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Among pathogens colonizing the CF lung,\n                  <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                  is predominant, infecting over 50% of adults with CF, and becoming antibiotic-resistant over time. Current therapies for CF, while providing tremendous benefits, fail to eliminate persistent bacterial infections, chronic inflammation, and irreversible lung damage, necessitating novel therapeutic strategies. Our group engineered mesenchymal stromal cell derived extracellular vesicles (MSC EVs) to carry the microRNA let-7b-5p as a dual anti-infective and anti-inflammatory treatment. MSC EVs are low-immunogenicity platforms with innate antimicrobial and immunomodulatory properties, while let-7b-5p reduces biofilm formation and inflammation. In a preclinical CF mice model, we reported that let-7b-5p-loaded MSC EVs reduced\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  burden, immune cells, and proinflammatory cytokines in the lungs. We hypothesize four complementary mechanisms for the observed\n                  <jats:italic>in-vivo<\/jats:italic>\n                  effects of the let-7b-5p loaded MSC EVs: antimicrobial activity, anti-inflammatory properties, inhibition of antibiotic-resistant\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  biofilm formation in CF airways, and stimulation of anti-inflammatory macrophage behaviors. This study focused on the second and third mechanisms and demonstrates that MSC EVs engineered to contain let-7b-5p effectively blocked the formation of antibiotic-resistant\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  biofilms on primary human bronchial epithelial cells (pHBECs) while also reducing\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  -induced inflammation. This approach holds promise for improving outcomes for people with CF. Future work will focus on optimizing delivery strategies and expanding the clinical applicability of MSC EVs to target other CF-associated pathogens.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>NEW AND NOTEWORTHY<\/jats:title>\n                  <jats:p>\n                    This is the first study demonstrating that let-7b-5p loaded Mesenchymal Stromal Cell Extracellular Vesicles (MSC EVs) block antibiotic-resistant\n                    <jats:italic>P. aeruginosa<\/jats:italic>\n                    biofilm formation and reduce inflammation in CF primary human bronchial epithelial cells.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>GRAPHICAL ABSTRACT<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"656674v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>","DOI":"10.1101\/2025.05.28.656674","type":"posted-content","created":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T00:55:20Z","timestamp":1748480120000},"source":"Crossref","is-referenced-by-count":3,"title":["Let-7b-5p loaded Mesenchymal Stromal Cell Extracellular Vesicles reduce\n                  <i>Pseudomonas<\/i>\n                  - biofilm formation and inflammation in CF Bronchial Epithelial Cells"],"prefix":"10.64898","author":[{"given":"Sharanya","family":"Sarkar","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roxanna","family":"Barnaby","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amanda","family":"Nymon","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lily A.","family":"Charpentier","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lily","family":"Taub","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew J.","family":"Wargo","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, VT, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel J.","family":"Weiss","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Vermont, Burlington, VT, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tracey L.","family":"Bonfield","sequence":"additional","affiliation":[{"name":"National Center for Regenerative Medicine, Case Western Reserve University, Cleveland, OH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruce A.","family":"Stanton","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026041123481675000_2025.05.28.656674v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2715"},{"key":"2026041123481675000_2025.05.28.656674v1.2","doi-asserted-by":"publisher","DOI":"10.1152\/ajplung.00326.2014"},{"key":"2026041123481675000_2025.05.28.656674v1.3","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-016-2562-z"},{"key":"2026041123481675000_2025.05.28.656674v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcf.2018.05.006"},{"key":"2026041123481675000_2025.05.28.656674v1.5","doi-asserted-by":"publisher","DOI":"10.3390\/antibiotics10030338"},{"key":"2026041123481675000_2025.05.28.656674v1.6","first-page":"e01828","article-title":"Competition in Biofilms between Cystic Fibrosis Isolates of Pseudomonas aeruginosa Is Shaped by R-Pyocins","volume":"10","year":"2019","journal-title":"mBio"},{"key":"2026041123481675000_2025.05.28.656674v1.7","first-page":"759348","article-title":"The formation of biofilms by Pseudomonas aeruginosa: a review of the natural and synthetic compounds interfering with control mechanisms","volume":"2015","year":"2015","journal-title":"Biomed Res Int"},{"key":"2026041123481675000_2025.05.28.656674v1.8","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.55.8.1813-1818.1987"},{"key":"2026041123481675000_2025.05.28.656674v1.9","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.175.11.7512"},{"key":"2026041123481675000_2025.05.28.656674v1.10","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.00275-18"},{"key":"2026041123481675000_2025.05.28.656674v1.11","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00373.2016"},{"key":"2026041123481675000_2025.05.28.656674v1.12","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1902734116"},{"key":"2026041123481675000_2025.05.28.656674v1.13","unstructured":"Smith, W. 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