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However, their potential to deliver bioactive lipid mediators remains largely unexplored. Here, we present a novel synthetic biology-based strategy to selectively load EVs with proresolving lipid mediators of the resolvin D- and E-series by coexpressing the resolvin biosynthetic enzymes cyclooxygenase 2, 5-lipoxygenase, and 15-lipoxygenase using a custom-designed multigene expression vector. Human embryonic kidney 293\u2009T cells transfected with the multigene expression vector and cultured in the presence of fatty acid free bovine serum albumen-complexed docosahexaenoic acid, eicosapentaenoic acid, and aspirin produced multiple members of the resolvin D, aspirin-triggered resolvin D-series, and resolvin E1 and E2, along with their biosynthetic precursors, which were subsequently packaged into EVs (referred to as resolvin EVs). Resolvin EVs attenuated neutrophil adhesion to endothelial cells both under static and flow conditions and preserved endothelial barrier integrity by upregulating VE-cadherin. In macrophages, resolvin EVs suppressed nuclear factor \u03baB reporter activity and the release of IL6 and TNF\u03b1. Effects of resolvin EVs on endothelial permeability and macrophage activation were abrogated by pharmacologic inhibition of EV uptake using nystatin and cytochalasin D. Furthermore, resolvin EVs enhanced efferocytosis in THP-1-derived macrophages compared to control EVs. Notably, postinjury administration of resolvin EVs attenuated pulmonary inflammation in lipopolysaccharide-treated mice without inducing systemic or pulmonary toxicity. Together, these findings establish a novel, scalable platform for generating resolvin-loaded EVs and highlight their therapeutic potential for acute lung injury and other chronic inflammatory disorders.<\/jats:p>","DOI":"10.1165\/rcmb.2025-0398oc","type":"journal-article","created":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T17:58:37Z","timestamp":1761155917000},"page":"661-674","update-policy":"https:\/\/doi.org\/10.1164\/crossmarkpolicies","source":"Crossref","is-referenced-by-count":2,"title":["Specialized proresolving mediator-loaded extracellular vesicles mitigate pulmonary inflammation"],"prefix":"10.1093","volume":"74","author":[{"given":"Manjula","family":"Karpurapu","sequence":"first","affiliation":[{"name":"Pulmonary, Critical Care and Sleep Medicine, Ohio State University Wexner Medical Center, Davis Heart and Lung Research Institute , Columbus, OH,","place":["United 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