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Cytoprotective effect of hyaluronic acid and hydroxypropyl methylcellulose against DNA damage induced by thimerosal in Chang conjunctival cells

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Abstract

Background

To investigate genotoxicity of the preservative thimerosal (Thi), and the cytoprotective and antioxidant effects of hyaluronic Acid (HA) and hydroxypropyl methylcellulose (HPMC) on Chang conjunctival cells.

Method

Cells were divided into three groups. One group was exposed to Thi at various concentrations (0.00001 %∼0.001 %) for 30 min; the other two groups were pretreated with 0.3 % HA or 0.3 % HPMC for 30 min before the Thi exposure. After cell viability was evaluated, alkaline comet assay and detection of the phosphorylated form of the histone variant H2AX (γH2AX) foci were used to determine DNA damage. Reactive oxygen species (ROS) production was assessed by the fluorescent probe, 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA).

Results

A significant change of cell viability was observed after exposure to 0.001 % Thi for 30 min. DNA single- and double-strand breaks were significantly increased in a dose-dependent manner with Thi exposure. In addition, intracellular ROS induced by Thi was dose-dependent, except at 0.001 % less ROS was induced than at 0.0005 %. However, cells pretreated with 0.3 % HA or 0.3 % HPMC showed significantly increased cell survival, decreased DNA damage, and decreased ROS production compared to cells exposed to Thi alone. Pretreatment with 0.3 % HA was found to be even more protective than 0.3 % HPMC.

Conclusion

Thi can induce DNA damage in human conjunctival epithelial cells, probably due to oxidative stress. HA and HPMC are protective agents that have antioxidant properties and can decrease DNA damage induced by Thi. Pretreatment of 0.3 % HA may be more protective of the ocular surface than 0.3 % HPMC.

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Acknowledgements

The work was supported by project foundation : 1.Zhejiang province Key Innovation Team Project of China (No.2009R50039) 2.Zhejiang Key Laboratory Fund of China (No.2011E10006) 3. Natural Science Foundation of China (30973270) 4. The Natural Science Foundation of China (81070756) 5. National “Twelfth Five-Year” Plan for Science & Technology Support of China (2012BAI08B01).

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Authors and Affiliations

  1. Department of Ophthalmology, The 2nd Affiliated Hospital of Zhejiang University, College of Medicine, Hangzhou, Zhejiang, China

    Juan Ye, Huina Zhang, Han Wu, Changjun Wang, Xin Shi, Jiajun Xie & Jinjing He

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

    Jun Yang

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

    Jun Yang

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  1. Juan Ye
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Correspondence to Juan Ye.

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Ye, J., Zhang, H., Wu, H. et al. Cytoprotective effect of hyaluronic acid and hydroxypropyl methylcellulose against DNA damage induced by thimerosal in Chang conjunctival cells. Graefes Arch Clin Exp Ophthalmol 250, 1459–1466 (2012). https://doi.org/10.1007/s00417-012-2087-4

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