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Published in: Cardiovascular Drugs and Therapy 4/2009

01-08-2009 | SHORT COMMUNICATION

One Hour Reperfusion is Enough to Assess Function and Infarct Size With TTC Staining in Langendorff Rat Model

Authors: R. Ferrera, S. Benhabbouche, J. C. Bopassa, B. Li, M. Ovize

Published in: Cardiovascular Drugs and Therapy | Issue 4/2009

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Abstract

Background

There is not general agreement concerning the optimal time of reperfusion necessary to assess myocardial function and necrosis on isolated perfused heart model. Nevertheless, the study of cardioprotection (especially, pre– and postconditioning) requires a reliable and standardized assessment of myocardial necrosis.

Objective

The objective of this study was thus to evaluate whether 1 h of reperfusion was sufficient to assess rat heart viability on Langendorff preparation. Isolated rat hearts (n = 30) underwent 40 min of global normothermic ischemia followed by 60 or 120 min Langendorff reperfusion. In each group, hearts were also randomly assigned into the 2 following sub-groups: postconditioning (PostC, consisting in 2 episodes of 30 s ischemia and 30 s reperfusion at the onset of reperfusion), and control (no intervention). Coronary flow, heart rate, dP/dt and rate-pressure-product were measured. Myocardial necrosis was assessed by TTC staining and LDH, CK release analysis.

Results

Our results indicated that heart function tended to slightly decrease between 60 min and 120 min reperfusion. Infarct size was identical at 60 min and 120 min reperfusion, averaging 33–34% of total LV area in controls versus 17% in PostC (p < 0.001 between control and PostC groups). Similarly, the maximum of enzymatic releases (CK and LDH) measured in coronary effluents was at 60 min of reperfusion, followed by a progressive decrease at 90 min and 120 min. As expected, postconditioning limited enzymatic releases whatever the studied time of reperfusion.

