Biostimulation of dermal fibroblast by sublethal Q-switched Nd:YAG 532 nm laser: collagen remodeling and pigmentation

The application of medical lasers in treating pigmented lesions has rapidly developed over the past decade. In both clinical and cosmetic application, melanin is targeted in pigmented areas and destroyed by the mechanism of selective photothermolysis. When laser radiation passes through superficial pigmented tissue, energy will be further reduced by dermal collagen scattering and absorption. Non-pigmented dermal fibroblasts will be exposed to co-incidental laser irradiation at lower energy levels. Biostimulation of dermal fibroblasts by low energy laser is reported in this paper. The Q-switched frequency doubled Nd:YAG 532nm laser used in clinical laser therapy was used in this study. Sublethal laser fluence was determined at 0.8J/cm(2) and used to stimulate normal human fibroblasts in monolayer culture. The results showed that there was no significant difference in collagen synthesis between the stimulated fibroblasts and controls. However, significant delay in collagen remodeling activity was demonstrated in the irradiated group by measuring fibroblast populated collagen lattice (FPCL) contraction. The stimulation of SCF, HGF and b-FGF gene expression was determined by RT-PCR analysis and demonstrated to vary between cases. Two out of six cell lineages that showed stronger responses to laser stimulation on SCF, HGF and b-FGF gene expressions were used to prepare conditioned media. The conditioned media from irradiated groups showed significant increase in SCF and b-FGF content and stimulated SK-mel-3 melanoma cells to synthesize more melanin in vitro. These results suggest that sublethal laser stimulation of fibroblasts may cause post-laser hyperpigmentation through production of melanogenic stimulatory cytokines. The degree of stimulation of SCF, HGF and b-FGF production varied between individual cell lineages, which may reflect the true variation of post-laser hyperpigmentation in clinical practice.