接近牙髓的深蛀牙可能发展成牙髓炎;为了防止这种情况,牙髓细胞需要平衡快速免疫反应,以避免牙髓快速肿胀。目前对接近牙髓的深度衰变的治疗涉及在牙髓中诱导低水平炎症以促进其修复的药物的应用,但是这种治疗有时是不够的。然而,不成功的治疗通常会导致牙髓炎。C5a-C5aR是免疫级联反应的初始阶段。阻断C5a-C5aR的结合可以减缓狭窄牙髓腔内的免疫反应,使牙髓细胞有足够的时间增殖,migrate,和区分。在这项研究中,我们比较了不同浓度和时间点的脂磷壁酸(LTA)和脂多糖(LPS),并使用C5aR拮抗剂W54011阻断C5a-C5aR轴.通过分析C5a的表达检测其阻断作用,C5aR,白细胞介素(IL)-6和Toll样受体2和4(TLR-2,4)。接下来,我们确定了LTA和LPS联合W54011治疗的最佳浓度和持续时间。根据我们的结果,我们选择1.0μg/mLLPS处理48h以产生人牙髓细胞的炎症模型。然后我们将细胞重新分组,并进行表达分析以监测C5a的表达,C5aR,IL-6和TLR-4在蛋白质和mRNA水平。LPS刺激48h和W54011治疗48h有效抑制炎症,不影响C5a表达。本研究为W54011在牙髓细胞中的后续研究提供了依据。
Deep tooth decay approaching the pulp may develop into pulpitis; to prevent this, pulp cells need to balance the rapid immune response to avoid rapid swelling of the pulp. Current treatment of deep decay that approaches the pulp involves the application of drugs that induce low-level inflammation in the dental pulp to promote its repair, but this treatment is sometimes insufficient. However, the unsuccessful treatment often resulted in pulpitis. The C5a-C5aR is the initial stage of the immune cascade response. Blocking the binding of C5a-C5aR can slow the immune response in the narrow pulp cavity, so that dental pulp cells have enough time to proliferate, migrate, and differentiate. In this study, we compared lipoteichoic acid (LTA) and lipopolysaccharides (LPS) at different concentrations and time points and used the C5aR antagonist W54011 to block the C5a-C5aR axis. The blocking effect was detected by analyzing the expression of C5a, C5aR, interleukin (IL)-6, and Toll-like receptors 2 and 4 (TLR-2, 4). Next, we determined the optimal concentration and duration of LTA and LPS treatment in combination with W54011. Based on our results, we selected 1.0 μg·mL-1 LPS treatment for 48 h to generate an inflammatory model of human dental pulp cells. We then regrouped the cells and conducted expression analyses to monitor the expression of C5a, C5aR, IL-6, and TLR-4 at the protein and mRNA levels. LPS stimulation for 48 h and treatment with W54011 for 48 h effectively inhibited inflammation and did not affect C5a expression. This study provides a basis for follow-up studies of W54011 in dental pulp cells.