Conclusion

In conclusion, we showed that prolonged reperfusion beyond 60 min was not useful for function assessment and did not change infarct size measurement, on Langendorff rat model of ischemia-reperfusion.
Literature
1.
go back to reference Munujos P, Coll-Canti J, González-Sastre F, Gella FJ. Assay of succinate dehydrogenase activity by a colorimetric-continuous method using iodonitrotetrazolium chloride as electron acceptor. Anal Biochem. 1993;212:506–9.PubMedCrossRef Munujos P, Coll-Canti J, González-Sastre F, Gella FJ. Assay of succinate dehydrogenase activity by a colorimetric-continuous method using iodonitrotetrazolium chloride as electron acceptor. Anal Biochem. 1993;212:506–9.PubMedCrossRef
2.
go back to reference Seidler E. The tetrazolium-formazan system: design and histochemistry. Prog Histochem Cytochem. 1991;24:1–86.PubMed Seidler E. The tetrazolium-formazan system: design and histochemistry. Prog Histochem Cytochem. 1991;24:1–86.PubMed
3.
go back to reference Ferrera R, Larese A, Berthod F, Guidollet J, Rodriguez C, Dureau G, et al. Quantitative reduction of MTT by heart biopsies in vitro is an index of viability. J Mol Cell Cardiol. 1993;25:1091–9.PubMedCrossRef Ferrera R, Larese A, Berthod F, Guidollet J, Rodriguez C, Dureau G, et al. Quantitative reduction of MTT by heart biopsies in vitro is an index of viability. J Mol Cell Cardiol. 1993;25:1091–9.PubMedCrossRef
4.
go back to reference Ferrera R, Bopassa JC, Angoulvant D, Ovize M. Post-conditioning protects from cardioplegia and cold ischemia via inhibition of mitochondrial permeability transition pore. J Heart Lung Transplant. 2007;26:604–9.PubMedCrossRef Ferrera R, Bopassa JC, Angoulvant D, Ovize M. Post-conditioning protects from cardioplegia and cold ischemia via inhibition of mitochondrial permeability transition pore. J Heart Lung Transplant. 2007;26:604–9.PubMedCrossRef
5.
go back to reference Ito WD, Schaarschmidt S, Klask R, Hansen S, Schafer HJ, Mathey D, et al. Infarct size measurement by triphenyltetrazolium chloride staining versus in vivo injection of propidium iodide. J Mol Cell Cardiol. 1997;29:2169–75.PubMedCrossRef Ito WD, Schaarschmidt S, Klask R, Hansen S, Schafer HJ, Mathey D, et al. Infarct size measurement by triphenyltetrazolium chloride staining versus in vivo injection of propidium iodide. J Mol Cell Cardiol. 1997;29:2169–75.PubMedCrossRef
6.
go back to reference Birnbaum Y, Hale SL, Kloner RA. Differences in reperfusion length following 30 min of ischemia in the rabbit influence infarct size, as measured by triphenyltetrazolium chloride staining. J Mol Cell Cardiol. 1997;29:657–66.PubMedCrossRef Birnbaum Y, Hale SL, Kloner RA. Differences in reperfusion length following 30 min of ischemia in the rabbit influence infarct size, as measured by triphenyltetrazolium chloride staining. J Mol Cell Cardiol. 1997;29:657–66.PubMedCrossRef
7.
go back to reference Schwarz ER, Somoano Y, Hale SL, Kloner RA. What is the Required Reperfusion Period for Assessment of Myocardial Infarct Size Using Triphenyltetrazolium Chloride Staining in the Rat? J Thromb Thrombolysis. 2000;10:181–7.PubMedCrossRef Schwarz ER, Somoano Y, Hale SL, Kloner RA. What is the Required Reperfusion Period for Assessment of Myocardial Infarct Size Using Triphenyltetrazolium Chloride Staining in the Rat? J Thromb Thrombolysis. 2000;10:181–7.PubMedCrossRef
8.
go back to reference Redel A, Jazbutyte V, Smul TM, Lange M, Eckle T, Eltzschig H, et al. Impact of Ischemia and Reperfusion Times on Myocardial Infarct Size in Mice In Vivo. Exp Biol Med. 2008;233:84–93.CrossRef Redel A, Jazbutyte V, Smul TM, Lange M, Eckle T, Eltzschig H, et al. Impact of Ischemia and Reperfusion Times on Myocardial Infarct Size in Mice In Vivo. Exp Biol Med. 2008;233:84–93.CrossRef
9.
go back to reference Freeman I, Grunwald MA, Robin B, Rao FS, Bodenheimer MM. Effect of early reperfusion on use of triphenyltetrazolium chloride to differentiate viable from non-viable myocardium in area of risk. Cardiovasc Res. 1990;24:109–14.PubMedCrossRef Freeman I, Grunwald MA, Robin B, Rao FS, Bodenheimer MM. Effect of early reperfusion on use of triphenyltetrazolium chloride to differentiate viable from non-viable myocardium in area of risk. Cardiovasc Res. 1990;24:109–14.PubMedCrossRef
10.
go back to reference Pitts KR, Stiko A, Buetow B, Lott F, Guo P, Virca D, et al. Washout of heme-containing proteins dramatically improves tetrazolium-based infarct staining. J Pharmacol Toxicol Methods. 2007;55:201–8.PubMedCrossRef Pitts KR, Stiko A, Buetow B, Lott F, Guo P, Virca D, et al. Washout of heme-containing proteins dramatically improves tetrazolium-based infarct staining. J Pharmacol Toxicol Methods. 2007;55:201–8.PubMedCrossRef
11.
go back to reference Benedek A, Moricz K, Juranyi Z, Gigler G, Levay G, Harsing LG, et al. Use of TTC staining for the evaluation of tissue injury in the early phases of reperfusion after focal cerebral ischemia in rats. Brain Res. 2006;1116:159–65.PubMedCrossRef Benedek A, Moricz K, Juranyi Z, Gigler G, Levay G, Harsing LG, et al. Use of TTC staining for the evaluation of tissue injury in the early phases of reperfusion after focal cerebral ischemia in rats. Brain Res. 2006;1116:159–65.PubMedCrossRef
12.
go back to reference Bopassa JC, Michel P, Gateau-Roesch O, Ovize M, Ferrera R. Low-pressure reperfusion alters mitochondrial permeability transition. Am J Physiol Circ Physiol. 2005;288:H2750–5.CrossRef Bopassa JC, Michel P, Gateau-Roesch O, Ovize M, Ferrera R. Low-pressure reperfusion alters mitochondrial permeability transition. Am J Physiol Circ Physiol. 2005;288:H2750–5.CrossRef
Metadata
Title
One Hour Reperfusion is Enough to Assess Function and Infarct Size With TTC Staining in Langendorff Rat Model
Authors
R. Ferrera
S. Benhabbouche
J. C. Bopassa
B. Li
M. Ovize
Publication date
01-08-2009
Publisher
Springer US
Published in
Cardiovascular Drugs and Therapy / Issue 4/2009
Print ISSN: 0920-3206
Electronic ISSN: 1573-7241
DOI
https://doi.org/10.1007/s10557-009-6176-5

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