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However, the progressive and mechanistic basis of blood\u2013brain barrier (BBB) injury following acute and sub-chronic 0% exposure remains poorly researched. In this study, we investigated whether short-term and prolonged 0% e-cigarette exposure alters BBB integrity using complementary in-vitro and in-vivo models. Using a well-established astrocyte\u2013bEnd.3 co-culture BBB model, 24-h exposure significantly reduced transendothelial electrical resistance (TEER) without increasing sodium fluorescein (NaF) permeability. Following prolonged 5-day exposure, TEER remained significantly reduced and was accompanied by increased NaF permeability. In contrast, permeability to 4 and 70\u2009kDa Fluorescein Isothiocyanate\u2013Dextran tracers remained unchanged. Immunocytochemistry (ICC) demonstrated reduced claudin-5 mean fluorescence intensity (MFI) after 24\u2009h, along with reduced zonula occludens-1 (ZO-1) and claudin-5 MFI and disrupted junctional morphology following 5-day exposure. Western blot analysis additionally demonstrated a significant reduction in claudin-5 expression after 5\u2009days. To assess translational relevance, male and female CD-1 mice were exposed to 0% aerosols for 7 or 14\u2009days, with 7-day exposures producing no significant changes in BBB tight junction proteins, inflammatory cytokines, or plasma cardiac troponin-l levels. Fourteen-day exposures reduced ZO-1 expression in both sexes, occludin expression in males, and elevated pro-inflammatory cytokines IL-17A, GM-CSF, MCP-1, TNF-\u03b1, and IL-1\u03b1 without affecting plasma troponin-1. Collectively, these findings demonstrate that repeated 0% e-cigarette exposure promotes progressive BBB dysfunction and a neuroinflammatory environment, highlighting the need to consider BBB vulnerability as a critical endpoint in 0% e-cigarette exposure toxicology.<\/jats:p>","DOI":"10.1093\/toxsci\/kfag105","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T11:38:28Z","timestamp":1787053108000},"source":"Crossref","is-referenced-by-count":0,"title":["Prolonged zero-nicotine e-cigarette exposure disrupts blood\u2013brain barrier integrity and promotes neuroinflammation: evidence from in-vitro and in-vivo models"],"prefix":"10.1093","volume":"209","author":[{"given":"David","family":"Mara","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center , Amarillo, TX 79106,","place":["United States"]},{"name":"Brain Drug Discovery Center, Texas Tech University Health Sciences Center , Amarillo, TX 79106,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khondker Ayesha","family":"Akter","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center , Amarillo, TX 79106,","place":["United States"]},{"name":"Brain Drug Discovery Center, Texas Tech University Health Sciences Center , Amarillo, TX 79106,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sejal Rajesh","family":"Jadhav","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center , Amarillo, TX 79106,","place":["United States"]},{"name":"Brain Drug Discovery Center, Texas Tech University Health Sciences Center , Amarillo, TX 79106,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas 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