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KMID : 0360220070480101399
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Journal of the Korean Ophthalmological Society
2007 Volume.48 No. 10 p.1399 ~ p.1409
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Effect of Cyclosporine A 0.05% on Human Corneal Epithelial Cells
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Lee Ji-Eun
Shin Wook-Bum
Lee Jong-Soo
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Abstract
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Purpose: To evaluate the toxic effect of topical cyclosporine on cultured human corneal epithelium and to investigate the apoptotic response and cellular morphologic changes associated with cyclosporine in vitro.
Methods: Human corneal epithelial cells were exposed to a concentration of cyclosporine A (0.05%) for a period of 3, 5, and 10 minutes. MTT-based calorimetric assay was performed to assess the metabolic activity of cellular proliferation and lactate dehydrogenase (LDH) leakage assay for cytotoxicity. Apoptotic response was evaluated with flow cytometric analysis and fluorescence staining with Annexin V and propiodium iodide. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy.
Results: The inhibitory effect of human corneal epithelial cell proliferation and cytotoxicity showed a time-dependent response and had a significant effect when exposed for 10 minutes (P=0.04). The maximun response did not reach the leathal dose (LD)50. Apoptosis was seen in flow cytometry and apoptotic cells were demonstrated in fluorescent micrograph after being treated treating with cyclosporine A (0.05%). Human corneal epithelial cells were more detached from the bottom of the dish and damaged cells show degenerative changes like microvilli disappearance, vacuoles formation, and chromatin of the nuclear remnant condensed along the nuclear periphery.
Conclusions: Cyclosporine A (0.05%) could be used without any significant toxic effect on human corneal epithelial cells except for exposure times longer than 10 minutes. Induction of apoptosis modulation may be one of the cyclosporine`s mechanism for inhibiting cellular proliferation.
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KEYWORD
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Apoptosis, Corneal epithelium, Cyclosporoine, Toxicity
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