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Three-Dimensional Imaging Provides Detailed Atherosclerotic Plaque Morphology and Reveals Angiogenesis After Carotid Artery Ligation.
Circulation Research ( IF 16.5 ) Pub Date : 2020-01-09 , DOI: 10.1161/circresaha.119.315804
Tobias Becher 1, 2, 3 , Dario F Riascos-Bernal 4 , Daniel J Kramer 1 , Vanessa M Almonte 4 , Jingy Chi 1 , Tao Tong 5 , Gustavo H Oliveira-Paula 4 , Issam Koleilat 6 , Wei Chen 7, 8 , Paul Cohen 1 , Nicholas E S Sibinga 4
Affiliation  

RATIONALE Remodeling of the vessel wall and the formation of vascular networks are dynamic processes that occur during mammalian embryonic development and in adulthood. Plaque development and excessive neointima formation are hallmarks of atherosclerosis and vascular injury. As our understanding of these complex processes evolves, there is a need to develop new imaging techniques to study underlying mechanisms. OBJECTIVE We used tissue clearing and light-sheet microscopy for 3-dimensional (3D) profiling of the vascular response to carotid artery ligation and induction of atherosclerosis in mouse models. METHODS AND RESULTS Adipo-Clear and immunolabeling in combination with light-sheet microscopy were applied to image carotid arteries and brachiocephalic arteries, allowing for 3D reconstruction of vessel architecture. Entire 3D neointima formations with different geometries were observed within the carotid artery and scored by volumetric analysis. Additionally, we identified a CD31-positive adventitial plexus after ligation of the carotid artery that evolved and matured over time. We also used this method to characterize plaque extent and composition in the brachiocephalic arteries of ApoE-deficient mice on high-fat diet. The plaques exhibited inter-animal differences in terms of plaque volume, geometry, and ratio of acellular core to plaque volume. A 3D reconstruction of the endothelium overlying the plaque was also generated. CONCLUSIONS We present a novel approach to characterize vascular remodeling in adult mice using Adipo-Clear in combination with light-sheet microscopy. Our method reconstructs 3D neointima formation after arterial injury and allows for volumetric analysis of remodeling, in addition to revealing angiogenesis and maturation of a plexus surrounding the carotid artery. This method generates complete 3D reconstructions of atherosclerotic plaques and uncovers their volume, geometry, acellular component, surface, and spatial position within the brachiocephalic arteries. Our approach may be used in a number of mouse models of cardiovascular disease to assess vessel geometry and volume. Visual Overview: An online visual overview is available for this article.

中文翻译:

三维成像可提供详细的动脉粥样硬化斑块形态,并显示颈动脉结扎后的血管生成。

血管壁的重塑和血管网络的形成是哺乳动物胚胎发育和成年期发生的动态过程。斑块发育和新内膜过度形成是动脉粥样硬化和血管损伤的标志。随着我们对这些复杂过程的理解的发展,需要开发新的成像技术来研究潜在机制。目的我们使用组织清除和光片显微镜对小鼠模型中对颈动脉结扎和诱发动脉粥样硬化的血管反应进行三维(3D)分析。方法和结果将Adipo-Clear和免疫标记与光片显微镜相结合,应用于颈动脉和头颅动脉的成像,从而可以对血管结构进行3D重建。在颈动脉内观察到具有不同几何形状的整个3D新内膜形成,并通过体积分析对其评分。此外,我们发现结扎颈动脉的CD31阳性外膜丛随着时间的流逝而逐渐成熟。我们还使用这种方法来表征高脂饮食对ApoE缺陷型小鼠的头臂动脉中的斑块程度和组成。斑块在斑块体积,几何形状以及无细胞核与斑块体积的比率方面表现出动物间差异。还生成了覆盖斑块的内皮细胞的3D重建。结论我们提出了一种新的方法来表征成年小鼠使用Adipo-Clear与光片显微镜相结合的血管重塑。我们的方法重建了动脉损伤后的3D新内膜形成,并允许进行体积分析重塑,此外还揭示了血管生成和颈动脉周围神经丛的成熟。此方法生成动脉粥样硬化斑块的完整3D重建,并揭示它们在头臂动脉内的体积,几何形状,无细胞成分,表面和空间位置。我们的方法可用于许多心血管疾病的小鼠模型中,以评估血管的几何形状和体积。视觉概述:本文提供了在线视觉概述。头臂动脉内的无细胞成分,表面和空间位置。我们的方法可用于许多心血管疾病的小鼠模型中,以评估血管的几何形状和体积。视觉概述:本文提供了在线视觉概述。头臂动脉内的无细胞成分,表面和空间位置。我们的方法可用于许多心血管疾病的小鼠模型中,以评估血管的几何形状和体积。视觉概述:本文提供了在线视觉概述。
更新日期:2020-02-28
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