Distinct macrophage phenotype and collagen organization within the intraluminal thrombus of abdominal aortic aneurysm Jayashree Rao, MS, Bryan N. Brown,

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Presentation transcript:

Distinct macrophage phenotype and collagen organization within the intraluminal thrombus of abdominal aortic aneurysm Jayashree Rao, MS, Bryan N. Brown, PhD, Justin S. Weinbaum, PhD, Emily L. Ofstun, BS, Michel S. Makaroun, MD, Jay D. Humphrey, PhD, David A. Vorp, PhD Journal of Vascular Surgery Volume 62, Issue 3, Pages 585-593 (September 2015) DOI: 10.1016/j.jvs.2014.11.086 Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 1 A and B, Cross sections show the gross morphologic appearance of the shoulder (S), luminal aspect (Lu), abluminal aspect (Ab) and midsection (M) of the intraluminal thrombus (ILT) at time of surgery. Note the layered structure. Samples were cut in two and then further dissected for microscopic investigation. C, A schematic view is shown of the ILT aspects depicted in panel B. The boxes represent the areas sampled. Journal of Vascular Surgery 2015 62, 585-593DOI: (10.1016/j.jvs.2014.11.086) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 2 Asymmetric composition of the intraluminal thrombus (ILT) is seen in representative histologic appearance of samples. Samples were stained with (A) hematoxylin and eosin, (B) Herovici's polychrome, and (C) picrosirius red (PSR). Note the distinct appearances of the abluminal (Ab), medial, and luminal (Lu) regions of the full-thickness ILT sample. B and C, Herovici's polychrome and PSR staining suggest the presence of collagen within the luminal region of the ILT. Scale bar = 250 μm. Journal of Vascular Surgery 2015 62, 585-593DOI: (10.1016/j.jvs.2014.11.086) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 3 Representative images show the histologic and immunohistochemical evaluation of the luminal (Lu) aspect of the intraluminal thrombus (ILT). Staining with (A and C) picrosirius red (PSR) and (B and D) Herovici's polychrome suggest the presence of collagen with alignment parallel to the luminal surface within the (A and B) shoulder region of the ILT. C and D, Collagen (ColI) is present but less organized within the midsection of the luminal surface. Immunolabeling (red) demonstrated (E) collagen I and (F) collagen III within the extracellular matrix and within the cytoplasm of the cells of the luminal aspect, and there was no distinct difference between midsection or shoulder regions. Scale bar = 100 μm. Journal of Vascular Surgery 2015 62, 585-593DOI: (10.1016/j.jvs.2014.11.086) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 4 Representative images of immunolabeling performed to examine the phenotype and location of cells within the intraluminal thrombus (ILT). A, A significant population of nucleated cells was observed within a “nuclear stratus” (NS) of 4′,6-diamidino-2-phenylindole-stained sections of the lumen (Lu) of the sample, with few cells found within the medial or abluminal regions. Nucleated cells within the lumen express a heterogeneous phenotype including (B) cluster of differentiation (CD) 45, (C) CD68, (D) human leukocyte antigen (HLA)-DR, (E) CD105, (F) smooth muscle actin (SMA), and (G) matrix metalloproteinase 9 (MMP9). Scale bar = 100 μm. Journal of Vascular Surgery 2015 62, 585-593DOI: (10.1016/j.jvs.2014.11.086) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 5 Ratio of nuclear stratus to total intraluminal thrombus (ILT) width. Data from six patients are shown. Two samples from each patient were used. The nuclear stratus (red) was not found to be proportional to the total ILT thickness (red + blue). Journal of Vascular Surgery 2015 62, 585-593DOI: (10.1016/j.jvs.2014.11.086) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 6 Spatial distribution of cells positive for (A) cluster of differentiation (CD) 45, (B) CD68, (C) CD105, (D) human leukocyte antigen (HLA)-DR, and (E) α-smooth muscle actin (α-SMA) within the intraluminal thrombus (ILT). Data from six patients are displayed. Two samples per patient were examined. Results demonstrate presence of cells expressing myeloid, mesenchymal, and progenitor cell markers, which are largely located within the luminal region. Few cells of any type were observed within the medial or abluminal portions of the ILT. Journal of Vascular Surgery 2015 62, 585-593DOI: (10.1016/j.jvs.2014.11.086) Copyright © 2015 Society for Vascular Surgery Terms and Conditions

Fig 7 Coexpression of fibrocyte markers in cells from the intraluminal thrombus (ILT). Cells within the ILT express a unique “fibrocyte-like” phenotype. Cells colocalizing expression of (A) cluster of differentiation (CD) 68+, (B) α-smooth muscle actin (α-SMA)+, (C) CD68/α-SMA; (D) CD68+, (E) collagen (Col) I+, and (F) CD68+/collagen I; and (G) CD68+, (H) collagen (Col) III+ and (I) CD68+/Col III were all observed. Red, CD68; blue, 4′,6-diamidino-2-phenylindole; (B) green, SMA, (E) collagen I, (H) collagen III. Scale bar = 100 μm. Journal of Vascular Surgery 2015 62, 585-593DOI: (10.1016/j.jvs.2014.11.086) Copyright © 2015 Society for Vascular Surgery Terms and Conditions