Impaired graft healing due to hypercholesterolemia is prevented by dietary supplementation with α-tocopherol Keiko Miyazaki, MD, Scott M. Colles, PhD,

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Impaired graft healing due to hypercholesterolemia is prevented by dietary supplementation with α-tocopherol Keiko Miyazaki, MD, Scott M. Colles, PhD, Linda M. Graham, MD Journal of Vascular Surgery Volume 48, Issue 4, Pages 986-993 (October 2008) DOI: 10.1016/j.jvs.2008.04.010 Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 1 Lipids were extracted from aortic and graft samples to measure levels of tissue cholesterol and α-tocopherol. Levels of (A) cholesterol or (B) α-tocopherol were determined using high-performance liquid chromatography and reported as the ng/mg tissue (wet weight). Values are expressed as the mean ± standard error for each tissue segment and rabbit dietary group: regular rabbit chow (chow, n = 3), high-cholesterol diet (HC, n = 3), high-cholesterol diet plus α-tocopherol (HC+AT, n = 3), and chow plus α-tocopherol (AT, n = 5). *P < .05, *∪P < .01, #P < .005, ##P < .0001. Journal of Vascular Surgery 2008 48, 986-993DOI: (10.1016/j.jvs.2008.04.010) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 2 Endothelialization and total luminal area of expanded polytetrafluoroethylene (ePTFE) grafts were determined by scanning electron microscopy using the Scion Image analysis program (Scion Corp, Frederick, Md). Endothelialization of ePTFE graft was reported as the percentage of vascular graft covered by endothelium relative to the total luminal area of the graft and expressed as the mean ± standard error for rabbits fed regular rabbit chow (chow, n = 10), a high-cholesterol diet (HC, n = 10), a high-cholesterol diet plus α-tocopherol (HC+AT, n = 5), and chow plus α-tocopherol (AT, n = 5). *P < .05. Journal of Vascular Surgery 2008 48, 986-993DOI: (10.1016/j.jvs.2008.04.010) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 3 Anastomotic intimal hyperplasia was measured on histologic sections between 1.5 mm and 2.5 mm from the graft end and expressed as the mean ± standard error for rabbits fed regular rabbit chow (chow, n = 7), a high-cholesterol diet (HC, n = 4), a high-cholesterol diet plus α-tocopherol (HC+AT, n = 3), and chow plus α-tocopherol (AT, n = 5). *P < .05, **P < .01. Journal of Vascular Surgery 2008 48, 986-993DOI: (10.1016/j.jvs.2008.04.010) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 4 Panels show macrophage staining. Grafts were perfusion-fixed with 4% paraformaldehyde and processed for immunohistochemistry using mouse antirabbit macrophage antibody (RAM-11). Representative sections are shown from rabbits fed regular rabbit chow (chow; n = 4), a high-cholesterol diet (HC; n = 4), a high-cholesterol diet plus α-tocopherol (HC+AT; n = 2), and chow plus α-tocopherol (AT, n = 2). Original magnification ×100. Journal of Vascular Surgery 2008 48, 986-993DOI: (10.1016/j.jvs.2008.04.010) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 5 Panels show nitrotyrosine staining. Grafts were perfusion-fixed with 4% paraformaldehyde and processed for immunohistochemistry using mouse antihuman nitrotyrosine antibody. Representative sections are shown from rabbits fed regular rabbit chow (chow; n = 4), a high-cholesterol diet (HC; n = 4), a high-cholesterol diet plus α-tocopherol (HC+AT; n = 2), and chow plus α-tocopherol (AT, n = 2). Original magnification ×100. Journal of Vascular Surgery 2008 48, 986-993DOI: (10.1016/j.jvs.2008.04.010) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions