• Carey Balaban, Ronald L Jackson, Jianzhong Liu, Wei Gao, and Michael E Hoffer.
• University of Pittsburgh, Department of Otolaryngology, United States. Electronic address: cbalaban@pitt.edu.
• J. Neurosci. Methods. 2016 Oct 15; 272: 56-68.

BackgroundMany previous experimental studies of blast wave effects have reported vascular and parenchymal injury in brains extracted from the skulls prior to histopathological assessment. Brain removal disrupts vasculature and structural features of the meninges that may be sources of signs and symptoms of mild traumatic brain injury, particular at lower blast overpressures (<5psi peak).New MethodImmunohistochemical and histopathological studies have been conducted in sections from decalcified, paraffin embedded, histologically sectioned whole rat heads. These sections preserve the entire cranial contents in situ, and permit evaluation of the inner ear, central nervous system and associated vasculature. The findings could also be correlated with mRNA expression patterns from whole brains subjected to similar treatment.ResultsLower levels of blast wave exposure produce primarily vascular effects in rats. Messenger RNA profiles of the whole brains showed evidence of both blast intensity and time dependent effects on vascular wound healing markers. The rats exposed to 10-11psi overpressure tended to show a similar pattern of mRNA expression changes in these vascular repair and inflammatory pathways as rats exposed to approximately 5psi overpressure, but the changes were greater. The changes in mRNA expression after a 14-15psi exposure were different and suggestive of more severe injury, particularly for DNA repair, lymphocyte activation and lymphocyte migration pathways. Histopathological examination of decalcified heads revealed that even 2.5-7.9psi blast exposures produced a high prevalence of mild venous hemorrhage and thrombosis (accompanied by inflammatory markers) in the inner ear, vertebrobasilar circulation, hippocampal choroidal fissure and the veins associated with velum interpositum.Comparison With Existing Method(S)The sites of vascular injury would not have been included in specimens extracted from the skull prior to processing.ConclusionsThe isolated regions of intravascular coagulation in small veins and the isolated, very small venous hemorrhages in the subarachnoid space are worthy of consideration as factors in both healing and chronic sequelae of mild blast concussion. Although small, remnants persisted in the subarachnoid space even 42 days after a single blast exposure. The high prevalence of very mild subdural and subarachnoid hemorrhage may be a target for clinical management.Copyright © 2016 Elsevier B.V. All rights reserved.

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