The role of urokinase plasminogen activator and plasmin activator inhibitor-1 on vein wall remodeling in experimental deep vein thrombosis Joe F. Baldwin,

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The role of urokinase plasminogen activator and plasmin activator inhibitor-1 on vein wall remodeling in experimental deep vein thrombosis Joe F. Baldwin, BS, Vikram Sood, BS, Megan A. Elfline, MS, Cathy E. Luke, LVT, Nicholas A. Dewyer, MD, Jose A. Diaz, MD, Dan D. Myers, DVM, Thomas Wakefield, MD, Peter K. Henke, MD Journal of Vascular Surgery Volume 56, Issue 4, Pages 1089-1097 (October 2012) DOI: 10.1016/j.jvs.2012.02.054 Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 1 A, Thrombus size was larger in urokinase-plasminogen activator (uPA) −/− as compared with wild type (WT). B, Plasmin activity was decreased in uPA −/− at 8 days as compared with WT; C, Despite larger thrombi, the fibrinogen content was lower, as was thrombin-antithrombin complex (TAT) at 8 days in uPA −/− mice compared with WT. D, At 8 and 21 days, venous thromboses (VT) were smaller in the plasminogen activator inhibitor-1 (PAI-1) −/− mice as compared with WT. E, Thrombus size was smaller in the PAI-1−/−as compared with WT. F, Plasmin activity was increased in PAI-1−/− at 8 days as compared with WT. *P < .05. Journal of Vascular Surgery 2012 56, 1089-1097DOI: (10.1016/j.jvs.2012.02.054) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 2 Hematoxylin and eosin stained inferior vena cava (IVC) sections of 21-day wild type (WT) (A) and urokinase-plasminogen activator (uPA) −/− (B) examples. Note the large and relative acellularity of the uPA −/− thrombus. The 21-day WT (C) and plasminogen activator inhibitor-1 (PAI-1) −/− (D) are shown with similar thrombus morphology and cellularity. T, Thrombus. Journal of Vascular Surgery 2012 56, 1089-1097DOI: (10.1016/j.jvs.2012.02.054) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 3 Gene expression of the endothelial marker CD31 was reduced in 21-day urokinase-plasminogen activator (uPA) −/− compared with wild type (WT) (A). Conversely, CD31 gene expression was increased at 8 days in plasminogen activator inhibitor-1 (PAI-1) −/− (B) as compared with WT. Vein wall intimal thickness was reduced in 8-day uPA −/− vein wall as compared with WT (C). Mild neointimal thickening is present in 8-day WT vein walls (D), but little in the uPA −/− vein wall (E). Note again the marked cellularity of 8-day WT thrombi, but few were present in uPA −/− thrombi. T, Thrombus; W, wall. Journal of Vascular Surgery 2012 56, 1089-1097DOI: (10.1016/j.jvs.2012.02.054) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 4 Vein wall procollagen I (A) and III (B) gene expression was elevated in 8-day plasminogen activator inhibitor-1 (PAI-1) −/− as compared with wild type (WT). By Picosirius red analysis, collagen content was elevated at 8 days in PAI-1 −/− compared with WT, and a trend at 21 days while no difference in collagen content was found in urokinase-plasminogen activator (uPA) −/− at 8 or 21 days (C). Birefringence images show a thin vein wall collagen in WT (D), but a greater amount in PAI-1 −/− vein wall (E). P < .05. T, Thrombus. Journal of Vascular Surgery 2012 56, 1089-1097DOI: (10.1016/j.jvs.2012.02.054) Copyright © 2012 Society for Vascular Surgery Terms and Conditions

Fig 5 Vascular smooth muscle cell (VSMC) gene marker showed increased SM22 (A) and alpha smooth muscle actin (αSMA) (B) expression in plasminogen activator inhibitor-1 (PAI-1) −/− vein wall compared to wild type (WT) at 8 and 21 days. Medial αSMA + cells were increased in PAI-1 −/− at 8 days as compared with WT (C). Immunohistology showed the mononuclear medial location of these positive cells in WT (D) and PAI-1 −/− (E) inferior vena cava (IVC) sections. *P < .05; arrows denote (+) cells. T, Thrombus; W, wall. Journal of Vascular Surgery 2012 56, 1089-1097DOI: (10.1016/j.jvs.2012.02.054) Copyright © 2012 Society for Vascular Surgery Terms and Conditions