Research on Biomedical Engineering
http://www.rbejournal.periodikos.com.br/article/doi/10.1590/2446-4740.00115
Research on Biomedical Engineering
Original Article

Comparative analysis of different doses of coherent light (laser) and non-coherent light (light-emitting diode) on cellular necrosis and apoptosis: a study in vitro

Vanessa dos Santos Silva; Elizângela Márcia de Carvalho Abreu; Renata Amadei Nicolau; Cristina Pacheco Soares

Downloads: 1
Views: 883

Abstract

Introduction: Threshold doses of electromagnetic radiation can initiate necrosis and apoptosis in cells. The purpose of this study was to evaluate cellular apoptosis and necrosis immediately (t0) and 24 hours (t24) after irradiation with different doses of coherent light (laser) or non-coherent light (LED).

Methods: CHO-K1 lineage cells were irradiated with laser (810nm) or LED (945±20nm), with 24mW, contact area of 1cm2 and doses of 10, 20, 30, 40 and 50J/cm2 for 300, 660, 960, 1230 and 1620s, respectively, at both wavelengths. Cells were evaluated by fluorescence microscopy, differentiating viable, apoptotic and necrotic cells immediately and 24 hours after irradiation.

Results: The number of necrotic cells at t0 was higher in the LED 40 and 50J/cm2 groups (86±14 and 84±16% respectively, p <0.05), than in the 10 and 20J/cm2 laser (5±2 and 5±3%, p<0.05) and LED (5±3 and 4±1%, p<0.05) conditions. At t24, the LED 40J/cm2 (80±20%, p<0.05) group also showed more necrosis than the control and lower dose groups (laser 10, 20, and 30J/cm2 percentage of 6±4, 10±3 and 7±3%, p<0.05; LED 10 and 20J/cm2 percentage of 3±1 and 17±10%, p<0.05). A decrease in apoptotic cells was observed in the laser group with doses of 10, 40, and 50J/cm2 (6±4, 3±1 and 1±1% respectively, not significant), as well as in the LED 40J/cm2 (2±2%, not significant) group versus control. The cells had a higher percentage of apoptosis cells in the control group and with laser doses of 10 and 30J/cm2 (percentage of 20±1 and 20±4%, not significant), while only the LED 40J/cm2 (10±10%, not significant) had a lower percentage compared the control group.

Conclusion: Laser or LED stimulation promoted an increase in cell necrosis in a high energy density condition as characterized in a dose-dependent inhibition therapy. Laser or LED infrared irradiation in low doses (up to 20J/cm2) reduced the percentage of apoptosis in CHO-K1 cells, while high doses (30J/cm2) elevated apoptosis.

Keywords

Culture cells, Laser, LED, Necrosis, Apoptosis

References

Alexandratou E, Yova D, Handris P, Kletsas D, Loukas S. Human fibroblast alterations induced by low power laser irradiation at the single cell level using confocal microscopy. Photochemical & Photobiological Sciences. 2002; 1(8):547-52. PMid:12659495. http://dx.doi.org/10.1039/b110213n.

Belletti S, Uggeri J, Mergoni G, Vescovi P, Merigo E, Fornaini C, Nammour S, Manfredi M, Gatti R. Effects of 915 nm GaAs diode laser on mitochondria of human dermal fibroblasts: analysis with confocal microscopy. Lasers in Medical Science. 2015; 30(1):375-81. PMid:25351448. http://dx.doi.org/10.1007/s10103-014-1651-z.

Chaves MEA, Araújo AR, Piancastelli ACC, Pinotti M. Effects of low-power light therapy on wound healing: LASER x LED. Anais Brasileiros de Dermatologia. 2014; 89(4):616-23. PMid:25054749. http://dx.doi.org/10.1590/abd1806-4841.20142519.

Hawkins D, Abrahamse H. Laboratory methods for evaluating the effect of low level laser therapy (LLLT) in wound healing. African Journal of Biomedical Research. 2005; 8:1-14. http://dx.doi.org/10.4314/ajbr.v8i1.35752.

Huang L, Wu S, Xing D. High fluence low-power laser irradiation induces apoptosis via inactivation of Akt/GSK3β signaling pathway. Journal of Cellular Physiology. 2011; 226(3):588-601. PMid:20683916. http://dx.doi.org/10.1002/jcp.22367.

Jeon BW, Kim KT, Chang S, Kim HY. Phosphoinositide 3-OH kinase/protein kinase b inhibits apoptotic cell death induced by reactive oxygen species in saccharomyces cerevisiae. Journal of Biochemistry. 2002; 131(5):693-9. PMid:11983076. http://dx.doi.org/10.1093/oxfordjournals.jbchem.a003153.

