J Korean Ophthalmol Soc > Volume 52(10); 2011 > Article
Journal of the Korean Ophthalmological Society 2011;52(10):1215-1221.
DOI: https://doi.org/10.3341/jkos.2011.52.10.1215    Published online October 15, 2011.
Effect of Mitomycin C, Dexamethasone, and Cyclosporine A 0.05% on the Proliferation of Human Corneal Keratocytes.
Jong Hoon Shin, Soo Jin Kim, Ji Eun Lee, Jong Soo Lee
1Department of Ophthalmology, Pusan National University College of Medicine, Busan, Korea. jongsool@pusan.ac.kr
2Department of Ophthalmology, Pusan National University Yangsan Hospital, Busan, Korea.
마이토마이신, 덱사메타존, 사이클로스포린 0.05%가 인간 각막실질세포에 미치는 영향
신종훈1⋅김수진1⋅이지은2⋅이종수1
Department of Ophthalmology, Pusan National University College of Medicine1, Busan, Korea Department of Ophthalmology, Pusan National University Yangsan Hospital2, Busan, Korea
Abstract
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.
Key Words: Cyclosporine;Dexamethasone;Keratocyte;Mitomycin C


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