In Vitro Construction And Identification Of Tissue-Engineered Human Anterior Hemi-Cornea Based On Bovine Collagen Composite Scaffold

Cornea is the anterior transparent part of the eyeball, acting as the protector of eyeball content. Corneal disease, injuries and infections may cause impaired vision or even blindness. In some cases, corneal epithelium and a portion of corneal stroma are injured and can only be cured by transplantation of anterior hemi-cornea nowadays.However, its severe shortage has caused blindness of those who have no access to healthy donated cornea. As the development of tissue engineering,stem cell technology and nanotechnology engineering,tissue-engineered human anterior hemi-comea(TE-aHC) constructed in vitro can repair injuries and resthre tissue functions. TE-aHC has been proved to be an effective equivalent.But lack of sufficient human comeal cells has hampered its development. Therefore, this study was intended to Construct a functional TE-aHC with human comeal stromal(HCS)cells and epithelial(HCEP) cells, as well as bovine collagen-chondroitin sulfate(BCOL-CS) composite scaffolds. Before the construction,morphological, functional and other properties of HCEP and HCS cells were proved to be similar to those in vivo. The BCOL-CS composite scaffolds exhibit tightly and orderly alignment of collagen fibers inside. To construct a TE-aHC, BCOL-CS composite scaffolds were folded and HCS cells mixed with fibrinogen and thrombin were distributed between each scaffold. HECP cells was seeded on the top of the uppermost composite scaffold and cultured fbr 5 days under air-liquid interface condition. The structural and functional properties of TE-aHC were characterized,which showed that the constructed TE-aHC had an inthgrated histhlogical structure similar to that of native anterior hemi-cornea,and the cells still maintained positive expression of marker and function proteins, indicating that the construethd TE-aHC posseses a native comea-like structure, property and fuction, with pothntial to be used as a promising anterior hemi-corneal equivalent.