Karu TI. Multiple roles of cytochrome C oxidase in mammalian cells under action of red and IR-A radiation. International Union of Biochemistry and Molecular Biology Life. 2010; 62(8):607-10. PMid:20681024. http://dx.doi.org/10.1002/iub.359.

Koutná M, Janisch R, Veselská R. Effects of low-power laser irradiation on cell proliferation. Scripta Medica. 2003; 76(3):163-72.

Lev-Tov H, Brody N, Siegel D, Jagdeo J. Inhibition of fibroblast proliferation in vitro using low-level infrared light-emitting diodes. Dermatologic Surgery. 2013a; 39(3 Pt1):422-5. PMid:23279196. http://dx.doi.org/10.1111/dsu.12087.

Lev-Tov H, Mamalis A, Brody N, Siegel D, Jagdeo J. Inhibition of fibroblast proliferation in vitro using red light-emitting diodes. Dermatologic Surgery. 2013b; 39(8):1167-70. PMid:23590233. http://dx.doi.org/10.1111/dsu.12212.

Magrini TD, Santos NV, Milazzotto MP, Cerchiaro G, Martinho HS. Low-level laser therapy on MCF-7 cells: a micro-Fourier transform infrared spectroscopy study. Journal of Biomedical Optics. 2012; 17(10):101516. PMid:23223992. http://dx.doi.org/10.1117/1.JBO.17.10.101516.

Mamalis A, Garcha M, Jagdeo J. Light emitting diode-generated blue light modulates fibrosis characteristics: fibroblast proliferation, migration speed, and reactive oxygen species generation. Lasers in Surgery and Medicine. 2015; 47(2):210-5. PMid:25655579. http://dx.doi.org/10.1002/lsm.22293.

McGahon AJ, Martin SJ, Bissonnette RP, Mahboubi A, Shi Y, Mogil RJ, Nishioka WK, Green DR. The end of the (cell) line: Methods for the study of apoptosis in vitro. Methods in Cell Biology. 1995; 46:153-84. PMid:7541883. http://dx.doi.org/10.1016/S0091-679X(08)61929-9.

Park JT, Hong KS. Effect of light-emitting-diode irradiation on the proliferation and migration in human gingival fibroblasts. Tissue Engineering and Regenerative Medicine. 2015; 12(1):37-42. http://dx.doi.org/10.1007/s13770-014-9061-7.

Poersch A, Santos FV, Maciel MAM, Camara JK, Dantas TN, Cólus IMS. Protective effect of DCTN (trans-dehydrocrotonin) against induction of micronuclei and apoptosis by different mutagenic agents in vitro. Mutation Research. 2007; 629(1):14-23. PMid:17276134. http://dx.doi.org/10.1016/j.mrgentox.2007.01.001.

Ribble D, Goldstein NB, Norris DA, Shellman YG. A simple technique for quantifying apoptosis in 96-well plates. BMC Biotechnology. 2005; 5(12):1-7. http://dx.doi.org/10.1186/1472-6750-5-12. PMid:15885144.

Rimessi A, Giorgi C, Pinton P, Rizzuto R. The versatility of mitochondrial calcium signals: from stimulation of cell metabolism to induction of cell death. Biochimica et Biophysica Acta. 2008; 1777:808-16. http://dx.doi.org/10.1016/j.bbabio.2008.05.449.

Sun X, Wu S, Xing D. The reactive oxygen species–Src–Stat3 pathway provokes negative feedback inhibition of apoptosis induced by high-fluence low-power laser irradiation. Federation of European Biochemical Societies Journal. 2010; 277(22):4789-802. http://dx.doi.org/10.1111/j.1742-4658.2010.07884.x. PMid:20977672.

Wu S, Xing D, Gao X, Chen W. High fluence low-power laser irradiation induces mitochondrial permeability transition mediated by reactive oxygen species. Journal of Cellular Physiology. 2009; 218(3):603-11. PMid:19006121. http://dx.doi.org/10.1002/jcp.21636.

Zhang H, Xing D, Wu S, Sun X. Protein kinase C δ promotes cell apoptosis induced by high fluence low-power laser irradiation. In: Proceedings of SPIE 7519. 8th International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2009); 2009 Aug 8; Wuhan, China. SPIE; 2009. http://dx.doi.org/10.1117/12.841566.

Zhang J, Xing D, Gao X. Low-power laser irradiation activates Src tyrosine kinase through reactive oxygen species-mediated signalling pathway. Journal of Cellular Physiology. 2008; 217(2):518-28. PMid:18615581. http://dx.doi.org/10.1002/jcp.21529.
 

589892820e8825320fb92415 rbejournal Articles
Links & Downloads

Res. Biomed. Eng.

Share this page
Page Sections