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L.-K. Yeh, Y.-H. Chen, C.-S. Chiu, T.-H. Young, I.-J. Wang; The Phenotype of Bovine Corneal Epithelial Cells on Chitosan Membrane. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1875. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To explore the issue of tissue engineering in ocular surfaces, we evaluated the possibility of applying the synthetic chitosan membrane (CM) as a substrate carrier for cultured bovine corneal epithelial cells in vitro. The aim of the present work is to evaluate the ability of chitosan membrane to maintain phenotypes of corneal epithelial cells.
We cultivated bovine corneal epithelial cells on chitosan membrane and evaluated cell viability and cytotoxicity by using the 3-[4,5-dimethylrhiazol-2-yl] -2,5-diphenyl tetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) assay. The morphology of cultivated corneal epithelial cells was observed by scanning electron microscopy (SEM). Immunocytochemical stainings (keratin 3,anti-p63, anti-Ki67, anti-panCK, anti-connexin43, and anti-ZO-1 )were used to confirm the phenotype of corneal epithelial cells Additionally, an in vitro organ culture system was used to compare the ability of promoting corneal epithelial healing between amniotic membrane and chitosan membrane.
In the MTT and LDH assays we found that the CM can support the growth of cultured corneal epithelial cells with minimal toxicity. For morphological study, SEM showed the similar morphology like corneal epitheial cells cultured on amniotic membrane. Immunohistocytochemistry showed that the phenotype of corneal epithelial cells is preserved and compatible with that of AM. In addition, CM was found to be able to promote corneal wound healing in an organ culture system.
We conclude that CM has the potential to be a suitable biomaterial for treating ocular surface disorders.
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