Skip to main content

Advertisement

SpringerLink
Log in

Evaluation of photobiomodulation therapy (117 and 90s) on pain, regeneration, and epigenetic factors (HDAC 2, DNMT3a) expression following spinal cord injury in a rat model

  • Original Papers
  • Published:
Photochemical & Photobiological Sciences Aims and scope Submit manuscript

Abstract

Background

Photobiomodulation therapy (PBMT), due to its anti-inflammatory, analgesic effects, and most importantly as a non-invasive procedure, has currently gained a special setting in pain relief and the treatment of Spinal cord injuries (SCI). However, the mechanism of action of the PBM is not yet completely understood.

Methods

In this study, SCI is induced by an aneurysm clip, and PBM therapy was applied by a continuous-wave (CW) laser with a wavelength of 660 nm. Adult male rats were divided into four groups: Control, SCI, SCI + PBMT 90s, and SCI + PBMT 117s. After 7 weeks, hyperalgesia, allodynia, and functional recovery were assessed. Fibroblasts infiltrating the spinal cord were counted after H&E staining. The expression of epigenetic factors (HDAC2, DNMT3a), protein relevant for pain (GAD65), and astrocytes marker (GFAP) after 4 weeks of daily PBMT (90 and 117s) was probed by western blotting.

Results

Both PBMTs (90 and 117s) significantly improved the pain and ability to move and fibroblast invasion was reduced. SCI + PBMT 90s, increased GAD65, HDAC2, and DNMT3a expression. However, PBMT 117s decreased GFAP, HDAC2, and DNMT3a.

Conclusion

PBMT 90 and 117s improved the pain, and functional recovery equally. The regulation of epigenetic mechanisms appears to be a significant effect of PBMT117s, which emphasizes on impact of radiation duration and accumulative energy.

Graphical abstract

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

Data availability

The data that support the findings of this study are available from the corresponding authors upon reasonable request.

References

  1. Leibinger, M., et al. (2019). GSK3-CRMP2 signaling mediates axonal regeneration induced by Pten knockout. Communications Biology, 2(1), 1–13.

    CAS  Google Scholar 

  2. Chambel, S. S., Tavares, I., & Cruz, C. D. (2020). Chronic pain after spinal cord injury: Is there a role for neuron-immune dysregulation? Frontiers in Physiology, 11, 748.

    PubMed  PubMed Central  Google Scholar 

  3. Van Gorp, S., et al. (2015). Pain prevalence and its determinants after spinal cord injury: A systematic review. European Journal of Pain, 19(1), 5–14.

    PubMed  Google Scholar 

  4. Hagen, E. M., & Rekand, T. (2015). Management of neuropathic pain associated with spinal cord injury. Pain and therapy, 4(1), 51–65.

    PubMed  PubMed Central  Google Scholar 

  5. Austin, P. J., & Moalem-Taylor, G. (2010). The neuro-immune balance in neuropathic pain: Involvement of inflammatory immune cells, immune-like glial cells and cytokines. Journal of Neuroimmunology, 229(1–2), 26–50.

    CAS  PubMed  Google Scholar 

  6. Zhao, H., et al. (2017). The role of microglia in the pathobiology of neuropathic pain development: what do we know? BJA: British Journal of Anaesthesia, 118(4), 504–516.

    CAS  PubMed  Google Scholar 

  7. Descalzi, G., et al. (2015). Epigenetic mechanisms of chronic pain. Trends in Neurosciences, 38(4), 237–246.

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Luo, D., et al. (2021). Epigenetic modifications in neuropathic pain. Molecular Pain, 17, 17448069211056768.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Moore, L. D., Le, T., & Fan, G. (2013). DNA methylation and its basic function. Neuropsychopharmacology, 38(1), 23–38.

    CAS  PubMed  Google Scholar 

  10. Wang, Y., et al. (2011). Intrathecal 5-azacytidine inhibits global DNA methylation and methyl-CpG-binding protein 2 expression and alleviates neuropathic pain in rats following chronic constriction injury. Brain Research, 1418, 64–69.

