Effect of therapeutic ultrasound of low intensity pulsed and continuous in fibroblastic cells in vitro
Oliveira, Priscila Daniele de; Pires-Oliveira, Deise Aparecida Almeida; Pacheco-Soares, Cristina; Szezerbaty, Stheace Kelly Fernandes; Spadaccini, Ana Flávia; Bertin, Larissa Dragonetti; Silva, Jéssica Lucio da; Santos, Adriele Flausino dos; Oliveira, Rodrigo Franco de
Abstract
Introduction: The ultrasound is widely used clinical practice focused on tissue repair, because it is a secure resource, minimally-invasive and low investment. However, despite the effects of ultrasound have been well described, the ideal relationship between dose and effect, still needs to be better elucidated and bounded. Objective: In this way, the objective of this study was to evaluate the influence of irradiation pulsed ultrasound and continuous in the viability of fibroblastic cells L929 in vitro. Method: The cells were distributed in TPP plates 12 wells in the concentration of 5x105 cells/µL and subdivided into the following groups: G1: control (not received radiation), G2: irradiated to 0.5 W/cm2 - 30% and G3: irradiated to 0.5 W/cm2 - 100%. Then received ultrasonic irradiation with intervals of 24, 48 and 72 hours, and after 24 hours of each irradiation was performed MTT cytotoxicity cell. Results: Among the three groups analyzed, only the G2 group showed a significant difference between the time 48 and 72 hours (p=0.05). In other times, despite variation in the percentage of viable cells, were not significant. When compared to peers at post-test, it was possible to observe a difference of 29% of viable cells between the groups G1 and G2 (p=0.05). However, the biggest difference was observed between the groups G2 and G3 (p=0.05). Conclusion: In this way, it can be concluded that irradiation pulsed ultrasound showed higher proliferation of fibroblasts L929 in time 48 hours, whereas in group irradiated in continuous mode, there was no statistically significant difference between the times.
Keywords
References
1. Vásquez B, Navarrete J, Farfán E, Cantín M. Effect of pulsed and continuous therapeutic ultrasound on healthy skeletal muscle in rats. Int J Clin Exp Pathol. 2014;7(2):779-783.
2. Ghorayeb SR, Patel US, Walmsley AD, Scheven BA. Biophysical characterization of low-frequency ultrasound interaction with dental pulp stem cells. Journal of Therap Ultrasound. 2013;1(12):1-9.
3. Nussbaum EL, Locke M. Heat shock protein expression in rat skeletal muscle after repeated applications of pulsed and continuous ultrasound. Arch Phys Med Rehabil. 2007;88(6):785-90.
4. Watson T. Ultrasound in contemporary physiotherapy practice. Ultrasonics. 2008;48(4):321-329.
5. Watson T. Tissue repair: The current state of the art. Sportex-Medicine. 2006;28:8-12.
6. Watson T. Electrotherapy and tissue repair. Sportex-Medicine. 2006;29:7- 13.
7. Oliveira PD, Pires-Oliveira DAA, Martinago CC, Poli-Frederico RC, Soares CP, Oliveira RF. Effect of low-intensity pulsed ultrasound therapy on fibroblasts cell culture Phys Ther Research. 2015;22(2):112-118.
8. Bertin LD, Pires-Oliveira DAA, Oliveira PD, Serpeloni JA, Szezerbaty SKF, Santos JPM, Oliveira FR. A influência da irradiação ultrassônica de baixa intensidade em cultura de células fibroblásticas. Revista Terapia Manual. 2015;13:306-15.
9. Oliveira RF, Oliveira DAAP, Machado AHA, Silva NS, Magini M, PachecoSoares C. Assessment of fibroblast cell ssubmitted ultrasonic irradiation. Cell Biol Int. 2008;32(10):1329-35.
10. Domenici F, Giliberti C, Bedini A, Palomba R, Udroiu I, Di Giambattista L, et al. Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro. Ultrasonics. 2014, 54:1020–1028.
11. Noriega S, Tarlan MS, Turner JA, Subramanian A. Intermittent Applications of Continuous Ultrasound on the Viability, Proliferation, Morphology, and Matrix Production of Chondrocytes in 3D Matrices. Tissue Engineering. 2007;13(3):611-8.
12. Hasanova GI, Noriega SE, Mamedov TG, Thakurta SG, Turner JA, Anuradha S. The effect of ultrasound stimulation on the gene and protein expression of chondrocytes seeded in chitosan scaffolds. J Tissue Eng Regen Med. 2011;5: 815–822.
