Skip to main content

Advertisement

SpringerLink
Log in

Analysis of the variation in low-level laser energy density on the crushed sciatic nerves of rats: a morphological, quantitative, and morphometric study

  • Original Article
  • Published:
Lasers in Medical Science Aims and scope Submit manuscript

Abstract

The objective of this study was to evaluate three energy densities of low-level laser therapy (LLLT, GaAlAs, 780 nm, 40 mW, 0.04 cm2) for the treatment of lesions to peripheral nerves using the sciatic nerve of rats injured via crushing model (15 kgf, 5.2 MPa). Thirty Wistar rats (♂, 200–250 g) were divided into five groups (n = 6): C—control, not injured, and irradiated; L0—injured nerve without irradiation; L4—injured nerve irradiated with LLLT 4 J/cm2 (0.16 J); L10—injured nerve irradiated with LLLT 10 J/cm2 (0.4 J); and L50—injured nerve irradiated with LLLT 50 J/cm2 (2 J). The animals were sacrificed 2 weeks after the injury via perfusion with glutaraldehyde (2.5%, 0.1 M sodium cacodylate buffer). The nerve tissue was embedded in historesin, cut (3 μm), mounted on slides, and stained (Sudan black and neutral red). The morphological and quantitative analysis (myelin and blood capillary densities) and morphometric parameters (maximum and minimum diameters of nerve fibers, axon diameter, G-ratio, myelin sheath thickness) were assessed using the ImageJ software. ANOVA (parametric) or Kruskal-Wallis (nonparametric) tests were used for the statistical analysis. Groups L0, L4, L10, and L50 exhibited diminished values of all the quantitative and morphometric parameters in comparison to the control group. The morphological, quantitative, and morphometric data revealed improvement after injury in groups L4, L10, and L50 (irradiated groups) compared to the injured-only group (L0); the best results, in general, were observed for the L10 group after 15 days of nerve injury.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Lundborg G (1987) Nerve regeneration and repair. A review. Acta orthop scand 58:145–169

    Article  CAS  PubMed  Google Scholar 

  2. Evans GR (2001) Peripheral nerve injury: a review and approach to tissue engineered constructs. Anat rec 263:396–404

    Article  CAS  PubMed  Google Scholar 

  3. Raimondo S, Fornaro M, Tos P et al (2011) Perspectives in regeneration and tissue engineering of peripheral nerves. Ann anat 193:334–340

    Article  PubMed  Google Scholar 

  4. Martínez De Albornoz P, Delgado PJ, Forriol F, Maffulli N (2011) Non-surgical therapies for peripheral nerve injury. Br med bull 100:73–100

    Article  PubMed  Google Scholar 

  5. Shen CC, Yang YC, Liu BS (2011) Large-area irradiated low-level laser effect in a biodegradable nerve guide conduit on neural regeneration of peripheral nerve injury in rats. Injury 42:803–813

    Article  PubMed  Google Scholar 

  6. Dubový P (2011) Wallerian degeneration and peripheral nerve conditions for both axonal regeneration and neuropathic pain induction. Ann anat 193:267–275

    Article  PubMed  Google Scholar 

  7. Rovak JM, Mungara AK, Aydin MA et al (2004) Effects of vascular endothelial growth factor on nerve regeneration in acellular nerve grafts. J reconstr microsurg 20:53–58

    Article  PubMed  Google Scholar 

  8. Morries LD, Cassano P, Henderson TA (2015) Treatments for traumatic brain injury with emphasis on transcranial near-infrared laser phototherapy. Neuropsychiatr dis treat 11:2159–2175

    PubMed  PubMed Central  Google Scholar 

  9. Hashmi JT, Huang YY, Osmani BZ et al (2010) Role of low-level laser therapy in neurorehabilitation. PM R 12:s292–s305

    Article  Google Scholar 

  10. Gomes LE, Dalmarco EM, André ES (2012) The brain-derived neurotrophic factor, nerve growth factor, neurotrophin-3, and induced nitric oxide synthase expressions after low-level laser therapy in an axonotmesis experimental model. Photomed laser surg 30:642–647

    Article  CAS  PubMed  Google Scholar 

  11. Yazdani SO, Golestaneh AF, Shafiee A et al (2012) Effects of low level laser therapy on proliferation and neurotrophic factor gene expression of human Schwann cells in vitro. J photochem photobiol b 107:9–13

