Neutrophil Adherence to Activated Saphenous Vein and Mammary Endothelium After Graft Preparation Axel Schlitt, MD, Diethard Pruefer, MD, Ute Buerke,

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Neutrophil Adherence to Activated Saphenous Vein and Mammary Endothelium After Graft Preparation Axel Schlitt, MD, Diethard Pruefer, MD, Ute Buerke, MD, Martin Russ, MD, Manfred Dahm, MD, Hellmut Oelert, MD, Karl Werdan, MD, Michael Buerke, MD The Annals of Thoracic Surgery Volume 81, Issue 4, Pages 1262-1268 (April 2006) DOI: 10.1016/j.athoracsur.2005.09.073 Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 (A) Bar graph showing adherence of unstimulated human polymorphonuclear neutrophils either to unstimulated human saphenous vein graft endothelium (control) or to thrombin (0.5 to 2 U/mL) -stimulated vascular graft endothelium. Fucoidin, as a selectin-blocking carbohydrate, significantly attenuated polymorphonuclear neutrophil adherence to thrombin-stimulated saphenous vein endothelium. (B) Bar graph showing adherence of unstimulated human polymorphonuclear neutrophils either to unstimulated human mammary artery graft endothelium (control) or to thrombin (0.5 to 2 U/mL) -stimulated arterial endothelium. An antiselectin carbohydrate (fucoidin) significantly attenuated polymorphonuclear neutrophil adherence to thrombin-stimulated human mammary artery graft endothelium. Values are mean ± standard error of the mean. Twenty-four to 29 saphenous vein graft segments of 12 patients were studied. Twenty-six to 30 human mammary artery graft segments of 14 patients were studied (*p < 0.05; **p < 0.01; ***p < 0.001). The Annals of Thoracic Surgery 2006 81, 1262-1268DOI: (10.1016/j.athoracsur.2005.09.073) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Bar graph showing adherence of unstimulated human polymorphonuclear neutrophils either to unstimulated saphenous vein endothelium (control) or to Bretschneider solution–dilated saphenous vein endothelium. An antiselectin carbohydrate (fucoidin) significantly attenuated polymorphonuclear neutrophil adherence to dilation-stimulated saphenous vein endothelium. Values are mean ± standard error of the mean. Twenty-five to 34 saphenous vein graft segments of 14 patients were studied (*p < 0.05; **p < 0.01; ***p < 0.001). The Annals of Thoracic Surgery 2006 81, 1262-1268DOI: (10.1016/j.athoracsur.2005.09.073) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Photomicrograph showing fluorescence microscopy analysis of unstimulated human polymorphonuclear neutrophils (PMNs) adhering to either unstimulated saphenous vein endothelium (control; A) or to Bretschneider solution–dilated saphenous vein endothelium (B). Only a few fluorescence-labeled neutrophils are adhering to normal venous graft endothelium. In contrast, increased neutrophil adherence is detectable in dilated saphenous vein graft. Arrows indicate adhering neutrophils. The Annals of Thoracic Surgery 2006 81, 1262-1268DOI: (10.1016/j.athoracsur.2005.09.073) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Scanning electron micrograph of unstimulated human polymorphonuclear neutrophils (PMNs) adhering to either unstimulated saphenous vein graft endothelium (control; A) or to Bretschneider solution–dilated saphenous vein graft endothelium (B). In control graft, well-preserved endothelium is visible. Endothelial cell nuclei are visible protruding into the lumen. Interendothelial cell junctions are intact. In contrast, endothelial disruption is detectable, as well as increased neutrophil adherence in dilated saphenous vein graft. Endothelium has been lost, exposing the subendothelial connective tissue. Arrows indicate adhering neutrophils. The Annals of Thoracic Surgery 2006 81, 1262-1268DOI: (10.1016/j.athoracsur.2005.09.073) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Bar graph showing adherence of unstimulated human polymorphonuclear neutrophils either to unstimulated human mammary artery graft endothelium (control) or to papaverine-dilated mammary artery graft endothelium. An antiselectin carbohydrate (fucoidin) significantly attenuated polymorphonuclear neutrophil adherence to papaverine-stimulated human mammary artery graft endothelium. Values are mean ± standard error of the mean. Eighteen to 22 human mammary artery graft endothelium segments of 9 patients were studied (**p < 0.01). The Annals of Thoracic Surgery 2006 81, 1262-1268DOI: (10.1016/j.athoracsur.2005.09.073) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions

Fig 6 Relaxation of unstimulated saphenous vein rings (normal) or Bretschneider solution–dilated saphenous vein rings in vitro to A23187 after precontraction with U46619 (A). Endothelial dependent-relaxation after A23187 was significantly reduced after dilation procedure when compared with normal venous grafts. In contrast, endothelial-independent relaxation after sodium nitroprusside did not demonstrate any significant differences when normal venous grafts were compared with grafts after high-pressure distention (B). Clearly, dilated saphenous vein rings demonstrated significant endothelial dysfunction when compared with normal venous graft rings, indicating loss of nitric oxide release after dilation procedure. Values are mean ± standard error of the mean. Twenty-eight to 34 saphenous vein graft segments of 15 patients were studied (*p < 0.05). The Annals of Thoracic Surgery 2006 81, 1262-1268DOI: (10.1016/j.athoracsur.2005.09.073) Copyright © 2006 The Society of Thoracic Surgeons Terms and Conditions