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Near infrared laser irradiation induces NETosis via oxidative stress and autophagy

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A Correction to this article was published on 22 June 2018

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Abstract

NETosis is a novel immune defense strategy in which neutrophil activation results in the formation of extracellular DNA/protein network which is able to kill microbial populations. NETosis can be induced in vitro by lipopolysaccharide (LPS) or phorbol myristate acetate (PMA). Due to the importance of NETosis in different physiological and pathological processes, photobiostimulation effect on this neutrophil activation mechanism has been investigated. Human granulocytes, isolated from venous blood of healthy donors, were stimulated with a diode laser emitting at 980 nm with an energy intensity ranging from 0 to 75 joules. After 3 h of laser stimulation, granulocytes were fixed and colored with crystal violet in order to assess the NETosis morphology while extracellular DNA produced has been quantified using Sytox Green fluorescent dye. To evaluate ROS production and autophagy role in photobiostimulation-induced NETosis, granulocytes were pre-treated with ROS scavengers (vitamin C, sodium pyruvate, l-NAME, sodium azide), and an autophagy inhibitor (wortmannin). Laser stimulation induced an energy-dependent neutrophil extracellular trap (NET) production in human granulocytes starting from 50-J laser intensity. ROS scavengers and the autophagy inhibitor were able to abrogate both morphological features of NETosis and extracellular DNA production without modifying the basal level of NETosis. Photobiostimulation induced an increase in NET production due to an increase in ROS levels and autophagy activation.

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Change history

  • 22 June 2018

    The published online version contains mistake on the author names. The first names and family names were interchanged. Corrected names are shown in the author group section above.

  • 22 June 2018

    The published online version contains mistake on the author names. The first names and family names were interchanged. Corrected names are shown in the author group section above.

References

  1. Zhang Y, Song S, Fong CC, Tsang CH, Yang Z, Yang M (2003) cDNA microarray analysis of gene expression profiles in human fibroblast cells irradiated with red light. J Invest Dermatol 120:849–857

    Article  CAS  Google Scholar 

  2. Peplow PV, Chung TY, Ryan B, Baxter GD (2011) Laser photobiomodulation of gene expression and release of growth factors and cytokines from cells in culture: a review of human and animal studies. Photomed Laser Surg 29:285–304

    Article  CAS  Google Scholar 

  3. Hawkins DH, Abrahamse H (2006) The role of laser fluence in cell viability, proliferation, and membrane integrity of wounded human skin fibroblasts following helium-neon laser irradiation. Lasers Surg Med 38:74–83

    Article  Google Scholar 

  4. Chen CH, Hung HS, Hsu SH (2008) Low-energy laser irradiation increases endothelial cell proliferation, migration, and eNOS gene expression possibly via PI3K signal pathway. Lasers Surg Med 40:46–54

    Article  Google Scholar 

  5. Fushimi T, Inui S, Nakajima T, Ogasawara M, Hosokawa K, Itami S (2012) Green light emitting diodes accelerate wound healing: characterization of the effect and its molecular basis in vitro and in vivo. Wound Repair Regen 20:226–235

    Article  Google Scholar 

  6. Pereira AN, Eduardo Cde P, 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

    Article  Google Scholar 

  7. Sharma SK, Kharkwal GB, Sajo M, Huang YY, De Taboada L, McCarthy T, Hamblin MR (2011) Dose response effects of 810 nm laser light on mouse primary cortical neurons. Lasers Surg Med 43:851–859

    Article  Google Scholar 

  8. Chung H, Dai T, Sharma SK, Huang YY, Carroll JD, Hamblin MR (2012) The nuts and bolts of low level laser (light) therapy. Ann Biomed Eng 40:516–533

    Article  Google Scholar 

  9. Karu TI, Kolyakov SF (2005) Exact action spectra for cellular responses relevant to phototherapy. Photomed Laser Surg 23:355–361

    Article  CAS  Google Scholar 

  10. Lane N (2006) Cell biology: power games. Nature 443:901–903

    Article  CAS  Google Scholar 

  11. Biermann MH, Podolska MJ, Knopf J, Reinwald C, Weidner D, Maueröder C, Hahn J, Kienhöfer D, Barras A, Boukherroub R, Szunerits S, Bilyy R, Hoffmann M, Zhao Y, Schett G, Herrmann M, Munoz LE (2016) Oxidative Burst-Dependent NETosis is implicated in the resolution of necrosis-associated sterile inflammation. Front Immunol 1:7–557

