containing GDNF in a concentration of 50 ng/ml, 250 ng/ml and 500 ng/ml, respectively; in group 4 - silk fibroin-based scaffolds without GDNF; in group 5 - a solution of GDNF with concentration of 500 ng/ml; group 6- control). The area of the corneal epithelial defect in groups 2, 3, and 5 was less than in the other groups. The most pronounced positive immunohistochemical reaction with antibodies to Bcl2, Bax, phosphoERK1/2 and phospho-JNK1/2, Ki67, Gap43 was observed in groups 2 and 3. Thus, silk fibroin-based scaffolds with GDNF stimulate the epithelialization process, proliferative activity of epithelial cells and keratocytes, accelerate the formation of the stromal nerve plexus and exhibit anti-apoptotic activity.

Objective is to study the effect of modified glial-derived neurotrophic factor (GDNF) on healing of epithelial and epithelial-stromal corneal lesions in mice C57BL / 6J. After corneal damage the instillations of supernatant conditioned by HEK293 cells expressed GDNF gene construction without pre- and pro- sequences were produced. For control, a medium conditioned by not transfected cells was used. We assessed the area of corneal epithelial defect and corneal erosion rate, developing after the epithelial defect. The immunohistochemical study using antibodies against cytokeratin 5/18, c-Met, collagen IV, phospho-ERK1/2, phospho-JNK1/2, Ki67, Bcl2, GAP43, TIMR-1, TGF-p, Bax, and MMP 9 was performed. The area of corneal epithelial defect in the eyes of experimental animals within one day after damage was smaller than in the control. Frequencies of corneal erosions formed in the eyes of experimental animals after damage was observed in 30-35% and 80-85% of cases in the experimental and control groups, respectively. Immunohistochemical studies using these antibodies showed that GDNF stimulated the proliferative activity of epithelial cells and keratinocytes, contributed to active migration and adhesion of epithelial cells, had anti-apoptotic and antifibrotic effects, took an active part in the formation of stromal nerve plexus. The results indicate the hopefulness of therapeutic application of the modified GDNF after corneal injury and the need for further research to develop and test methods for the therapeutic use of drugs on the basis of this neurotrophic factor.