Endothelial cells are susceptible to rapid siRNA transfection and gene silencing ex vivo Nicholas D. Andersen, MD, Atish Chopra, Thomas S. Monahan, MD,

Slides:

Advertisements

Similar presentations

Professionalism and the work-life balance Thomas S. Huber, MD, PhD Journal of Vascular Surgery Volume 60, Issue 4, Pages (October 2014) DOI:

Advertisements

Ultrasound-guided percutaneous delivery of tissue-engineered endothelial cells to the adventitia of stented arteries controls the response to vascular.

Eddie L. Hoover, M. D. , Margery Ross, Ph. D. , Kazam Fani, M. D

Without prejudice Journal of Vascular Surgery

Vein wall re-endothelialization after deep vein thrombosis is improved with low- molecular-weight heparin Daria K. Moaveni, MD, Erin M. Lynch, MS, Cathy.

Mesenchymal stem cells attenuate angiotensin II-induced aortic aneurysm growth in apolipoprotein E-deficient mice Ryotaro Hashizume, MD, Aika Yamawaki-Ogata,

Decellularized vein as a potential scaffold for vascular tissue engineering Patrick J Schaner, MD, Niels D Martin, MD, Thomas N Tulenko, PhD, Irving M.

Wei Zhou, MD, Hong Chai, MD, PhD, Andy Courson, BS, Peter H

Undifferentiated mesenchymal stem cells seeded on a vascular prosthesis contribute to the restoration of a physiologic vascular wall Alain Mirza, MD,

Differential expression of Hedgehog/Notch and transforming growth factor-β in human abdominal aortic aneurysms Adam J. Doyle, MD, Eileen M. Redmond,

Early hypercholesterolemia contributes to vasomotor dysfunction and injury associated atherogenesis that can be inhibited by nitric oxide Kathleen G.

Growth hormone–releasing peptide ghrelin inhibits homocysteine-induced endothelial dysfunction in porcine coronary arteries and human endothelial cells

Improved vessel preservation after 4 days of cold storage: Experimental study in rat arteries Birgit Zatschler, Peter Dieterich, PhD, Bianca Müller,

Celiprolol reduces the intimal thickening of autogenous vein grafts via an enhancement of nitric oxide function through an inhibition of superoxide production

David G. Cable, MD, James A. Caccitolo, MD, Eric A

Wei Zhou, MD, Hong Chai, MD, PhD, Peter H. Lin, MD, Alan B

VCAM-1 siRNA reduces neointimal formation after surgical mechanical injury of the rat carotid artery Yanming Qu, MD, Xiangen Shi, MD, Hongwei Zhang,

Tissue engineering applications to vascular bypass graft development: The use of adipose-derived stem cells Paul DiMuzio, MD, Thomas Tulenko, PhD Journal.

Gene transduction into aortic wall using plasmid-loaded cationized gelatin hydrogel- coated polyester stent graft Hongshan Zhong, MD, Osamu Matsui, MD,

Inhibitory Effects of Calcitonin Gene-Related Peptides on Experimental Vein Graft Disease Xiaoning Zhang, MD, Jian Zhuang, MD, Hongsui Wu, MB, Zhihong.

Gene transduction into aortic wall using plasmid-loaded cationized gelatin hydrogel- coated polyester stent graft Hongshan Zhong, MD, Osamu Matsui, MD,

Inflammation-related induction of absent in melanoma 2 (AIM2) in vascular cells and atherosclerotic lesions suggests a role in vascular pathogenesis

Rapid recurrence of cystic adventitial disease in femoral artery and an etiologic consideration for the cyst Keiji Oi, MD, PhD, Tetsuya Yoshida, MD,

Association of smooth muscle cell phenotypes with extracellular matrix disorders in thoracic aortic dissection Lixin Wang, MD, PhD, Jing Zhang, MD, PhD,

Lucas P. Neff, MD, Bryan W. Tillman, MD, PhD, Saami K

A novel function for cadherin 11/osteoblast-cadherin in vascular smooth muscle cells: Modulation of cell migration and proliferation Thomas S. Monahan,

Hemagglutinating virus of Japan-liposome–mediated gene transfer of endothelial cell nitric oxide synthase inhibits intimal hyperplasia of canine vein.

