Aparecido Monteiro Duque da Fonseca Guilherme, Carvalho Roxo Denise, Moreira Figueira Mariana, Scherer Rodrigo, da Silva Brum Igor, Frigo Lucio, Fronza Marcio
Guarulhos University, Guarulhos, Brazil.
Universidade Vila Velha, Vila Velha, Brazil.
Lasers Med Sci. 2025 Sep 18;40(1):366. doi: 10.1007/s10103-025-04614-5.
Macrophages are pivotal cells in the inflammatory process, and their functions can be modulated by laser irradiation. However, there is no consensus in the literature about Laser-photobiomodulation (L-PBM) parameters that are more suited to stimulate or inhibit them. The goal of this research is to contribute to the understanding of the effects of different L-PBM protocols on inflammatory mediators' production/inhibition and cell viability of macrophage cells in vitro.
The macrophages cells (RAW 264.7) were divided into two groups: LPS-stimulated (Lipopolysaccharide from Escherichia coli) and non-LPS-stimulated. Each group was subdivided into non-irradiated and irradiated groups. The irradiated group was further divided into: 660 nm, 1 J, 10s; 660 nm, 2 J, 20s; 660, 3 J, 30s; 808 nm 1 J, 10s; 808 nm, 2 J, 20s; 808, 3 J, 30s groups. To all groups and subgroups, a negative (non-irradiated) group was added. All groups and subgroups were evaluated for cell viability, Nitric oxide (NO), interleukin-6 (IL-6) and Tumor Necrosis Factor- α (TNF-α) production.
The results indicated that macrophages irradiated at 660 nm and 808 nm, operating in 1 J, 10s; 2 J, 20s and 3 J, 30s did not change NO, TNF-α and IL-6 production nor cell Viability in non-LPS groups. However, in LPS-stimulated macrophages, a significant stimulatory effect on NO production was observed after laser-irradiation with 660 nm for 2 J, 20s. This NO-stimulatory effect was not observed in the 880 nm irradiated group. LPS-activated macrophages and L-PBM irradiation at 660 nm and 808 nm also resulted in significant inhibitory effects on TNF-α and IL-6 production after 2 J, 20s irradiation. However, TNF-α and IL-6 inhibition using 1 J, 10s was achieved only in 660 nm group.
Our findings suggested that, if other parameters are fixed, time and related fluence/energy delivery are important dimensions to consider in L-PBM-production/inhibition on the inflammatory mediators tested. The NO synthesis was more prone to be modulated by a specific wavelength (660 nm) and a broader time-range inhibition of TNF-α and IL-6 was observed in 660 nm groups. Cell viability was not changed in any parameter tested.
巨噬细胞是炎症过程中的关键细胞，其功能可通过激光照射进行调节。然而，关于更适合刺激或抑制巨噬细胞的激光光生物调节（L-PBM）参数，文献中尚无定论。本研究的目的是有助于理解不同L-PBM方案对体外巨噬细胞炎症介质产生/抑制及细胞活力的影响。
巨噬细胞（RAW 264.7）分为两组：脂多糖刺激组（来自大肠杆菌的脂多糖）和非脂多糖刺激组。每组再细分为未照射组和照射组。照射组进一步分为：660nm，1J，10s；660nm，2J，20s；660，3J，30s；808nm 1J，10s；808nm，2J，20s；808，3J，30s组。给所有组和亚组添加一个阴性（未照射）组。对所有组和亚组评估细胞活力、一氧化氮（NO）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的产生。
结果表明，在非脂多糖刺激组中，660nm和808nm照射、能量为1J、10s；2J、20s和3J、30s时，巨噬细胞的NO、TNF-α和IL-6产生及细胞活力均未改变。然而，在脂多糖刺激的巨噬细胞中，660nm、2J、20s激光照射后对NO产生有显著的刺激作用。在880nm照射组中未观察到这种NO刺激作用。脂多糖激活巨噬细胞以及660nm和808nm的L-PBM照射在2J、20s照射后对TNF-α和IL-6产生也有显著抑制作用。然而，仅在660nm组中，1J、10s照射可实现对TNF-α和IL-6的抑制。
我们的研究结果表明，如果其他参数固定，时间和相关的通量/能量传递是L-PBM对所测试的炎症介质产生/抑制作用中需要考虑的重要因素。NO合成更容易受到特定波长（660nm）的调节，并且在660nm组中观察到对TNF-α和IL-6更广泛的时间范围抑制。在所测试的任何参数下细胞活力均未改变。
