Effects of 915 nm laser irradiation on human osteoblasts: a preliminary in vitro study

  • Giovanni Mergoni
  • Paolo Vescovi
  • Silvana Belletti
  • Jacopo Uggeri
  • Samir Nammour
  • Rita Gatti
Original Article
  • 12 Downloads

Abstract

Photobiomodulation (PBM) is a non-invasive treatment that uses laser or led devices making its effects a response to light and not to heat. The possibility of accelerating dental implant osteointegration and orthodontic movements and the need to treat refractory bone lesions, such as bisphosphonate related osteonecrosis of the jaws, has led researchers to consider the effects of PBM on bone for dentistry purposes. The aim of our study was to investigate the effects of 915 nm light supplied with a GaAs diode laser on human osteoblasts in vitro. Osteoblasts were isolated from mandibular cortical bone of a young healthy donor. The irradiation parameters were as follows: doses = 5, 15 and 45 J/cm2; power densities = 0.12 and 1.25 W/cm2; and irradiation times = 41.7, 125 and 375 s. We performed one irradiation per day for 3 and 6 days to study proliferation and differentiation, respectively. Microscopic analysis showed a greater amount of bone nodules in samples treated with 5 J/cm2 and 0.12 W/cm2 compared to controls (56.00 ± 10.44 vs 19.67 ± 7.64, P = 0.0075). Cell growth and quantification of calcium deposition did not show any differences when comparing irradiated and non-irradiated samples. Photobiomodulation, with the parameters investigated in the present study, positively modulated the mineralization process in human osteoblasts, inducing the formation of a greater amount of bone nodules, but did not increase cell proliferation.

Keywords

Photobiomodulation Diode laser Osteoblast Proliferation Differentiation 

Notes

Compliance with ethical standards

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.

