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Navin K. Kapur, MD, Ce Bian, MD, Edward Lin, MD, Clayton B

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Presentation on theme: "Navin K. Kapur, MD, Ce Bian, MD, Edward Lin, MD, Clayton B"— Presentation transcript:

1 Inhibition of transforming growth factor-β restores endothelial thromboresistance in vein grafts 
Navin K. Kapur, MD, Ce Bian, MD, Edward Lin, MD, Clayton B. Deming, MS, Jason L. Sperry, MD, Baranda S. Hansen, MS, Nikolaos Kakouros, MBBS, PhD, Jeffrey J. Rade, MD  Journal of Vascular Surgery  Volume 54, Issue 4, Pages e1 (October 2011) DOI: /j.jvs Copyright © 2011 Society for Vascular Surgery Terms and Conditions

2 Fig 1 Effects of cyclic stretch on thrombomodulin (TM) and transforming growth factor-β (TGF-β) isoform expression. A, Human saphenous endothelial cells (HSVECs) were subjected to cyclic stretch for 16 hours and the change in TM mRNA expression was determined by quantitative polymerase chain reaction (PCR). Values shown are the mean ± SEM of n = 4/group. *P < .01 versus unstretched controls. B, Human saphenous vein smooth muscle cells (HSVSMCs) were subjected to cyclic stretch for 16 hours and the change in TGF-β isoform mRNA expression was determined by quantitative PCR. Values shown are the mean ± SEM of n = 8 to 11/group. Panel insert is a Western blot showing stretch-induced change in TGF-β1 protein expression. *P < .001 versus unstretched controls. C, HSVECs were co-cultured on stationary filters suspended in the media of wells containing HSVSMCs subjected to 10% cyclic stretch for 16 hours and the changes in TM and PAI-1 mRNA expression were determined by quantitative PCR. Values shown are the mean ± SEM of n = 3/group. D, HSVEC were co-cultured on stationary filters suspended in the media of wells containing HSVSMCs subjected to 10% cyclic stretch for 16 hours in the presence of a pan-neutralizing anti-TGF-β antibody (1D11) or nonneutralizing isotype control antibody (13C4). Changes in TM and PAI-1 mRNA expression were determined by quantitative PCR. Values are the mean ± SEM of n = 3/group. *P = .02 versus static controls. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

3 Fig 2 Transforming growth factor-β (TGF-β) expression in vein grafts. A, Rabbit vein grafts (VGs) were harvested 7 days after implantation, and relative changes in the mRNA expression of the three major TGF-β isoforms and PAI-1 were determined by quantitative polymerase chain reaction (PCR). Values are the mean ± SEM of n = 4 to 6/group. *P < .001 and #P = .02 versus ungrafted vein controls. B, Immunostaining for TGF-β1 (left column, brown color), PAI-1 (middle column, brown color) and thrombomodulin (TM) (right column, red color) in rabbit VGs harvested 7 days and 6 weeks after implantation. Staining in a control jugular vein and carotid artery are shown for comparison. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

4 Fig 3 Effect of external stenting on transforming growth factor-β1 (TGF-β1) and thrombomodulin (TM) mRNA expression in vein grafts. TGF-β1 and TM mRNA expression was measured in rabbit vein grafts (VGs) with and without external stents 3 days after implantation. Values are the mean ± SEM of n = 3 to 4/group. *P < .001 and #P = .02 versus ungrafted vein controls. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

5 Fig 4 Effect of transforming growth factor-β (TGF-β) inhibition on vein graft (VG) thromboresistance. Rabbits were administered 10 mg/kg of the anti-TGF-β 1D11 antibody or the isotype control 13C4 antibody before VG implantation with two subsequent doses prior to graft harvest on day 7. A, Thrombomodulin (TM) mRNA expression was determined by quantitative polymerase chain reaction (PCR) analysis. B, TM protein expression was determined by densitometric analysis of Western blots (a representative Western blot shown in right panel). C, Activated protein C (APC) generating capacity and D, bound thrombin activity, an indicator of the degree of thrombus formation, were measured in situ using chromogenic assays. E, PAI-1 mRNA expression was determined by quantitative PCR analysis. Values shown are the mean ± SEM for n = 6/group. *P < .05 versus 13C4-treated VGs. Journal of Vascular Surgery  , e1DOI: ( /j.jvs ) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

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