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An in vitro Method to Test the Safety and Efficacy of Low-Level Laser Therapy (LLLT) in the Healing of a Canine Skin Model

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dc.contributor.advisor Gibson, Thomas Gagnon, Dominique 2015-09-10T12:39:04Z 2015-09-10T12:39:04Z 2015-07 2015-07-27 2015-09-10
dc.description.abstract Low-level laser therapy (LLLT) aims to photoactivate cellular mechanisms that have the potential to improve healing of the treated regions, and reduce local edema, inflammation and pain. Low-level laser therapy has been used clinically as a treatment modality in humans for a variety of medical conditions, including the improvement of wound-healing processes. The results obtained appear to be closely related to parameters such as dose, time of exposure and wavelength. Although LLLT is gaining popularity, a universally accepted theory that explains the cellular effects and beneficial biological processes seen in wound healing has not been described. This research was designed to primarily evaluate the effect of LLLT on cellular migration and proliferation of cultured canine epidermal keratinocytes (CPEK), in an in vitro wound healing model. A secondary objective was to perform a pilot study with this model in order to evaluate the effect of LLLT on the expression of microRNA-21 (miR-21), a microRNA associated with wound healing, using quantitative real time polymerase chain reaction (qRT-PCR). Keratinocyte migration and proliferation were assessed using a scratch migration assay and a proliferation assay, respectively. The proliferation assay was performed using water-soluble tetrazolium (WST-1), a proliferation reagent. Canine keratinocytes were cultured in tissue culture flasks and then seeded in 6-well tissue culture plates at different cellular concentrations. Cells between the third- and fifth-passage were used for all experiments. Laser treatment of monolayer cultures was performed using a Helium-Neon Class IV laser system. Sham irradiation and different energy doses of 0.1, 0.2, 1.2 and 10 J/cm2 were evaluated. In this in vitro wound healing model, keratinocytes exposed to doses of 0.1, 0.2, and 1.2 J/cm2 migrated significantly more rapidly (p < 0.03) and showed significantly higher rates of proliferation (p < 0.0001) compared to non-irradiated cells cultured in the same medium and cells exposed to the higher energy dose of 10 J/cm2. Irradiation with 10 J/cm2 was characterized by decreased cellular migration and proliferation. These data suggest that the beneficial effects of LLLT in vivo may be due, in part, to effects on keratinocyte behavior and high doses may be detrimental to wound healing. en_US
dc.description.sponsorship Ontario Veterinary College Pet Trust Fund, LiteCure—Companion Therapy Laser System en_US
dc.language.iso en en_US
dc.subject Wound healing en_US
dc.subject Canine epidermal keratinocyte progenitors en_US
dc.subject Low-level laser therapy en_US
dc.subject Helium-neon laser en_US
dc.subject Laser basics en_US
dc.subject Laser physics en_US
dc.subject Scratch migration assay en_US
dc.subject Water-soluble tetrazolium en_US
dc.subject Proliferation assay en_US
dc.subject MicroRNA en_US
dc.title An in vitro Method to Test the Safety and Efficacy of Low-Level Laser Therapy (LLLT) in the Healing of a Canine Skin Model en_US
dc.type Thesis en_US Veterinary Science en_US Doctor of Veterinary Science en_US Department of Clinical Studies en_US

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