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The aim of this study was to determine adequate energy doses using specific parameters of LLLT to produce biostimulatory effects on human gingival fibroblast culture. Cells (3 × 10 4  cells/cm 2 ) were seeded on 24-well acrylic plates using plain DMEM supplemented with 10% fetal bovine serum. After 48-hour incubation with 5% CO 2  at 37°C, cells were irradiated with a InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm; 40 mW) with energy doses of 0.5, 1.5, 3, 5, and 7 J/cm 2 . Cells were irradiated every 24 h totalizing 3 applications. Twenty-four hours after the last irradiation, cell metabolism was evaluated by the MTT assay and the two most effective doses (0.5 and 3 J/cm 2 ) were selected to evaluate the cell number (trypan blue assay) and the cell migration capacity (wound healing assay; transwell migration assay). Data were analyzed by the Kruskal-Wallis and Mann-Whitney nonparametric tests with statistical significance of 5%. Irradiation of the fibroblasts with 0.5 and 3 J/cm 2  resulted in significant increase in cell metabolism compared with the nonrradiated group ( P  < 0.05). Both energy doses promoted significant increase in the cell number as well as in cell migration ( P  < 0.05). These results demonstrate that, under the tested conditions, LLLT promoted biostimulation of fibroblasts in vitro.

In Vitro Wound Healing Improvement by Low-Level Laser Therapy Application in Cultured Gingival Fibroblasts