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Evaluation of Cytotoxicity of Various Ophthalmic Drugs, Eye Drop Excipients and Cyclodextrins in an Immortalized Human Corneal Epithelial Cell Line

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Abstract

Purpose. An immortalized human corneal epithelial cell line (HCE) was tested as a screening tool for prediction of topical ocular irritation/ toxicity by pharmaceuticals.

Methods. Effects of various drugs, excipients and cyclodextrins (CDs) on viability of HCE cells were evaluated using two in vitrocytotoxicity tests, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) dye reduction assay and propidium iodide assay.

Results. Mitochondrion-based MTT test was a more sensitive indicator of cytotoxicity than the plasma membrane-based propidium iodide test. The tests revealed following cytotoxic rankings for ophthalmic drugs: dipivefrin > timolol > pilocarpine ≈ dexamethasone; for excipients: benzalkonium chloride (BAC) > sodium edetate (NA2EDTA) > poly-vinyl alcohol (PVA) > methylparaben; and for CDs: α-CD > dimethyl-β-cyclodextrin (DM-β-CD) > sulfobutyl ether (β-cyclodextrin ((SBE)7m-β-CD) ≈ hydroxypropyl-β-cyclodextrin (HP-β-CD) > γ-CD. In consideration of the in vivoclinical situation, the short exposure time (5 min) is more relevant even though toxic effects of some test substances were seen only after longer exposure times (30 and 60 min).

Conclusions. Immortalized HCE cells are a promising tool for rapid cytotoxicity assays of ocular medications. The cell line is potentially useful in predicting the in vivocorneal toxicity of ocularly applied compounds.

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Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, University of Kuopio, P.O. Box 1627, FIN-70211, Kuopio, Finland

    Paula Saarinen-Savolainen & Tomi Järvinen

  2. Toyanaka Municipal Hospital, Osaka, Japan

    Kaoru Araki-Sasaki

  3. Department of Ophthalmology, Osaka University Medical School, Osaka, Japan

    Hitoshi Watanabe

  4. Department of Pharmaceutics, University of Kuopio, P.O. Box 1627, FIN-70211, Kuopio, Finland

    Arto Urtti

Authors
  1. Paula Saarinen-Savolainen
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  3. Kaoru Araki-Sasaki
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  4. Hitoshi Watanabe
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  5. Arto Urtti
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Saarinen-Savolainen, P., Järvinen, T., Araki-Sasaki, K. et al. Evaluation of Cytotoxicity of Various Ophthalmic Drugs, Eye Drop Excipients and Cyclodextrins in an Immortalized Human Corneal Epithelial Cell Line. Pharm Res 15, 1275–1280 (1998). https://doi.org/10.1023/A:1011956327987

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