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 fibroblasts (PDL), 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, LTB₄, 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 (0.27-27nM) reduced cytokine induced production of PGE₂, and up-regulated 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 pro-resolution actions of RvD1 with up-regulation of arachidonic acid derived endogenous resolution pathways (LXA₄) and suggest resolution pathway synergy establishing a novel mechanism for the pro-resolution activity of -3 DHA-derived resolution agonist RvD1.