Department of Pathology and Parasitology, Institute of Biomedical Sciences, Federal University of Alfenas, 700 Gabriel Monteiro da Silva Street, Alfenas, MG, 37130-000, Brazil.
Department of Clinic and Surgery, School of Dentistry, Federal University of Alfenas, 700 Gabriel Monteiro da Silva Street, Alfenas, MG, 37130-000, Brazil.
Lasers Med Sci. 2022 Feb;37(1):135-148. doi: 10.1007/s10103-020-03179-9. Epub 2020 Nov 6.
This study aims to evaluate the effects of photobiomodulation (PBM) on human monocytes, assessing the oxidative burst and ultimate fungicidal potential of these cells, as well as the gene expression at the mRNA level of CD68, CD80, CD163, CD204, IL-6, TNF-α and IL-10 in derived macrophages. Primary cultures of human monocytes were irradiated with an InGaAlP (660 nm)/GaAlAs (780 nm) diode laser (parameters: 40 mW, 0.04 cm, 1 W/cm; doses: 200, 400 and 600 J/cm). Cells were submitted to the chemiluminescence assay, and a microbicidal activity assay against Candida albicans was performed. Reactive oxygen species (ROS) and nitric oxide (NO) production were measured, and cell viability was assessed by the exclusion method using 0.2% Trypan blue reagent. Irradiated monocytes were cultured for 72 h towards differentiation into macrophages. Total RNA was extracted, submitted to reverse transcription and real-time PCR. The results were analysed by ANOVA and the Tukey test (α = 0.05). Irradiated monocytes revealed a significant increase in their intracellular and extracellular ROS (P < 0.001). The 660 nm wavelength and 400 J/cm dose were the most relevant parameters (P < 0.001). The fungicidal capacity of the monocytes was shown to be greatly increased after PBM (P < 0.001). PBM increased the expression of TNF-α (P = 0.0302) and the production of NO (P < 0.05) and did not impair monocyte viability. PBM induces a pro-inflammatory Th1-driven response in monocytes and macrophages.
本研究旨在评估光生物调节（PBM）对人类单核细胞的影响，评估这些细胞的氧化爆发和最终杀菌潜能，以及衍生巨噬细胞中 CD68、CD80、CD163、CD204、IL-6、TNF-α 和 IL-10 的 mRNA 水平的基因表达。用人单核细胞原代培养物照射 InGaAlP（660nm）/GaAlAs（780nm）二极管激光（参数：40mW、0.04cm、1W/cm；剂量：200、400 和 600J/cm）。对细胞进行化学发光检测，并对白色念珠菌进行杀菌活性检测。测量活性氧（ROS）和一氧化氮（NO）的产生，并通过使用 0.2%台盼蓝试剂排除法评估细胞活力。照射后的单核细胞培养 72 小时以分化为巨噬细胞。提取总 RNA，进行逆转录和实时 PCR。结果通过方差分析和 Tukey 检验（α=0.05）进行分析。照射后的单核细胞显示其细胞内和细胞外 ROS 显著增加（P<0.001）。660nm 波长和 400J/cm 剂量是最相关的参数（P<0.001）。PBM 后单核细胞的杀菌能力大大增加（P<0.001）。PBM 增加了 TNF-α 的表达（P=0.0302）和 NO 的产生（P<0.05），且不损害单核细胞活力。PBM 诱导单核细胞和巨噬细胞中促炎 Th1 驱动的反应。
