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Biological effects of low-level laser irradiation (LLLI) on stem cells from human exfoliated deciduous teeth (SHED)

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Abstract

Objectives

To investigate the effects of low-level laser irradiation (LLLI) on viability/proliferation, migration, osteo/odontogenic differentiation, and in vitro biomineralization of stem cells from human exfoliated deciduous teeth (SHED).

Materials and methods

SHED cultures were established by enzymatic dissociation from pulps of deciduous teeth. SHED were irradiated with a diode laser (InGaAsP; 940 nm; 0.2 W, continuous mode) at energy fluences 4, 8, and 16 J/cm2 in the dark, while non-irradiated SHED served as control. Cell viability/proliferation was evaluated by MTT assay and cell mobilization by Transwell™ migration assay. Expression of osteo/odontogenesis-related genes (ALP, BMP-2, BGLAP, DSPP, MSX2, RUNX2) was assessed by real-time PCR, while in vitro biomineralization by Alizarin Red staining. Statistical analysis was performed by two-way ANOVA and Tukey’s post hoc tests (*p < 0.05, **p < 0.01).

Results

Statistically significant stimulation of cell viability/proliferation was observed at all energy fluences, reaching the highest effect for the 4 and 16 J/cm2. Although the 8 J/cm2 fluence showed the lowest stimulatory effect on cell viability/proliferation, it was the most effective in inducing SHED migration, upregulation of odontogenesis-related genes (DSPP, ALP, BMP-2) at specific time-points, and the in vitro biomineralization potential of SHED compared to the other two energy fluences.

Conclusions

LLLI proved beneficial in promoting SHED biological processes critical for pulp repair in deciduous teeth. Overall, the 8 J/cm2 energy fluence showed the most beneficiary effects.

Clinical relevance

These results provide insights on a narrow “therapeutic window” of LLLI application in vital pulp therapies of deciduous teeth, paving the way for the establishment of effective clinical protocols.

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Acknowledgements

We would like to thank Prof. E. Kriezis, Professor of Optical and Microwave Communications at the Department of Electrical and Computer Engineering, Aristotle University of Thessaloniki (A.U.Th), Greece, for his scientific advices, as well as for the supply of relevant to LLLI measurement equipment.

Funding

The work was supported by institutional funds of the Departments Pediatric Dentistry and Prosthodontics, School of Dentistry, Faculty of Health Science, Aristotle University of Thessaloniki (A.U.Th).

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Authors and Affiliations

  1. Department of Paediatric Dentistry, School of Dentistry, Faculty of Health Sciences, Aristotle University of Thessaloniki (A.U.Th), 541 24, Thessaloniki, Greece

    M. Paschalidou, E. Athanasiadou, K. Arapostathis & N. Kotsanos

  2. Department of Prosthodontics, School of Dentistry, Faculty of Health Sciences, Aristotle University of Thessaloniki (A.U.Th), 541 24, Thessaloniki, Greece

    P. T. Koidis, Athina Bakopoulou & Anna Theocharidou

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  1. M. Paschalidou
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Correspondence to Athina Bakopoulou or Anna Theocharidou.

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All procedures performed in studies involving human biological material (dental pulp tissue from extracted deciduous teeth) were in accordance with the ethical standards of the Institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Paschalidou, M., Athanasiadou, E., Arapostathis, K. et al. Biological effects of low-level laser irradiation (LLLI) on stem cells from human exfoliated deciduous teeth (SHED). Clin Oral Invest 24, 167–180 (2020). https://doi.org/10.1007/s00784-019-02874-4

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