Investigating Mechanisms of Injury and Intervention in a Novel In Vitro Model of Traumatic Brain Injury in Organotypic Hippocampal Slice Cultures
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Doctor of Philosophy (PhD)
Arts and Sciences
Dr. Mark Prendergast
Traumatic brain injuries (TBIs) impact millions of individuals each year and can pose long term consequences. Despite numerous attempts, no medication has been approved by the FDA to treat TBIs. The causes of these failed trials are multifaceted, but in part can be attributed to the complex nature of TBIs, as well as a lack of sufficient pre-clinical data. In vitro models of TBI are an important tool to help understand the cellular changes seen following the injury, in a highly controlled environment. For the following experiments, a novel model of TBI was used to injure organotypic hippocampal slice cultures, harvested from eight-day old Sprague Dawley rat pups. The focus of experiment 1 was to begin characterizing a new model as a valid model of TBI. A spinal cord injury impactor was used to administer a stretch injury to the tissue. To assess the damage, slices were stained with propidium iodide (PI), NeuN, or DCFH-DA. Generally, injured tissue showed an increase in PI and DCF fluorescence while NeuN fluorescence was unaffected. The purpose of experiment 2 was to determine the effect of ethanol pre-exposure on the injury. Between 25-50% of people are intoxicated at the time of injury, but the effects of alcohol are unknown. In the current experiment, tissue was administered ethanol for two days prior to the injury. Post injury, tissue was stained with PI and NeuN. Generally, the effect of ethanol pre-exposure on these outcome measures was dependent on the severity of the injury. The purpose of experiment 3 was to reduce the effect of the injury. N-acetylcysteine amide (NACA) works by increasing levels of glutathione to reduce ROS related damage. NACA was given immediately post injury and tissue was assessed with PI, NeuN, and DCFH-DA. There was no benefit of NACA, as the timing and concentrations of NACA used were not optimal. The data from these studies suggest the model can be a beneficial tool for studying TBIs. However, further research is necessary to determine the biochemical workings of the injuries.
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This project was supported in part by National Institute on Alcohol Abuse and Alcoholism training grant T32 AA027488 and in part by National Institute of Health grant R44AA025804-03S1 from 2019-2022.
Jagielo-Miller, Julia Elaine, "Investigating Mechanisms of Injury and Intervention in a Novel In Vitro Model of Traumatic Brain Injury in Organotypic Hippocampal Slice Cultures" (2022). Theses and Dissertations--Psychology. 215.