Department of Prosthodontics, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China.
Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research Center for Oral Diseases, Shanghai 200011, China.
Oxid Med Cell Longev. 2019 Aug 14;2019:1529520. doi: 10.1155/2019/1529520. eCollection 2019.
To investigate whether methylene blue-mediated photodynamic therapy (MB-PDT) can affect the "fate" of macrophages or in periodontitis tissues and to explore the potential mechanism.
For treatments, THP-1 macrophages were divided into three experimental groups: C/control, no treatment; MB, methylene blue treatment; and MB-PDT, MB and laser irradiation treatment. Then, apoptosis and apoptosis-related proteins were detected in each group. For treatments, periodontitis was ligature-induced in the first molars of the bilateral maxilla in 12 Sprague Dawley (SD) rats. After six weeks, the ligatures were removed and all the induced molars underwent scaling and root planning (SRP). Then, the rats were divided into three groups according to the following treatments: SRP, saline solution; MB, phenothiazinium dye; and MB-PDT, MB and laser irradiation. Apoptotic macrophages, inflammation levels, and alveolar bone resorption in the periodontal tissues of rats were analyzed in each group.
, flow cytometry analysis demonstrated that 10 M MB and 40 J/cm laser irradiation maximized the apoptosis rate (34.74%) in macrophages. Fluorescence probe and Western blot analyses showed that MB-PDT induced macrophage apoptosis via reactive oxygen species (ROS) and the mitochondrial-dependent apoptotic pathway. Conversely, the addition of exogenous antioxidant glutathione (GSH) and the pan-caspase inhibitor Z-VAD-FMK markedly reduced the apoptotic response in macrophages. , immunohistochemistry, histology, radiographic, and molecular biology experiments revealed fewer infiltrated macrophages, less bone loss, and lower IL-1 and TNF- levels in the MB-PDT group than in the SRP and MB groups ( < 0.05). Immunohistochemistry analysis also detected apoptotic macrophages in the MB-PDT group.
MB-PDT could induce macrophage apoptosis and in rats with periodontitis. This may be another way for MB-PDT to relieve periodontitis in addition to its antimicrobial effect. Meanwhile, MB-PDT induced apoptosis in THP-1 macrophages via the mitochondrial caspase pathway.
研究亚甲蓝介导的光动力疗法（MB-PDT）是否能影响牙周炎组织中巨噬细胞的“命运”，并探讨其潜在机制。
对于 处理，将 THP-1 巨噬细胞分为三组实验：C/对照，无处理；MB，亚甲蓝处理；MB-PDT，亚甲蓝和激光照射处理。然后，检测各组细胞凋亡及凋亡相关蛋白。对于 处理，在双侧上颌第一磨牙的结扎诱导牙周炎 12 只 Sprague Dawley（SD）大鼠。六周后，去除结扎线，所有诱导磨牙均进行洁治和根面平整（SRP）。然后，根据以下处理将大鼠分为三组：SRP，生理盐水；MB，吩噻嗪染料；MB-PDT，亚甲蓝和激光照射。分析各组大鼠牙周组织中凋亡的巨噬细胞、炎症水平和牙槽骨吸收情况。
体外实验结果显示，10 μM MB 和 40 J/cm2 激光照射可使巨噬细胞凋亡率达到最大值（34.74%）。荧光探针和 Western blot 分析表明，MB-PDT 通过活性氧（ROS）和线粒体依赖的凋亡途径诱导巨噬细胞凋亡。相反，添加外源性抗氧化剂谷胱甘肽（GSH）和泛半胱天冬酶抑制剂 Z-VAD-FMK 可显著降低巨噬细胞的凋亡反应。体内实验结果显示，免疫组织化学、组织学、影像学和分子生物学实验表明，MB-PDT 组的浸润巨噬细胞较少，骨损失较少，IL-1 和 TNF-α 水平较低（<0.05）。免疫组织化学分析还检测到 MB-PDT 组中有凋亡的巨噬细胞。
MB-PDT 可诱导牙周炎大鼠的巨噬细胞凋亡，这可能是 MB-PDT 除抗菌作用外缓解牙周炎的另一种方式。同时，MB-PDT 通过线粒体半胱天冬酶途径诱导 THP-1 巨噬细胞凋亡。
