Ventilator-induced lung injury causes platelet-dependent NET formation. Glycol- and busulfan-treated WT mice were ventilated with sham or VILI settings. (A) The paO2/FiO2 ratio, (B) neutrophil recruitment into the alveoli, and (C) protein concentration in the BAL were analyzed after 2 hours (n = 4). (D) The formation of circulating platelet-neutrophil aggregates was measured by flow cytometry 30 minutes after induction of sham or VILI ventilation (n = 4). (E) Ultrathin cross-sectioned vessel/capillary imaged by transmission electron microscopy from lung tissue of WT mice 2 hours after induction of VILI ventilation showing a neutrophil (asterisk) surrounded by platelets. Inserts at higher magnification illustrate the close contact between platelets (arrows) and neutrophil. NET formation in lung sections was visualized by immunofluorescence staining. Exemplary images from (F) sham-ventilated and (G) VILI-ventilated animals. (H) Circulating NET structures were quantified by MPO-DNA-ELISA (n = 4). *P < .05.
