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Effect of laser therapy on gene expression and production of cytokines, chemokines and growth factors involved in muscle repair

Grant number: 13/07502-1
Support type:Regular Research Grants
Duration: August 01, 2014 - January 31, 2017
Field of knowledge:Health Sciences - Physiotherapy and Occupational Therapy
Principal Investigator:Kristianne Porta Santos Fernandes
Grantee:Kristianne Porta Santos Fernandes
Home Institution: Universidade Nove de Julho (UNINOVE). Campus Vergueiro. São Paulo , SP, Brazil
Assoc. researchers:Fabio Daumas Nunes ; Raquel Agnelli Mesquita Ferrari

Abstract

Effect of laser therapy on gene expression and production of cytokines, chemokines and growth factors involved in muscle repairThe interactions between muscle and the macrophages that invade this tissue after the occurrence of injury are crucial to the repair process. Low-level laser therapy (LLLT) has been widely used in clinical and experimental settings for the treatment of muscle injuries. However, little is known regarding the effects of laser irradiation on macrophages. The objectives of this project are to evaluate the effect of LLLT (2 parameters) on: the gene expression and protein synthesis of 12 products (that may play an important role in the muscle repair process) of macrophages (activated into M1, M2a or M2c phenotypes); the differentiation and fusion of myogenic precursor cells (C2C12) cultivated with activated macrophages supernatants; the phenotype of macrophages in the tibialis anterior muscle of rats after acute injury; and the immunohistochemical expression of two proteins (the most modulated during in vitro evaluations) in the tibialis anterior muscle of rats after acute injury. The 2 LBI dosimetric parameters will be chosen for its ability to modulate the production of IL-6 and TNF-alpha in a dose-response experiment. After this, J774 macrophage cultures will be activated, washed and will receive two different laser irradiation parameters (chosen taking into account the dose response curve). After the incubation periods (4 and 24h), the gene expression of the chemokines cxcl2 (MIP-2a), ccl 2 (MCP-1), ccl 3 (MIP-1a) and CCl4 (MIP-1b), cytokines IL-1 beta, IFN gamma, IL-6, TNF-alpha and IL-13 and growth factors TGF beta1, IGF1 and Vegfa will be evaluated using polymerase chain reaction arrays. The supernatant of the cultures from the different experimental groups (24h after irradiations) will be used to assess protein production from macrophages and also to treat myogenic precursor cells (C2C12) in order to analyze their differentiation and fusion. In each experimental situation, non-irradiated cells will serve as controls. Three independent experiments will be performed under each condition. For in vivo analysis, Wistar rats will be allocated into four groups: control (n=5); injury group (n=15); injury + LLLT (parameter 1) group (n=15); and injury + LLLT (parameter 2) group (n=15). After 2, 4, and 7 days, the animals will be sacrificed and the muscles will be removed and processed for immunohistochemistry. M1(CD68 and CD80 labeling), M2a (CD206 labeling) and M2c (CD163 labeling) macrophage profiles and two proteins whose production has been more clearly modulated by LLLT in vitro study will be identified. The immunohistochemical expression data will be correlated with the different stages of the skeletal muscle repair process. All results will be statistically analyzed. (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)
ANDREO, LUCAS; SOLDERA, CARLA A.; RIBEIRO, BEATRIZ GUIMARAES; VICENTE DE MATOS, PATRICK RAOUL; SOUSA, PAOLA BINDES; DE ALCANTARA ARAUJO AMORIM, WESLLEY WALLACE; RATTO TEMPESTINI HORLIANA, ANNA CAROLINA; BUSSADORI, SANDRA KALIL; SANTOS FERNANDES, KRISTIANNE PORTA; MESQUITA-FERRARI, RAQUEL AGNELLI. Effects of Photobiomodulation on Functionality in Wistar Rats with Sciatic Nerve Injury. Photochemistry and Photobiology, v. 95, n. 3, p. 879-885, MAY 2019. Web of Science Citations: 0.
SCHALCH, TATIANA DIAS; FERNANDES, MARIA HELENA; SETUBAL DESTRO RODRIGUES, MARIA FERNANDA; GUIMARAES, DOUGLAS MAGNO; NUNES, FABIO DAUMAS; RODRIGUES, JOAO COSTA; GARCIA, MONICA PEREIRA; MESQUITA FERRARI, RAQUEL AGNELLI; BUSSADORI, SANDRA KALIL; SANTOS FERNANDES, KRISTIANNE PORTA. Photobiomodulation is associated with a decrease in cell viability and migration in oral squamous cell carcinoma. Lasers in Medical Science, v. 34, n. 3, p. 629-636, APR 2019. Web of Science Citations: 1.
SOUZA, NADHIA H. C.; MESQUITA-FERRARI, RAQUEL A.; RODRIGUES, MARIA FERNANDA S. D.; DA SILVA, DANIELA F. T.; RIBEIRO, BEATRIZ G.; ALVES, AGNELO N.; GARCIA, MONICA P.; NUNES, FABIO D.; DA SILVA JUNIOR, EVALDO M.; FRANCA, CRISTIANE M.; BUSSADORI, SANDRA K.; FERNANDES, KRISTIANNE P. S. Photobiomodulation and different macrophages phenotypes during muscle tissue repair. JOURNAL OF CELLULAR AND MOLECULAR MEDICINE, v. 22, n. 10, p. 4922-4934, OCT 2018. Web of Science Citations: 1.
ANDREO, L.; SOLDERA, C. B.; RIBEIRO, B. G.; DE MATOS, P. R. V.; BUSSADORI, S. K.; FERNANDES, K. P. S.; MESQUITA-FERRARI, R. A. Effects of photobiomodulation on experimental models of peripheral nerve injury. Lasers in Medical Science, v. 32, n. 9, p. 2155-2165, DEC 2017. Web of Science Citations: 5.
DA SILVA JUNIOR, EVALDO MOREIRA; MESQUITA-FERRARI, RAQUEL AGNELLI; FRANCA, CRISTIANE MIRANDA; ANDREO, LUCAS; BUSSADORI, SANDRA KALIL; SANTOS FERNANDES, KRISTIANNE PORTA. Modulating effect of low intensity pulsed ultrasound on the phenotype of inflammatory cells. BIOMEDICINE & PHARMACOTHERAPY, v. 96, p. 1147-1153, DEC 2017. Web of Science Citations: 2.
DO VALE, KATIA LLANOS; MARIA, DURVANEI AUGUSTO; PICOLI, LARA CRISTINA; DEANA, ALESSANDRO MELO; MASCARO, MARCELO BETTI; MESQUITA FERRARI, RAQUEL AGNELLI; BUSSADORI, SANDRA KALIL; SANTOS FERNANDES, KRISTIANNE PORTA. The Effects of Photobiomodulation Delivered by Light-Emitting Diode on Stem Cells from Human Exfoliated Deciduous Teeth: A Study on the Relevance to Pluripotent Stem Cell Viability and Proliferation. Photomedicine and Laser Surgery, v. 35, n. 12, p. 659-665, DEC 2017. Web of Science Citations: 0.

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