Nilima Shukla, PhD, Gianni D. Angelini, FRCS, Jamie Y. Jeremy, PhD

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Presentation transcript:

Interactive Effects of Homocysteine and Copper on Angiogenesis in Porcine Isolated Saphenous Vein Nilima Shukla, PhD, Gianni D. Angelini, FRCS, Jamie Y. Jeremy, PhD The Annals of Thoracic Surgery Volume 84, Issue 1, Pages 43-49 (July 2007) DOI: 10.1016/j.athoracsur.2007.03.087 Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Diagrammatic representation model of angiogenesis in the pig saphenous vein: (a) thrombin plus fibrinogen are placed in the bottom of the well, creating a fibrin bed; (b) rings of saphenous vein are placed on the fibrin bed, and thrombin plus fibrinogen are placed over the ring, effectively embedding the ring in fibrin; (c) fibrin is covered with serum free medium, and (d) tubules grow into fibrin which are then counted over time. The Annals of Thoracic Surgery 2007 84, 43-49DOI: (10.1016/j.athoracsur.2007.03.087) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Tubule formation by porcine saphenous vein rings embedded and incubated in fibrin over time: control (●); 1 μg/mL angiostatin (▲); 10 nM thapsigargin (♦); 1 μM copper chloride (■); 10 μM copper chloride (○). (See also Figures 3 and 5.) Each point = mean ± standard error of the mean; n = 6 saphenous vein rings. *p < 0.01 comparing effect at individual time points with controls (untreated). The Annals of Thoracic Surgery 2007 84, 43-49DOI: (10.1016/j.athoracsur.2007.03.087) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Representative photomicrographs demonstrate tubule formation in porcine saphenous vein explants assessed at 7 days after embedding in fibrin: (a) control, (b) 10 nM thapsigargin, (c) 1 μg/mL angiostatin (see Fig 2). The Annals of Thoracic Surgery 2007 84, 43-49DOI: (10.1016/j.athoracsur.2007.03.087) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Immunocytochemical staining of (left) lectin (endothelial cells), (right) endothelial nitric oxide synthase (eNOS) in pig saphenous veins Positive staining indicated by large arrows. Note staining is at the lumen endothelium and on the outer surface (adventitial) of the medial layer. The Annals of Thoracic Surgery 2007 84, 43-49DOI: (10.1016/j.athoracsur.2007.03.087) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Effect of homocysteine at 100 μM (■) and 1 mM (○) on tubule formation in porcine saphenous vein explants compared to controls (●) and the effect of 1 μM copper chloride + 10 μM homocysteine; (▲) and 10 μM copper chloride + 100 μM homocysteine over a 14 day time course. Each point = mean ± standard error of the mean; n = 6 experiments. *p < 0.01 comparing effect at individual time points with controls (untreated). The Annals of Thoracic Surgery 2007 84, 43-49DOI: (10.1016/j.athoracsur.2007.03.087) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions

Fig 6 Representative photomicrographs demonstrate tubule formation assessed at 7 days after embedding in fibrin. (A) Control, (B) 1 μM copper chloride, (C) 10 μM copper chloride, (D) 10 μM homocysteine plus 1μM copper chloride, (E) 100 μM homocysteine plus 1 μM copper chloride; (F) 100 μM homocysteine + 10 μM copper chloride. (Bar = 500 μm.) The Annals of Thoracic Surgery 2007 84, 43-49DOI: (10.1016/j.athoracsur.2007.03.087) Copyright © 2007 The Society of Thoracic Surgeons Terms and Conditions