Use of TTC staining for the evaluation of tissue injury in the early phases of reperfusion after focal cerebral ischemia in rats

Angéla Benedek, Krisztina Móricz, Zsolt Jurányi, Gábor Gigler, György Lévay, L. Hársing, P. Mátyus, G. Szénási, Mihály Albert

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Background and purpose: 2,3,5-Triphenyltetrazolium chloride (TTC) staining measures tissue viability used to evaluate infarct size. The goal of this study was to compare viability of neuronal tissue during the early phases of ischemia-reperfusion assessed either by perfusion of the brain with TTC solution transcardially, in vivo, or by staining brain slices, in vitro. Methods: The middle cerebral artery was occluded for 1 h in male SPRD rats and then reperfused for 0, 1, 4, 8, 16 and 24 h. Ischemic damage was evaluated by TTC staining, in vivo and in vitro, and by histology (Luxol Fast Blue and Fluoro-Jade staining, electron microscopy). Results: Core volume of tissue injury measured in vivo was large at 0 h and steadily decreased by 50% (p <0.001) up to 16 h, but substantially increased from 16 to 24 h of reperfusion. In contrast, a significant core volume appeared at 4 h only, in vitro, and gradually increased up to 24 h. Core volume was larger in vivo than in vitro at all times except at 16 h when the opposite was observed. Evans blue administered intracardially stained TTC-negative areas at 1 and 24 h. Histology covered the evolution of serious tissue injury but also demonstrated some morphologically preserved neurons in the infracted area at 24 h. Conclusions: Formation of formazan from TTC can depend on both the staining method and the metabolic burden of the brain tissue causing uncertainties in the volume of ischemia-induced brain injury measured by TTC staining.

Original languageEnglish
Pages (from-to)159-165
Number of pages7
JournalBrain Research
Volume1116
Issue number1
DOIs
Publication statusPublished - Oct 20 2006

Fingerprint

Brain Ischemia
Reperfusion
Staining and Labeling
Wounds and Injuries
Tissue Survival
Histology
Brain
Ischemia
Formazans
Evans Blue
Middle Cerebral Artery
Brain Injuries
Uncertainty
triphenyltetrazolium
Electron Microscopy
Perfusion
Neurons
In Vitro Techniques

Keywords

  • Brain infarction
  • Cerebral ischemia
  • Fluoro-Jade
  • Rat

ASJC Scopus subject areas

  • Neuroscience(all)
  • Clinical Neurology
  • Developmental Biology
  • Molecular Biology

Cite this

Use of TTC staining for the evaluation of tissue injury in the early phases of reperfusion after focal cerebral ischemia in rats. / Benedek, Angéla; Móricz, Krisztina; Jurányi, Zsolt; Gigler, Gábor; Lévay, György; Hársing, L.; Mátyus, P.; Szénási, G.; Albert, Mihály.

In: Brain Research, Vol. 1116, No. 1, 20.10.2006, p. 159-165.

Research output: Contribution to journalArticle

Benedek, Angéla ; Móricz, Krisztina ; Jurányi, Zsolt ; Gigler, Gábor ; Lévay, György ; Hársing, L. ; Mátyus, P. ; Szénási, G. ; Albert, Mihály. / Use of TTC staining for the evaluation of tissue injury in the early phases of reperfusion after focal cerebral ischemia in rats. In: Brain Research. 2006 ; Vol. 1116, No. 1. pp. 159-165.
@article{d5f66ff3b121471b96dee5c64e30ea30,
title = "Use of TTC staining for the evaluation of tissue injury in the early phases of reperfusion after focal cerebral ischemia in rats",
abstract = "Background and purpose: 2,3,5-Triphenyltetrazolium chloride (TTC) staining measures tissue viability used to evaluate infarct size. The goal of this study was to compare viability of neuronal tissue during the early phases of ischemia-reperfusion assessed either by perfusion of the brain with TTC solution transcardially, in vivo, or by staining brain slices, in vitro. Methods: The middle cerebral artery was occluded for 1 h in male SPRD rats and then reperfused for 0, 1, 4, 8, 16 and 24 h. Ischemic damage was evaluated by TTC staining, in vivo and in vitro, and by histology (Luxol Fast Blue and Fluoro-Jade staining, electron microscopy). Results: Core volume of tissue injury measured in vivo was large at 0 h and steadily decreased by 50{\%} (p <0.001) up to 16 h, but substantially increased from 16 to 24 h of reperfusion. In contrast, a significant core volume appeared at 4 h only, in vitro, and gradually increased up to 24 h. Core volume was larger in vivo than in vitro at all times except at 16 h when the opposite was observed. Evans blue administered intracardially stained TTC-negative areas at 1 and 24 h. Histology covered the evolution of serious tissue injury but also demonstrated some morphologically preserved neurons in the infracted area at 24 h. Conclusions: Formation of formazan from TTC can depend on both the staining method and the metabolic burden of the brain tissue causing uncertainties in the volume of ischemia-induced brain injury measured by TTC staining.",
keywords = "Brain infarction, Cerebral ischemia, Fluoro-Jade, Rat",
author = "Ang{\'e}la Benedek and Krisztina M{\'o}ricz and Zsolt Jur{\'a}nyi and G{\'a}bor Gigler and Gy{\"o}rgy L{\'e}vay and L. H{\'a}rsing and P. M{\'a}tyus and G. Sz{\'e}n{\'a}si and Mih{\'a}ly Albert",
year = "2006",
month = "10",
day = "20",
doi = "10.1016/j.brainres.2006.07.123",
language = "English",
volume = "1116",
pages = "159--165",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Use of TTC staining for the evaluation of tissue injury in the early phases of reperfusion after focal cerebral ischemia in rats

