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Comparative assessment of the cytotoxicity of six anti-inflammatory eyedrops in four cultured ocular surface cell lines, as determined by cell viability scores

Authors Ayaki M , Iwasawa, Niwano

Received 13 August 2012

Accepted for publication 3 October 2012

Published 13 November 2012 Volume 2012:6 Pages 1879—1884

DOI https://doi.org/10.2147/OPTH.S36968

Checked for plagiarism Yes

Review by Single anonymous peer review

Peer reviewer comments 3



Masahiko Ayaki,1 Atsuo Iwasawa,2 Yoshimi Niwano3

1Department of Ophthalmology, International University of Health and Welfare, Mita Hospital, Tokyo, 2Department of Bioengineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, 3Laboratory for Redox Regulation, Tohoku University Graduate School of Dentistry, Sendai, Japan

Purpose: Anti-inflammatory eyedrops are often used in the treatment of corneal epithelial disorders. In the present study, we evaluated the cytotoxicity of six anti-inflammatory eyedrops in four ocular surface cell lines.
Methods: The cytotoxicity of six commercially available anti-inflammatory ophthalmic solutions (ie, diclofenac, bromfenac, pranoprofen, betamethasone, and fluoromethorone) was assessed in three corneal cell lines and one conjunctival cell line. Cell viability was determined by the 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide and neutral red assays after exposing the cells to 10, 30, and 60 minutes of onefold, twofold, and tenfold dilutions of the drugs. Cytotoxicity was compared using the cell viability score (CVS), an integrated cytotoxic parameter that takes various factors into account, such as dilution by tear fluid or concentration by evaporation, drug exposure time, and ocular surface cell type.
Results: Based on the CVS scores, the order of the anti-inflammatory eyedrops tested from least to most cytotoxic, with the active ingredient %CVS50, and %CVS40/80 for each solution given in parentheses, was as follows: Rinderon® (betamethasone, 100%, 100%) >0.02% Flumethoron® (fluoromethorone, 68%, 22%) = 0.1% Flumethoron® (fluoromethorone, 76%, 22%) >Bronuck® (0.1% bromfenac, 53%, −8%) = Diclod® (0.1% diclofenac, 44%, −15%) = Niflan® (pranoprofen, 50%, −19%). Rinderon® exhibited the least toxicity of all the anti-inflammatory eyedrops tested. Eyedrops containing non-steroidal anti-inflammatory drugs exhibited greater cytotoxicity than those containing steroids with benzalkonium at comparable concentrations. Concentration was the most significant factor affecting cell viability.
Conclusion: The cytotoxicity of the anti-inflammatory eyedrops evaluated in the present study depended on both the pharmaceutical components and preservatives. The CVS is a concise indicator of drug cytotoxicity.

Keywords: benzalkonium chloride, toxicity, cornea, cell viability score, anti-inflammatory drug

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