Establishment of novel technical methods for evaluating brain edema and lesion volume in stroked rats: A standardization of measurement procedures

Boyko, Matthew; Zvenigorodsky, Vladislav; Grinshpun, Julia; Shiyntum, Honore N.; Melamed, Israel; Kutz, Ruslan; Shelef, Ilan; Brotfain, Evgeni; Frank, Dmitry; Zlotnik, Alexander

doi:10.1016/j.brainres.2019.04.022

Cited by 10 publications

(22 citation statements)

References 39 publications

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“…Additionally, reproducibility may also be an issue. It seems that the novel approach using magnetic resonance imaging for stroke evaluation may enhance reproducibility [120].…”

Section: Potential Dm-induced Mechanisms Leading To Diabetic Strokementioning

confidence: 99%

Potential Biochemical Mechanisms of Brain Injury in Diabetes Mellitus

Xing¹,

Tang²,

Lei³

et al. 2020

Aging and disease

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The goal of this review was to summarize current biochemical mechanisms of and risk factors for diabetic brain injury. We mainly summarized mechanisms published in the past three years and focused on diabetes induced cognitive impairment, diabetes-linked Alzheimer's disease, and diabetic stroke. We think there is a need to conduct further studies with increased sample sizes and prolonged period of follow-ups to clarify the effect of DM on brain dysfunction. Additionally, we also think that enhancing experimental reproducibility using animal models in conjunction with application of advanced devices should be considered when new experiments are designed. It is expected that further investigation of the underlying mechanisms of diabetic cognitive impairment will provide novel insights into therapeutic approaches for ameliorating diabetes-associated injury in the brain.

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“…Additionally, reproducibility may also be an issue. It seems that the novel approach using magnetic resonance imaging for stroke evaluation may enhance reproducibility [120].…”

Section: Potential Dm-induced Mechanisms Leading To Diabetic Strokementioning

confidence: 99%

Potential Biochemical Mechanisms of Brain Injury in Diabetes Mellitus

Xing¹,

Tang²,

Lei³

et al. 2020

Aging and disease

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“…In order to compare the original histologic to MRI imaging techniques, we analyzed data obtained from our previous work (Boyko et al, 2019b; Frank et al, 2019). The experimental procedure was carried out on a 3T MRI clinical scanner (Ingenia, Philips Medical Systems, Best, Netherlands), fitted out with a gradient system of 45 mT/m at 225 μs ramp time, using a commercial eight-channel receive-only wrist coil.…”

Section: Methodsmentioning

confidence: 99%

“…An expert blinded to the experimental procedure carried out image analysis using a Philips software package and an ImageJ software, performed calculations, and analyzed results. The Philips software package (Ingenia, Philips Medical Systems, Best, Netherlands) was used to individual ROI’s for assessment BBB breakdown and to produce a map for the brain images generate quantitative ADC maps, in mm 2 /s, which were subsequently analyzed using the Image J software 1.50i version 1 , described previously (Boyko et al, 2019b; Frank et al, 2019). The viability thresholds used for the identification of every pixel showing irregular ADC features on the slide were 0.53 × 10 –3 mm 2 /s for ADC (Bardutzky et al, 2005).…”

Section: Methodsmentioning

confidence: 99%

“…When determining the extent of neurological injury in the MCAO model, measured outcomes include cerebral edema (O’Brien et al, 1974; Chen et al, 2006; Durukan and Tatlisumak, 2007), infarct zone (Wang-Fischer, 2008; Liu et al, 2009) and blood-brain barrier (BBB) permeability (Belayev et al, 1996; Sifat et al, 2017; Jiang et al, 2018). Following MCAO, the most popular techniques for measuring brain edema are drying (Chen et al, 2006) or calculating hemispheric volumes (Boyko et al, 2010, 2019b). The infarct zone is mostly determined by a 2,3,5-triphenyltetrazolium chloride (TTC) staining method (Liu et al, 2009) that differentiates between infarcted and viable tissue (Joshi et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

“…For example, a combination of TTC staining and immunofluorescent staining methods (Li et al, 2018) as well as other molecular and biochemical analyses after TTC staining (Kramer et al, 2010) have been described. Calculating brain hemisphere volumes to assess brain edema has been performed in our laboratory together with TTC staining to calculate infarct zone using the same brain set (Boyko et al, 2019b). However, measuring BBB permeability in the same set of brain samples remains a challenge and has yet to be described in the literature.…”

Section: Introductionmentioning

confidence: 99%

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A Novel Method for Assessing Cerebral Edema, Infarcted Zone and Blood-Brain Barrier Breakdown in a Single Post-stroke Rodent Brain

et al. 2019

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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.

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