AU - Benedek, Angéla

AU - Móricz, Krisztina

AU - Jurányi, Zsolt

AU - Gigler, Gábor

AU - Lévay, György

AU - Hársing, L.

AU - Mátyus, P.

AU - Szénási, G.

AU - Albert, Mihály

PY - 2006/10/20

Y1 - 2006/10/20

N2 - Background and purpose: 2,3,5-Triphenyltetrazolium chloride (TTC) staining measures tissue viability used to evaluate infarct size. The goal of this study was to compare viability of neuronal tissue during the early phases of ischemia-reperfusion assessed either by perfusion of the brain with TTC solution transcardially, in vivo, or by staining brain slices, in vitro. Methods: The middle cerebral artery was occluded for 1 h in male SPRD rats and then reperfused for 0, 1, 4, 8, 16 and 24 h. Ischemic damage was evaluated by TTC staining, in vivo and in vitro, and by histology (Luxol Fast Blue and Fluoro-Jade staining, electron microscopy). Results: Core volume of tissue injury measured in vivo was large at 0 h and steadily decreased by 50% (p <0.001) up to 16 h, but substantially increased from 16 to 24 h of reperfusion. In contrast, a significant core volume appeared at 4 h only, in vitro, and gradually increased up to 24 h. Core volume was larger in vivo than in vitro at all times except at 16 h when the opposite was observed. Evans blue administered intracardially stained TTC-negative areas at 1 and 24 h. Histology covered the evolution of serious tissue injury but also demonstrated some morphologically preserved neurons in the infracted area at 24 h. Conclusions: Formation of formazan from TTC can depend on both the staining method and the metabolic burden of the brain tissue causing uncertainties in the volume of ischemia-induced brain injury measured by TTC staining.

AB - Background and purpose: 2,3,5-Triphenyltetrazolium chloride (TTC) staining measures tissue viability used to evaluate infarct size. The goal of this study was to compare viability of neuronal tissue during the early phases of ischemia-reperfusion assessed either by perfusion of the brain with TTC solution transcardially, in vivo, or by staining brain slices, in vitro. Methods: The middle cerebral artery was occluded for 1 h in male SPRD rats and then reperfused for 0, 1, 4, 8, 16 and 24 h. Ischemic damage was evaluated by TTC staining, in vivo and in vitro, and by histology (Luxol Fast Blue and Fluoro-Jade staining, electron microscopy). Results: Core volume of tissue injury measured in vivo was large at 0 h and steadily decreased by 50% (p <0.001) up to 16 h, but substantially increased from 16 to 24 h of reperfusion. In contrast, a significant core volume appeared at 4 h only, in vitro, and gradually increased up to 24 h. Core volume was larger in vivo than in vitro at all times except at 16 h when the opposite was observed. Evans blue administered intracardially stained TTC-negative areas at 1 and 24 h. Histology covered the evolution of serious tissue injury but also demonstrated some morphologically preserved neurons in the infracted area at 24 h. Conclusions: Formation of formazan from TTC can depend on both the staining method and the metabolic burden of the brain tissue causing uncertainties in the volume of ischemia-induced brain injury measured by TTC staining.

KW - Brain infarction

KW - Cerebral ischemia

KW - Fluoro-Jade

KW - Rat

UR - http://www.scopus.com/inward/record.url?scp=33749239959&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749239959&partnerID=8YFLogxK

U2 - 10.1016/j.brainres.2006.07.123

DO - 10.1016/j.brainres.2006.07.123

M3 - Article

VL - 1116

SP - 159

EP - 165

JO - Brain Research

JF - Brain Research

SN - 0006-8993

IS - 1

ER -