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Photobiomodulation effect on angiogenic proteins produced and released by dental pulp cells

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Abstract

Objective

To verify the photobiomodulation effect on angiogenic proteins produced and released by dental human pulpal fibroblasts (HPFs).

Material and methods

HPFs were irradiated with 660-nm low-level laser at fluences of 2.5 J/cm2 and 3.7 J/cm2. The control group was not irradiated. MTT, crystal violet, and ELISA assays respectively verified viability, proliferation, and angiogenic protein (supernatant/lysate) at 6 h, 12 h, and 24 h after photobiomodulation. Capillary-like structure formation assay verified functional role. Two-way ANOVA/Tukey’s test and ANOVA/Bonferroni’s multiple comparisons test respectively verified cell viability/proliferation and intragroup and intergroup comparisons of protein synthesis (p < 0.05).

Results

Irradiated and non-irradiated HPFs showed statistically similar cell viability and proliferation pattern. Intragroup comparisons showed similar patterns of protein synthesis for all groups: VEGF-A, VEGF-C, and vascular endothelial growth factor receptor 1 (VEGFR1) increased significantly in the supernatant, while FGF-2 and VEGF-A increased significantly in the lysate. The lower fluence significantly increased BMP-9 (6 h) in the supernatant and VEGFR1 (6 h and 12 h) and VEGF-D (24 h) in the lysate, while the higher fluence significantly increased BMP-9 (6 h) in the supernatant and VEGFR1 (12 h) in the lysate. Regardless of the time, both fluences statistically downregulated placental growth factor (PLGF) and PDGF secretion. Both fluences statistically decreased VEGF-A secretion (24 h) and PLGF production (6 h).

Conclusion

Photobiomodulation produced stimulatory effects on angiogenic protein secretion by pulp fibroblasts. In terms of photobiomodulation, over time, both fluences significantly increased the secretion of VEGF-A, VEGF-C, and VEGFR1 and significantly upregulated BMP-9 (6 h) in the supernatant; for capillary-like structure formation, the fluence of 2.5 J/cm2 was better than the fluence of 3.7 J/cm2.

Clinical relevance

This study results addressed effective photobiomodulation parameters tailored for pulp angiogenesis.

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Acknowledgments

The authors thank Evandro José Dionisio (in memoriam), Thiago Dionisio, and Cintia Kazuko Tokuhara for their laboratorial assistance.

Funding

This study was funded by Sao Paulo Research Foundation (FAPESP) (grant nos. 2018/20316-6 and 2017/11396-3).

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Authors and Affiliations

  1. Department of Pediatric Dentistry, University of Sacred Heart, Rua Irmã Arminda, 10-50, Bauru, São Paulo, 17011-160, Brazil

    Luciana Lourenço Ribeiro Vitor

  2. Department of Pediatric Dentistry, Orthodontics, and Public Health, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil

    Mariel Tavares Oliveira Prado Bergamo, Natalino Lourenço-Neto, Thiago Cruvinel, Daniela Rios, Maria Aparecida Andrade Moreira Machado & Thais Marchini Oliveira

  3. Department of Clinics and Surgery, School of Dentistry, Federal University of Alfenas, Alfenas, Minas Gerais, Brazil

    Vivien Thiemy Sakai

  4. Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil

    Rodrigo Cardoso Oliveira, Gustavo Pompermaier Garlet & Carlos Ferreira Santos

Authors
  1. Luciana Lourenço Ribeiro Vitor
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Contributions

Luciana Lourenço Ribeiro Vitor participated in the study design, performed the analyses, and drafted the manuscript.

Mariel Tavares Oliveira Prado Bergamo performed the analyses and drafted the manuscript.

Natalino Lourenço-Neto participated in the study design, helped in analyzing the results, and drafted the manuscript.

Vivien Thiemy Sakai participated in the study design, helped in analyzing the results, and drafted the manuscript.

Rodrigo Cardoso Oliveira participated in the study design, helped in analyzing the results, and drafted the manuscript.

Thiago Cruvinel helped in the analyses and drafted the manuscript.

Daniela Rios helped in the analyses and drafted the manuscript.

Gustavo Pompermaier Garlet participated in the study design, helped in analyzing the results, and drafted the manuscript.

Carlos Ferreira Santos helped in analyzing the results and drafted the manuscript.

Maria Aparecida Andrade Moreira Machado participated in the study design and drafted the manuscript.

Thais Marchini Oliveira conceived the study, participated in the study design, and drafted the manuscript.

Corresponding author

Correspondence to Luciana Lourenço Ribeiro Vitor.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards (protocol CAAE 54859816.1.0000.5417).

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Informed consent was obtained from all individual participants included in the study.

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Vitor, L.L.R., Bergamo, M.T.O.P., Lourenço-Neto, N. et al. Photobiomodulation effect on angiogenic proteins produced and released by dental pulp cells. Clin Oral Invest 24, 4343–4354 (2020). https://doi.org/10.1007/s00784-020-03298-1

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