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Jae Lim Chung, Young A Kwon, Sang Wroul Song, Byung Yeop Kim, Joon H Lee, Kyoung Yul Seo, Christopher Ta; Cytotoxicity of Ophthalmic Antimicrobial Solutions on SV40-immortalized Human Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5786.
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To enhance the pharmacokinetic properties of ophthalmic antimicrobials, high-concentration of active ingredient or surface-retentive delivery system are being used. However, ocular toxicity is a concern when using these agents. In this study, we evaluated the cytotoxicity of various ophthalmic antibiotics on SV40-immortalized human corneal epithelial cells using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay.
We tested original and 10-fold diluted solutions of Cravit (levofloxacin 0.5%), Cravit 1.5 (levofloxacin 1.5%), Quixin (levofloxacin 0.5%, Benzalkonium chloride (BAK) 0.005%), Iquix (levofloxacin 1.5%), Vigamox (moxifloxacin 0.5%), Moxeza (moxifloxacin 0.5%, xanthan gum), Gatiflo (gatifloxacin 0.3%), Zymaxid (gatifloxacin 0.5%, BAK 0.005%), Besivance (besifloxacin, BAK 0.01%, durasite), Azasite (azithromycin 1%, BAK 0.003%, durasite), Tobrex (tobramycin 0.3%, BAK 0.01%). We also tested using standard material powders of each drug to rule out the effect of other components in ophthalmic solutions. The MTS assay was performed after 5 to 120 minutes of exposure to each solution. According to the cell survival score, we graded the cytotoxicity as mild(over 75%), moderate(50~75%) and marked(below 50%).
Following exposure to undiluted solutions, cell viabilities were decreased to 9~73% after 5 minutes. After 30 minutes Tobrex, Zymaxid and Quixin showed marked toxicity, Cravit 1.5, Iquix and Vigamox showed moderate toxicity and Cravit and Gatiflo showed mild toxicity. When exposed to 10-fold diluted solutions, Tobrex, Besivance, Zymaxid, Quixin and Azasite, which were BAK containing products, showed moderate toxicity. After exposure to standard powder solutions, 0.6% besifloxacin, 0.5% gatifloxacin, 0.5% moxifloxacin, 1.5% levofloxacin and 0.5% levofloxacin showed moderate toxicity in decreasing order. However, 1% azithromycin and 0.3% tobramycin standard powder solutions showed the least toxicity.
High concentration alone did not further increase cytotoxicity. The most important factor for the determination of cytotoxicity was the concentration of BAK in antimicrobial solutions.
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