    CAS  PubMed  Google Scholar 

  11. Tochiki, K. K., et al. (2012). The expression of spinal methyl-CpG-binding protein 2, DNA methyltransferases and histone deacetylases is modulated in persistent pain states. Molecular Pain. https://doi.org/10.1186/1744-8069-8-14

    Article  PubMed  PubMed Central  Google Scholar 

  12. Liang, L., & Tao, Y.-X. (2018). Expression of acetyl-histone H3 and acetyl-histone H4 in dorsal root ganglion and spinal dorsal horn in rat chronic pain models. Life Sciences, 211, 182–188.

    CAS  PubMed  Google Scholar 

  13. Li, K., et al. (2014). Epigenetic upregulation of Cdk5 in the dorsal horn contributes to neuropathic pain in rats. NeuroReport, 25(14), 1116–1121.

    CAS  PubMed  Google Scholar 

  14. Cherng, C.-H., et al. (2014). Baicalin ameliorates neuropathic pain by suppressing HDAC1 expression in the spinal cord of spinal nerve ligation rats. Journal of the Formosan Medical Association, 113(8), 513–520.

    CAS  PubMed  Google Scholar 

  15. Zhang, Z., et al. (2011). Epigenetic suppression of GAD65 expression mediates persistent pain. Nature Medicine, 17(11), 1448–1455.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Kanao-Kanda, M., et al. (2020). Viral vector-mediated gene transfer of glutamic acid decarboxylase for chronic pain treatment: A literature review. Human Gene Therapy, 31(7–8), 405–414.

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Dima, R., et al. (2017). Review of literature on low-level laser therapy benefits for nonpharmacological pain control in chronic pain and osteoarthritis. Trials, 5, 6.

    Google Scholar 

  18. Janzadeh, A., et al. (2020). The effect of chondroitinase ABC and photobiomodulation therapy on neuropathic pain after spinal cord injury in adult male rats. Physiology & Behavior, 227, 113141.

    CAS  Google Scholar 

  19. Ramezani, F., et al. (2020). Photobiomodulation for spinal cord injury: A systematic review and meta-analysis. Physiology & Behavior, 224, 112977.

    CAS  Google Scholar 

  20. Mojarad, N., et al. (2018). The role of low level laser therapy on neuropathic pain relief and interleukin-6 expression following spinal cord injury: an experimental study. Journal of Chemical Neuroanatomy, 87, 60–70.

    CAS  PubMed  Google Scholar 

  21. Neshasteh-Riz, A., et al. (2022). Optimization of the duration and dose of photobiomodulation therapy (660 nm laser) for spinal cord injury in rats. Photobiomodulation, Photomedicine, and Laser Surgery, 40(7), 488–498.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Janzadeh, A., et al. (2016). Photobiomodulation therapy reduces apoptotic factors and increases glutathione levels in a neuropathic pain model. Lasers in Medical Science, 31, 1863–1869.

    PubMed  Google Scholar 

  23. Azim, K., & Butt, A. M. (2011). GSK3β negatively regulates oligodendrocyte differentiation and myelination in vivo. Glia, 59(4), 540–553.

    PubMed  Google Scholar 

  24. Yousefifard, M., et al. (2016). Human bone marrow-derived and umbilical cord-derived mesenchymal stem cells for alleviating neuropathic pain in a spinal cord injury model. Stem Cell Research & Therapy, 7(1), 1.

    Google Scholar 

  25. Janzadeh, A., et al. (2017). Combine effect of Chondroitinase ABC and low level laser (660ánm) on spinal cord injury model in adult male rats. Neuropeptides, 65, 90–99.

    CAS  PubMed  Google Scholar 

  26. Hosseini, M., et al. (2020). Simultaneous intrathecal injection of muscimol and endomorphin-1 alleviates neuropathic pain in rat model of spinal cord injury. Brain and Behavior, 10(5), e01576.

