title: In vivo analysis of microvascular injury after myocardial cryothermia.
abstract: We studied microvascular injury after myocardial cryothermia in rats using intravital fluorescence microscopic techniques. Cryolesions were induced to the right ventricle by freezing with -160 degrees C (probe diameter: 5 mm) for a total of 5 min. Fluorescence microscopy was performed at 15, 30, 60, 90, and 120 min as well as at 3 and 7 days after cryothermia. Analysis of the epicardial microvasculature 15 min after cryothermia revealed an area of 24.6 +/- 3.8 mm2 of nonperfused tissue, which was reduced to 5.3 +/- 1.5 mm2 (P < 0.05) after the initial 2-h observation period. Vital microscopic images of reperfused tissue characteristically demonstrated extravasation of the macromolecular fluorescent tracer FITC-dextran (21.7 +/- 3.4 mm2), suggesting substantial loss of endothelial integrity. In vivo propidium iodide staining confirmed membrane damage of microvascular endothelial cells. Three days after cryoinjury the area of nonperfused tissue was reduced further to 1.1 +/- 0.4 mm2 in the center of the lesion, while the area of perfused tissue with disruption of endothelial integrity was found significantly increased to 47.4 +/- 5.9 mm2 (P < 0.05) toward the periphery. Analysis at 7 days revealed endothelial repair at the periphery of the cryolesion, but now a central necrotic area was found demarcated (nonperfused), presenting with a size (26.0 +/- 3.5 mm2) similar to that shown during the very early (15 min) reperfusion period. Our study demonstrates recovery of microvascular perfusion during the first hours and days after myocardial cryothermia. This is, however, associated with endothelial injury, i.e., damage of plasma membrane and loss of barrier function. Infarction with capillary perfusion failure is evident at 7 days with a size which strikingly corresponds to the sizeof nonperfused tissue observed immediately after cryointervention.
authors.affiliation: Department of Thoracic and Cardiovascular Surgery, University of Saarland, Homburg/Saar, D-66421, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Huwer
authors.forename: N
authors.initials: N
authors.lastname: Nikoloudakis
authors.forename: J
authors.initials: J
authors.lastname: Rissland
authors.forename: B
authors.initials: B
authors.lastname: Vollmar
authors.forename: M D
authors.initials: MD
authors.lastname: Menger
authors.forename: H J
authors.initials: HJ
authors.lastname: Schäfers
mesh_terms: D000818:Animals; D003080:Cold Temperature / Q000009:adverse effects*; D003331:Coronary Vessels / Q000473:pathology*; D004195:Disease Models, Animal; D008833:Microcirculation; D008856:Microscopy, Fluorescence; D009203:Myocardial Infarction / Q000209:etiology*; D051381:Rats; D017207:Rats, Sprague-Dawley