Photobiomodulation therapy can change actin filaments of 3T3 mouse fibroblast
- 30 Downloads
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.
KeywordsPhotobiomodulation therapy Fibroblasts Actin filaments Low-level light therapy
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.
This article does not contain any studies with human participants or animals performed by any of the authors.
- 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
- 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
- 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
- 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.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.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.The MathWorks I (2013) MATLAB and Statistics Toolbox Release 2013bGoogle Scholar
- 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
- 25.Ballestrem C, Wehrle-Haller B, Imhof BA (1998) Actin dynamics in living mammalian cells. J Cell Sci 111:1649–1658Google Scholar