    PubMed  PubMed Central  Google Scholar 

  27. Ramezani, F., et al. (2021). Mechanistic aspects of photobiomodulation therapy in the nervous system. Lasers in Medical Science. https://doi.org/10.1007/s10103-021-03277-2

    Article  PubMed  Google Scholar 

  28. Behroozi, Z., et al. (2023). Evaluation of epigenetic (HDAC, DNMT) and pain (Gad65, TGF) factors following photobiomodulation therapy in a neuropathic pain model. Photochemistry and Photobiology. https://doi.org/10.1111/php.13824

    Article  PubMed  Google Scholar 

  29. Basso, D. M., Beattie, M. S., & Bresnahan, J. C. (1995). A sensitive and reliable locomotor rating scale for open field testing in rats. Journal of Neurotrauma, 12(1), 1–21.

    CAS  PubMed  Google Scholar 

  30. Chaplan, S. R., et al. (1994). Quantitative assessment of tactile allodynia in the rat paw. Journal of Neuroscience Methods, 53(1), 55–63.

    CAS  PubMed  Google Scholar 

  31. Yoon, C., et al. (1994). Behavioral signs of ongoing pain and cold allodynia in a rat model of neuropathic pain. Pain, 59(3), 369–376.

    PubMed  Google Scholar 

  32. Hargreaves, K., et al. (1988). A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain, 32(1), 77–88.

    CAS  PubMed  Google Scholar 

  33. Maximow, A. A. (1927). Development of non-granular leucocytes (lymphocytes and monocytes) into polyblasts (macrophages) and fibroblasts in vitro. Proceedings of the Society for Experimental Biology and Medicine, 24(6), 570–572.

    Google Scholar 

  34. Nimmerjahn, A., Kirchhoff, F., & Helmchen, F. (2005). Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo. Science, 308(5726), 1314–1318.

    CAS  PubMed  Google Scholar 

  35. Behroozi, Z., et al. (2021). Platelet-rich plasma in umbilical cord blood reduces neuropathic pain in spinal cord injury by altering the expression of ATP receptors. Physiology & Behavior, 228, 113186.

    CAS  Google Scholar 

  36. Fischer, A. H., et al. (2008). Hematoxylin and eosin staining of tissue and cell sections. Cold Spring Harbor Protocols, 2008(5), p.pdb.prot4986.

    Google Scholar 

  37. Park, J., et al. (2018). Reducing inflammation through delivery of lentivirus encoding for anti-inflammatory cytokines attenuates neuropathic pain after spinal cord injury. Journal of Controlled Release, 290, 88–101.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Madrid, A., et al. (2021). DNA methylation and hydroxymethylation have distinct genome-wide profiles related to axonal regeneration. Epigenetics, 16(1), 64–78.

    PubMed  Google Scholar 

  39. Saha, R., & Pahan, K. (2006). HATs and HDACs in neurodegeneration: A tale of disconcerted acetylation homeostasis. Cell Death & Differentiation, 13(4), 539–550.

    CAS  Google Scholar 

  40. Zamani, A. R. N., et al. (2020). Modulatory effect of photobiomodulation on stem cell epigenetic memory: A highlight on differentiation capacity. Lasers in Medical Science, 35, 299–306.

    PubMed  Google Scholar 

  41. Martins, M. D., et al. (2021). Photobiomodulation therapy drives massive epigenetic histone modifications, stem cells mobilization and accelerated epithelial healing. Journal of Biophotonics, 14(2), e202000274.

    CAS  PubMed  Google Scholar 

  42. Zaccara, I. M., et al. (2020). Photobiomodulation therapy improves human dental pulp stem cell viability and migration in vitro associated to upregulation of histone acetylation. Lasers in Medical Science, 35, 741–749.

    PubMed  Google Scholar 

  43. Li, G., Tian, Y., & Zhu, W.-G. (2020). The roles of histone deacetylases and their inhibitors in cancer therapy. Frontiers in Cell and Developmental Biology, 8, 576946.

    PubMed  PubMed Central  Google Scholar 

  44. Wang, G., et al. (2015). HDAC inhibition prevents white matter injury by modulating microglia/macrophage polarization through the GSK3β/PTEN/Akt axis. Proceedings of the National Academy of Sciences, 112(9), 2853–2858.

