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Effects of 915 nm GaAs diode laser on mitochondria of human dermal fibroblasts: analysis with confocal microscopy

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Abstract

Low-level laser therapy (LLLT) is widely used in tissue regeneration and pain therapy. Mitochondria are supposed to be one of the main cellular targets, due to the presence of cytochrome C oxidase as photo-acceptor. Laser stimulation could influence mitochondria metabolism affecting mainly transmembrane mitochondrial potential (Δψm). The aim of our study is to evaluate “in vitro” the early mitochondrial response after irradiation with a 915 GaAs laser. Since some evidences suggest that cellular response to LLLT can be differently modulated by the mode of irradiation, we would like to evaluate whether there are changes in the mitochondrial potential linked to the use of the laser treatments applied with continuous wave (CW) in respect to those applied with pulsed wave (PW). In this study, we analyzed effects of irradiation with a 915-nm GaAs diode laser on human dermal fibroblast. We compared effects of irradiation applied with either CW or PW at different fluences 45-15-5 J/cm2 on Δψm. Laser scanning microscopy (LSM) was used in living cells to detect ROS (reactive oxygen species) using calcein AM and real-time changes of and Δψm following distribution of the potentiometric probe tetramethylrhodamine methyl ester (TMRM). At higher doses (45–15 J/cm2), fibroblasts showed a dose-dependent decrement of Δψm in either the modalities employed, with higher amplitudes in CW-treated cells. This behavior is transient and not followed by any sign of toxicity, even if reactive oxygen species generation was observed. At 5 J/cm2, CW irradiation determined a little decrease (5 %) of the baseline level of Δψm, while opposite behavior was shown when cells were irradiated with PW, with a 10 % increment. Our results suggest that different responses observed at cellular level with low doses of irradiation, could be at the basis of efficacy of LLLT in clinical application, performed with PW rather than CW modalities.

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Authors and Affiliations

  1. Unit of Anatomy Histology and Embryology—Department of Biomedical, Biotechnological and Translational Sciences (S.Bi.Bi.T), University of Parma, Parma, Italy

    Silvana Belletti, Jacopo Uggeri & Rita Gatti

  2. Unit of Oral Pathology, Medicine and Laser Surgery, Department of Biomedical, Biotechnological and Translational Sciences (S.Bi.Bi.T), University of Parma, Parma, Italy

    Giovanni Mergoni, Paolo Vescovi, Elisabetta Merigo, Carlo Fornaini & Maddalena Manfredi

  3. Department of Dental Sciences, Faculty of Medicine, University of Liège, Liège, Belgium

    Samir Nammour

  4. Unit of Oral Pathology, Medicine and Laser Surgery, Department of Biomedical, Biotechnological and Translational Sciences (S.Bi.Bi.T), University of Parma, via Gramsci, 14, 43126, Parma, Italy

    Elisabetta Merigo

Authors
  1. Silvana Belletti
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  2. Jacopo Uggeri
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  3. Giovanni Mergoni
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  4. Paolo Vescovi
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  6. Carlo Fornaini
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  8. Maddalena Manfredi
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  9. Rita Gatti
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Correspondence to Elisabetta Merigo.

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Belletti, S., Uggeri, J., Mergoni, G. et al. Effects of 915 nm GaAs diode laser on mitochondria of human dermal fibroblasts: analysis with confocal microscopy. Lasers Med Sci 30, 375–381 (2015). https://doi.org/10.1007/s10103-014-1651-z

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  • DOI: https://doi.org/10.1007/s10103-014-1651-z

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