Efeitos da fotobiomodulação com diferentes dosimetrias associada ao plasma rico em plaquetas na artrite reumatoide aguda induzida em ratas Wistar
Data
2024-04-05
Autores
Gomes, Bruna Silva 
Orientadores
Lopes Filho, Gaspar de Jesus
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Objetivo: Diante da incidência mundial acometida pela artrite reumatoide (AR), o objetivo deste estudo foi avaliar os efeitos da fotobiomodulação (FBM) com laser de baixa intensidade com diferentes dosimetrias associada ao plasma rico em plaquetas (PRP) em ratas submetidas a artrite reumatoide aguda induzida. Métodos: Oitenta e quatro ratas Wistar foram distribuídas em sete grupos (n=12): controle; artrite induzida (sham); artrite induzida e tratadas com PRP; artrite induzida e tratada com FBM 0,8J; artrite induzida e tratadas com PRP e FBM 0,8J; artrite induzida e tratadas com FBM 1,4J e artrite induzida e tratadas com PRP e FBM 1,4J. Os grupos que receberam fotobiomodulação com laser Arseneto de Gálio e Alumínio foram irradiados com comprimento de onda = 808nm, potência = 25mW, fluência = 20J/cm2 ou 50J/cm2, diâmetro de feixe = 0,02mm, energia total = 0,8J ou 1,4J e tempo = 33 ou 56 segundos, enquanto o grupo PRP recebeu injeção intra-articular com concentração igual a 8x105 plaquetas. Após três dias os animais foram eutanasiados e divididos para as análises. As análises morfológicas foram realizadas a partir da medição das áreas inflamadas da região sinovial, cada uma das lâminas teve 5 campos aferidos com o programa Image J. A qPCR foi utilizada para analisar a expressão gênica através da técnica de Ct comparativo (2-ΔΔCt). Para análises de expressão proteica foi utilizada a técnica imuno-histoquímica, empregando a contagem de células positivas a partir da reação imuno-histoquímica formada, seja ela celular ou extracelular. Em seguida, os dados quantitativos foram realizados em média±DP com teste ANOVA e pós-teste de Tukey com nível de significância de 5% (p<0,05). Resultados: As análises morfológicas evidenciaram um aumento significativo no número de células inflamatórias nos grupos sham, FBM 0,8J, PRP+FBM 0,8J, FBM 1,4J e PRP+FBM 1,4J, enquanto o grupo PRP não apresentou diferença. A expressão gênica revelou que os genes IL-6 e C3 tiveram diminuição estatística (p<0,0001 e p=0,0085 respectivamente) nos grupos irradiados com FBM e/ou PRP+FBM, contudo, o grupo PRP não apresentou diferença considerável para C3. As análises de expressão proteica revelaram que tanto IL-6 quanto C3 tiveram aumento elevado de células inflamatórias no grupo sham quando comparado ao controle; os grupos irradiados apresentaram diminuição significativa com relação ao sham, sendo que para IL-6 eles apresentaram um p<0,0001 e para C3 apresentaram p=0,0028. Novamente sem alterações consideráveis no grupo PRP. Conclusão: Neste estudo, observamos que a fotobiomodulação com laser de baixa intensidade cuja dosimetria foi 808nm, 20J/cm2, 0,8J e 33 segundos associada ou não ao plasma rico em plaquetas apresentou os melhores resultados nos aspectos avaliados.
