Department of Bioengineering, Stanford University, Stanford, California 94305, USA.
J Histochem Cytochem. 2012 Feb;60(2):97-109. doi: 10.1369/0022155411433066. Epub 2011 Dec 1.
This study investigated the spatial and temporal remodeling of blood vessel wall microarchitecture and cellular morphology during abdominal aortic aneurysm (AAA) development using immunofluorescent array tomography (IAT), a high-resolution three-dimensional (3D) microscopy technology, in the murine model. Infrarenal aortas of C57BL6 mice (N=20) were evaluated at 0, 7, and 28 days after elastase or heat-inactivated elastase perfusion. Custom algorithms quantified volume fractions (VF) of elastin, smooth muscle cell (SMC) actin, and adventitial collagen type I, as well as elastin thickness, elastin fragmentation, non-adventitial wall thickness, and nuclei amount. The 3D renderings depicted elastin and collagen type I degradation and SMC morphological changes. Elastin VF decreased 37.5% (p<0.01), thickness decreased 48.9%, and fragmentation increased 449.7% (p<0.001) over 28 days. SMC actin VF decreased 78.3% (p<0.001) from days 0 to 7 and increased 139.7% (p<0.05) from days 7 to 28. Non-adventitial wall thickness increased 61.1%, medial nuclei amount increased 159.1% (p<0.01), and adventitial collagen type I VF decreased 64.1% (p<0.001) over 28 days. IAT and custom image analysis algorithms have enabled robust quantification of vessel wall content, microstructure, and organization to help elucidate the dynamics of vascular remodeling during AAA development.
本研究使用免疫荧光层切术（IAT），一种高分辨率三维（3D）显微镜技术，在小鼠模型中研究了腹主动脉瘤（AAA）发展过程中血管壁微观结构和细胞形态的空间和时间重塑。在弹性蛋白酶或热失活弹性蛋白酶灌注后 0、7 和 28 天，评估 C57BL6 小鼠的肾下主动脉（N=20）。定制算法量化了弹力蛋白、平滑肌细胞（SMC）肌动蛋白和外膜胶原 I 的体积分数（VF），以及弹力蛋白厚度、弹力蛋白碎片化、非外膜壁厚度和核数量。3D 渲染图描绘了弹力蛋白和胶原 I 的降解以及 SMC 形态变化。28 天内，弹力蛋白 VF 降低了 37.5%（p<0.01），厚度降低了 48.9%，碎片化增加了 449.7%（p<0.001）。SMC 肌动蛋白 VF 从第 0 天到第 7 天降低了 78.3%（p<0.001），从第 7 天到第 28 天增加了 139.7%（p<0.05）。非外膜壁厚度增加了 61.1%，中膜核数量增加了 159.1%（p<0.01），外膜胶原 I VF 降低了 64.1%（p<0.001），28 天内。IAT 和定制的图像分析算法能够对血管壁含量、微观结构和组织进行强大的定量分析，有助于阐明 AAA 发展过程中血管重塑的动力学。
