Comparative effect of low-level laser therapy and low intensity ultrasound on myofiber phenotypes, oxidative stress and proteins during muscle repair

• Grant number: 13/21540-3
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: October 01, 2014
• End date: July 26, 2016
• Field of knowledge: Health Sciences - Physiotherapy and Occupational Therapy
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Raquel Agnelli Mesquita Ferrari
• Grantee: Agnelo Neves Alves
• Host Institution: Universidade Nove de Julho (UNINOVE). Campus Vergueiro. São Paulo , SP, Brazil

Abstract

Muscle injuries represent a major clinical conditions found in the rehabilitation center. Among the major therapeutic resources used in physical therapy clinics for the treatment of muscle injuries are the Low-Level Laser Therapy (LLLT) and Low-Intensity Pulsed Ultrasound (LIPUS). Both therapeutic resources have presented good results with respect to modulation of the muscle repair process, but there is not description on how it could influence the determination of the phenotype of the newly formed fibers in the type of myosin heavy chain (MyHC) expresses literature and little is known about its effects on oxidative stress. Furthermore, there is a need to determine the molecular mechanisms involved, as well as the methodology and suitable dosimetric parameters. The objective of this proposal is to evaluate the effects of LLLT and LIPUS on the expression of isoforms of MyHC, the expression and synthesis of proteins involved in muscle repair (myostatin and calcineurin) and oxidative stress during the repair process of skeletal muscle of rats. Wistar rats will be divided into four groups: 1) control; 2) cryoinjured without treatment; 3) cryoinjured treated with LLLT and 4) cryoinjured treated with LIPUS. Analyses will be performed at 3, 6, 12 and 24 hours for analysis of oxidative stress, and 3, 7 and 14 days for analysis of MyHC, myostatin and calcineurin. Cryoinjury will be performed on the belly of the tibialis anterior (TA) muscle. LLLT will be performed daily using the MMOptics GaAlAs laser (wavelength of 780 nm) using an energy density of 10 J/cm², an output power of 40 mW, a beam spot of 0.4 cm² and time of 10 seconds. The treatment with LIPUS will be performed using the Sonomaster® equipment with the frequency of 1 MHz, an intensity of 0.4 W/cm², a pulsed model cycles 1:4 and stationary application of 3 minutes. For the analysis of gene expression will be extracted total RNA of the TA muscle and will obtain cDNA for performing Real-time PCR using specific primers for each gene and proteins analysis will be performed using enzyme-linked immunosorbent assays (ELISA) and Western blotting, the stress oxidative will be performed by lipid peroxidation (LPO) and the damage to proteins, and the analysis of antioxidant enzymes: catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx). (AU)

Scientific publications

(6)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

VIANA DE MELO, CLAUDIA APARECIDA; ALVES, AGNELO NEVES; LINS TERENA, STELLA MARIS; SANTOS FERNANDES, KRISTIANNE PORTA; NUNES, FABIO DAUMAS; TEIXEIRA DA SILVA, DANIELA DE FATIMA; BUSSADORI, SANDRA KALIL; DEANA, ALESSANDRO MELO; MESQUITA-FERRARI, RAQUEL AGNELLI. Light-emitting diode therapy increases collagen deposition during the repair process of skeletal muscle. Lasers in Medical Science, v. 31, n. 3, p. 531-538, APR 2016. (14/12381-1, 13/21540-3)

MESQUITA-FERRARI, RAQUEL AGNELLI; ALVES, AGNELO NEVES; CARDOSO, VINICIUS DE OLIVEIRA; ARTILHEIRO, PAOLA PELEGRINELI; BUSSADORI, SANDRA KALIL; ROCHA, LILIA ALVES; NUNES, FABIO DAUMAS; SANTOS FERNANDES, KRISTIANNE PORTA. Low-level laser irradiation modulates cell viability and creatine kinase activity in C2C12 muscle cells during the differentiation process. Lasers in Medical Science, v. 30, n. 8, p. 2209-2213, NOV 2015. (13/21540-3, 10/09191-5)

ALVES, AGNELO NEVES; RIBEIRO, BEATRIZ GUIMARES; SANTOS FERNANDES, KRISTIANNE PORTA; COSTA SOUZA, NADHIA HELENA; ROCHA, LILIA ALVES; NUNES, FABIO DAUMAS; BUSSADORI, SANDRA KALIL; MESQUITA-FERRARI, RAQUEL AGNELLI. Comparative effects of low-level laser therapy pre- and post-injury on mRNA expression of MyoD, myogenin, and IL-6 during the skeletal muscle repair. Lasers in Medical Science, v. 31, n. 4, p. 679-685, MAY 2016. (14/12381-1, 13/21540-3, 12/11461-6, 11/04452-8, 11/17638-2)

RIBEIRO, BEATRIZ GUIMARAES; ALVES, AGNELO NEVES; DIAS DOS SANTOS, LUCAS ANDREO; CANTERO, TATIANE MATARAZZO; SANTOS FERNANDES, KRISTIANNE PORTA; DIAS, DANIELLE DA SILVA; BERNARDES, NATHALIA; DE ANGELIS, KATIA; MESQUITA-FERRARI, RAQUEL AGNELLI. Red and Infrared Low-Level Laser Therapy Prior to Injury with or without Administration after Injury Modulate Oxidative Stress during the Muscle Repair Process. PLoS One, v. 11, n. 4, APR 15 2016. (14/12381-1, 13/21540-3)

RODRIGUES, DANIELLE DE LIMA; ALVES, AGNELO NEVES; GUIMARAES, BEATRIZ RIBEIRO; DE ALCANTARA ARAUJO AMORIM, WESLLEY WALLACE; BUSSADORI, SANDRA KALIL; SANTOS FERNANDES, KRISTIANNE PORTA; MESQUITA-FERRARI, RAQUEL AGNELLI. Effect of prior application with and without post-injury treatment with low-level laser on the modulation of key proteins in the muscle repair process. Lasers in Medical Science, v. 33, n. 6, p. 1207-1213, AUG 2018. (17/06461-0, 14/12381-1, 13/21540-3)

RIBEIRO, BEATRIZ GUIMARAES; ALVES, AGNELO NEVES; DIAS DOS SANTOS, LUCAS ANDREO; SANTOS FERNANDES, KRISTIANNE PORTA; CANTERO, TATIANE MATARAZZO; GOMES, MARIANA TEIXEIRA; FRANCA, CRISTIANE MIRANDA; TEIXEIRA DA SILVA, DANIELA DE FATIMA; BUSSADORI, SANDRA KALIL; MESQUITA-FERRARI, RAQUEL AGNELLI. The effect of low-level laser therapy (LLLT) applied prior to muscle injury. Lasers in Surgery and Medicine, v. 47, n. 7, p. 571-578, SEP 2015. (14/12381-1, 12/11461-6, 12/25198-5, 13/21540-3)