Culturing and Characterization of Primary Corneal Epithelial Cells on PGS Films for Cornea Tissue Engineering

The cornea is the transparent, avascular, watch-glass-shaped tissue at the outermost part of the eye. It is effective in good vision as it allows the light coming into the eye to be transmitted to the lens. The cornea consists of five distinct layers, which are the epithelium, Bowman's membrane, stroma, Descemet's membrane, and endothelium. The epithelium layer is the outermost layer of the cornea and is in direct contact with the outer environment. It has important functions such as water intake from tear and protection of the cornea from external factors. The corneal epithelium layer consists of 5-7 layers of corneal epithelial cells connected by tight junctions. Thus, selectivity is ensured in the intake of water and minerals from outside. The limbal epithelial stem cells in the limbus layer, which are located next to the cornea differentiate, and form corneal epithelium. By this stem cell source, corneal epithelium layer can be regenerated. The corneal epithelium is located on a thin layer consisted of collagen fibrils called Bowman's membrane. Bowman's membrane is a cell-free corneal layer and does not regenerate. Due to reasons such as keratoplasty being the only treatment method for corneal diseases, limited donor for transplantation and high donor age, corneal tissue engineering studies are of great importance.<br/>In this study, it was aimed to mimic the corneal epithelium layer. Accordingly, corneal epithelial cells immortalized after being isolated from the human donor cornea were cultured on PGS films. The most suitable PGS films produced by different length pre-polymers were selected and the ability of the cells to show their properties on Bowman's membrane was examined. According to the MTS assay results, PGS membrane with longer pre-polymer chains provided better corneal epithelial viability. In addition to this, cytokeratin-3, which is the main corneal epithelial marker, was expressed in higher amounts in the cells cultured on PGS membrane produced with longer pre-polymers. This study presents a recent innovation for cornea tissue engineering and makes a significant contribution to the literature.<br/><br/><i>This work is funded by 2232 International Fellowship for Outstanding Researchers Program of TUBITAK (Grant number 118C371). The funder had no role in study design, data collection or analysis, the decision to publish, or the preparation of this abstract.</i>