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Photobiomodulation therapy can change actin filaments of 3T3 mouse fibroblast

  • Ana Carolina de MagalhãesEmail author
  • Zwinglio Guimarães-Filho
  • Elisabeth Mateus Yoshimura
  • Lothar Lilge
Original Article
  • 18 Downloads

Abstract

The purpose of this study was to investigate the effects that photobiomodulation therapy might produce in cells, in particular, related to their structure. Thus, this paper presents the results of morphological changes in fibroblasts following low-intensity light illumination. Mouse fibroblasts were grown on glass coverslips on either 4 kPa or 16 kPa gels, to mimic normal tissue conditions. Cells were photo-irradiated with laser light at either 625 nm or 808 nm (total energies ranging from 34 to 47 J). Cells were fixed at 5 min, 1 h, or 24 h after photo-irradiation, stained for both actin filaments and the cell nucleus, and imaged by confocal microscopy. A non-light exposed group was also imaged. A detailed analysis of the images demonstrated that the total polymerized actin and number of actin filaments decrease, while the nucleus area increases in treated cells shortly after photo-irradiation, regardless of substrate and wavelength. This experiment indicated that photobiomodulation therapy could change the morphological properties of cells and affect their cytoskeleton. Further investigations are required to determine the specific mechanisms involved and how this phenomenon is related to the photobiomodulation therapy mechanisms of action.

