May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Comparative Study on the Cytotoxic Effects of Benzalkonium Chloride on Wong-Kilbourne Derivative of Chang Conjunctival and IOBA-NHC Cell Lines
Author Affiliations & Notes
  • E. Brasnu
    Department of Ophthalmology III, Quinze-Vingts National Ophthalmology Hospital, Paris, France
    Department of Toxicology, Faculty of Biological and Pharmacological Sciences, Paris Descartes University, Paris, France
  • F. Brignole-Baudouin
    Department of Toxicology, Faculty of Biological and Pharmacological Sciences, Paris Descartes University, Paris, France
    Inserm UMR-S 872, Cordeliers Biomedical Institute, Paris Descartes University, Paris, France
  • A. Pauly
    Department of Toxicology, Faculty of Biological and Pharmacological Sciences, Paris Descartes University, Paris, France
    Inserm UMR-S 872, Cordeliers Biomedical Institute, Paris Descartes University, Paris, France
  • H. Liang
    Department of Toxicology, Faculty of Biological and Pharmacological Sciences, Paris Descartes University, Paris, France
    Inserm UMR-S 872, Cordeliers Biomedical Institute, Paris Descartes University, Paris, France
  • L. Riancho
    Department of Toxicology, Faculty of Biological and Pharmacological Sciences, Paris Descartes University, Paris, France
    Inserm UMR-S 872, Cordeliers Biomedical Institute, Paris Descartes University, Paris, France
  • J.-M. Warnet
    Department of Toxicology, Faculty of Biological and Pharmacological Sciences, Paris Descartes University, Paris, France
    Inserm UMR-S 872, Cordeliers Biomedical Institute, Paris Descartes University, Paris, France
  • C. Baudouin
    Department of Ophthalmology III, Quinze-Vingts National Ophthalmology Hospital, Paris, France
    Inserm UMR-S 872, Cordeliers Biomedical Institute, Paris Descartes University, Paris, France
  • Footnotes
    Commercial Relationships  E. Brasnu, None; F. Brignole-Baudouin, None; A. Pauly, None; H. Liang, None; L. Riancho, None; J. Warnet, None; C. Baudouin, None.
  • Footnotes
    Support  Unrestricted grants from INSERM (UMRS 872)
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 6040. doi:https://doi.org/
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      E. Brasnu, F. Brignole-Baudouin, A. Pauly, H. Liang, L. Riancho, J.-M. Warnet, C. Baudouin; Comparative Study on the Cytotoxic Effects of Benzalkonium Chloride on Wong-Kilbourne Derivative of Chang Conjunctival and IOBA-NHC Cell Lines. Invest. Ophthalmol. Vis. Sci. 2008;49(13):6040. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : The Wong-Kilbourne derivative of Chang conjunctiva-derived cell line has been widely used for toxicological and functional in vitro studies on ocular surface, but is contaminated with HeLa cells. Recently, the IOBA-NHC spontaneously immortalized conjunctival epithelial cell line has been developed and did not show other cell type contamination. In the present study, we compared in the two cell lines the toxic effects of benzalkonium chloride (BAC), a preservative which toxicity has been often reported on conjunctival in vivo and in vitro models, in order to assess whether both cell lines would be equally suitable for in vitro toxicological studies.

Methods: : Microplate cytofluorometry tests (neutral red, H2DCF-DA, hydroethidine and Yopro-1), flow cytometry tests (Annexin V/7-AAD and DNA content) and standard immunofluorescence stainings were used to assess the necrotic, apoptotic and oxidative effects of BAC on Chang and IOBA-NHC cell lines. Cells were exposed to five concentrations of BAC (10-2%, 5.10-3%, 10-3%, 10-4% and 10-5%) during 15 minutes and 15 minutes followed by 24 hours of cell recovery in complete medium.

Results: : With the microplate cytofluorometry and the flow cytometry tests, the lowest cellular viability, the highest reactive oxygen species production and the highest apoptotic effects were obtained with the highest BAC concentrations and varied in a concentration-dependent manner on both IOBA-NHC and Chang cell lines. Standard immunofluorescence stainings showed increasing cell shrinkage with high concentrations of BAC on either Chang or IOBA-NHC cells.

Conclusions: : Our findings validated the use of both IOBA-NHC and Chang cell lines for toxicological in vitro studies. However, their respective disadvantages have to be known and considered in any further study using IOBA-NHC or Chang cells.

Keywords: conjunctiva • apoptosis/cell death • oxidation/oxidative or free radical damage 
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