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Title: UV-mediated DNA damage in corneal epithelial cells assessed using the comet assay procedure
Authors: Choy, CKM
Benzie, IFF 
Cho, P 
To, T 
Keywords: Aging
Cornea: basic science
Cornea: tears/tear film/dry eye
Issue Date: 2004
Publisher: Association for Research in Vision and Ophthalmology
Source: Investigative ophthalmology and visual science, May 2004, v. 45, no. 13, 1452 (Abstracts) How to cite?
Journal: Investigative ophthalmology and visual science 
Abstract: Purpose:To develop a comet assay (single cell gel electrophoresis) corneal epithelial cell model for evaluation of DNA damage in single cells, and to assess effect of ultraviolet (UV) irradiation on DNA damage in corneal epithelial cells.
Methods: Baseline DNA damage of three types of porcine corneal epithelial cells – ‘superficial–layer’, ‘middle–layer’, and ‘deeper–layer’ cells – were assessed. ‘Deeper–layer’ cells were selected and exposed to ambient room light (control) or an energy–standardized dose (0.216 J/cm2) of UVA, UVB or UVC to determine the effect of UV irradiation.
Results: Baseline DNA damage (%DNA in comet tail) in non–irradiated ‘superficial–layer’, ‘middle–layer’ and ‘deeper–layer’ corneal epithelial cells were (Mean±SD) 30.4±3.2%, 10.5±1.0% and 4.0±0.3%, respectively. Exposure to UVA, UVB and UVC caused significant (P<0.001) increases in DNA damage in ‘deeper–layered’ cells: mean±SD %DNA scores (10 gels per treatment, with 100 irradiated cells scored per gel) were 27.4±4.6% for UVB, 10.2±1.4% for UVA, and 14.7±1.8% for UVC (P<0.001), compared to 4.2±0.5% in the control cells.
Conclusions:The comet assay can be used successfully with corneal epithelial cells, and will support future studies investigating environmental influences on ocular health and assessment of possible protective strategies. UVB radiation damages DNA in corneal epithelial cells, while UVC causes less damage, probably due to UVC absorption by ascorbic acid in the corneal cells. Results highlight the possible need for additional protection from UVB in high–risk individuals due to either high levels of solar radiation or corneal thinning due to photorefractive keratectomy.
Description: ARVO Annual Meeting, Florida, US, 25-29 April 2004
ISSN: 0146-0404
EISSN: 1552-5783
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