    Article  CAS  PubMed  Google Scholar 

  12. Belchior AC, Dos Reis FA, Nicolau RA et al (2009) Influence of laser (660 nm) on functional recovery of the sciatic nerve in rats following crushing lesion. Lasers med sci 24:893–899

    Article  PubMed  Google Scholar 

  13. Dos Reis FA, Belchior AC, De Carvalho PT et al (2009) Effect of laser therapy (660 nm) on recovery of the sciatic nerve in rats after injury through neurotmesis followed by epineural anastomosis. Lasers med sci 24:741–747

    Article  PubMed  Google Scholar 

  14. Barbosa RI, Marcolino AM, De Jesus Guirro RR et al (2010) Comparative effects of wavelengths of low-power laser in regeneration of sciatic nerve in rats following crushing lesion. Lasers med sci 25:423–430

    Article  PubMed  Google Scholar 

  15. Gigo-Benato D, Russo TL, Tanaka EH et al (2010) Effects of 660 and 780 nm low-level laser therapy on neuromuscular recovery after crush injury in rat sciatic nerve. Lasers surg med 42:673–862

    Article  PubMed  Google Scholar 

  16. Medalha CC, Di Gangi GC, Barbosa CB et al (2012) Low-level laser therapy improves repair following complete resection of the sciatic nerve in rats. Lasers med sci 27:629–635

    Article  PubMed  Google Scholar 

  17. Marcolino AM, Barbosa RI, Das Neves LM, Mazzer N, De Jesus Guirro RR, De Cássia Registro Fonseca M (2013) Assessment of functional recovery of sciatic nerve in rats submitted to low-level laser therapy with different fluences. An experimental study: laser in functional recovery in rats. J hand microsurg 5:49–53

    Article  PubMed  PubMed Central  Google Scholar 

  18. Sene GA, Souza FF, Fazan VS, E Barbieri CH (2013) Effects of laser therapy in peripheral nerve regeneration. Acta ortop bras 21:266–270

    Article  PubMed  PubMed Central  Google Scholar 

  19. Wang CZ, Chen YJ, Wang YH, Yeh ML, Huang MH, Ho ML, Liang JI, Chen CH (2014) Low-level laser irradiation improves functional recovery and nerve regeneration in sciatic nerve crush rat injury model. Plos one 9:e103348

    Article  PubMed  PubMed Central  Google Scholar 

  20. Takhtfooladi MA, Jahanbakhsh F, Takhtfooladi HA, Yousefi K, Allahverdi A (2015) Effect of low-level laser therapy (685 nm, 3 J/cm2) on functional recovery of the sciatic nerve in rats following crushing lesion. Lasers med sci 30:1047–1052

    Article  PubMed  Google Scholar 

  21. Dias FJ, Issa JPM, Coutinho-Netto J et al (2015) Morphometric and high resolution scanning electron microscopy analysis of low-level laser therapy and latex protein (Hevea brasiliensis) administration following a crush injury of the sciatic nerve in rats. J neurol sci 349:129–137

    Article  CAS  PubMed  Google Scholar 

  22. Muniz KL, Dias FJ, Coutinho-Netto J et al (2015) Properties of the tibialis anterior muscle after treatment with laser therapy and natural latex protein following sciatic nerve crush. Muscle nerve 52:869–875

    Article  CAS  PubMed  Google Scholar 

  23. Schiaveto De Souza A, Da Silva CA, Del Bel EA (2004) Methodological evaluation to analyze functional recovery after sciatic nerve injury. J neurotrauma 21:627–635

    Article  CAS  PubMed  Google Scholar 

  24. Behmer OA, Tolosa EMC, Freitas Neto AG (1976) Manual of techniques for normal and pathological histology [Portuguese] 1 ed. EDART, Editora da Universidade de São Paulo, São Paulo

  25. Arino H, Brandt J, Dahlin LB (2008) Implantation of Schwann cells in rat tendon autografts as a model for peripheral nerve repair: long term effects on functional recovery. Scand j plast reconstr surg hand surg 42:281–285

    Article  PubMed  Google Scholar 

  26. Chomiak T, Hu B (2009) What is the optimal value of the g-ratio for myelinated fibers in the rat CNS? A theoretical approach. Plos one 4:e7754

