Photobiomodulation effects on synovial morphology, iNOS gene, and protein expression in a model of acute inflammation.

Acta cirurgica brasileira Pub Date : 2024-03-15 eCollection Date: 2024-01-01 DOI:10.1590/acb392024
Fernando Russo Costa do Bomfim, Bruna Silva Gomes, Sabrina Zanchetta Lanza, Marcelo Augusto Marretto Esquisatto, Gaspar de Jesus Lopes-Filho
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Abstract

Purpose: To evaluate morphological aspects and inducible nitric oxide synthase (iNOS) gene and protein expression in a model of acute inflammation.

Methods: Thirty-six female Wistar rats were assigned into three groups: control (saline, n = 12), sham (arthritis, n = 12), and PBM (arthritis and photobiomodulation, n = 12). Arthritis induction was performed with 200 μg of intra-articular Zymosan in sham and PBM animals. PBM was performed 24 h after induction with a laser device (λ = 808 nm, 25 mW of nominal power, fluence of 20 J/cm2, beam area of 0.02 mm2, time of 33 s, total energy of 0.825 J) with punctual and single dose application. Morphological analysis of joint structure (HE) and immunohistochemistry (anti-iNOS antibody) were performed on knee samples, and synovial tissue was submitted to RNA extraction, cDNA synthesis and gene expression analysis by quantitative polymerase chain reaction. Statistical analyses were performed with p < 0.05.

Results: It was observed an increase in the thickness of the synovial lining epithelium and inflammatory infiltrate in sham compared to PBM. Gene expression analysis showed higher iNOS expression in PBM, and iNOS protein expression decreased in PBM compared to sham.

Conclusions: Photobiomodulation decreased inflammation in PBM animals, upregulated iNOS gene expression, however down egulated protein expression compared to sham.

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光生物调节对急性炎症模型中滑膜形态、iNOS 基因和蛋白表达的影响。
目的:评估急性炎症模型的形态学方面以及诱导型一氧化氮合酶(iNOS)基因和蛋白的表达:将 36 只雌性 Wistar 大鼠分为三组:对照组(生理盐水,n = 12)、假组(关节炎,n = 12)和 PBM 组(关节炎和光生物调节,n = 12)。用 200 μg 关节内注射 Zymosan 诱导假性和 PBM 动物关节炎。诱导24小时后,使用激光设备(λ = 808 nm,标称功率25 mW,通量20 J/cm2,光束面积0.02 mm2,时间33 s,总能量0.825 J)以单剂量准时进行PBM。对膝关节样本进行关节结构形态学分析(HE)和免疫组化(抗 iNOS 抗体),并对滑膜组织进行 RNA 提取、cDNA 合成和定量聚合酶链反应基因表达分析。统计分析以 p < 0.05 为限:结果:与 PBM 相比,观察到假关节滑膜上皮厚度增加,炎症浸润增加。基因表达分析表明,与假体相比,PBM 的 iNOS 表达更高,而 iNOS 蛋白表达降低:结论:与假体相比,光生物调节减少了 PBM 动物的炎症反应,上调了 iNOS 基因表达,但降低了 iNOS 蛋白表达。
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