Genotoxic effects of 1064-nm Nd:YAG and 532-nm KTP lasers on fibroblast cell cultures

摘要

Background. Several different laser types are used in cutaneous surgery. The neodymium:yttrium-aluminium-garnet (Nd:YAG) and frequency-doubled Nd:YAG (KTP, potassium titanyl phosphate) lasers are widely used in dermatology. Objectives. To investigate the possible genotoxic effects on fibroblasts of irradiation with a 1064-nm Nd:YAG laser and a 532-nm KTP laser.Methods. Fibroblast cell cultures were exposed to each of the lasers, using 10-mm spot size at 60 ms pulse duration with 10, 20, 40 J/cm~2and 3, 6, 12 J/cm~2 fluences, respectively. Fibroblasts in passages 1-6 were used. During laser irradiation, 96-well microplate cultures were kept on a cooling block and transported on ice and in the dark, and processed immediately for single-cell gel electrophoresis (SCGE) assay (also known as a comet assay).Results. DNA damage was determined by computerized assessment of comet assay. There was increasing damage with increasing numbers of passages. For the Nd:YAG laser, the greatest damage occurred on passages 5 and 6, whereas the greatest damage appeared at passages 3 and 4 for KTP and returned to baseline at passages 5 and 6. Damage also increased with each dose increment for both wavelengths. At the highest dose for both wavelengths (Nd:YAG 40 J/cm~2 and KTP 12 J/cm~2), damage was higher with the Nd:YAG laser.Conclusions. Different patterns of cellular damage were seen for different cell-culture passages, treatment doses, and laser wavelengths. These dose ranges are generally used for the treatment of vascular and pigmented lesions and for rejuvenation purposes. As replicative ageing or cell senescence is one of the critical factors determining the extent of cell damage induced by laser therapy, these results may have important implications for clinical practice.