The Development And Application Of Tissue Engineering Corneal Epithelium Derived From The Plant

Purpose:Onion Epithelial Membrane(OEM)was used as the scaffold material for cultured corneal epithelial cells to explore the feasibility of plant-derived materials as tissue engineering corneal materials in this paper.Methods:The optical properties of the OEM are evaluated by a transparency experiment.The surface features and ultrastructure of OEM were observed by scanning electron microscopy and transmission electron microscopy.The cells were cultured with different types of cell lines and primary cells,their adherent growth status and cell morphology were compared with the culture dish.The isolated rabbit corneal epithelial cells were seeded onto OEM,OEM coated with Collagen? and PETM to observe the continuous growth status of RCEs at 3d,5d and 7d;and then the method of air-liquid interface culture was used to construct tissue engineered corneal epithelium.The ultrastructure of RCEs was observed by scanning electron microscopy.The histological structure of tissue engineering corneal epithelium was observed by HE staining,the expression of K3/K12,Cytokeratin and E-cadherin was detected by immunofluorescence staining.Results:Transparency experiments show that both wet and dry conditions have good optical transparency compared with PETM materials.The results of scanning electron microscopy showed that the surface of the OEM was smooth and no other substances,there was no changes observed before and after sterilization,the OEM coated with Collagen? was observed in the collagen epidermis.The cell lines and primary cells cultured with OEM were not significantly different from the cell morphology and growth status on the culture dish;There were cell morphology better,cell structure more stereotactic,cell connections clearer and more microvilli of RCEs with OEM;Tissue engineering corneal epithelium with OEM as carrier has a tightly organized histological structure;The expression of cell tight junction markers(?-catenin and E-cadherin)and corneal epithelial differentiation protein(K3/K12)was positive,it shows that OEM can form cell connections and maintain corneal epithelial phenotype.Conclusion:Tissue-engineered corneal epithelium based on OEM has a certain histological structure of corneal epithelium,it can maintain the normal differentiation of corneal epithelial cells and the formation of cell connections.OEM is expected to become a new carrier material of tissue engineering cornea.