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Endothelial Cell Recovery Following Histone-induced Damage from Trauma and Traumatic Brain Injury
Wentzel, Kara
Wentzel, Kara
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Attribution-NonCommercial-NoDerivatives 4.0 International
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Abstract
The vascular endothelium lines all blood vessels in the body and plays a central role in regulating blood flow and barrier function. Following traumatic brain injury (TBI) or other forms of trauma, circulating histone proteins are released as damage-associated molecular patterns that disrupt endothelial integrity. Resulting endotheliopathy is associated with increased morbidity and mortality, yet there is a knowledge gap in understanding the extent to which endothelial cells (ECs) can recover after acute injury, limiting therapeutic advances.
This study examines the regenerative capacity of endothelial cells in vitro using a live-cell imaging model. Endothelial monolayers were stimulated with histones to induce acute injury, and recovery was evaluated under different recovery mediums. Cell viability and permeability were assessed using fluorescent dyes and time-point microscopy to visualize changes at each interval. Quantitative image analysis was performed to assess the proportion of cells that remained intact, became permeable, or showed signs of recovery. Findings suggest a subset of endothelial cells are capable of restoring membrane integrity following acute histone exposure, indicating the potential for recovery rather than irreversible injury. The extent of recovery was enhanced by treatment with components derived from healthy human donor blood platelets, but not the plasma. This supports a role for platelet components to support endothelial cell repair mechanisms as a novel therapeutic strategy for improving transfusion medicine in critical illness.
Description
This thesis evaluates the regenerative capacity of vascular ECs after damage by histone proteins. Additionally, we use our model system to demonstrate that healthy human donor platelet components, and not plasma, optimizes this recovery. Determining the extent and timing of EC recovery after trauma, and mechanisms that could enhance this restoration, is the critical and necessary next step to developing therapeutic targets to improve patient recovery and survival after TBI.
Specific Aims
Aim 1. To determine the extent and timing of vascular endothelial recovery after an acute injury.
We hypothesize that ECs will have a threshold of acute injury beyond which they are unable to recover, and that the timing and extent to which EC restoration occurs depends on the fraction of cells that have exceeded this threshold. We will test this hypothesis in a model of EC damage by exposure to injurious trauma factors, using live cell video microscopy to record both injury and recovery after application of fresh serum.
Aim 2. To establish mechanisms of EC regeneration after an acute injury.
We hypothesize that a subset of ‘sentinel’ ECs sensitive to trauma factors serve a role in activation of innate immune responses after injury; other ECs in the same tissue bed or field resist these stimuli and instead serve to regenerate the local endothelial layer after an insult. We will identify sensitive and resistant cell subtypes using microscopy imaging methods in cultured endothelial cells stimulated with histone proteins. This will provide the basis for future work using flow cytometry and genetic approaches to characterize and identify subgroups of regenerative ECs.
Aim 3. To improve the recovery of endothelial cells with additional methods of recovery
We hypothesize that EC recovery can be increased with the use of human plasma. We will quantify recovery by comparing fluorescence of cultured endothelial cells recovered in media, plasma, and platelet components using our imaging microscopy model and quantitative analysis in ImageJ.
Date
2026-01-10
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Wentzel_HonorsThesis.pdf
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