    CAS  Google Scholar 

  45. Ouyang, B., et al. (2019). Normalizing HDAC2 levels in the spinal cord alleviates thermal and mechanical hyperalgesia after peripheral nerve injury and promotes GAD65 and KCC2 expression. Frontiers in Neuroscience, 13, 346.

    PubMed  PubMed Central  Google Scholar 

  46. Demyanenko, S., & Uzdensky, A. (2019). Epigenetic alterations induced by photothrombotic stroke in the rat cerebral cortex: Deacetylation of histone H3, upregulation of histone deacetylases and histone acetyltransferases. International Journal of Molecular Sciences, 20(12), 2882.

    CAS  PubMed  PubMed Central  Google Scholar 

  47. Jacob, C., et al. (2011). HDAC1 and HDAC2 control the transcriptional program of myelination and the survival of Schwann cells. Nature Neuroscience, 14(4), 429–436.

    CAS  PubMed  Google Scholar 

  48. Gomez-Sanchez, J. A., et al. (2022). Emerging role of hDACs in regeneration and ageing in the peripheral nervous system: Repair schwann cells as pivotal targets. International Journal of Molecular Sciences, 23(6), 2996.

    CAS  PubMed  PubMed Central  Google Scholar 

  49. Zhang, S., et al. (2018). Class I histone deacetylase (HDAC) inhibitor CI-994 promotes functional recovery following spinal cord injury. Cell Death & Disease, 9(5), 460.

    CAS  Google Scholar 

  50. Lee, J. Y., et al. (2014). Valproic acid protects motor neuron death by inhibiting oxidative stress and endoplasmic reticulum stress-mediated cytochrome C release after spinal cord injury. Journal of Neurotrauma, 31(6), 582–594.

    PubMed  PubMed Central  Google Scholar 

  51. Kim, H. J., et al. (2007). Histone deacetylase inhibitors exhibit anti-inflammatory and neuroprotective effects in a rat permanent ischemic model of stroke: Multiple mechanisms of action. Journal of Pharmacology and Experimental Therapeutics, 321(3), 892–901.

    CAS  PubMed  Google Scholar 

  52. Zhao, J.-Y., et al. (2017). DNA methyltransferase DNMT3a contributes to neuropathic pain by repressing Kcna2 in primary afferent neurons. Nature Communications, 8(1), 14712.

    PubMed  PubMed Central  Google Scholar 

  53. Wang, Y., et al. (2016). Abnormal DNA methylation in the lumbar spinal cord following chronic constriction injury in rats. Neuroscience Letters, 610, 1–5.

    CAS  PubMed  Google Scholar 

  54. Xu, B., et al. (2017). Role of MicroRNA-143 in nerve injury-induced upregulation of Dnmt3a expression in primary sensory neurons. Frontiers in Molecular Neuroscience, 10, 350.

    PubMed  PubMed Central  Google Scholar 

  55. Wu, Z., et al. (2012). Dnmt3a regulates both proliferation and differentiation of mouse neural stem cells. Journal of Neuroscience Research, 90(10), 1883–1891.

    CAS  PubMed  PubMed Central  Google Scholar 

  56. Nguyen, S., et al. (2007). Ablation of de novo DNA methyltransferase Dnmt3a in the nervous system leads to neuromuscular defects and shortened lifespan. Developmental Dynamics: An Official Publication of the American Association of Anatomists, 236(6), 1663–1676.

    CAS  PubMed  Google Scholar 

  57. Feng, J., et al. (2005). Dynamic expression of de novo DNA methyltransferases Dnmt3a and Dnmt3b in the central nervous system. Journal of Neuroscience Research, 79(6), 734–746.

    CAS  PubMed  Google Scholar 

  58. Nakamura, M., et al. (2003). Differences in cytokine gene expression profile between acute and secondary injury in adult rat spinal cord. Experimental Neurology, 184(1), 313–325.

