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DNA damage after phototherapy in wounded fibroblast cells irradiated with 16 J/cm2

Paper ID Volume ID Publish Year Pages File Format Full-Text
30399 44476 2009 7 PDF Available
Title
DNA damage after phototherapy in wounded fibroblast cells irradiated with 16 J/cm2
Abstract

Background and objectivePhototherapy or biomodulation is a remarkable therapy that has become more popular and widely used in the treatment of a variety of medical conditions, such as slow to heal wounds, pain, soft tissue injuries and skin trauma. It has been shown to induce DNA damage; however this damage appears to be repairable. This study aimed to determine the effects of phototherapy induced DNA damage and activation of the DNA repair gene methylpurine DNA glycosylase (MPG).Materials and methodsDNA integrity was assessed using the comet assay, with and without formamidopyrimidine glycosylase (Fpg). For the comet assay, wounded human skin fibroblast cells (WS1) were irradiated twice, once at 30 min and again at 72 h with 5 or 16 J/cm2 using a diode laser at 636 nm and cellular responses were assessed 1 or 24 h post-irradiation. Real time reverse transcriptase polymerase chain reaction (RT-PCR) assessed MPG expression and three reference genes namely; beta Actin (ACTB), Glyceraldehyde three phosphate dehydrogenase (GAPDH) and Ubiquitin c (UBC). Wounded cells were irradiated once (30 min) with 16 J/cm2, and MPG expression was assessed at 0, 3 and 8 h post-laser irradiation.ResultsAt both 1 and 24 h, wounded cells irradiated with 5 J/cm2 showed insignificant DNA damage compared to control cells, while irradiation with 16 J/cm2 showed significant damage. However, 24 h post-irradiation these cells showed a significant decrease in damage compared to cells left to incubate for 1 h. This observation was attributed to activation of DNA repair mechanisms. Real time RT-PCR showed that ACTB was not influenced by cell culture conditions or laser irradiation, and MPG expression was not detected.ConclusionIn conclusion, irradiation with 5 J/cm2 did not produce additional DNA damage, while damage to cells irradiated with 16 J/cm2 was repairable by mechanisms other than MPG. This study also showed that ACTB can be used as a reference gene in laser experiments, using parameters set out in this study.

Keywords
Phototherapy; DNA repair; Comet assay; MPG
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DNA damage after phototherapy in wounded fibroblast cells irradiated with 16 J/cm2
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Photochemistry and Photobiology B: Biology - Volume 94, Issue 2, 9 February 2009, Pages 131–137
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us