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KMID : 0360220110520101215
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Journal of the Korean Ophthalmological Society
2011 Volume.52 No. 10 p.1215 ~ p.1221
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Effect of Mitomycin C, Dexamethasone, and Cyclosporine A 0.05% on the Proliferation of Human Corneal Keratocytes
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Shin Jong-Hoon
Kim Su-Jin
Lee Ji-Eun
Lee Jong-Soo
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Abstract
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Purpose:To investigate the biologic effect of mitomycin C, dexamethasone and cyclosporine A 0.05% on cultured human keratocytes in vitro.
Methods: Human corneal keratocytes were exposed to a concentration of mitomycin C (0.05%), dexamethasone (0.05%) and cyclosporine A (0.05%) for a period of 3, 5, and 10 minutes. MTT-based colorimetric assay was performed to assess the metabolic activity of cellular proliferation and the concentration of type I procollagen COOH-terminal peptide (PIP) and laminin were measured. Cell damage was determined by using the lactate dehydrogenase (LDH) assay. Apoptotic response was evaluated utilizing flow cytometric analysis with Annexin V and propiodium iodide.
Results: The inhibitory effect of cellular proliferation and cytotoxicity in cultured human keratocytes showed a time-dependent response in all drugs. The production of PIP and laminin showed a time-dependent response in cultured cells. Apoptosis was observed in flow cytometry after being treated with mitomycin C, dexamethasone and cyclosporine A. Cyclosporin A resulted in less apoptosis of keratocytes than mitomycin C and dexamethasone.
Conclusions: The apoptotic response of mitomycin C, dexamethasone and cyclosporine A is associated with the inhibitory effect of human corneal keratocyte proliferation. To decrease corneal opacity, mitomycin C and dexamethasone were more effective than cyclosporine A in the present study. Additionally, a high concentration of cyclosporine A greater than 0.05% is necessary to lower corneal opacity. J Korean Ophthalmol Soc 2011;52(10):1215-1221
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KEYWORD
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Cyclosporine, Dexamethasone, Keratocyte, Mitomycin C
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