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Photobiomodulation of inflammatory-cytokine-related effects in a 3-D culture model with gingival fibroblasts

Abstract

The aim of this study was to assess the effects of IL-6 and IL-8 cytokines on human gingival fibroblasts (HGF) cultured in a 3-D model and the possible photobiomodulation (PBM) of such effects by low-level laser therapy. In complete culture medium (DMEM), HGF from a healthy patient were seeded in a type I collagen matrix inserted into 24-well plates. After 5 days of incubation, the cytokines were added or not to serum-free DMEM, which was applied to the cell-enriched matrices. Then, PBM was performed: three consecutive irradiations using LaserTable diode device (780 nm, 0.025 W) at 0.5 J/cm2 were delivered or not to the cells. Twenty-four hours after the last irradiation, cell viability and morphology, gene expression, and synthesis of inflammatory cytokines and growth factors were assessed. The histological evaluation demonstrated that, for all groups, matrices presented homogeneous distribution of cells with elongated morphology. However, numerous cytokine-exposed cells were rounded. IL-6 and IL-8 decreased cell viability, synthesis of VEGF, and gene expression of collagen type I. PBM enhanced cell density in the matrices and stimulated VEGF expression, even after IL-6 challenge. Reduced TNF-α synthesis occurred in those cells subjected to PBM. In conclusion, PBM can penetrate collagen matrix and stimulate HGF, highlighting the relevance of this research model for further phototherapy studies and in vitro biomodulation of the healing process.

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Funding

This study received financial support from the Coordination of Superior Level Staff Improvement (CAPES) and the Brazilian National Council for Scientific and Technological Development (CNPq).

Author information

Correspondence to Fernanda Gonçalves Basso.

Ethics declarations

This study was performed in accordance with the local Ethics Committee (CAAE: 74823317.0.0000.5416).

Conflict of interest

The authors declare that they have no conflict of interest.

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Cardoso, L.M., Pansani, T.N., Hebling, J. et al. Photobiomodulation of inflammatory-cytokine-related effects in a 3-D culture model with gingival fibroblasts. Lasers Med Sci (2020). https://doi.org/10.1007/s10103-020-02974-8

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Keywords

  • Cytokines
  • Low-level laser therapy
  • 3-D cell culture
  • Gingival fibroblasts