    Article  PubMed  PubMed Central  Google Scholar 

  27. Santos AP, Suaid CA, Xavier M, Yamane F (2012) Functional and morphometric differences between the early and delayed use of phototherapy in crushed median nerves of rats. Lasers med sci 27:479–486

    Article  PubMed  Google Scholar 

  28. Holm S (1979) A simple sequentially rejective multiple test procedure. Scand j stat 6:65–70

    Google Scholar 

  29. Rochkind S, Leider-Trejo L, Nissan M et al (2007) Efficacy of 780-nm laser phototherapy on peripheral nerve regeneration after neurotube reconstruction procedure (double-blind randomized study). Photomed laser surg 25:137–143

    Article  PubMed  Google Scholar 

  30. Sousa FF, Ribeiro TL, Fazan VP, Barbieri CH (2013) Lack of effectiveness of laser therapy applied to the nerve course and the correspondent medullary roots. Acta ortop bras 21:92–97

    Article  PubMed  PubMed Central  Google Scholar 

  31. Wood MD, Kemp SW, Weber C, Borschel GH, Gordon T (2011) Outcome measures of peripheral nerve regeneration. Ann anat 193:321–333

    Article  PubMed  Google Scholar 

  32. Mendonça RJ, Maurício VB, Teixeira Lde B, Lachat JJ, Coutinho-Netto J (2010) Increased vascular permeability, angiogenesis and wound healing induced by the serum of natural latex of the rubber tree Hevea brasiliensis. Phytother res 24:764–768

    PubMed  Google Scholar 

  33. de Campos D, Heck L, Jotz GP (2014) Degree of myelination (G-ratio) of the human recurrent laryngeal nerve. Eur arch otorhinolaryngol 271:1277–1281

    Article  PubMed  Google Scholar 

  34. Rushton WA (1951) A theory of the effects of fibre size in medullated nerve. J physiol 115:101–122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Raimondo S, Fornaro M, Di Scipio F et al (2009) Chapter 5: methods and protocols in peripheral nerve regeneration experimental research: part II-morphological techniques. Int rev neurobiol 87:81–103

    Article  PubMed  Google Scholar 

  36. Byrnes KR, Waynant RW, Ilev IK et al (2005) Light promotes regeneration and functional recovery and alters the immune response after spinal cord injury. Lasers surg med 36:171–185

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors are grateful to FAPESP (processes no. 2014/07254-3 and 2014/22360-1) for their financial support of this study.

Author information

Authors and Affiliations

  1. School of Dentistry, University of São Paulo, USP, Ribeirão Preto, Brazil

    Eduardo Keiske Mastuda Ziago, Mamie Mizusaki Iyomasa, Luiz Gustavo Sousa & Paula Yumi Yamauchi

  2. School of Medicine, University of São Paulo, USP, Ribeirão Preto, Brazil

    Valéria Paula Sassoli Fazan, Eunice Aparecida da Silva & Fernando José Dias

  3. Department of Integral Dentistry, CICO – Research Centre in Dental Sciences, Dental School, Universidad de La Frontera, Temuco, Chile

    Eduardo Borie, Ramón Fuentes & Fernando José Dias

Authors
  1. Eduardo Keiske Mastuda Ziago
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Valéria Paula Sassoli Fazan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mamie Mizusaki Iyomasa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Luiz Gustavo Sousa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Paula Yumi Yamauchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Eunice Aparecida da Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Eduardo Borie
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ramón Fuentes
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Fernando José Dias
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fernando José Dias.

Ethics declarations

Ethical approval

The project was approved by the Ethics Committee on Animal Use of Ribeirão Preto Dental School, University of São Paulo (Protocol. 2014.1162.58.1) and followed the international laws on animal experimentation, performed according to the Helsinki Declaration.

Conflict of interest

The authors declare that they have no conflict of interest.

Role of funding source

The entire study was funded by FAPESP (São Paulo Research Foundation), a public foundation, funded by the taxpayer in the State of São Paulo (Brazil), with the mission to support research projects in higher education and research institutions, in all fields of knowledge. FAPESP is not linked to any company and/or product cited in this study.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ziago, E.K.M., Fazan, V.P.S., Iyomasa, M.M. et al. Analysis of the variation in low-level laser energy density on the crushed sciatic nerves of rats: a morphological, quantitative, and morphometric study. Lasers Med Sci 32, 369–378 (2017). https://doi.org/10.1007/s10103-016-2126-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10103-016-2126-1

Keywords

Search

Navigation