Administration of FVIIa-released EEVs to mice protects against LPS-induced inflammation and barrier disruption. (A-F) EVs were isolated from the supernatant medium of bEND.3 cells that were transfected with scr miR or anti-miR10a, and then treated with a control vehicle or FVIIa. An equal number of EVs (2 × 10⁸) were administered to C57WT/6J mice via the tail-vein. Four hours later, mice were given an intraperitoneal injection of LPS (5 mg/kg). Twelve hours after administration of LPS, blood was obtained from the mice, and the levels of TNF-α (A), IL-6 (B), and IL-1β (C) in the plasma were measured. In a subset of the same group of mice, vascular leakage into the lung (D) and heart (E) was evaluated. In another subset, mice were euthanized 6 hours after administration of LPS, and the lungs were collected. Lung tissue sections were stained for neutrophil infiltration and imaged at 40× magnification (F), and the number of neutrophils was counted (G). (H-J) EEVs (2 × 10⁸), isolated as described for panel A, were administered into the peritoneum of C57WT/6J mice. Four hours later, LPS (5 mg/kg) was administered to the mice IP. Two hours after administration, the mice were euthanized, and peritoneal macrophages were isolated. mRNA expression levels of TNF-α (H), IL-6 (I), and IL-1β (J) were determined by qRT-PCR. *P < .05; **P < .01; ***P < .001; ****P < .0001; ns, not significant.