Keywords

Photobiomodulation therapy Fibroblasts Actin filaments Low-level light therapy 

Notes

Funding

This study was funded by Coordination for the Improvement of Higher Education Personnel, CAPES (Proc. No. BEX 3481/14-0), and Brazilian National Council for Scientific and Technological Development, CNPq, Brazilian funding agencies. Additional support was provided by the Ontario Ministry of Health and Long-Term Care through operational funding of the Princess Margaret Cancer Centre.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. 1.
    Chung H, Dai T, Sharma SK, Huang Y-Y, Carroll JD, Hamblin MR (2012) The nuts and bolts of low-level laser (light) therapy. Ann Biomed Eng 40:516–533.  https://doi.org/10.1007/s10439-011-0454-7 CrossRefGoogle Scholar
  2. 2.
    Tata DB, Waynant RW (2011) Laser therapy: a review of its mechanism of action and potential medical applications. Laser Photon Rev 5:1–12.  https://doi.org/10.1002/lpor.200900032 CrossRefGoogle Scholar
  3. 3.
    Karu T (1999) Primary and secondary mechanisms of action of visible to near-IR radiation on cells. J Photochem Photobiol B 49:1–17.  https://doi.org/10.1016/S1011-1344(98)00219-X CrossRefGoogle Scholar
  4. 4.
    Amat A, Rigau J, Waynant RW, Ilev IK, Anders J (2006) The electric field induced by light can explain cellular responses to electromagnetic energy: a hypothesis of mechanism. J Photochem Photobiol B Biol 82:152–160CrossRefGoogle Scholar
  5. 5.
    Karu TI, Pyatibrat LV, Afanasyeva NI (2004) A novel mitochondrial signaling pathway activated by visible-to-near infrared radiation. Photochem Photobiol 80:366–372.  https://doi.org/10.1562/2004-03-25-RA-123 CrossRefGoogle Scholar
  6. 6.
    Chang HY, Chi J-T, Dudoit S, Bondre C, van de RM, Botstein D, Brown PO (2002) Diversity, topographic differentiation, and positional memory in human fibroblasts. Proc Natl Acad Sci 99:12877–12882.  https://doi.org/10.1073/pnas.162488599 CrossRefGoogle Scholar
  7. 7.
    Rieske P, Krynska B, Azizi SA (2005) Human fibroblast-derived cell lines have characteristics of embryonic stem cells and cells of neuro-ectodermal origin. Differentiation 73:474–483.  https://doi.org/10.1111/j.1432-0436.2005.00050.x CrossRefGoogle Scholar
  8. 8.
    Peplow PV, Chung T-Y, Baxter GD (2010) Laser photobiomodulation of proliferation of cells in culture: a review of human and animal studies. Photomed Laser Surg 28:S–3-S-40.  https://doi.org/10.1089/pho.2010.2771 CrossRefGoogle Scholar
  9. 9.
    Ragoonanan V, Less R, Aksan A (2013) Response of the cell membrane-cytoskeleton complex to osmotic and freeze/thaw stresses. Part 2: the link between the state of the membrane-cytoskeleton complex and the cellular damage. Cryobiology 66:96–104.  https://doi.org/10.1016/j.cryobiol.2012.10.008 CrossRefGoogle Scholar
  10. 10.
    Liu L, Luo Q, Sun J, Song G (2016) Nucleus and nucleus-cytoskeleton connections in 3D cell migration. Exp Cell Res 348:56–65.  https://doi.org/10.1016/j.yexcr.2016.09.001 CrossRefGoogle Scholar
  11. 11.
    Toh KC, Ramdas NM, Shivashankar GV (2015) Actin cytoskeleton differentially alters the dynamics of lamin A, HP1α and H2B core histone proteins to remodel chromatin condensation state in living cells. Integr Biol 7:1309–1317.  https://doi.org/10.1039/C5IB00027K CrossRefGoogle Scholar
  12. 12.
    Chow RT, David MA, Armati PJ (2007) 830 nm laser irradiation induces varicosity formation, reduces mitochondrial membrane potential and blocks fast axonal flow in small and medium diameter rat dorsal root ganglion neurons: implications for the analgesic effects of 830 nm laser. J Peripher Nerv Syst 12:28–39.  https://doi.org/10.1111/j.1529-8027.2007.00114.x CrossRefGoogle Scholar
  13. 13.
    Ricci R, Pazos MC, Borges RE, Pacheco-Soares C (2009) Biomodulation with low-level laser radiation induces changes in endothelial cell actin filaments and cytoskeletal organization. J Photochem Photobiol B 95:6–8.  https://doi.org/10.1016/j.jphotobiol.2008.11.007 CrossRefGoogle Scholar
  14. 14.
    Houreld NN, Ayuk SM, Abrahamse H (2014) Expression of genes in normal fibroblast cells (WS1) in response to irradiation at 660nm. J Photochem Photobiol B 130:146–152.  https://doi.org/10.1016/j.jphotobiol.2013.11.018 CrossRefGoogle Scholar
  15. 15.
    Avila R, Medina-Villalobos N, Tamariz E, Chiu R, Lopez-Marín LM, Acosta A, Castaño V (2014) Optical tweezers experiments for fibroblast cell growth stimulation. Proccedings SPIE 9129:91291U.  https://doi.org/10.1117/12.2064939 CrossRefGoogle Scholar
  16. 16.
    Sassoli C, Chellini F, Squecco R, Tani A, Idrizaj E, Nosi D, Giannelli M, Zecchi-Orlandini S (2016) Low intensity 635 nm diode laser irradiation inhibits fibroblast-myofibroblast transition reducing TRPC1 channel expression/activity: new perspectives for tissue fibrosis treatment. Lasers Surg Med 48:318–332.  https://doi.org/10.1002/lsm.22441 CrossRefGoogle Scholar
  17. 17.
    Fischer RS, Myers KA, Gardel ML, Waterman CM (2012) Stiffness-controlled three-dimensional extracellular matrices for high-resolution imaging of cell behavior. Nat Protoc 7:2056–2066.  https://doi.org/10.1038/nprot.2012.127 CrossRefGoogle Scholar
  18. 18.
    Niu CJ, Fisher C, Scheffler K, Wan R, Maleki H, Liu H, Sun Y, Simmons C A, Birngruber R, Lilge L (2015) Polyacrylamide gel substrates that simulate the mechanical stiffness of normal and malignant neuronal tissues increase protoporphyin IX synthesis in glioma cells. J Biomed Opt 20:098002-1–7.  https://doi.org/10.1117/1.JBO.20.9.098002 CrossRefGoogle Scholar
  19. 19.
    Rotsch C, Jacobson K, Radmacher M (1999) Dimensional and mechanical dynamics of active and stable edges in motile fibroblasts investigated by using atomic force microscopy. PNAS Cell Biol 96:921–926Google Scholar
  20. 20.
    Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez J-Y, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682.  https://doi.org/10.1038/nmeth.2019 CrossRefGoogle Scholar
  21. 21.
    The MathWorks I (2013) MATLAB and Statistics Toolbox Release 2013bGoogle Scholar
  22. 22.
    Kreutzer J, Viehrig M, Maki AJ, Kallio P, Rahikainen R, Hytönen V (2017) Pneumatically actuated elastomeric device for simultaneous mechanobiological studies & live-cell fluorescent microscopy. Int Conf Manip Autom Robot Small Scales, MARSS 2017 - Proc.  https://doi.org/10.1109/MARSS.2017.8001929
  23. 23.
    Lee TC, Kashyap RL, Chu CN (1994) Building skeleton models via 3-D medial surface axis thinning algorithms. CVGIP Graph Model Image Process 56:462–478.  https://doi.org/10.1006/cgip.1994.1042 CrossRefGoogle Scholar
  24. 24.
    Arganda-Carreras I, Fernández-González R, Muñoz-Barrutia A, Ortiz-De-Solorzano C (2010) 3D reconstruction of histological sections: application to mammary gland tissue. Microsc Res Tech 73:1019–1029.  https://doi.org/10.1002/jemt.20829 CrossRefGoogle Scholar
  25. 25.
    Ballestrem C, Wehrle-Haller B, Imhof BA (1998) Actin dynamics in living mammalian cells. J Cell Sci 111:1649–1658Google Scholar
  26. 26.
    Rotsch C, Radmacher M (2000) Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: an atomic force microscopy study. Biophys J 78:520–535.  https://doi.org/10.1016/S0006-3495(00)76614-8 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of PhysicsUniversity of Sao PauloSão PauloBrazil
  2. 2.InsperSão PauloBrazil
  3. 3.Department of Medical BiophysicsUniversity of TorontoTorontoCanada

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