Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Angiogenic protein synthesis after photobiomodulation therapy on SHED: a preliminary study


This study evaluated the viability, proliferation, and protein expression after photobiomodulation (PBM) of stem cell from human exfoliated deciduous teeth (SHED). The groups were the following: G1 (2.5 J/cm2), G2 (3.7 J/cm2), and control (not irradiated). According to the groups, cells were irradiated with InGaAlP diode laser at 660 nm wavelength, continuous mode, and single time application. After 6 h, 12 h, and 24 h from irradiation, the cell viability and proliferation, and the protein expression were analyzed by MTT, crystal violet, and ELISA multiplex assay, respectively. Twenty-four hours after PBM, SHED showed better proliferation. Over time in the supernatant, all groups had an increase at the levels of VEGF-C, VEGF-A, and PLGF. In the lysate, the control and G2 exhibited a decrease of the VEGF-A, PECAM-1, and PLGF expression, while control and G3 decreased VEGF-C, VEGF-A, and PDGF expression. The dosimetries of 2.5 J/cm2 and 3.7 J/cm2 maintained viability, improved proliferation, and synthesis of the angiogenic proteins in the supernatant in the studied periods on SHED.

This is a preview of subscription content, log in to check access.

Fig. 1


  1. 1.

    Rosa V, Dubey N, Islam I, Min KS, Nör JE (2016) Pluripotency of stem cells from human exfoliated deciduous teeth for tissue engineering. Stem Cells Int 2016:5957806.

  2. 2.

    Bottino MC, Pankajakshan D, Nör JE (2017) Advanced scaffolds for dental pulp and periodontal regeneration. Dent Clin N Am 61:689–711.

  3. 3.

    Chalisserry EP, Nam SY, Park SH, Anil S (2017) Therapeutic potential of dental stem cells. J Tissue Eng 8:2041731417702531.

  4. 4.

    Arany PR (2016) Photobiomodulation therapy: communicating with stem cells for regeneration. Photomed Laser Surg 34:497–499.

  5. 5.

    Sakai VT, Zhang Z, Dong Z, Neiva KG, Machado MAAM, Shi S, Santos CF, Nör JE (2010) SHED differentiate into functional odontoblasts and endothelium. J Dent Res 89:791–796.

  6. 6.

    Bento LW, Zhang Z, Imai A, Nör F, Dong Z, Shi S, Araujo FB, Nör JE (2013) Endothelial differentiation of SHED requires MEK1/ERK signaling. J Dent Res 92:51–57.

  7. 7.

    Zhang Z, Nör F, Oh M, Cucco C, Shi S, Nör JE (2016) Wnt/β-catenin signaling determines the vasculogenic fate of postnatal mesenchymal stem cells. Stem Cells 34:1576–1587.

  8. 8.

    Gronthos S, Mankani M, Brahim J, Robey PG, Shi S (2000) Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A 97:13625–13630.

  9. 9.

    Miura M, Gronthos S, Zhao M, Lu B, Fisher LW, Robey PG, Shi S (2003) SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci U S A 100:5807–5812.

  10. 10.

    Wang H, Zhong Q, Yang T, Qi Y, Fu M, Yang X, Qiao L, Ling Q, Liu S, Zhao Y (2018) Comparative characterization of SHED and DPSCs during extended cultivation in vitro. Mol Med Rep 17:6551–6559.

  11. 11.

    Nakamura S, Yamada Y, Katagiri W, Sugito T, Ito K, Ueda M (2009) Stem cell proliferation pathways comparison between human exfoliated deciduous teeth and dental pulp stem cells by gene expression profile from promising dental pulp. J Endod 35:1536–1542.

  12. 12.

    Kaneko T, Gu B, Sone PP, Zaw SYM, Murano H, Zaw ZCT, Okiji T (2018) Dental pulp tissue engineering using mesenchymal stem cells: a review with a protocol. Stem Cell Rev 14:668–676.

  13. 13.

    Marques MM, Diniz IM, de Cara SP, Pedroni AC, Abe GL, D’Almeida-Couto RS, Lima PL, Tedesco TK, Moreira MS (2016) Photobiomodulation of dental derived mesenchymal stem cells: a systematic review. Photomed Laser Surg 34:500–508.

  14. 14.

    Marques NCT, Neto NL, Prado MTO, Vitor LLR, Oliveira RC, Sakai VT, Santos CF, Machado MAAM, Oliveira TM (2017) Effects of PBM in different energy densities and irradiance on maintaining cell viability and proliferation of pulp fibroblasts from human primary teeth. Lasers Med Sci 32:1621–1628.

  15. 15.

