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Lung-Kun Yeh, Shih-Chun Huang, Hsin-Ta Wang; The Cell Fade Of Cultured Corneal Endothelial Cells On The Genipin-crosslinking Descemet Membrane. Invest. Ophthalmol. Vis. Sci. 2012;53(14):1525.
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Genipin, a nature crosslinking material extracted from the fruits of Gardenia jasminoides, has been widely used in many biological applications. The Descement membrane is easy rolled and easy teard after peeling from corneal tissue. In this study, we try to study the phenotypes of cultured endothelial cells on the genipin-crosslinker Descement membrane.
The crosslinking process between Descement membrane.and genipin was performed before cell plating. The crosslinking characteristics of the genipin-fixed Descemet membrane was investigated. Furthermore, the mechanical strength and resistance were compared between the genipin-fixed Descemet membrane and non-genipin-fixed Descemet membrane. The proliferation rate of cultured bovine corneal endothelial cells were study by MTT assay. Furthermore, the immunohistochemistry study and Scanning EM were also investigated.
The mechanical strength and resistance were increased in the group of the genipin-fixed Descemet membrane as compared with the group of non-genipin-fixed Descemet membrane. The genipin-fixed Descemet membrane was easy handled, more smooth surface, and not easy teared. The SEM analysis showed the normal endothelial cells phenotype between the two groups. The MTT assay showed the proliferation rate was increased in the group of genipin-fixed Descemet membrane. The immunohistochemistry staining showed the ZO-1 positive and Na-K ATPase positive cultured cells on the genipin-fixed Descemet membrane.
The results demonstrated that genipin can form stable crosslinked products and is an effective crosslinking agent for Descemet membrane fixation. Therefore, the non-toxic nature crosslinker genipin may be useful for the application on the reconstruction of the ocular surface.
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