Lasers in Medical Science

, Volume 32, Issue 7, pp 1621–1628 | Cite as

Effects of PBM in different energy densities and irradiance on maintaining cell viability and proliferation of pulp fibroblasts from human primary teeth

  • Nádia Carolina Teixeira Marques
  • Natalino Lourenço Neto
  • Mariel Tavares Oliveira Prado
  • Luciana Lourenço Ribeiro Vitor
  • Rodrigo Cardoso Oliveira
  • Vivien Thiemy Sakai
  • Carlos Ferreira Santos
  • Maria Aparecida Andrade Moreira Machado
  • Thais Marchini Oliveira
Original Article
First Online:
Received:
Accepted:

Abstract

This study aimed to compare the effects of photobiomodulation (PBM) in different energy densities and irradiances on maintaining cell viability, and proliferation of pulp fibroblasts from human primary teeth (HPF) were cultured in DMEM and used between the fourth and eighth passages. Then, HPF were irradiated with the following different energy densities: 1.25 J/cm2 (a), 2.50 J/cm2 (b), 3.75 J/cm2 (c), 5.00 J/cm2 (d), and 6.25 J/cm2 (e); but varying either the time of irradiation (groups 1a–1e) or the output power (groups 2a–2e). Positive (groups 1f and 2f) and negative controls (groups 1g and 2g), respectively, comprised non-irradiated cells grown in regular nutritional conditions (10% fetal bovine serum [FBS]) and under nutritional deficit (1% FBS). Cell viability and proliferation were respectively assessed through MTT and crystal violet (CV) assays at 24, 48, and 72 h after irradiation. Statistical analysis was performed by two-way ANOVA, followed by Tukey test (P < 0.05). The negative controls showed significantly lower viability in relation to most of the corresponding subgroups, both for MTT and CV assays. For both assays, the intragroup comparison showed that the periods of 24 h exhibited lower viability than the periods of 48 and 72 h for most of the subgroups, except the negative controls with lower viability. The different irradiation protocols (equal energy densities applied with different irradiances) showed no statistically significant differences on cell viability and proliferation at the evaluated periods. The proposed PBM in different energy densities and irradiance did not affect the viability and proliferation of pulp fibroblasts from human primary teeth.

Keywords

Culture techniques Cell survival Cell proliferation Dental pulp Low-level light therapy Tooth, deciduous 
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Notes

Acknowledgments

The authors would like to thank all the volunteers and their guardians for consenting to participate in this study, and the São Paulo Research Foundation (FAPESP) for the financial support.

Compliance with ethical standards

Funding

This study received financial support from the São Paulo Research Foundation (FAPESP) (grant nos. 2013/16156-0, 2013/18886-5, and 2015/19696-0).

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This study was approved by the Institutional Review Board regarding ethical aspects (protocol CAAE 21032913.0.0000.5417), and the procedures were performed according to the Helsinki Declaration.

Informed consent

Parents and guardians of the children signed informed consent forms after receiving information about the research.

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Copyright information

© Springer-Verlag London Ltd. 2017

Authors and Affiliations

  • Nádia Carolina Teixeira Marques
    • 1
  • Natalino Lourenço Neto
    • 1
  • Mariel Tavares Oliveira Prado
    • 1
  • Luciana Lourenço Ribeiro Vitor
    • 1
  • Rodrigo Cardoso Oliveira
    • 2
  • Vivien Thiemy Sakai
    • 3
  • Carlos Ferreira Santos
    • 2
  • Maria Aparecida Andrade Moreira Machado
    • 1
  • Thais Marchini Oliveira
    • 1
  1. 1.Department of Pediatric Dentistry, Orthodontics and Public Health, Bauru School of DentistryUSP—University of São PauloBauruBrazil
  2. 2.Department of Biology Science, Bauru School of DentistryUSP—University of São PauloBauruBrazil
  3. 3.Department of Clinics and SurgeryFederal University of Alfenas—UNIFAL-MGAlfenasBrazil

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