Advertisement

Lasers in Medical Science

, Volume 32, Issue 4, pp 833–840 | Cite as

Neuropeptide expression and morphometric differences in crushed alveolar inferior nerve of rats: Effects of photobiomodulation

  • Daniel Oliveira MartinsEmail author
  • Fabio Martinez dos Santos
  • Adriano Polican Ciena
  • Ii-sei Watanabe
  • Luiz Roberto G. de Britto
  • José Benedito Dias Lemos
  • Marucia Chacur
Original Article

Abstract

Inferior alveolar nerve (IAN) injuries may occur during various dental routine procedures, especially in the removal of impacted lower third molars, and nerve recovery in these cases is a great challenge in dentistry. Here, the IAN crush injury model was used to assess the efficacy of photobiomodulation (PBM) in the recovery of the IAN in rats following crushing injury (a partial lesion). Rats were divided into four experimental groups: without any procedure, IAN crush injury, and IAN crush injury with PBM and sham group with PBM. Treatment was started 2 days after surgery, above the site of injury, and was performed every other day, totaling 10 sessions. Rats were irradiated with GaAs Laser (Gallium Arsenide, Laserpulse, Ibramed Brazil) emitting a wavelength of 904 nm, an output power of 70 mWpk, beam spot size at target ∼0.1 cm2, a frequency of 9500 Hz, a pulse time 60 ns, and an energy density of 6 J/cm2. Nerve recovery was investigated by measuring the morphometric data of the IAN using TEM and by the expression of laminin, neurofilaments (NFs), and myelin protein zero (MPZ) using Western blot analysis. We found that IAN-injured rats which received PBM had a significant improvement of IAN morphometry when compared to IAN-injured rats without PBM. In parallel, all MPZ, laminin, and NFs exhibited a decrease after PBM. The results of this study indicate that the correlation between the peripheral nerve ultrastructure and the associated protein expression shows the beneficial effects of PBM.

Keywords

Molecular changes Alveolar inferior nerve Photobiomodulation Myelin sheath Rat 

Notes

Acknowledgements

The authors are grateful to Sônia Regina Yokomizo for her valuable help.

Authors’ contributions

All authors made substantial contributions to the following tasks of research: initial conception (Martins D.O., Britto L.R.G., Lemos J.B.D., Chacur M.), design (Martins D.O., Britto L.R.G., Lemos J.B.D., Chacur M), provision of resources (Chacur M), collection of data (Martins D.O, Santos F.M.,, Ciena A.P., Watanabe I.), analysis and interpretation of data (Martins D.O., Chacur M., Ciena A.P., Watanabe I.), writing the first draft of the paper or important intellectual content (Martins D.O, Santos F.M.), and revision of paper (Martins D.O., Chacur M., Britto L.R.G.).

Compliance with ethical standards

Ethics approval and consent to participate

All procedures were approved by the Institutional Animal Care Committee of the University of São Paulo (protocol number 150/2010) and performed in accordance with the guidelines for the ethical use of conscious animals in pain study published by the international association for the study of pain.

Competing interests

The authors declare that they have no competing interests.

