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Role of benzalkonium chloride in DNA strand breaks in human corneal epithelial cells

  • Basic Science
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Abstract

Purpose

To investigate the toxic effects of benzalkonium chloride (BAC), a preservative commonly used in ophthalmic preparations, on DNA single- and double-strand breaks in immortalized human corneal epithelial cells (HCEs).

Methods

HCEs were treated with BAC in concentrations ranging from 0.00005% to 0.001% for 30 min. Cells were examined immediately after BAC exposure and after 24-h recovery. Alkaline comet assay was used to detect DNA single-strand breaks (SSBs). Immunofluorescence microscope detection of the phosphorylated form of histone variant H2AX (γH2AX) foci indicated DNA double-strand breaks (DSBs). Cell viability was measured by the MTT test.

Results

A significant increase of SSBs, detected by alkaline comet assay, was observed in a dose-dependent manner with BAC exposure in HCEs at concentrations of 0.00005% and higher. Such BAC treatment also exhibited a dose-dependent increase in DSBs, evaluated by number of γH2AX foci. In addition, a significant change in the relative cell survival rate of HCEs was observed after exposure to 0.001% BAC for 30 min. Although the toxic effects of BAC could be partly repaired after 24 h of cell recovery, SSBs and DSBs in HCEs were still present after BAC removal.

Conclusions

The results demonstrated that exposure to BAC in HCEs, even at low concentrations, could induce DNA strand breaks, which were present after BAC removal. Cell survival analysis indicated that BAC-induced DNA damage was correlated with the cytotoxic effects.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (NOS. 81070756); the Natural Science Foundation of Zhejiang Province of China (NOS. Y208396); and the International Science and Technology Cooperation Project of Zhejiang Province of China (NOS. 2008C14099).

Financial Interest

No authors have any type of financial interest related to this manuscript.

Author information

Authors and Affiliations

  1. Department of Ophthalmology, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310009, China

    Juan Ye, Han Wu, Huina Zhang, Xiuming Jin & Xin Shi

  2. Department of Toxicology, Zhejiang University School of Public Health, Hangzhou, Zhejiang, 310058, China

    Yihua Wu & Jun Yang

  3. Department of Toxicology, Hangzhou Normal University School of Public Health, Hangzhou, Zhejiang, 310036, China

    Jun Yang

Authors
  1. Juan Ye
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  2. Han Wu
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  3. Huina Zhang
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  4. Yihua Wu
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  5. Jun Yang
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  6. Xiuming Jin
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  7. Xin Shi
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Correspondence to Juan Ye.

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The authors have full control of all primary data, and all the authors agree to allow Graefe's Archive for Clinical and Experimental Ophthalmology to review our data upon request.

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Ye, J., Wu, H., Zhang, H. et al. Role of benzalkonium chloride in DNA strand breaks in human corneal epithelial cells. Graefes Arch Clin Exp Ophthalmol 249, 1681–1687 (2011). https://doi.org/10.1007/s00417-011-1755-0

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  • DOI: https://doi.org/10.1007/s00417-011-1755-0

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