Local treatment with recombinant tissue factor pathway inhibitor reduces the development of intimal hyperplasia in experimental vein grafts Tam T.T.

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Presentation transcript:

Local treatment with recombinant tissue factor pathway inhibitor reduces the development of intimal hyperplasia in experimental vein grafts Tam T.T. Huynh, MDa, Mark G. Davies, MD, PhDa, Michael A. Thompson, b, Michael D. Ezekowitz, MDc, Per-Otto Hagen, PhDa,d, Brian H. Annex, MDb Journal of Vascular Surgery Volume 33, Issue 2, Pages 400-407 (February 2001) DOI: 10.1067/mva.2001.111989 Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 1 Immunohistochemistry of representative sections from ungrafted contralateral jugular vein (A and B) , 3-day gel-control vein graft (C and D), and 3-day rTFPI-in-gel vein graft (E and F). Endothelial cell staining (red) was assessed by use of murine antibody raised against human Tie-2, endothelial cell–specific tyrosine kinase receptor. Murine monoclonal anti-rabbit CD-18, which detects β2 chain of leukocyte adherence complex in activated neutrophils and monocytes (red) , was used to determine inflammatory infiltrate. In jugular veins, Tie-2 staining was uniform in endothelial cells (A) and CD-18–positive leukocytes were infrequently present (B). At 3 days after operation, Tie-2 staining was virtually absent (C) and CD-18–positive leukocytes were markedly increased (D) in gel-control vein grafts. Treatment with rTFPI-in-gel preserved endothelial Tie-2 staining (E) and decreased the presence of CD-18 positive leukocytes (F) in vein grafts at 3 days. Note that Tie-2 staining in rTFPI-in-gel vein grafts was similar to that in ungrafted jugular veins (E versus A, respectively). Vessel lumen is at top in all sections. Original magnification × 50 in all panels. Journal of Vascular Surgery 2001 33, 400-407DOI: (10.1067/mva.2001.111989) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 2 Representative Western blot analysis of total TF protein in ungrafted jugular veins and 3-day vein grafts. TF protein was detected as band just above 43-kDa molecular weight marker, by use of AP-1, antibody that has been shown to block TF-mediated thrombosis both in vitro and in vivo.28,29 Equal amounts (10 μg) of protein were loaded in each lane. A, Compared with their respective contralateral ungrafted jugular veins (lanes 1 and 3 ), total TF proteins were similar in 3-day control vein grafts (lane 2 ) and 3-day gel-control vein grafts (lane 4 ). B, In contrast, there is marked reduction in total TF protein in 3-day rTFPI-in-gel vein grafts (lanes 2 and 4 ), compared with corresponding contralateral ungrafted jugular veins (lanes 1 and 3 ). Journal of Vascular Surgery 2001 33, 400-407DOI: (10.1067/mva.2001.111989) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 3 Densitometric analysis of total TF protein expression in 3-day rTFPI-in-gel and gel-control vein grafts. For each vein graft, total TF protein was expressed as densitometric ratio (densitometric ratio = integrated density of TF band in vein graft/integrated density of TF band in corresponding contralateral jugular vein). Combined ex vivo and in vivo treatment with rTFPI reduced total TF protein by 6.2-fold as compared with gel-control (densitometric ratio: 0.12 ± 0.02 versus 0.74 ± 0.07, respectively; P <.05). Values are means ± SEM (n = 3 animals per group). Statistical differences between two groups were compared by use of Mann-Whitney rank sum test. Journal of Vascular Surgery 2001 33, 400-407DOI: (10.1067/mva.2001.111989) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 4 Composite photomicrograph of representative cross-sections from wall of 28-day rTFPI (A) , control (B) , rTFPI-in-gel (C) , and gel-control (D) vein grafts. Arrowheads delineate demarcation between intima and media. Vessel lumen is at top. IH, Intimal hyperplasia; M, media. Original magnification × 250. Journal of Vascular Surgery 2001 33, 400-407DOI: (10.1067/mva.2001.111989) Copyright © 2001 Mosby, Inc. Terms and Conditions

Fig. 5 Cross-sectional thickness of intima and media of 28-day rTFPI, control, rTFPI-in-gel, and gel-control vein grafts. Ex vivo incubation treatment with rTFPI reduced intimal thickness by 21% in 28-day vein grafts, compared with control incubation (69 ± 4 versus 87 ± 5 μm, respectively; P <.05). Combined ex vivo and in vivo rTFPI treatment decreased intimal thickness by 30% (60 ± 4 vs 86 ± 5 μm, respectively; P <.01), compared with gel-controls. Medial thickness of 28-day rTFPI vein grafts was also reduced by 20%, compared with controls (73 ± 3 versus 91 ± 5 μm, respectively; P >.05), and by 15% in rTFPI-in-gel vein grafts compared with gel-controls (73 ± 4 versus 86 ± 4μm, respectively; P >.05), although these findings were not statistically significant. Journal of Vascular Surgery 2001 33, 400-407DOI: (10.1067/mva.2001.111989) Copyright © 2001 Mosby, Inc. Terms and Conditions