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Measuring Post-Stroke Cerebral Edema, Infarct Zone and Blood-Brain Barrier Breakdown in a Single Set of Rodent Brain Samples
Published on: October 23, 2020
A Novel Method for Assessing Cerebral Edema, Infarcted Zone and Blood-Brain Barrier Breakdown in a Single Post-stroke
Ruslan Kuts1, Dmitry Frank1, Benjamin F Gruenbaum2
1Division of Anesthesiology and Critical Care, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, Beersheba, Israel.
Abstract:
Stroke is a major cause of global morbidity and mortality. Middle cerebral artery occlusion (MCAO) has historically been the most common animal model of simulating ischemic stroke. The extent of neurological injury after MCAO is typically measured by cerebral edema, infarct zone, and blood-brain barrier (BBB) permeability. A significant limitation of these methods is that separate sets of brains must be used for each measurement. Here we examine an alternative method of measuring cerebral edema, infarct zone and BBB permeability following MCAO in the same set of brain samples. Ninety-six rats were randomly divided into three experimental groups. Group 1 (n = 27) was used for the evaluation of infarct zone and brain edema in rats post-MCAO (n = 17) vs. sham-operated controls (n = 10). Group 2 (n = 27) was used for the evaluation of BBB breakdown in rats post-MCAO (n = 15) vs. sham-operated controls (n = 10). In Group 3 (n = 42), all three parameters were measured in the same set of brain slices in rats post-MCAO (n = 26) vs. sham-operated controls (n = 16). The effect of Evans blue on the accuracy of measuring infarct zone by 2,3,5-triphenyltetrazolium chloride (TTC) staining was determined by measuring infarct zone with and without an applied blue filter. The effects of various concentrations of TTC (0, 0.05, 0.35, 0.5, 1, and 2%) on the accuracy of measuring BBB permeability was also assessed. There was an increase in infarct volume (p < 0.01), brain edema (p < 0.01) and BBB breakdown (p < 0.01) in rats following MCAO compared to sham-operated controls, whether measured separately or together in the same set of brain samples. Evans blue had an effect on measuring infarct volume that was minimized by the application of a blue filter on scanned brain slices. There was no difference in the Evans blue extravasation index for the brain tissue samples without TTC compared to brain tissue samples incubated in TTC. Our results demonstrate that measuring cerebral edema, infarct zone and BBB permeability following MCAO can accurately be measured in the same set of brain samples.
Insights
This study demonstrates that measuring cerebral edema, infarct zone, and blood-brain barrier permeability after middle cerebral artery occlusion (MCAO) in the same rat brain samples is accurate. This method streamlines stroke research by reducing the need for separate brain analyses.
Area of Science:
- Neuroscience
- Stroke Research
- Animal Models
Background:
- Stroke is a leading cause of death and disability worldwide.
- Middle cerebral artery occlusion (MCAO) is a common animal model for ischemic stroke.
- Current methods require separate brain samples for measuring edema, infarct zone, and blood-brain barrier (BBB) permeability, limiting efficiency.
Purpose of the Study:
- To evaluate an alternative method for measuring cerebral edema, infarct zone, and BBB permeability using the same set of brain samples after MCAO.
- To assess the accuracy of this combined measurement technique.
Main Methods:
- Ninety-six rats were divided into experimental and sham-operated control groups.
- Measurements of infarct volume, brain edema, and BBB permeability were performed on separate sets of brains and then on the same set of brain slices.
- The influence of Evans blue and 2,3,5-triphenyltetrazolium chloride (TTC) concentrations on measurement accuracy was investigated.
Main Results:
- MCAO significantly increased infarct volume, brain edema, and BBB breakdown compared to controls (p < 0.01).
- These increases were consistent whether parameters were measured separately or together in the same brain samples.
- Evans blue interference with infarct volume measurement was minimized by using a blue filter.
Conclusions:
- Measuring cerebral edema, infarct zone, and BBB permeability in the same brain samples following MCAO is accurate and feasible.
- This integrated approach can enhance the efficiency of stroke research.
- The findings support a streamlined methodology for evaluating neurological injury in MCAO models.
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