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Analysis of the effects of photobiomodulation on the activity of chondrocytes and articular cartilage in experimental model of arthritis induced by collagen: in vivo and in vitro study

Grant number: 18/26062-6
Support type:Regular Research Grants
Duration: May 01, 2019 - July 31, 2022
Field of knowledge:Health Sciences - Physiotherapy and Occupational Therapy
Principal researcher:Rodrigo Labat Marcos
Grantee:Rodrigo Labat Marcos
Home Institution: Universidade Nove de Julho (UNINOVE). Campus Vergueiro. São Paulo , SP, Brazil
Assoc. researchers:Andrey Jorge Serra ; Rodrigo Labat Marcos

Abstract

Rheumatoid arthritis (RA) is a chronic inflammatory, systemic and progressive autoimmune disease that leads to irreversible destruction of cartilage and bone. Cytokines are essential in inflammation, and responsible for the synthesis and excessive release of metalloproteinases (MMPs). In vivo and in vitro experimental studies using photobiomodulation therapy have demonstrated positive effects on the modulation of the inflammatory process. Objective: To analyze the in vitro effects of photobiomodulation therapy on inflammatory cytokines IL-1, IL-6, TNF-±, metalloproteinases 2 and 9 (MMPs) and collagen II on cultured chondrocytes implanted in alginate hydrogel obtained from experimental model of Collagen-induced Rheumatoid Arthritis. Materials and Methods: Twenty-four male Wistar rats will be divided into two groups, control and collagen induced arthritis (AIC), by means of 2 immunizations of Freund's Complete Adjuvant associated with Collagen II on days 0 and 7 and after 28 will be applied 3rd dose in the knee joint, after the last induction, the culture will be performed chondrocytes extracted from the articular cartilages. The cells will be irradiated by the laser in 2 stages: 1-stage, four chondrocyte culture plates will receive preconditioning with PBMT for 3 days in the chondrocyte cultures, with a power of 50 mW, wavelength of 660 nm, area beam of 0.028 cm 2, irradiance of 1.78 W / cm 2, in order to construct a dose response curve the energies used will be 0.3; 0.6; 0.9; and 1.2 joules of energy, and radiant exposure of 10.7; 21.4; 32.1; and 42.8 J / cm2. The cells that show better viability will be used for implantation in hydrogel of Alginate and submitted to growth in bioreactor, where the 2nd stage of the irradiation with the same parameters used in the culture of chondrocytes will occur. The protein and gene expression analyzes of the cytokines IL-1, IL-6, TNF-±, metalloproteinases 1, 2, 9 and 13, collagen II, Runx-2, Sox-9 and ADAMTS -4 and 5 and HIF-Hypoxia -Inducible Factor will be performed by Western Blotting, RT / PCR and ELISA techniques. The data found will be submitted to normality test (Shapiro Wilk test), and if normal one-way ANOVA was performed with the Tukey post hoc test, considered P <0.05 to indicate a significant difference. (AU)

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Scientific publications
(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)
AMADIO, ELIANE MARTINS; MARCOS, RODRIGO LABAT; SERRA, ANDREY JORGE; DOS SANTOS, SOLANGE ALMEIDA; CAIRES, JHENIPHE ROCHA; FERNANDES, GUILHERME HENRIQUE CARDOSOS; LEAL-JUNIOR, ERNESTO CESAR; FERRARI, JOAO CARLOS CORREA; DE TARSO CAMILLO DE CARVALHO, PAULO. Effect of photobiomodulation therapy on the proliferation phase and wound healing in rats fed with an experimental hypoproteic diet. Lasers in Medical Science, NOV 2020. Web of Science Citations: 0.

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