Techniques to harvest diseased human peripheral arteries and measure endothelial function in an ex vivo model Ryan O. Lakin, MD, Weifei Zhu, PhD, Lindsay.

Increased connexin43 expression in human saphenous veins in culture is associated with intimal hyperplasia Sébastien Déglise, MD, David Martin, MS, Hervé.

Metformin stimulates ischemia-induced revascularization through an eNOS dependent pathway in the ischemic hindlimb mice model Noriko Takahashi, MD, Rei.

Decellularized vein as a potential scaffold for vascular tissue engineering Patrick J Schaner, MD, Niels D Martin, MD, Thomas N Tulenko, PhD, Irving M.

Simvastatin pretreatment enhances ischemia-induced neovascularization and blood flow recovery in streptozotocin-treated mice Po-Hsun Huang, MD, PhD,

Daniel J. Wong, MD, Daniel Y. Lu, MD, Clinton D

Overexpression of transforming growth factor–β1 correlates with increased synthesis of nitric oxide synthase in varicose veins Theresa Jacob, PhD, Anil.

Mohamed A. Zayed, MD, PhD, Ronald L. Dalman, MD, Jason T. Lee, MD

Ambient pressure upregulates nitric oxide synthase in a phosphorylated-extracellular regulated kinase– and protein kinase C–dependent manner Angela G.

Endothelium-dependent vasorelaxations in response to aggregating platelets are impaired in reversed vein grafts Kimihiro Komori, MD, Peter Gloviczki,

Desmuslin gene knockdown causes altered expression of phenotype markers and differentiation of saphenous vein smooth muscle cells Ying Xiao, PhD, Zhibin.

Mechanosensitive transient receptor potential vanilloid type 1 channels contribute to vascular remodeling of rat fistula veins Yih-Sharng Chen, MD, PhD,

Sarpogrelate hydrochloride reduced intimal hyperplasia in experimental rabbit vein graft Akio Kodama, MD, Kimihiro Komori, MD, PhD, Keisuke Hattori, MD,

The first operation on the profunda femoris artery

Spatiotemporal expression and localization of matrix metalloproteinas-9 in a murine model of thoracic aortic aneurysm Jeffrey A. Jones, PhD, John R.

Without prejudice Journal of Vascular Surgery

Ultrasound-guided percutaneous delivery of tissue-engineered endothelial cells to the adventitia of stented arteries controls the response to vascular.

Harry M. Salinas, MD, Saiqa I. Khan, MD, Michael C

Role of hemodynamic forces in the ex vivo arterialization of human saphenous veins Xavier Berard, MD, PhD, Sébastien Déglise, MD, Florian Alonso, PhD,

Surgical marking pen dye inhibits saphenous vein cell proliferation and migration in saphenous vein graft tissue Shinsuke Kikuchi, MD, Richard D. Kenagy,

Gene delivery to in situ veins: Differential effects of adenovirus and adeno-associated viral vectors Mohammad H. Eslami, MD, Sidhu P. Gangadharan, MD,

IP161. Development of a Hybrid Cryogel-Coated Prosthetic Vascular Graft for Delivery of Targeted Gene Therapies Cindy Huynh, MD, Ting-Yu Shih, MS, Amruta.

Adventitial versus intimal liposome-mediated ex vivo transfection of canine saphenous vein grafts with endothelial nitric oxide synthase gene Manju Kalra,

Gene expression changes evoked in a venous segment exposed to arterial flow Deborah Abeles, MD, Stephanie Kwei, MD, George Stavrakis, MS, Yuzhi Zhang,

The effects of ginsenoside Rb1 on endothelial damage and ghrelin expression induced by hyperhomocysteine Zhiwei Xu, PhD, Taohua Lan, MD, Weikang Wu,

The future of vascular surgery

Aortic wall cell proliferation via basic fibroblast growth factor gene transfer limits progression of experimental abdominal aortic aneurysm Katsuyuki.

Correction Journal of Vascular Surgery

Expression of Endothelin-1 and Constitutional Nitric Oxide Synthase Messenger RNA in Saphenous Vein Endothelial Cells Exposed to Arterial Flow Shear Stress

Retaining perivascular tissue of human saphenous vein grafts protects against surgical and distension-induced damage and preserves endothelial nitric.