    Google Scholar 

  12. Bullard DC, Kunkel EJ, Kubo H, Hicks MJ, Lorenzo I, Doyle NA, Doerschuk CM, Ley K, Beaudet AL (1996) Infectious susceptibility and severe deficiency of leukocyte rolling and recruitment in E-selectin and P-selectin double mutant mice. J Exp Med 183:2329–2336

    Article  CAS  Google Scholar 

  13. Savill J (1997) Apoptosis in resolution of inflammation. J Leukoc Biol 61:375–380

    Article  CAS  Google Scholar 

  14. Rocha Junior AM et al, (2007) Effects of low-level laser therapy in wound healing in humans: the contribution of in vitro and in vivo experimental studies. J Vasc Bras 6:258-266

  15. Baum CL, Arpey CJ (2005) Normal cutaneous wound healing: clinical correlation with cellular and molecular events. Dermatol Surg 31:674–686

    Article  CAS  Google Scholar 

  16. Ebaid H (2014) Neutrophil depletion in the early inflammatory phase delayed cutaneous wound healing in older rats: improvements due to the use of un-denatured camel whey protein. Diagnostic Pathol 9:46

    Article  Google Scholar 

  17. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, Weinrauch Y, Zychlinsky A (2014) Neutrophil extracellular traps kill bacteria. Science 303:1532–1535

    Article  Google Scholar 

  18. Fuchs TA, Abed U, Goosmann C, Hurwitz R, Schulze I, Wahn V, Weinrauch Y, Brinkman V, Zychlinsky A (2007) Novel cell death program leads to meutrophils to neutrophil extracellular traps. J Cell Biol 176:231–241

    Article  CAS  Google Scholar 

  19. Mòcsai A (2013) Diverse novel function of neutrophils in immunity, inflammation, and beyond. J Exp Med 210:1283–1299

    Article  Google Scholar 

  20. Kaplan MJ, Radic M (2012) Neutrophil extracellular traps: double-edged swords of innate immunity. J Immunol 189:2689–2695

    Article  CAS  Google Scholar 

  21. Remijsen Q, Vanden Berghe T, Wirawan E, Asselbergh B, Parthoens E, De Rycke R, Noppen S, Delforge M, Willems J, Vandenabeele P (2011) Neutrophil extracellular trap cell death requires both autophagy and superoxide generation. Cell Res 21:290–304

    Article  CAS  Google Scholar 

  22. Hahn J, Knopf J, Maueröder C, Kienhöfer D, Leppkes M, Herrmann M (2016) Neutrophils and neutrophil extracellular traps orchestrate initiation and resolution of inflammation. Clin Exp Rheumatol 34:6–8

    PubMed  Google Scholar 

  23. Papayannopoulos V, Zychlinsky A (2009) NETs: a new strategy for using old weapons. Trends Immunol 30:513–521

    Article  CAS  Google Scholar 

  24. Rizzi M, Migliario M, Rocchetti V, Tonello S, Renò F (2016) Near-infrared laser increases MDPC-23 odontoblast-like cells proliferation by activating redox sensitive pathways. Photochem Photobiol B 164:283–288

    Article  CAS  Google Scholar 

  25. Molinari C, Rizzi M, Rocchetti V, Tonello S, Renò F (2016) Near-infrared laser increases MDPC-23 odontoblast-lik cells proliferation by activating redox sensitive pathways. Photochem Photobiol B 164:283–288

    Article  Google Scholar 

  26. Vong L, Sherman PM, Glogauer M (2013) Quantification and visualization of neutrophil extracellular traps (NETs) from murine bone marrow-derived neutrophils. Methods Mol Biol 1031:41–50

    Article  CAS  Google Scholar 

  27. Itakura A, McCarty OJ (2013) Pivotal role for the mTOR pathway in the formation of neutrophil extracellular traps via regulation of autophagy. Am J Physiol Cell Physiol 305:C348–C354

    Article  CAS  Google Scholar 

  28. Dunnill C, Patton T, Brennan J, Barrett J, Dryden M, Cooke J, Leaper D, Georgopoulos NT (2017) Reactive oxygen species (ROS) and wound healing: the functional role of ROS and emerging ROS-modulating technologies for augmentation of the healing process. Int Wound J 14:89–96

    Article  Google Scholar 

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Acknowledgments

The authors would like to thank Dr. N. Trivero for the technical help in the revision of this manuscript.

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Correspondence to Filippo Renò.

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The original version of this article was revised:Given names and family names were interchanged.

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Migliario, M., Tonello, S., Rocchetti, V. et al. Near infrared laser irradiation induces NETosis via oxidative stress and autophagy. Lasers Med Sci 33, 1919–1924 (2018). https://doi.org/10.1007/s10103-018-2556-z

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