Introduction: In the face of the global incidence of rheumatoid arthritis (RA), the aim of this study was to evaluate the effects of low-intensity laser photobiomodulation (LLLT) with different dosages associated with platelet-rich plasma (PRP) in rats with induced acute rheumatoid arthritis. Methods: Eighty-four Wistar rats were divided into seven groups (n=12): control; induced arthritis (sham); induced arthritis and treated with PRP; induced arthritis and treated with LLLT 0.8J; induced arthritis and treated with PRP and LLLT 0.8J; induced arthritis and treated with LLLT 1.4J and induced arthritis and treated with PRP and LLLT 1.4J. The groups receiving photobiomodulation with Gallium Arsenide and Aluminum Laser were irradiated with a wavelength of 808nm, power of 25mW, fluence of 20J/cm2 or 50J/cm2, beam diameter of 0.02mm, total energy of 0.8J or 1.4J, and time of 33 or 56 seconds, while group C received intra-articular injection with a concentration of 8x105 platelets. After three days, the animals were euthanized and divided for analysis. Morphological analyses were performed by measuring the inflamed areas of the synovial region, with 5 fields assessed per slide using Image J software. qPCR was used to analyze gene expression through comparative Ct technique (2-ΔΔCt). Immunohistochemical technique was used for protein expression analysis, counting positive cells from the formed immunohistochemical reaction, whether cellular or extracellular. Quantitative data were then analyzed using mean±SD with ANOVA test and Tukey's post-test with a significance level of 5% (p<0.05). Results: Morphological analyses showed a significant increase in the number of inflammatory cells in groups sham, LLLT 0,8J, PRP+LLLT 0,8J, LLLT 1,4J and PRP+LLLT 1,4J, while group PRP showed no difference. Gene expression revealed statistical decrease (p<0.0001 and p=0.0085 respectively) in IL-6 and C3 genes in groups irradiated with LLLT and/or PRP+LLLT, however, group PRP showed no considerable difference for C3. Protein expression analyses showed that both IL-6 and C3 had significantly increased inflammatory cells in group sham compared to control; irradiated groups showed significant decrease compared to sham, with p<0.0001 for IL-6 and p=0.0028 for C3. Once again, no considerable changes were observed in group PRP. Conclusion: In this study, we observed that low-intensity laser photobiomodulation with a dosage of 808nm, 20J/cm2, 0.8J, and 33 seconds, whether associated with platelet-rich plasma or not, yielded the best results in the evaluated aspects.
Introduction: In the face of the global incidence of rheumatoid arthritis (RA), the aim of this study was to evaluate the effects of low-intensity laser photobiomodulation (LLLT) with different dosages associated with platelet-rich plasma (PRP) in rats with induced acute rheumatoid arthritis. Methods: Eighty-four Wistar rats were divided into seven groups (n=12): control; induced arthritis (sham); induced arthritis and treated with PRP; induced arthritis and treated with LLLT 0.8J; induced arthritis and treated with PRP and LLLT 0.8J; induced arthritis and treated with LLLT 1.4J and induced arthritis and treated with PRP and LLLT 1.4J. The groups receiving photobiomodulation with Gallium Arsenide and Aluminum Laser were irradiated with a wavelength of 808nm, power of 25mW, fluence of 20J/cm2 or 50J/cm2, beam diameter of 0.02mm, total energy of 0.8J or 1.4J, and time of 33 or 56 seconds, while group C received intra-articular injection with a concentration of 8x105 platelets. After three days, the animals were euthanized and divided for analysis. Morphological analyses were performed by measuring the inflamed areas of the synovial region, with 5 fields assessed per slide using Image J software. qPCR was used to analyze gene expression through comparative Ct technique (2-ΔΔCt). Immunohistochemical technique was used for protein expression analysis, counting positive cells from the formed immunohistochemical reaction, whether cellular or extracellular. Quantitative data were then analyzed using mean±SD with ANOVA test and Tukey's post-test with a significance level of 5% (p<0.05). Results: Morphological analyses showed a significant increase in the number of inflammatory cells in groups sham, LLLT 0,8J, PRP+LLLT 0,8J, LLLT 1,4J and PRP+LLLT 1,4J, while group PRP showed no difference. Gene expression revealed statistical decrease (p<0.0001 and p=0.0085 respectively) in IL-6 and C3 genes in groups irradiated with LLLT and/or PRP+LLLT, however, group PRP showed no considerable difference for C3. Protein expression analyses showed that both IL-6 and C3 had significantly increased inflammatory cells in group sham compared to control; irradiated groups showed significant decrease compared to sham, with p<0.0001 for IL-6 and p=0.0028 for C3. Once again, no considerable changes were observed in group PRP. Conclusion: In this study, we observed that low-intensity laser photobiomodulation with a dosage of 808nm, 20J/cm2, 0.8J, and 33 seconds, whether associated with platelet-rich plasma or not, yielded the best results in the evaluated aspects.
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Citação
GOMES, Bruna Silva. Efeitos da fotobiomodulação com diferentes dosimetrias associadas ao plasma rico em plaquetas na artrite reumatoide aguda induzida em ratas wistar. 2024. 65 f. Dissertação (Mestrado em Ciência Cirúrgica Interdisciplinar) – Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, 2024.