    Marques NP, Lopes CS, Marques NCT, Cosme-Silva L, Oliveira TM, Duque C, Sakai VT, Hanemann JAC (2019) A preliminary comparison between the effects of red and infrared laser irradiation on viability and proliferation of SHED. Lasers Med Sci 34:465–471.

  16. 16.

    Fernandes AP, Junqueira MA, Marques NC, Machado MAAM, Santos CF, Oliveira TM, Sakai VT (2016) Effects of low-level laser therapy on stem cells from human exfoliated deciduous teeth. J Appl Oral Sci 24:332–337.

  17. 17.

    Ferreira LS, Diniz IMA, Maranduba CMS, Miyagi SPH, Rodrigues MFSD, Moura-Netto C, Marques MM (2019) Short-term evaluation of photobiomodulation therapy on the proliferation and undifferentiated status of dental pulp stem cells. Lasers Med Sci 34(4):659–666.

  18. 18.

    Souza L, Rinco U, Aguiar D, Almeida Junior L, Cosme-Silva L, Oliveira T, Marques N, Sakai V (2018) Effect of photobiomodulation on viability and proliferation of stem cells from exfoliated deciduous teeth under different nutritional conditions. Laser Phys 28:1–5.

  19. 19.

    Almeida-Lopes L, Rigau J, Zângaro RA, Guidugli-Neto J, Jaeger MM (2001) Comparison of the low level laser therapy effects on cultured human gingival fibroblasts proliferation using different irradiance and same fluence. Lasers Surg Med 29:179–184.

  20. 20.

    Basso FG, Pansani TN, Turrioni AP, Bagnato VS, Hebling J, de Souza Costa CA (2012) In vitro wound healing improvement by low-level laser therapy application in cultured gingival fibroblasts. Int J Dent 2012:719452.

  21. 21.

    Kreisler M, Christoffers AB, Al-Haj H, Willershausen B, d’Hoedt B (2002) Low level 809-nm diode laser-induced in vitro stimulation of the proliferation of human gingival fibroblasts. Lasers Surg Med 30:365–369.

  22. 22.

    Pereira AN, Eduardo CP, Matson E, Marques MM (2002) Effect of low-power laser irradiation on cell growth and procollagen synthesis of cultured fibroblasts. Lasers Surg Med 31:263–267.

  23. 23.

    AlGhamdi KM, Kumar A, Moussa NA (2012) Low-level laser therapy: a useful technique for enhancing the proliferation of various cultured cells. Lasers Med Sci 27:237–249.

  24. 24.

    Peplow PV, Chung TY, Baxter GD (2010) Laser photobiomodulation of proliferation of cells in culture: a review of human and animal studies. Photomed Laser Surg 28(Suppl 1):S3–S40.

  25. 25.

    Marques MM, Pereira AN, Fujihara NA, Nogueira FN, Eduardo CP (2004) Effect of low-power laser irradiation on protein synthesis and ultrastructure of human gingival fibroblasts. Lasers Surg Med 34:260–265.

  26. 26.

    Ginani F, Soares DM, de Oliveira Rocha HA, de Souza LB, Barboza CAG (2018) Low-level laser irradiation induces in vitro proliferation of stem cells from human exfoliated deciduous teeth. Lasers Med Sci 33:95–102.

  27. 27.

    Ginani F, Soares DM, Barreto MP, Barboza CA (2015) Effect of low-level laser therapy on mesenchymal stem cell proliferation: a systematic review. Lasers Med Sci 30:2189–2194.

  28. 28.

    Vasconcelos RG, Ribeiro RA, Vasconcelos MG, Lima KC, Barboza CA (2012) In vitro comparative analysis of cryopreservation of undifferentiated mesenchymal cells derived from human periodontal ligament. Cell Tissue Bank 13:461–469.

  29. 29.

    Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, Horwitz E (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317.

  30. 30.

    Prado MTO, Vitor LLR, Neto NL, Marques NCT, Sakai VT, Rios D, Cruvinel T, Oliveira RC, Santos CF, Machado MAAM, Oliveira TM (2019) Effects of different culture media, cell densities and adhesion periods on stem cells from human exfoliated deciduous teeth after photobiomodulation. Laser Phys Lett 16:095601

  31. 31.

    Vitor LLR, Prado MTO, Lourenço Neto N, Oliveira RC, Santos CF, Machado MAAM, Oliveira TM (2018) Photobiomodulation changes type 1 collagen gene expression by pulp fibroblasts. Laser Phys 2018:1–5.

  32. 32.

    Farivar S, Malekshahabi T, Shiari R (2014) Biological effects of low level laser therapy. J Lasers Med Sci 5:58–62

  33. 33.

