Impact of severe traumatic brain injury (TBI) on cerebral blood flow and the endothelial cell phenotype

TBI causes the disruption of blood vessels in brain leading to hemorrhage, edema, and changes in cerebral blood flow. How such altered vascular and flow condition affect endothelial cells, the cellular unit of blood vessels, is not known. We used single cell RNA sequencing (scRNAseq) to analyze the heterogenity in endothelial cell clusters in brain, and how key cellular and signaling signatures of these clusters are temporally changed post-TBI at the site of injury. We used closed skull cortical impact (CCI) mouse model. The brain cortex specimens were isolated from the site of injury at day 1, 7 and 28 post-TBI. Site-matched specimens were also harvested from uninjured control mice. Single cells were harvested from the collected tissue specimens. Myelin debris was removed from the cells and the specimens were immediately fixed in 4% paraformaldehyde. Fixed samples were subjected to scRNAseq as per manufacturer's protocol.