Human Fibroblast Gene Expression Modulation Using 940 NM Diode Laser

Rebeca Illescas-Montes,Concepción Ruiz,Francisco Javier Manzano-Moreno,Olga García-Martínez,Elvira de Luna-Bertos,Lucía Melguizo-Rodríguez,Javier Ramos-Torrecillas

doi:10.1038/s41598-019-48595-2

Rebeca Illescas-Montes, Concepción Ruiz + Show 5 more

Open Access

https://doi.org/10.1038/s41598-019-48595-2

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Abstract

Low-Level Laser Therapy is used as regenerative therapy in different clinical fields. This is due to its photobiomodulation effect via cell signaling on different cell populations, Including fibroblasts, cells involved in tissue regeneration and healing. The aim was to analyze the effect of 940 nm diode laser on the gene expression of different markers involved in fibroblast growth, differentiation, and migration. Real-time polymerase chain reaction (q-RT-PCR) was used to quantify the expression of fibroblast growth factor (FGF), connective tissue growth factor (CTGF), vascular-endothelial growth factor (VEGF), transforming growth factor β1 (TGF-β1), TGFβ-receptors (TGFβR1, TGFβR2, and TGFβR3), discoidin-domain receptor-2 (DDR2), matrix metalloproteinase-2 (MMP2), α-actin, fibronectin, decorin, and elastin on human fibroblast, treated with single dose (T1) or two doses (T2) of diode laser at 0.5 Watts and 4 J/cm2. A significant increase in the expression of FGF, TGF-β1, TGFβR1, TGFβR2, α-actin, fibronectin, decorin, DDR2 and MMP2 was observed after both treatments. A decrease was observed in expression of elastin (T1 and T2), and CTGF (T2). These changes underlie the biostimulatory effect of laser on fibroblasts, which translates into an increase in short-term proliferation and in long-term differentiation to myofibroblasts. These data support the therapeutic potential of diode laser for wound repair.

Highlights

Low-Level Laser Therapy (LLLT) is applied in multiple clinical fields[5]
The expression of α-actin, fibronectin, decorin, discoidin-domain receptor-2 (DDR2) and MMP-2 was increased versus controls with both treatments (T1 and T2), whereas the expression of elastin was reduced with T1 and T2 treatments
The present study demonstrates that in vitro treatment with this laser modulates the expression of the human fibroblast markers fibroblast growth factor (FGF), connective tissue growth factor (CTGF), vascular-endothelial growth factor (VEGF), transforming growth factor β1 (TGF-β1), TGFβR1, TGFβR2, TGFβR3, α-actin, fibronectin, decorin, elastin, DDR2, and matrix metalloproteinase-2 (MMP2), the effect depended on the treatment protocol

Summary

Introduction

Low-Level Laser Therapy (LLLT) is applied in multiple clinical fields[5]. It is based on the application of low-power amplified light radiation capable of promoting biochemical, bioelectric, and structural changes that produce analgesic, anti-inflammatory, and/or microcirculation-stimulating effects[6,7,8]. The 940 nm diode laser has biostimulatory effect on various cell populations involved in regenerative processes, including osteoblasts in hard tissues[10,11,12] and fibroblasts in soft tissues[13]. Treatment with this laser for 24 or 72 h increased the proliferative capacity of cultured human fibroblasts as a function of the energy dose applied and induced their expression of α-actin, a marker of fibroblast differentiation[13]. The objective of this study was to determine the effects on fibroblasts of treatment with 940 nm diode laser, analyzing the expression of fibroblast growth and differentiation markers and exploring the usefulness of LLLT in soft connective tissue healing

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