    Borzabadi-Farahani A (2016) Effect of low-level laser irradiation on proliferation of human dental mesenchymal stem cells; a systemic review. J Photochem Photobiol B 162:577–582.

  34. 34.

    Marques M, Patricia S, Miyagi H, Abe G, Clara A, Pedroni F, Márcia I, Diniz A, Moreira M (2017) Effects of photobiomodulation therapy in dentoalveolar-derived mesenchymal stem cells : a review of literature. Lasers Dent Sci 1:1–7.

  35. 35.

    Zaccara IM, Mestieri LB, Moreira MS, Grecca FS, Martins MD, Kopper PMP (2018) Photobiomodulation therapy improves multilineage differentiation of dental pulp stem cells in three-dimensional culture model. J Biomed Opt 23:1–9.

  36. 36.

    da Silva PCS, Marques NP, Farina MT, Oliveira TM, Duque C, Marques NCT, Sakai VT (2019) Laser treatment contributes to maintain membrane integrity in stem cells from human exfoliated deciduous teeth (shed) under nutritional deficit. Lasers Med Sci 34:15–21.

  37. 37.

    Karu T (1999) Primary and secondary mechanisms of action of visible to near-IR radiation on cells. J Photochem Photobiol B 49:1–17.

  38. 38.

    George S, Hamblin MR, Abrahamse H (2018) Effect of red light and near infrared laser on the generation of reactive oxygen species in primary dermal fibroblasts. J Photochem Photobiol B 188:60–68.

  39. 39.

    Arany P, Cho A, Hunt T, Sidhu G, Shin K, Hahm E, Huang G, Weaver J, Chen A, Padwa B, Hamblin M, Barcellos-Hoff M, Kulkarni A, Mooney D (2014) Photoactivation of endogenous latent transforming growth factor–β1 directs dental stem cell differentiation for regeneration. Sci Transl Med 238ra69:01–22.

  40. 40.

    Claesson-Welsh L (2016) VEGF receptor signal transduction-a brief update. Vasc Pharmacol 86:14–17.

  41. 41.

    Osman A, Gnanasegaran N, Govindasamy V, Kathivaloo P, Wen AS, Musa S, Abu Kasim NH (2014) Basal expression of growth-factor-associated genes in periodontal ligament stem cells reveals multiple distinctive pathways. Int Endod J 47:639–651.

  42. 42.

    Autiero M, Waltenberger J, Communi D, Kranz A, Moons L, Lambrechts D, Kroll J, Plaisance S, De Mol M, Bono F, Kliche S, Fellbrich G, Ballmer-Hofer K, Maglione D, Mayr-Beyrle U, Dewerchin M, Dombrowski S, Stanimirovic D, Van Hummelen P, Dehio C, Hicklin DJ, Persico G, Herbert JM, Shibuya M, Collen D, Conway EM, Carmeliet P (2003) Role of PlGF in the intra- and intermolecular cross talk between the VEGF receptors Flt1 and Flk1. Nat Med 9:936–943.

  43. 43.

    Smith AJ, Duncan HF, Diogenes A, Simon S, Cooper PR (2016) Exploiting the bioactive properties of the dentin-pulp complex in regenerative endodontics. J Endod 42:47–56.

  44. 44.

    Duong T, Koltowska K, Pichol-Thievend C, Le Guen L, Fontaine F, Smith KA, Truong V, Skoczylas R, Stacker SA, Achen MG, Koopman P, Hogan BM, Francois M (2014) VEGFD regulates blood vascular development by modulating SOX18 activity. Blood 123:1102–1112.

  45. 45.

    Dou L, Yan Q, Liang P, Zhou P, Zhang Y, Ji P (2018) iTRAQ-based proteomic analysis exploring the influence of hypoxia on the proteome of dental pulp stem cells under 3D culture. Proteomics 18:3–4.

  46. 46.

    Yun YR, Won JE, Jeon E, Lee S, Kang W, Jo H, Jang JH, Shin US, Kim HW (2010) Fibroblast growth factors: biology, function, and application for tissue regeneration. J Tissue Eng 2010:218142.

Download references


This study was funded by the State of São Paulo Research Foundation (Grant #2017/11396-3; #2018/20316-6).

Author information

Correspondence to Thais Marchini Oliveira.

Ethics declarations

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 88330218.6.0000.5417).

Informed consent

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

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Oliveira Prado Bergamo, M.T., Vitor, L.L.R., Lopes, N.M. et al. Angiogenic protein synthesis after photobiomodulation therapy on SHED: a preliminary study. Lasers Med Sci (2020).

Download citation


  • Photobiomodulation therapy
  • Dental pulp
  • Angiogenic proteins
  • Stem cell