Development of corneal tissue substitutes

The cornea, a transparent, avascular, dome-shaped tissue at the eye's outermost layer, is vital for vision. Corneal transplantation, or keratoplasty, is the primary treatment for corneal diseases. However, the success of this method is significantly influenced by factors such as donor cornea health, donor age, and the risk of tissue rejection. Therefore, as presented in this thesis, the research in corneal tissue engineering is of utmost importance, offering potential solutions to these challenges.Within the scope of this thesis, corneal and limbal cells were isolated from human donor corneas using various methods. Characterization processes were conducted on the isolated primary cells to assess their suitability for corneal tissue engineering. Additionally, the impact of immortalization on primary corneal cells was evaluated, determining that immortalized passage 12 corneal cells are most suitable for corneal tissue engineering. Furthermore, the characterization of corneal cell lines confirmed their suitability for corneal tissue engineering studies.Corneal epithelial cells were cultured on transparent PGS membranes to mimic corneal layers and performed thorough characterization procedures. The obtained data strongly indicate the feasibility of mimicking epithelial and endothelial layers with PGS membrane and corneal cells, offering hope for future advancements in corneal tissue engineering.The thickest layer of the cornea, the stromal layer, was successfully mimicked using cross-linked RADA hydrogels containing PGS nanoparticles. The results from the 3D hydrogel culture of corneal keratocyte cells demonstrated the promise of the tissue scaffold created using this method, paving the way for potential future applications in corneal tissue engineering.