Funding

FAPESP 2010/20026-6; 2012/05840-4; 2014/24533-0

References

  1. 1.
    Raimondo S et al (2011) Perspectives in regeneration and tissue engineering of peripheral nerves. Ann Anat 193(4):334–40CrossRefPubMedGoogle Scholar
  2. 2.
    Mendonca AC, Barbieri CH, Mazzer N (2003) Directly applied low intensity direct electric current enhances peripheral nerve regeneration in rats. J Neurosci Methods 129(2):183–90CrossRefPubMedGoogle Scholar
  3. 3.
    Anders JJ, Geuna S, Rochkind S (2004) Phototherapy promotes regeneration and functional recovery of injured peripheral nerve. Neurol Res 26(2):233–9CrossRefPubMedGoogle Scholar
  4. 4.
    Tucker BA, Mearow KM (2008) Peripheral sensory axon growth: from receptor binding to cellular signaling. Can J Neurol Sci 35(5):551–66CrossRefPubMedGoogle Scholar
  5. 5.
    Wang X et al (2011) Schwann-like mesenchymal stem cells within vein graft facilitate facial nerve regeneration and remyelination. Brain Res 1383:71–80CrossRefPubMedGoogle Scholar
  6. 6.
    Thompson DM, Buettner HM (2001) Schwann cell response to micropatterned laminin surfaces. Tissue Eng 7(3):247–65CrossRefPubMedGoogle Scholar
  7. 7.
    Chen ZL, Strickland S (2003) Laminin gamma1 is critical for Schwann cell differentiation, axon myelination, and regeneration in the peripheral nerve. J Cell Biol 163(4):889–99CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Yuan A et al (2009) Neurofilaments form a highly stable stationary cytoskeleton after reaching a critical level in axons. J Neurosci 29(36):11316–29CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Al-Chalabi A, Miller CC (2003) Neurofilaments and neurological disease. Bioessays 25(4):346–55CrossRefPubMedGoogle Scholar
  10. 10.
    Walker KL et al (2001) Loss of neurofilaments alters axonal growth dynamics. J Neurosci 21(24):9655–66PubMedGoogle Scholar
  11. 11.
    Shao Y et al (2002) Effect of nerve growth factor on changes of myelin basic protein and functional repair of peripheral nerve following sciatic nerve injury in rats. Chin J Traumatol 5(4):237–40PubMedGoogle Scholar
  12. 12.
    Boyce VS et al (2007) Neurotrophic factors promote and enhance locomotor recovery in untrained spinalized cats. J Neurophysiol 98(4):1988–96CrossRefPubMedGoogle Scholar
  13. 13.
    Sasaki M et al (2009) BDNF-hypersecreting human mesenchymal stem cells promote functional recovery, axonal sprouting, and protection of corticospinal neurons after spinal cord injury. J Neurosci 29(47):14932–41CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Petruska JC, Mendell LM (2004) The many functions of nerve growth factor: multiple actions on nociceptors. Neurosci Lett 361(1–3):168–71CrossRefPubMedGoogle Scholar
  15. 15.
    Zochodne DW (2000) The microenvironment of injured and regenerating peripheral nerves. Muscle Nerve Suppl 9:S33–8CrossRefPubMedGoogle Scholar
  16. 16.
    de Oliveira Martins D et al (2013) Laser therapy and pain-related behavior after injury of the inferior alveolar nerve: possible involvement of neurotrophins. J Neurotrauma 30(6):480–6CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Zimmermann M (1983) Ethical guidelines for investigations of experimental pain in conscious animals. Pain 16(2):109–10CrossRefPubMedGoogle Scholar
  18. 18.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–54CrossRefPubMedGoogle Scholar
  19. 19.
    Ciena AP et al (2012) Fine structure of myotendinous junction between the anterior belly of the digastric muscle and intermediate tendon in adults rats. Micron 43(2–3):258–62CrossRefPubMedGoogle Scholar
  20. 20.
    da Silva JT et al (2015) Neural mobilization promotes nerve regeneration by nerve growth factor and myelin protein zero increased after sciatic nerve injury. Growth Factors 33(1):8–13CrossRefPubMedGoogle Scholar
  21. 21.
    Watanabe I, Yamada E (1983) The fine structure of lamellated nerve endings found in the rat gingiva. Arch Histol Jpn 46(2):173–82CrossRefPubMedGoogle Scholar
  22. 22.
    Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–5CrossRefPubMedGoogle Scholar
  23. 23.
    Snedecor GW, Sokal RR, Rohlf FJ (1946) Statistical methods biometry. 4 ed. Ames, ed. W.H. Freeman & Co. New York: Owa State University Press. p.859Google Scholar
