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D.H. Lee, T. Kim; The Effect Of Mitomycin–C On Keratocytes: An In Vitro Study . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2987.
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Mitomycin–C (MM–C) is a potent alkylating agent that cross links DNA and inhibits keratocyte proliferation. It is used to treatin the treatment of corneal haze after refractive surgery and and to prevent haze in primary surface ablation procedurescases. There has been no known complications associated with in ccorneal applications thus so far. However, the But long– term safety of MM–C still needs to be addressedshould be answered. The purpose of this study is to evaluate the long term effect of MM–CMitomycin–C on keratocytes of different species.
Primary keratocytes from porcine, chicken, human and rabbit corneas were cultured in DMEM media with 10% FBS after overnight digestion with collagenase and hyaluronidase. Keratocytes between 3rd and 5th passages were used. They were treated with 0.01%, 0.001%, and 0.0001% of mMitomycin–c solution for 100 minutes. The cell toxicity was measured using flow cytometric analysis after staining with aAnnexin V–FITC and pPropidium iIodide. Cell proliferation was assessed by MTT (dimethylthiazol diphenyl tetrazolium bromide) assay.
MTT assay showed that cell proliferation reduced to 32%, 12% and 10% in 0.01%, 0.001%, 0.0001% MMC solution treatment respectively. Flow cytometric analysis with annexin V–FITC /PI staining showed increased early apoptosis only in the 0.01% concentration groupof 0.01% , and there were no significant differences between other concentrations. There were nNo differences in late apoptosis were observed.
This study revealed that mMitomycin–C 0.01% shows keratocytecell toxicity in 0.01% and causes early apoptosis in vitro.
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