In vitro wound healing improvement by low-level laser therapy application in cultured gingival fibroblasts

Fernanda G Basso, Taisa N Pansani, Ana Paula S Turrioni, Vanderlei S Bagnato, Josimeri Hebling, Carlos A de Souza Costa, Fernanda G Basso, Taisa N Pansani, Ana Paula S Turrioni, Vanderlei S Bagnato, Josimeri Hebling, Carlos A de Souza Costa

Abstract

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.

Figures

Figure 1
Figure 1
Photomicrographs showing human gingival fibroblast cultures seeded in 24-well plates after LLLT. The control group exhibits a large cell-free area on acrylic surface. The group irradiated with 0.5 J/cm² exhibits cell proliferation and migration, with consequent reduction of the “in vitro wound” size. The group irradiated with 3.0 J/cm² presented more intense cell proliferation and migration, resulting in almost complete closure of the “in vitro wound.”
Figure 2
Figure 2
SEM micrograph showing cells with normal morphology that migrated through the transwell membrane. SEM ×500.

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Source: PubMed

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