  24. 24.
    Chaplan SR et al (1994) Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Methods 53:55–63Google Scholar
  25. 25.
    Yonehara N, Kudo C, Kamisaki Y (2003) Involvement of NMDA-nitric oxide pathways in the development of tactile hypersensitivity evoked by the loose-ligation of inferior alveolar nerves in rats. Brain Res 963(1–2):232–43Google Scholar
  26. 26.
    Chen ZL, Yu WM, Strickland S (2007) Peripheral regeneration. Annu Rev Neurosci 30:209–33CrossRefPubMedGoogle Scholar
  27. 27.
    Fortun J, Hill CE, Bunge MB (2009) Combinatorial strategies with Schwann cell transplantation to improve repair of the injured spinal cord. Neurosci Lett 456(3):124–32CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Toft A et al (2013) A comparative study of glial and non-neural cell properties for transplant-mediated repair of the injured spinal cord. Glia 61(4):513–28CrossRefPubMedGoogle Scholar
  29. 29.
    Webber CA et al (2011) Schwann cells direct peripheral nerve regeneration through the Netrin-1 receptors, DCC and Unc5H2. Glia 59(10):1503–17CrossRefPubMedGoogle Scholar
  30. 30.
    Podratz JL, Rodriguez E, Windebank AJ (2001) Role of the extracellular matrix in myelination of peripheral nerve. Glia 35(1):35–40CrossRefPubMedGoogle Scholar
  31. 31.
    Tsiper MV, Yurchenco PD (2002) Laminin assembles into separate basement membrane and fibrillar matrices in Schwann cells. J Cell Sci 115(Pt 5):1005–15PubMedGoogle Scholar
  32. 32.
    Mirsky R et al (2001) Regulation of genes involved in Schwann cell development and differentiation. Prog Brain Res 132:3–11CrossRefPubMedGoogle Scholar
  33. 33.
    Uziyel Y, Hall S, Cohen J (2000) Influence of laminin-2 on Schwann cell-axon interactions. Glia 32(2):109–21CrossRefPubMedGoogle Scholar
  34. 34.
    Rankin SL et al (2008) Neurotrophin-induced upregulation of p75NTR via a protein kinase C-delta-dependent mechanism. Brain Res 1217:10–24CrossRefPubMedGoogle Scholar
  35. 35.
    Luo L (2002) Actin cytoskeleton regulation in neuronal morphogenesis and structural plasticity. Annu Rev Cell Dev Biol 18:601–35CrossRefPubMedGoogle Scholar
  36. 36.
    Nakagawa M et al (2001) Schwann cell myelination occurred without basal lamina formation in laminin alpha2 chain-null mutant (dy3K/dy3K) mice. Glia 35(2):101–10CrossRefPubMedGoogle Scholar
  37. 37.
    Yang D et al (2005) Coordinate control of axon defasciculation and myelination by laminin-2 and -8. J Cell Biol 168(4):655–66CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Yu WM et al (2005) Schwann cell-specific ablation of laminin gamma1 causes apoptosis and prevents proliferation. J Neurosci 25(18):4463–72CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Rotenstein L et al (2008) Characterization of the shark myelin Po protein. Brain Behav Evol 72(1):48–58CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Varejao AS et al (2004) Functional and morphological assessment of a standardized rat sciatic nerve crush injury with a non-serrated clamp. J Neurotrauma 21(11):1652–70CrossRefPubMedGoogle Scholar
  41. 41.
    Perrot R et al (2008) Review of the multiple aspects of neurofilament functions, and their possible contribution to neurodegeneration. Mol Neurobiol 38(1):27–65CrossRefPubMedGoogle Scholar
  42. 42.
    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):S3–40PubMedGoogle Scholar

Copyright information

© Springer-Verlag London 2017

Authors and Affiliations

  • Daniel Oliveira Martins
    • 1
    Email author
  • Fabio Martinez dos Santos
    • 1
    • 2
  • Adriano Polican Ciena
    • 1
    • 3
  • Ii-sei Watanabe
    • 1
  • Luiz Roberto G. de Britto
    • 4
  • José Benedito Dias Lemos
    • 5
  • Marucia Chacur
    • 1
  1. 1.Department of Anatomy, Institute of Biomedical SciencesUniversity of São PauloSão PauloBrazil
  2. 2.University Nove de JulhoSão PauloBrazil
  3. 3.Institute of BiosciencesUniversity Estadual Paulista Júlio de Mesquita FilhoRio ClaroBrazil
  4. 4.Department of Physiology and Biophysics, Institute of Biomedical SciencesUniversity of São PauloSão PauloBrazil
  5. 5.Department of Surgery, School of DentistryUniversity of São PauloSão PauloBrazil

Personalised recommendations