p-Coumaric acid (PCA) is a kind of phenolic compound, and as one of the
cinnamic acid derivatives, it has many
biological functions such as
antioxidants, anti-inflammatory, antiplatelet, and anticancer activity. Low-level
laser irradiation has received increasing interest in the fields of tissue regeneration and wound healing. In this study, the effect of low-level
laser irradiation on human fibroblast cells (human dermal fibroblast) and human
melanoma cancer cells (A375 and SK-MEL-37) treated with PCA was investigated. The human dermal fibroblast, A375, and SK-MEL-37 cells were exposed to low-level
laser at 660-nm wavelength with 3 J/cm for 90 s, and then the cells were treated with different concentrations of PCA (0-1000 μg/ml for 24 h), separately. In another experiment, first the cells were treated by PCA and then irradiated with low-level
laser as described before. The effect of various irradiation energy (1-6 J/cm) on the
melanoma cells, which were then treated by PCA, was studied. The cell viability using MTT assay and
lactate dehydrogenase assay was determined. Morphological changes owing to apoptosis induction by irradiation and PCA were detected by fluorescence microscopy using
acridine orange/
ethidium bromide double staining. The results showed that pretreatment with low-level
laser irradiation and then PCA reduced the survival and growth of
melanoma cells more than the early treatment with PCA and then low-level
laser irradiation.
Lactate dehydrogenase activity was reduced significantly by preirradiation and then PCA treatment in comparison with the dark group in
melanoma cells. The cell cytotoxicity at different irradiation energy and then IC50 concentration of PCA was increased up to 3 J/cm and then decreased following increasing irradiation energy. The morphology study with light microscopy and apoptotic assay using
acridine orange/
ethidium bromide dual staining confirmed the MTT results. This study showed that low-level
laser irradiation alone is not able to kill human normal fibroblast and human
melanoma cancer cells. Preirradiation followed by treatment with PCA did not change the cell viability in human fibroblast significantly but reduced the cell viability in
melanoma cells presumably through the apoptosis pathway.