Department of Clinical Dentistry-Center for Clinical Dental Research, University of Bergen, Bergen, Norway;
Am J Physiol Cell Physiol. 2013 Sep 15;305(6):C673-9. doi: 10.1152/ajpcell.00242.2012. Epub 2013 Jul 17.
Resolution agonists are endogenous mediators that drive inflammation to homeostasis. We earlier demonstrated in vivo activity of resolvins and lipoxins on regenerative periodontal wound healing. The goal of this study was to determine the impact of resolvin D1 (RvD1) on the function of human periodontal ligament (PDL) fibroblasts, which are critical for wound healing during regeneration of the soft and hard tissues around teeth. Primary cells were cultured from biopsies obtained from three individuals free of periodontal diseases. Peripheral blood mononuclear cells were isolated by density gradient centrifugation from whole blood of healthy volunteers. PGE2, leukotriene B4 (LTB4), and lipoxin A4 (LXA4) in culture supernatants were measured by ELISA. The direct impact of RvD1 on PDL fibroblast proliferation was measured and wound closure was analyzed in vitro using a fibroblast culture "scratch assay." PDL fibroblast function in response to RvD1 was further characterized by basic FGF production by ELISA. IL-1β and TNF-α enhanced the production of PGE2. Treatment of PDL cells and monocytes with 0.1-10 ng/ml RvD1 (0.27-27 M) reduced cytokine induced production of PGE2 and upregulated LXA4 production by both PDL cells and monocytes. RvD1 significantly enhanced PDL fibroblast proliferation and wound closure as well as basic FGF release. The results demonstrate that anti-inflammatory and proresolution actions of RvD1 with upregulation of arachidonic acid-derived endogenous resolution pathways (LXA4) and suggest resolution pathway synergy establishing a novel mechanism for the proresolution activity of the ω-3 docosahexaenoic acid-derived resolution agonist RvD1.
内源性介质（resolution agonists）是促炎向稳态发展的内源性介质。我们先前在体内实验中证明了代谢产物（resolvins 和 lipoxins）在再生性牙周伤口愈合中的作用。本研究旨在确定内源性介质（resolution D1，RvD1）对人牙周韧带（PDL）成纤维细胞功能的影响，这些细胞在牙齿周围软组织和硬组织的再生过程中的伤口愈合中起关键作用。从三个无牙周病的个体的活检中培养原代细胞。通过密度梯度离心从健康志愿者的全血中分离外周血单核细胞。通过 ELISA 测量培养上清液中的 PGE2、白三烯 B4（LTB4）和脂氧素 A4（LXA4）。直接测量 RvD1 对 PDL 成纤维细胞增殖的影响，并通过体外成纤维细胞培养“划痕实验”分析伤口闭合。通过 ELISA 测量碱性成纤维细胞生长因子（basic FGF）的产生，进一步研究 RvD1 对 PDL 成纤维细胞功能的影响。IL-1β 和 TNF-α 增强 PGE2 的产生。用 0.1-10ng/ml RvD1（0.27-27μM）处理 PDL 细胞和单核细胞，可减少细胞因子诱导的 PGE2 产生，并上调 PDL 细胞和单核细胞产生的 LXA4。RvD1 可显著增强 PDL 成纤维细胞增殖、伤口闭合和碱性成纤维细胞生长因子释放。结果表明，RvD1 具有抗炎和促分解作用，可上调花生四烯酸衍生的内源性分解途径（LXA4），提示分解途径协同作用，为 ω-3 二十二碳六烯酸衍生的分解激动剂 RvD1 的促分解活性建立了一种新的机制。