    CAS  PubMed  Google Scholar 

  59. Gaudet, A. D., & Fonken, L. K. (2018). Glial cells shape pathology and repair after spinal cord injury. Neurotherapeutics, 15, 554–577.

    CAS  PubMed  PubMed Central  Google Scholar 

  60. Setoguchi, T., et al. (2004). Treatment of spinal cord injury by transplantation of fetal neural precursor cells engineered to express BMP inhibitor. Experimental Neurology, 189(1), 33–44.

    CAS  PubMed  Google Scholar 

  61. Abematsu, M., et al. (2010). Neurons derived from transplanted neural stem cells restore disrupted neuronal circuitry in a mouse model of spinal cord injury. The Journal of Clinical Investigation, 120(9), 3255–3266.

    CAS  PubMed  PubMed Central  Google Scholar 

  62. Yiu, G., & He, Z. (2006). Glial inhibition of CNS axon regeneration. Nature Reviews Neuroscience, 7(8), 617–627.

    CAS  PubMed  PubMed Central  Google Scholar 

  63. Liddelow, S. A., & Barres, B. A. (2017). Reactive astrocytes: Production, function, and therapeutic potential. Immunity, 46(6), 957–967.

    CAS  PubMed  Google Scholar 

  64. Park, C., et al. (2019). The landscape of myeloid and astrocyte phenotypes in acute multiple sclerosis lesions. Acta Neuropathologica Communications, 7(1), 1–13.

    CAS  Google Scholar 

  65. Brenner, M. (2014). Role of GFAP in CNS injuries. Neuroscience Letters, 565, 7–13.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This study was supported by a scientific project Grant (No: 98-4-68-16528) financed by the Iran University of Medical Science (IUMS) and Islamic Azad University.

Author information

Author notes

  1. Ali Motamed Nezhad and Zahra Behroozi have contributed equally to this work.

Authors and Affiliations

  1. College of Veterinary Medicine, Islamic Azad University, Karaj, Alborz, Iran

    Ali Motamed Nezhad

  2. Physiology Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

    Zahra Behroozi

  3. International Campus, Iran University of Medical Sciences, Tehran, Iran

    Keihan Kookli & Seyedalireza Moghaddas Fazeli

  4. Cancer Control Research Center, Cancer Control Foundation, Iran University of Medical Sciences, Tehran, Iran

    Keihan Kookli & Seyedalireza Moghaddas Fazeli

  5. Department of Biophysics, Medical School, University of Pécs, Pécs, 7622, Hungary

    Alireza Ghadaksaz

  6. Dr. Moshiri Veterinary Clinic, Tehran, Iran

    Ali Moshiri

  7. Physiology Research Center, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran

    Fatemeh Ramezani

  8. Department of Medical Physics, Scool of Medicine, Iran University of Medical Sciences, Tehran, Iran

    Molood Gooniband Shooshtari

  9. Radiation Biology Research Center (RBRC), Iran University of Medical Sciences (IUMS), Tehran, Iran

    Fatemeh Ramezani & Atousa Janzadeh

Authors
  1. Ali Motamed Nezhad
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zahra Behroozi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Keihan Kookli
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alireza Ghadaksaz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Seyedalireza Moghaddas Fazeli
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ali Moshiri
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Fatemeh Ramezani
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Molood Gooniband Shooshtari
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Atousa Janzadeh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Fatemeh Ramezani or Atousa Janzadeh.

Ethics declarations

Conflict of interest

The authors declared that they have no competing interests.

Ethical statement

The Institutional Animal Ethical Committee of Iran University of Medical Sciences confirmed all experimental tests and the procedures at present work (IR.IUMS.REC.1399.147).

Consent for publication

All authors consented to publication.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Motamed Nezhad, A., Behroozi, Z., Kookli, K. et al. Evaluation of photobiomodulation therapy (117 and 90s) on pain, regeneration, and epigenetic factors (HDAC 2, DNMT3a) expression following spinal cord injury in a rat model. Photochem Photobiol Sci 22, 2527–2540 (2023). https://doi.org/10.1007/s43630-023-00467-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43630-023-00467-5

Keywords

Search

Navigation