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Effects of green light photobiomodulation on Dental Pulp Stem Cells: enhanced proliferation and improved wound healing by cytoskeleton reorganization and cell softening

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Abstract

Photobiomodulation (PBM) has been shown to improve cell proliferation and cell migration. Many cell types have been investigated, with most studies using deep penetrating red light irradiation. Considering the interest of surface biostimulation of oral mesenchymal cells after surgical wound, the present study aimed to assess green light irradiation effects on Dental Pulp Stem Cells’ (DPSC) proliferation and migration. To understand the mechanisms underlying these effects, we investigated cytoskeleton organization and subsequent cell shape and stiffness. A 532-nm wavelength Nd:YAG laser (30 mW) was applied between 30 and 600 s on DPSC in vitro. Cell proliferation was analyzed at 24, 48, and 72 h after irradiation, by cell counting and enzymatic activity quantification (paranitrophenylphosphate phosphatase (pNPP) test). A wound healing assay was used to study cell migration after irradiation. Effects of PBM on cytoskeleton organization and cell shape were assessed by actin filaments staining. Elasticity changes after irradiation were quantified in terms of Young’s modulus measured using Atomic Force Microscopy (AFM) force spectroscopy. Green light significantly improved DPSC proliferation with a maximal effect obtained after 300-s irradiation (energy fluence 5 J/cm2). This irradiation had a significant impact on cell migration, improving wound healing after 24 h. These results were concomitant with a decrease of cells’ Young’s modulus after irradiation. This cell softening was explained by actin cytoskeleton reorganization, with diminution of cell circularity and more abundant pseudopodia. This study highlights the interest of green laser PMB for the proliferation and migration of mesenchymal stem cells, with encouraging results for clinical application, especially for surgical wound healing procedures.

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Authors and Affiliations

  1. LBN, University Montpellier, 545 Av Pr JL Viala, 34193 CEDEX 4, Montpellier, France

    Eve Malthiery, Batoul Chouaib, Ana María Hernandez-Lopez, Jacques-Henri Torres, Frédéric J Cuisinier & Pierre-Yves Collart-Dutilleul

  2. Universidad Autónoma de Nuevo León, San Nicolás de los Garza, Mexico

    Ana María Hernandez-Lopez

  3. L2C, CNRS, University Montpellier, Montpellier, France

    Marta Martin & Csilla Gergely

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  1. Eve Malthiery
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Correspondence to Pierre-Yves Collart-Dutilleul.

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Dental Pulp Stem Cells’ (DPSC) collection was declared to the Ministry of Higher Education, Research and Innovation: “Direction Générale de la Recherche et de l’Innovation,” Déclaration DC-2014-2198, Récépissé n°3119(bis). Clinical protocol for DPSC recovery was approved by the local ethical committee.

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Malthiery, E., Chouaib, B., Hernandez-Lopez, A.M. et al. Effects of green light photobiomodulation on Dental Pulp Stem Cells: enhanced proliferation and improved wound healing by cytoskeleton reorganization and cell softening. Lasers Med Sci 36, 437–445 (2021). https://doi.org/10.1007/s10103-020-03092-1

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