Michael A. Rosenbaum, MD, Keiko Miyazaki, MD, Scott M. Colles, Linda M

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Antioxidant therapy reverses impaired graft healing in hypercholesterolemic rabbits Michael A. Rosenbaum, MD, Keiko Miyazaki, MD, Scott M. Colles, Linda M. Graham, MD Journal of Vascular Surgery Volume 51, Issue 1, Pages 184-193 (January 2010) DOI: 10.1016/j.jvs.2009.08.061 Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 1 To measure tissue cholesterol content, lipids were extracted from (A) aortic and (B) graft samples and tissue cholesterol levels were determined using high-performance liquid chromatography. Levels are reported as μg/mg tissue (wet weight) and expressed as the mean ± standard error for each tissue segment and rabbit group: chow (n = 3), chow plus N-acetylcysteine (NAC; n = 2), chow plus probucol (n = 3), high cholesterol (HC, n = 3), HC plus NAC (n = 3), and HC plus probucol (n = 3). *P < .05. #P < .005. ††Data from previous study.10 Journal of Vascular Surgery 2010 51, 184-193DOI: (10.1016/j.jvs.2009.08.061) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 2 Plasma thiobarbituric acid reactive substance (TBARS) levels were determined 6 weeks postoperatively and reported as nmol MDA/mL. Values are expressed as the mean ± standard error for each rabbit group: chow (n = 7), chow plus N-acetylcysteine (NAC; n = 6), chow plus probucol (n = 8), high cholesterol (HC, n = 6), HC plus NAC (n = 7), and HC plus probucol (n = 8). *P < .05 between chow and HC groups. ††Data from previous study.10 Journal of Vascular Surgery 2010 51, 184-193DOI: (10.1016/j.jvs.2009.08.061) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 3 Endothelialization of expanded polytetrafluoroethylene (ePTFE) grafts and total luminal area were determined by scanning electron microscopy. A, Endothelialization 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 each group: chow (n = 10), chow plus N-acetylcysteine (NAC; n = 4), chow plus probucol (n = 5), high cholesterol (HC, n = 11), HC plus NAC (n = 4), and HC plus probucol (n = 5). *P < .05 between chow and chow plus probucol or HC groups. #P < .05 between HC and HC plus NAC groups. B, Anastomotic intimal hyperplasia was measured on histologic sections between 1.5 and 2.5 mm from the graft end and expressed as the mean ± standard error for each group: chow (n = 7), chow plus NAC (n = 4) chow plus probucol (n = 5), HC (n = 4), HC plus NAC (n = 4), and HC plus probucol (n = 5). *P < .05 between chow and HC groups. ††Data from previous study.10 Journal of Vascular Surgery 2010 51, 184-193DOI: (10.1016/j.jvs.2009.08.061) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 4 To show macrophage infiltration, graft sections underwent perfusion-fixation with 4% paraformaldehyde and were processed for immunohistochemistry and stained using mouse anti-rabbit macrophage antibody (RAM-11). Representative sections (original magnification ×100) are shown from each rabbit group: (A) chow diet (n = 6), (B) high cholesterol (HC) diet (n = 5), (C) chow plus N-acetylcysteine (NAC; n = 3), (D) HC plus NAC (n = 4), (E) chow plus probucol (n = 5), and (F) HC plus probucol (n = 5). ††Data from previous study.10 Journal of Vascular Surgery 2010 51, 184-193DOI: (10.1016/j.jvs.2009.08.061) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 5 Nitrotyrosine staining. After perfusion-fixation with 4% paraformaldehyde, graft sections were processed for immunohistochemistry and stained using mouse anti-human nitrotyrosine antibody. Representative sections (original magnification ×100) are shown from rabbits fed (A) regular chow diet (n = 3), (B) high cholesterol (HC) diet (n = 5), (C) chow plus N-acetylcysteine (NAC; n = 2), (D) HC plus NAC (n = 3), (E) chow plus probucol (n = 5), and (F) HC plus probucol (n = 5). Journal of Vascular Surgery 2010 51, 184-193DOI: (10.1016/j.jvs.2009.08.061) Copyright © 2010 Society for Vascular Surgery Terms and Conditions