Download or read book Three-dimensional Vessel Wall Normal Microarchitecture and Remodeling with Abdominal Aortic Aneurysms Quantified Using Immunofluorescent Array Tomography written by Sanaz Saatchi and published by . This book was released on 2011 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Abdominal aortic aneurysms (AAA) are described by a pathological dilation of at least 150% in the abdominal aorta. Aneurysm pathogenesis is characterized by extracellular matrix (ECM) remodeling, smooth muscle cell (SMC) apoptosis, and inflammatory cell infiltration. Overall, the structural organization and integrity of the vessel wall is lost. In order to better understand the mechanism of AAA development, a novel microscopy technology, in combination with an AAA animal model, were used to visualize and quantify the microstructural and cellular changes in the vessel wall. Therefore, the goals of this work were to introduce Immunofluorescent Array Tomography (IAT), a novel three-dimensional high resolution microscopy technology, to the field of cardiovascular research, and to apply IAT to describe the vessel wall microarchitecture of a healthy aorta, as well as to investigate the spatial and temporal changes in tissue and cellular content, structure, and organization during AAA development. The purpose of the initial studies was two-fold: to develop the methods needed to enable the application of IAT to murine blood vessels and to investigate the microarchitecture of the healthy murine aorta. The anterior and posterior regions of the infrarenal aorta of 8 to 10 week old C57BL6 mice were evaluated. Staining and custom image analysis methods were developed. Antibody selection, primary antibody concentration, co-staining with multiple primary antibodies, and the multi-cycle staining design were optimized to produce positive and specific staining of elastin, smooth muscle cell actin (SMCA), and collagen type I. Algorithms were developed and applied to the healthy murine aorta to quantify volume fractions (VF) of medial elastin (27.5 ± 0.99%), SMCA (18.4 ± 0.67%), and nuclei (6.1 ± 0.14%), as well as adventitial collagen type I (22.3 ± 1.7%). Elastin thickness (1.6 ± 0.35 [Mu]m), spacing between elastin lamellae (3.5 ± 0.13 [Mu]m), elastin fragmentation (4.2 x 10-3 ± 1.6 x 10-4 # of objects/elastin area ([Mu]m)), media wall thickness (20.4 ± 3.1 [Mu]m), nuclei aspect ratio (3.2 ± 0.21 [Mu]m), and nuclei amount (17.3 ± 0.69 nuclei) were also quantified. The 3D microstructure and cellular morphology of the anterior and posterior infrarenal murine aorta were qualitatively and quantitatively described using IAT. IAT was then used to investigate the spatial and temporal remodeling of vessel wall microarchitecture and cellular morphology during AAA development in the murine elastase perfusion 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 VFs of elastin, SMCA, and adventitial collagen type I, as well as elastin thickness, elastin fragmentation, media thickness, and nuclei amount. 3D renderings depicted elastin and collagen type I degradation, and dynamic changes in SMC phenotype, morphology, and amount. Elastin degradation was described by a 37.5% (p