Persistent dysmorphology and dysfunction of the brain microvasculature following repeated trauma

Mild traumatic brain injury typically produces no abnormalities on neuroimaging yet elicits symptoms that, in an increasing fraction of survivors, linger for years, particularly with recurring injuries. To elucidate the underlying biological substrates, we leveraged two-photon fluorescence microscopy resonant scanning and our recently developed deep-learning based pipeline to evaluate the cerebrovascular network in the subacute stage following three intact-skull cortical impacts in adult mice under low isoflurane. Microvascular volume density increased by 19.1 ± 18.6% after moderate impacts, while mean capillary diameters increased by 5.6 ± 3.2% for mild and by 1.1 ± 2.6% for moderate impact series; consistent with network remodeling and tone dysregulation, which limits cerebrovascular reserve. Mild impacts cohort showed a paradoxical hypercapnia response with decreases in red blood cell velocities (vRBC) (-20 ± 13%) in the majority (62%) of cortical penetrating arteries and a net ipsicontusional hypercapnia-induced arteriolar vRBC decrease, in stark contrast to the physiologically normal increase seen in sham mice (+27 ± 15%). Downstream, the mild impacts resulted in attenuation of hypercapnia-induced cortical capillaries' flow increase while the moderate impacts cohort showed no ipsicontusional capillary flow response. Sustained aberrations in brain microvasculature post-trauma may underlie the persistence of symptoms and mediate susceptibility to further injury.