Concussive injury elicits human cerebrovascular endothelial cell activation in vitro

Concussive brain injury may directly injure cerebrovascular endothelial cells (CVEC) of the blood‐brain barrier and/or CVEC may be altered by circulating trauma‐induced inflammatory mediators. Thus, we assessed for activation of CVEC by either a concussive injury or by application of plasma from trauma patients. Methods Plasma was obtained from pediatric multisystem trauma patients (injury severity score≥12) and age/sex‐matched controls. Human immortalized CVEC (hCMEC/D3; provided by Dr. P. Couraud, INSERM) were employed to assess for activation in vitro . Concussive injury was induced in hCMEC/D3 grown on flexible supports with a compressed air pulse at 4 psi/well. In parallel, uninjured CVEC were treated with 20% v/v blood plasma collected from trauma patients or healthy controls. Results A single concussive injury to hCMEC/D3 resulted in increased nitric oxide (NO; DAF‐FM nitrosation) at 15 and 30 minutes, increased reactive oxygen species (ROS; DHR‐123 oxidation) at 2 hours and increased 51 Cr‐PMN adhesion at 6 hours. In contrast, application of 20% v/v trauma patient plasma to hCMEC/D3 failed to induce oxidative stress, activate NF‐κB (ELISA) or elicit PMN adhesion assessed under “flow” (relative to control plasma‐treated hCMEC/D3). Conclusions Direct concussive injury induces hCMEC/D3 activation in vitro , whereas application of 20% v/v trauma plasma is not potent enough to activate CVEC.