References

  1. 1.
    Mester E (1966) The use of the laser beam in therapy. Orv Hetil 107:1012–1016PubMedGoogle Scholar
  2. 2.
    Karu T (1999) Primary and secondary mechanisms of action of visible to near-IR radiation on cells. J Photochem Photobiol B Biol 49:1–17CrossRefGoogle Scholar
  3. 3.
    Khadra M, Lyngstadaas SP, Haanaes HR, Mustafa K (2005) Effect of laser therapy on attachment, proliferation and differentiation of human osteoblast-like cells cultured on titanium implant material. Biomaterials 26:3503–3509CrossRefPubMedGoogle Scholar
  4. 4.
    Walsh LJ (1997) The current status of low level laser therapy in dentistry. Part 1. Soft tissue applications. Aust Dent J 42:247–254CrossRefPubMedGoogle Scholar
  5. 5.
    Walsh LJ (1997) The current status of low level laser therapy in dentistry. Part 2. Hard tissue applications. Aust Dent J 42:302–306CrossRefPubMedGoogle Scholar
  6. 6.
    Huang YY, Sharma SK, Carroll J, Hamblin MR (2011) Biphasic dose response in low level light therapy—an update. Dose Response 9:602–618CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Heymann PGB, Ziebart T, Kammerer PW et al (2016) The enhancing effect of a laser photochemotherapy with cisplatin or zolendronic acid in primary human osteoblasts and osteosarcoma cells in vitro. J Oral Pathol Med 45:803–809CrossRefPubMedGoogle Scholar
  8. 8.
    Karoussis IK, Kyriakidou K, Psarros C, Lang NP, Vrotsos IA (2017) Nd:YAG laser radiation (1.064 nm) accelerates differentiation of osteoblasts to osteocytes on smooth and rough titanium surfaces in vitro. Clin Oral Implants Res 28:785–790CrossRefPubMedGoogle Scholar
  9. 9.
    Oliveira FA, Matos AA, Santesso MR et al (2016) Low intensity lasers differently induce primary human osteoblast proliferation and differentiation. J Photochem Photobiol B Biol 163:14–21CrossRefGoogle Scholar
  10. 10.
    Crisan L, Soritau O, Baciut M, Baciut G, Crisan BV (2015) The influence of laser radiation on human osteoblasts cultured on nanostructured composite substrates. Clujul Med 88:224–232CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Stein A, Benayahu D, Maltz L, Oron U (2005) Low-level laser irradiation promotes proliferation and differentiation of human osteoblasts in vitro. Photomed Laser Surg 23:161–166CrossRefPubMedGoogle Scholar
  12. 12.
    Ueda Y, Shimizu N (2003) Effects of pulse frequency of low-level laser therapy (LLLT) on bone nodule formation in rat calvarial cells. J Clin Laser Med Surg 21:271–277CrossRefPubMedGoogle Scholar
  13. 13.
    Tschon M, Incerti Parenti S, Cepollaro S, Checchi L, Fini M (2015) Photobiomodulation with low-level diode laser promotes osteoblast migration in an in vitro micro wound model. J Biomed Opt 20:078002–078010CrossRefGoogle Scholar
  14. 14.
    Asai T, Suzuki H, Kitayama M et al (2014) The long-term effects of red light-emitting diode irradiation on the proliferation and differentiation of osteoblast-like MC3T3-E1 cells. Kobe J Med Sci 60:E12–E18PubMedGoogle Scholar
  15. 15.
    Migliario M, Pittarella P, Fanuli M, Rizzi M, Renò F (2014) Laser-induced osteoblast proliferation is mediated by ROS production. Lasers Med Sci 29:1463–1467CrossRefPubMedGoogle Scholar
  16. 16.
    Incerti Parenti S, Checchi L, Fini M, Tschon M (2014) Different doses of low-level laser irradiation modulate the in vitro response of osteoblast-like cells. J Biomed Opt 19:108002–108007CrossRefPubMedGoogle Scholar
  17. 17.
    Kanenari M, Zhao J, Abiko Y (2011) Enhancement of microtubule-associated protein-1 alpha gene expression in osteoblasts by low level laser irradiation. Laser Ther 20:47–51CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Kiyosaki T, Mitsui N, Suzuki N, Shimizu N (2010) Low-level laser therapy stimulates mineralization via increased Runx2 expression and ERK phosphorylation in osteoblasts. Photomed Laser Surg 28(Suppl 1):S167–S172PubMedGoogle Scholar
  19. 19.
    Hirata S, Kitamura C, Kukushima H et al (2010) Low-level laser irradiation enhances BMP-induced osteoblast differentiation by stimulating the BMP/Smad signaling pathway. J Cell Biochem 111:1445–1452CrossRefPubMedGoogle Scholar
  20. 20.
    Shimizu N, Mayahara K, Kiyosaki T et al (2007) Low-intensity laser irradiation stimulates bone nodule formation via insulin-like growth factor-I expression in rat calvarial cells. Lasers Surg Med 39:551–559CrossRefPubMedGoogle Scholar
  21. 21.