13. Nakamura T, Fujihara S, Katsura T, Yamamoto K, Inubushi T, Tanimoto K, et al. Effects of Low-Intensity Pulsed Ultrasound on the Expression and Activity of Hyaluronan Synthase and Hyaluronidase in IL-1b-Stimulated Synovial Cells. Annals of Biomedical Engineering. 2010;38(11):3363–3370.
14. Oliveira RF, Oliveira DAAP, Pacheco-Soares C. Efeito do ultrassom pulsado de baixa intensidade em culturas fibroblásticas L929. ConScienta e Saúde. 2008;7(3):315-321.
15. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1-2):55-63.
16. Piedade MCB, Ferreira MA, Galhardo MS, Caldini EG, Battlehner CN, Toledo OMS. Effect of ultrasound therapy on the repair of Gastrocnemius muscle injury in rats. Ultrasonics. 2008;48(5):403:411.
17. Silva JMN, Carvalho JP, Júnior MJM, Arisawa EALS, Martin AA, Sá HP, et al. Estudo da ação do ultrassom terapêutico em modelo experimental de tendinite em ratos Wistar. ConScientia e Saúde. 2010;9(4):625:632.
18. Silveira DS, PippiII NL, Costa FS, Vescovi LA, Conti LMC, Weiss A, et al. O ultra-som terapêutico de 1 MHz, na dose de 0,5 W cm2 , sobre o tecido ósseo de cães avaliado por densitometria óptica em imagens radiográficas. Ciência Rural. 2008;38(8):2225:2231.
19. Nakamura T, Fujihara S, Yamamoto-Nagata K, Katsura T, Inubushi T, Tanaka E. Low-Intensity Pulsed Ultrasound Reduces the Inflammatory Activity of Synovitis. Annals of Biomedical Engineering. 2011;39(12):2964–2971.
20. Uenaka K, Imai S, Ando K, Matsusue Y. Relation of low-intensity pulsed ultrasound to the cell density of scaffold-free cartilage in a high-density static semi-open culture system. J Orthop Sci. 2010;15:816–824.
21. Dubinsky TJ, Cuevas C, Dighe MK, Kolokythas O, Hwang JH. High intensity focused ultrasound: current potential and oncologic applications. Am J Roentgenol. 2008;190: 191-199.
22. Jens UQ, Desilets C, Martin BSP. High Intensity Focused Ultrasound. The European Aesthetic Guide Spring (2010).
23. Demir H, Yaray S, Kirnap M, Yaray K. Comparison of the effects of laser and ultrasoundtreatmentson experimental woundhealing in rats. Journal of Rehabilitation Research Development. 2004;41(5):721-728.
24. Deyne PGD, Kirsch-Volders M. In Vitro Effects of Therapeutic Ultrasound on the Nucleus of Human Fibroblasts. Phys Ther. 1995;75(7): 629-634.
25. Marks R, Ghanagaraja S, Ghassemi M. Ultrasound for osteoarthritis of the knee: a systematic review. Physiotherapy. 2000;86:452-463.
26. Persson J, Stromqvist B, Zanoli G, Mccarthy I, Lidgren L. Ultrasound nucleolysis: an in vitro study. Ultrasound Med Biol. 2002;28:1189-1197.
27. Carrer VM, Setti JAP, Veronez DL, Moser AD. Continuous therapeutic ultrasound inthe healing process in rat skin. Fisioter Mov. 2015;28(4):751-758.
28. Maeshige N, Terashi H, Aoyama M, Torii K, Sugimoto M, Usami M. Effect of Ultrasound Irradiation on α-SMA and TGF-β1 Expression in Human Dermal Fibroblasts. Kobe J. Med. Sci. 2010;56(6):E242-E252.
29. Doan N, Reher P, Meghji S, Harris M. In vitro effects of therapeutic ultrasound on cell proliferation, protein synthesis and cytokine production by human fibroblasts, osteoblasts and monocytes. J Oral Maxillofac Surg. 1999;57:409-19.
30. Pires-Oliveira DA, De Oliveira RF, Magini M, Zangaro RA, Soares CP. Assessment of cytoskeleton and endoplasmatic reticulum of fibroblast cells submitted to low-level laser therapy and low-intensity pulsed ultrasound. Photomed Laser Surg. 2009;27(3):461-466.
Facebook
Google+
Twitter
LinkedIn
Mendeley
StumbleUpon
CiteULike
Reddit
Email