Circ Cardiovasc Imaging

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Circ Cardiovasc Imaging Noninvasive Assessment of Hypoxia in Rabbit Advanced Atherosclerosis Using 18F-fluoromisonidazole Positron Emission Tomographic ImagingCLINICAL PERSPECTIVE by Jesus Mateo, David Izquierdo-Garcia, Juan J. Badimon, Zahi A. Fayad, and Valentin Fuster Circ Cardiovasc Imaging Volume 7(2):312-320 March 18, 2014 Copyright © American Heart Association, Inc. All rights reserved.

In vivo magnetic resonance imaging monitoring of plaque progression. In vivo magnetic resonance imaging monitoring of plaque progression. Representative axial T2-weighted magnetic resonance images of the abdominal aorta of a control (A), and an atherosclerotic rabbit scanned at 7 months (B) and at 16 months (C) after diet initiation. Images in B and C correspond to the same animal and at the same anatomic location. Red arrows indicate the abdominal aorta (magnified view provided at the bottom left of each image). Jesus Mateo et al. Circ Cardiovasc Imaging. 2014;7:312-320 Copyright © American Heart Association, Inc. All rights reserved.

In vivo 18F-fluoromisonidazole (18F-FMISO) positron emission tomography (PET) of hypoxia. In vivo 18F-fluoromisonidazole (18F-FMISO) positron emission tomography (PET) of hypoxia. A, Combined PET/computed tomography coronal views (top) of the abdominal aorta illustrating the 18F-FMISO uptake of 2 atherosclerotic rabbits after 8 months (middle) and 16 months (right) on atherogenic diet, compared with a control animal (left). The bottom shows axial views of the corresponding slices indicated with a green line in the coronal view. Green arrows denote the abdominal aorta (magnified view provided at the bottom of each image). L indicates left; and R, right. B, Dot plot comparing the 18F-FMISO uptake (standardized uptake values [SUV] mean) of the abdominal aorta of atherosclerotic rabbits versus control animals. Lines represent the mean. Jesus Mateo et al. Circ Cardiovasc Imaging. 2014;7:312-320 Copyright © American Heart Association, Inc. All rights reserved.

Ex vivo 18F-fluoromisonidazole (18F-FMISO) positron emission tomography (PET) of hypoxia. Ex vivo 18F-fluoromisonidazole (18F-FMISO) positron emission tomography (PET) of hypoxia. Representative ex vivo PET images (from PET/magnetic resonance imaging scans) demonstrating the uptake of 18F-FMISO along the abdominal aorta of an atherosclerotic rabbit after >12 months on diet (C), in comparison with the virtual absence of uptake in a healthy animal (A). B and D, Dissected aortas corresponding to the scans shown in A and C, respectively, and histological sections of the indicated aortic segments (boxed in red) stained with pimonidazole. Photomicrographs taken at 20× magnification. LK indicates left kidney; and RK, right kidney. Jesus Mateo et al. Circ Cardiovasc Imaging. 2014;7:312-320 Copyright © American Heart Association, Inc. All rights reserved.

In vivo 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) of metabolic rate/inflammation and correlation with 18F-fluoromisonidazole (18F-FMISO). In vivo 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) of metabolic rate/inflammation and correlation with 18F-fluoromisonidazole (18F-FMISO). A, Combined PET/computed tomography (CT) coronal views (top) of the abdominal aorta illustrating the 18F-FDG uptake of 2 atherosclerotic rabbits after 8 months (middle) and >12 months (right) on atherogenic diet, compared with a control animal (left). The bottom shows the axial views of the corresponding slices indicated with a green line in the coronal view. Green arrows denote the abdominal aorta (magnified view provided at the bottom of each image). L indicates left; and R, right. B, Dot plot comparing the 18F-FDG uptake (standardized uptake values [SUV] mean) of the abdominal aorta of atherosclerotic rabbits with that of control animals. Lines represent the mean. C, Representative 18F-FDG and 18F-FMISO PET/CT studies obtained within a few days of each other from an animal of the progression group. Green arrows highlight the abdominal aorta. D, Correlations between the uptake of 18F-FMISO and 18F-FDG. Jesus Mateo et al. Circ Cardiovasc Imaging. 2014;7:312-320 Copyright © American Heart Association, Inc. All rights reserved.

Detection of hypoxia and macrophages in atherosclerotic plaques by immunohistochemistry. Detection of hypoxia and macrophages in atherosclerotic plaques by immunohistochemistry. A, Staining of hypoxia (pimonidazole) and macrophages (RAM-11) in serial sections of abdominal aortas obtained from atherosclerotic rabbits after the induction period (top) and at progression (middle). In the images on the left, hematoxylin-eosin (H&E) staining showing plaque complexity. Photomicrographs taken at 20× magnification. Red arrows indicate areas of nonhypoxic macrophages. B, As in A, from a rabbit of the progression group showing only superficial macrophage infiltration not associated with hypoxia. C, Bar graphs showing the staining quantification of hypoxia and macrophages, as percentage of vessel wall area, in the atherosclerotic animals. The staining was negative in control animals (blank). Bars represent mean±SEM. D, Correlation between the extent of hypoxia and macrophage staining per animal. Each data point represents the average value for all the sections stained with pimonidazole and RAM-11 of a single abdominal aorta (≈25 sections/aorta). Jesus Mateo et al. Circ Cardiovasc Imaging. 2014;7:312-320 Copyright © American Heart Association, Inc. All rights reserved.

Association of neovascularization with hypoxia and inflammation in atherosclerotic plaques. Association of neovascularization with hypoxia and inflammation in atherosclerotic plaques. A, Detection of neovessels (CD31, right), hypoxia (pimonidazole, left), and macrophages (RAM-11, middle) in serial sections of abdominal aortas obtained from atherosclerotic rabbits after the induction period (top) and at progression (bottom). Photomicrographs taken at 20× magnification. B, Bar graph showing neovessel quantification, as number of CD31+-microvessels per section, in the atherosclerotic animals. Bars represent mean±SEM. C, Higher power magnification image (100×) showing abundant microvessels (CD31 staining) in the media and at the plaque base of an advanced lesion. D, Transmission electron microscopy detail of capillaries surrounded by lipid-laden foam cells in the media (right) and in the plaque base (left) of an advanced lesion. Jesus Mateo et al. Circ Cardiovasc Imaging. 2014;7:312-320 Copyright © American Heart Association, Inc. All rights reserved.

Plaque neovessels colocalize with hypoxia-inducible factor (HIF)-1α–positive hypoxic macrophages. Plaque neovessels colocalize with hypoxia-inducible factor (HIF)-1α–positive hypoxic macrophages. Representative staining of hypoxia (pimonidazole), HIF-1α, macrophages (RAM-11), neovessels (CD31), and H&E in serial sections of an advanced atherosclerotic lesion showing colocalization of intimal plaque neovessels with HIF-1α–positive macrophage-rich hypoxic areas (pimonidazole+/HIF-1α+/RAM-11+). Photomicrographs taken at 100× magnification. L indicates lumen. Jesus Mateo et al. Circ Cardiovasc Imaging. 2014;7:312-320 Copyright © American Heart Association, Inc. All rights reserved.