    Hamajima S, Hiratsuka K, Kiyama-Kishikawa M et al (2003) Effect of low-level laser irradiation on osteoglycin gene expression in osteoblasts. Lasers Med Sci 18:78–82CrossRefPubMedGoogle Scholar
  22. 22.
    Yamamoto M, Tamura K, Hiratsuka K, Abiko Y (2001) Stimulation of MCM3 gene expression in osteoblast by low level laser irradiation. Lasers Med Sci 16:213–217CrossRefPubMedGoogle Scholar
  23. 23.
    Huang TH, Lu YC, Kao CT (2012) Low-level diode laser therapy reduces lipopolysaccharide (LPS)-induced bone cell inflammation. Lasers Med Sci 27:621–627CrossRefPubMedGoogle Scholar
  24. 24.
    Bayat M, Virdi A, Rezaei F, Chien S (2017) Comparison of the in vitro effects of low-level laser therapy and low-intensity pulsed ultrasound therapy on bony cells and stem cells. Progress Biophys Molec Biol 1–13Google Scholar
  25. 25.
    Beresford JN, Gallagher JA, Gowen M et al (1984) The effects of monocyte-conditioned medium and interleukin 1 on the synthesis of collagenous and non-collagenous proteins by mouse bone and human bone cells in vitro. Biochim Biophys Acta 801:58–65CrossRefPubMedGoogle Scholar
  26. 26.
    Gregory CA, Gunn WG, Peister A, Prockop DJ (2004) An alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal Biochem 329:77–84CrossRefPubMedGoogle Scholar
  27. 27.
    Kaur G, Dufour JM (2012) Cell lines: valuable tools or useless artifacts. Spermatogenesis 2:1–5CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Incerti Parenti S, Panseri S, Gracco A et al (2013) Effect of low-level laser irradiation on osteoblast-like cells cultured on porous hydroxyapatite scaffolds. Ann Ist Super Sanita 49:255–260PubMedGoogle Scholar
  29. 29.
    Stein E, Koehn J, Sutter W et al (2008) Initial effects of low-level laser therapy on growth and differentiation of human osteoblast-like cells. Wien Klin Wochenschr 120:112–117CrossRefPubMedGoogle Scholar
  30. 30.
    Renno ACM, McDonnell PA, Parizotto NA, Laakso E-L (2007) The effects of laser irradiation on osteoblast and osteosarcoma cell proliferation and differentiation in vitro. Photomed Laser Surg 25:275–280CrossRefPubMedGoogle Scholar
  31. 31.
    Pagin MT, de Oliveira FA, Oliveira RC et al (2014) Laser and light-emitting diode effects on pre-osteoblast growth and differentiation. Lasers Med Sci 29:55–59CrossRefPubMedGoogle Scholar
  32. 32.
    Fujihara NA, Hiraki KRN, Marques MM (2006) Irradiation at 780 nm increases proliferation rate of osteoblasts independently of dexamethasone presence. Lasers Surg Med 38:332–336CrossRefPubMedGoogle Scholar
  33. 33.
    Ozawa Y, Shimizu N, Kariya G, Abiko Y (1998) Low-energy laser irradiation stimulates bone nodule formation at early stages of cell culture in rat calvarial cells. Bone 22:347–354CrossRefPubMedGoogle Scholar
  34. 34.
    Bloise N, Ceccarelli G, Minzioni P et al (2013) Investigation of low-level laser therapy potentiality on proliferation and differentiation of human osteoblast-like cells in the absence/presence of osteogenic factors. J Biomed Opt 18:128006CrossRefPubMedGoogle Scholar
  35. 35.
    Tang W, Tian J, Zheng Q et al (2015) Implantable self-powered low-level laser cure system for mouse embryonic osteoblasts’ proliferation and differentiation. ACS Nano 9:7867–7873CrossRefPubMedGoogle Scholar
  36. 36.
    Saracino S, Mozzati M, Martinasso G et al (2009) Superpulsed laser irradiation increases osteoblast activity via modulation of bone morphogenetic factors. Lasers Surg Med 41:298–304CrossRefPubMedGoogle Scholar
  37. 37.
    Kim IS, Cho TH, Kim K, Weber FE, Hwang SJ (2010) High power-pulsed Nd:YAG laser as a new stimulus to induce BMP-2 expression in MC3T3-E1 osteoblasts. Lasers Surg Med 42:510–518CrossRefPubMedGoogle Scholar
  38. 38.
    Belletti S, Uggeri J, Mergoni G et al (2015) Effects of 915 nm GaAs diode laser on mitochondria of human dermal fibroblasts: analysis with confocal microscopy. Lasers Med Sci 30:375–381CrossRefPubMedGoogle Scholar
  39. 39.
    Tunér J, Hode L (2004) The laser therapy handbook. Prima Books, GrängesbergGoogle Scholar

Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Unit of Oral Pathology and Laser-assisted Oral Surgery; Department of Medicine and SurgeryUniversity of ParmaParmaItaly
  2. 2.Unit of Anatomy Histology and Embryology, Department of Medicine and SurgeryUniversity of ParmaParmaItaly
  3. 3.Unità Operativa di Patologia ClinicaOspedale di VaioFidenzaItaly
  4. 4.Department of Dental Sciences, Faculty of MedicineUniversity of LiègeLiegeBelgium

Personalised recommendations