Altered Oxidative Stress Responses and Increased Type I Collagen Expression in Bicuspid Aortic Valve Patients Julie A. Phillippi, PhD, Michael A. Eskay,

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Altered Oxidative Stress Responses and Increased Type I Collagen Expression in Bicuspid Aortic Valve Patients Julie A. Phillippi, PhD, Michael A. Eskay, BS, Adam A. Kubala, BS, Bruce R. Pitt, PhD, Thomas G. Gleason, MD The Annals of Thoracic Surgery Volume 90, Issue 6, Pages 1893-1898 (December 2010) DOI: 10.1016/j.athoracsur.2010.07.069 Copyright © 2010 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Expression of vascular endothelial growth factor (VEGF) gene expression in primary human ascending aortic smooth muscle cells (SMC). Cells from nonaneurysmal patients (n = 11), aneurysmal with tricuspid aortic valve (TAV) patients (n = 8), and aneurysmal with bicuspid aortic valve (BAV) patients (n = 9) were cultured in the presence or absence of 600 μM t-butyl hydroperoxide (tBHP [black bars]) for 6 hours. For all experiments, bars represent mean ± SEM. *Significant from control (−tBHP [white bars]), p less than 0.01. The Annals of Thoracic Surgery 2010 90, 1893-1898DOI: (10.1016/j.athoracsur.2010.07.069) Copyright © 2010 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Cell viability in the presence of oxidative stress. Primary human ascending aortic smooth muscle cells (SMC) from bicuspid aortic valve (BAV) patients (dotted line) were cultured in the presence of 10 ng/mL vascular endothelial growth factor (VEGF) before exposure to 0 to 4,800 μM t-butyl hydroperoxide (tBHP) for 3 hours and compared with SMCs from nonaneurysmal patients (solid line). Lines represent mean ± SEM, n = 3 and n = 4 for nonaneurysmal and BAV, respectively. The Annals of Thoracic Surgery 2010 90, 1893-1898DOI: (10.1016/j.athoracsur.2010.07.069) Copyright © 2010 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Type I collagen gene expression. (A) Human ascending aorta from nonaneurysmal patients (n = 23), aneurysmal with tricuspid aortic valve (TAV) patients (n = 21), and aneurysmal with bicuspid aortic valve (BAV) patients (n = 40). Bars represent mean ± SEM. *Significant from nonaneurysmal for p less than 0.05. The Annals of Thoracic Surgery 2010 90, 1893-1898DOI: (10.1016/j.athoracsur.2010.07.069) Copyright © 2010 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Cell viability of primary ascending aortic smooth muscle cells (SMC) from wild-type mice (solid line) and MT−/− mice (dashed line). *Significant from wild-type, p less than 0.02. (MT = metallothionein; tBHP = t-butyl hydroperoxide.) The Annals of Thoracic Surgery 2010 90, 1893-1898DOI: (10.1016/j.athoracsur.2010.07.069) Copyright © 2010 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Expression of vascular endothelial growth factor (VEGF) gene expression in primary ascending aortic smooth muscle cells (SMC) from wild-type mice and MT−/− mice. The SMCs were cultured in the presence (solid bars) and absence (open bars) of 600 μM t-butyl hydroperoxide (tBHP) for 6 hours. *Significant from wild-type, p less than 0.05. (MT = metallothionein.) The Annals of Thoracic Surgery 2010 90, 1893-1898DOI: (10.1016/j.athoracsur.2010.07.069) Copyright © 2010 The Society of Thoracic Surgeons Terms and Conditions