Document Detail


Comparison of cumulative planimetry versus manual dissection to assess experimental infarct size in isolated hearts.
MedLine Citation:
PMID:  19732842     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
INTRODUCTION: Infarct size (IS) is an important variable to estimate cardiac ischemia/reperfusion injury in animal models. Triphenyltetrazolium chloride (TTC) stains viable cells red while leaving infarcted cells unstained. To quantify IS, infarcted and non-infarcted tissue is often manually dissected and weighed (IS-DW). An alternative is to measure infarcted areas by cumulative planimetry (IS-CP).
METHODS: We prospectively compared these two methods in 141 Langendorff-prepared guinea pig hearts (1.44+/-0.02 g) that were part of different studies on mechanisms of cardioprotection. Hearts were perfused with Krebs-Ringer's and subjected to 30 min global ischemia after various cardioprotective treatments. Two hours after reperfusion hearts were cut into 6-7 transverse sections (3mm) and stained for 5 min in 1% TTC and 0.1M KH2PO4 buffer (pH 7.4, 38 degrees C). Each slice was first scanned and its infarcted area measured with Image 1.62 software (NIH). Infarctions in individual slices of each heart were averaged (IS-CP) on the basis of their weight. After scanning, IS-DW was determined by careful manual dissection of infarcted from non-infarcted tissue and measuring their respective total weight.
RESULTS: We found limited tissue permeation of TTC in relation to the slice thickness leaving tissue in the center unstained, as well as significant cross-contamination of stained vs. unstained tissue after manual dissection. IS-CP and IS-DW ranged from 6.0 to 73.1% and 19.4 to 70.5%, respectively, and correlated as follows: IS-DW=(27.6+/-1.4)+(0.518+/-0.038) * IS-CP; r=0.75 (Pearson), p<0.001. In addition, IS-CP correlated better with return of function after reperfusion like developed left ventricular pressure, contractility and relaxation, and myocardial oxygen consumption.
DISCUSSION: Despite a good correlation between both methods, limited tissue permeation by TTC diffusion and limited precision in the ability to manually dissect stained from unstained tissue leads to an overestimation of infarct size by dissection and weighing compared to cumulative planimetry.
Authors:
Matthias L Riess; Samhita S Rhodes; David F Stowe; Mohammed Aldakkak; Amadou K S Camara
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2009-09-03
Journal Detail:
Title:  Journal of pharmacological and toxicological methods     Volume:  60     ISSN:  1873-488X     ISO Abbreviation:  J Pharmacol Toxicol Methods     Publication Date:    2009 Nov-Dec
Date Detail:
Created Date:  2009-11-25     Completed Date:  2010-07-30     Revised Date:  2013-12-25    
Medline Journal Info:
Nlm Unique ID:  9206091     Medline TA:  J Pharmacol Toxicol Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  275-80     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Disease Models, Animal*
Dissection / methods*,  standards
Female
Guinea Pigs
Heart
Image Processing, Computer-Assisted / methods*,  standards
Male
Myocardial Infarction / pathology*,  physiopathology,  radiography
Myocardial Ischemia / pathology,  physiopathology,  radiography
Organ Size / physiology
Prospective Studies
Surface Properties
Grant Support
ID/Acronym/Agency:
HL073246-01/HL/NHLBI NIH HHS; HL58691/HL/NHLBI NIH HHS; K01 HL073246-05/HL/NHLBI NIH HHS; R01 HL089514-04/HL/NHLBI NIH HHS
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