Volume 59, Issue 6, Pages (June 2001)

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Volume 59, Issue 6, Pages 2325-2334 (June 2001) Venous neointimal hyperplasia in polytetrafluoroethylene dialysis grafts  Prabir Roy-Chaudhury, Burnett S. Kelly, Mary Ann Miller, Anita Reaves, Janice Armstrong, Nuwan Nanayakkara, Sue C. Heffelfinger  Kidney International  Volume 59, Issue 6, Pages 2325-2334 (June 2001) DOI: 10.1046/j.1523-1755.2001.00750.x Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 1 Diagrammatic representation of the venous anastomosis to demonstrate how the sections were cut. Maximal neointimal hyperplasia occurred at the site of the graft-vein anastomosis (B) and in the downstream vein (A). Kidney International 2001 59, 2325-2334DOI: (10.1046/j.1523-1755.2001.00750.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 2 Scoring scale for angiogenesis (microvessel formation). (a) (vWF ×300, neointima) A single microvessel (score = 1+). (b) (vWF ×300, neointima) A few scattered microvessels (score 2+). (c) (vWF ×300, adventitia) A large number of microvessels (score 3+). (d) (vWF ×300, adventitia) Density of microvessels needed to have a score of 4+. Kidney International 2001 59, 2325-2334DOI: (10.1046/j.1523-1755.2001.00750.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 3 Normal vein. (a) Note the thin one- to two-layer intima (thin arrows) and three- to four-layer media (thick arrows) of normal vein (hematoxylin and eosin ×400). (b) Relative absence of endothelial cell staining in the adventitia and media of normal vein. Note the intense staining of the single layer endothelium (vWF ×1500). (c) Thickness of normal venous (between thin arrows, V) and arterial (double-headed arrow, A) intima media (SMA ×117). (d) Thickness of the venous neointima and media in a dialysis patient with venous stenosis (SMA ×117). At an identical magnification to (c), the venous neointima (d; double-headed arrow, N) is 20 times thicker than the intima media of normal vein (c; between thin arrows). Also, note that the venous media (d; M, length of bar) is as thick as the arterial media in (c), indicating arterialization of the vein. Kidney International 2001 59, 2325-2334DOI: (10.1046/j.1523-1755.2001.00750.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 4 Cell types and cytokines (adventitia to left and lumen to right). (a) PTFE graft (hematoxylin and eosin ×200). Note the significant venous neointimal hyperplasia (extent of arrow) between the graft (G) and the lumen (L). (b) Downstream vein (hematoxylin and eosin ×200). Note the presence of microvessels (thin arrows) within the adventitia (A). Also note the thickened (arterialized) media (M, double-headed arrow) and the significant amount of neointimal hyperplasia (N, bar). (c) Downstream vein; neointima (vWf ×400). Note the prominent angiogenesis within the neointima (arrows), as assessed by this endothelial cell marker (d) downstream vein; neointima (vWF + Ki67 ×800). High-power view of a microvessel within the neointima of downstream vein. Note the distinct colocalization of blue (endothelial) and brown (proliferating) cells indicating active endothelial cell proliferation (angiogenesis). (e) Upstream graft; neointima (PG-M1 ×2000). High-power view of a macrophage giant cell adjacent to the neointimal surface of PTFE graft (G). Also note the large number of macrophages in this area (thin arrows). (f) Downstream vein; neointima (SMA + Ki-67 ×1000). High-power view of a portion of the neointima stained for smooth muscle cells (brown) and proliferating cells (blue). Note that almost all the active cellular proliferation in this specimen (arrows) is occurring within the neointimal microvessels (angiogenesis). At the time that this specimen was harvested, there was no ongoing smooth muscle cell proliferation (g) PTFE graft; adventitia (bFGF ×500). Note the strong expression of bFGF in adventitial vessels (thick arrow) and by the macrophage giant cell layer (thin arrow) lining the graft. (f) Downstream vein; media and neointima (PDGF ×400). There is strong expression of this cytokine in the venous media (M) and by smooth muscle cells/myofibroblasts within the neointima (N, bar). Kidney International 2001 59, 2325-2334DOI: (10.1046/j.1523-1755.2001.00750.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 5 Matrix and structural proteins. (a) PTFE graft (tenascin ×200). There is strong expression of tenascin in the region of the macrophage giant cell layer (thin arrow) surrounding PTFE graft (G) and on the abluminal side of the neointima (thick arrow). (b) PTFE graft (collagen IV ×160). Collagen is present as expected within the walls of prominent adventitial blood vessels (thin arrows) and in this particular sample within the luminal portion of the neointima (thick arrow). (c) Downstream vein; neointima (fibronectin ×500). There is strong diffuse expression of fibronectin by ECM components. (d) Downstream vein; neointima (laminin ×500). Laminin is a prominent component of microvessels (arrows) within the neointima. Kidney International 2001 59, 2325-2334DOI: (10.1046/j.1523-1755.2001.00750.x) Copyright © 2001 International Society of Nephrology Terms and Conditions