Soft contact lenses as a basis for the cultivation of the fetal fibroblasts

Abstract

Severe damage and disease of the cornea, accompanied by a limbal stem cells deficiency, encourages ophthalmologists to look for alternative methods of treatment. The development of cellular and tissue engineering in ophthalmology based on the use of cultured corneal cells–fetal fi broblasts, which stimulates the processes of substitution and regeneration in various corneal injuries due to transplantation of cells grown in vitro from healthy tissues, is an actual problem of our time. The aim of the work was to study the possibility of using different on composition soft contact lenses as a basis for the cultivation of the fetal fibroblasts. Materials and methods: The source of fibroblast culture was fetal cells from the cornea of the eye of 8-week-old human embryos. Two types of soft contact lenses (Aqualan, Ukraine) were used for the substrate. The first group of lenses contained a hydrogel 87% and polyacrylamide–13%, second group is included hydrogel 87%, polyacrylamide 12% and 1% collagen (rat). The cultivation of fetal fibroblasts was carried out in accordance with international GMP standards. Cell proliferation was monitored using phase contrast microscopy. On the 5th day, some objects with cultured cells were fi xed in 10% formalin solution, stained with hematoxylin and eosin. Results: During cultivation, an increase in the number of cells by 100 or more times compared with the initial number was observed. As a result of cultivation on the first day in both groups, cells were attached to the inner surface of the lenses in the same way–from 40 min to 2 h. On the 5th day of cultivation in the first group of lenses containing 13% polyacrylamide, it was not possible to obtain a complete monolayer of fibroblasts. In the second group of hydrophilic lenses, which included 12% polyacrylamide and 1% collagen, a monolayer of fetal fibroblasts was obtained. It covered almost the entire area of the inner surface of the lens. Conclusions: When cultivating fibroblasts in vitro, it is possible to form a monolayer of fibroblasts on the inner surface of a soft contact lens containing 1% collagen. The possibility of culturing fibroblasts on contact lenses with subsequent transplantation of stem cells to the surface of the cornea can form a new direction in the treatment of severe corneal pathology accompanied by limbal stem cell deficiency.