Development of a Hybrid Cryogel-Coated Prosthetic Vascular Graft for Delivery of Targeted Gene Therapies Cindy Huynh, MD, Ting-Yu Shih, MS, Amruta Samant,

Reply Journal of Vascular Surgery

Recurrence of a popliteal venous aneurysm

Free radical attenuation prevents thrombosis and enables photochemical inhibition of vein graft intimal hyperplasia Giuseppe R Nigri, MD, PhD, Sylvie.

Reviewer Acknowledgment

Thrombotic mesenteric ischemia due to aortic dissection

Identification and characterization of microRNAs in vascular smooth muscle cells from patients with abdominal aortic aneurysms Bernice Lai Yee Cheuk,

Effect of Vein Graft Harvesting on Endothelial Nitric Oxide Synthase and Nitric Oxide Production Michael R. Dashwood, PhD, Kay Savage, PhD, Audrey Dooley,

Activation of transglutaminase type 2 for aortic wall protection in a rat abdominal aortic aneurysm formation Takashi Munezane, MD, Tomomi Hasegawa,

Bhagwan Satiani, MD, MBA, Thomas E. Williams, MD, PhD, Michael R

Application of a Cryogel-Coated Prosthetic Vascular Graft Material for Delivery of Targeted Gene Therapies in a Rabbit Model Cindy Huynh, Ting-Yu Shih,

Panagiotis Kougias, MD, Hong Chai, MD, PhD, Peter H. Lin, MD, Alan B

Gene expression changes evoked in a venous segment exposed to arterial flow Deborah Abeles, MD, Stephanie Kwei, MD, George Stavrakis, MS, Yuzhi Zhang,

Presentation transcript:

Endothelial cells are susceptible to rapid siRNA transfection and gene silencing ex vivo Nicholas D. Andersen, MD, Atish Chopra, Thomas S. Monahan, MD, Junaid Y. Malek, MD, Monica Jain, BS, Leena Pradhan, PhD, Christiane Ferran, MD, PhD, Frank W. LoGerfo, MD Journal of Vascular Surgery Volume 52, Issue 6, Pages 1608-1615 (December 2010) DOI: 10.1016/j.jvs.2010.06.169 Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 1 Custom pressure chamber designed to expose standard tissue culture plates to hyperbaric pressure. Journal of Vascular Surgery 2010 52, 1608-1615DOI: (10.1016/j.jvs.2010.06.169) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 2 Endothelial siRNA delivery exceeds medial and adventitial delivery and is increased using distending pressure transfection. (A) Vein segments transfected with Cy5 siRNA demonstrate discrete areas of blue fluorescence independent of the red/green autofluorescence of the elastic fibers (representative micrograph, ×600). Quantitation of blue pixels by tissue layer demonstrates (B) maximal Cy5 fluorescence in the endothelium after distending pressure (DP) transfection as compared with the soak and hyperbaric pressure (HBP) transfections (*P < .05 for both comparisons). (C-E) Cy5 fluorescence in the medial layer, adventitial layer, and total vessel wall was statistically equivalent between the three transfection conditions. (F) Cumulative analysis of images from all transfection conditions demonstrates greater Cy5 fluorescence in the endothelial layer as compared to the media and the adventitia (*P < .05 for both comparisons). For all data, n = 5 to 6 vein segments per condition. Journal of Vascular Surgery 2010 52, 1608-1615DOI: (10.1016/j.jvs.2010.06.169) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 3 Endothelial glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein knockdown exceeds medial and adventitial GAPDH knockdown and is increased using distending pressure transfection. (A) Immunohistochemistry demonstrates GAPDH knockdown within all tissue layers after distending pressure (DP) transfection with GAPDH siRNA (panel b) as compared to vein segments treated with control siRNA (panel a). Protein knockdown is specific to GAPDH as CD31 (panels c,d) and actin (panels e,f) levels are preserved. Quantitation demonstrates (B) greater GAPDH knockdown in the endothelium using distending pressure as compared with hyperbaric (HBP) and non-pressurized (soak) transfection. (C) Medial/adventitial GAPDH levels were significantly reduced after distending pressure transfection, but not with hyperbaric or non-pressurized transfection. (D) Cumulative analysis of all transfections revealed greater knockdown in the endothelium as compared with the media/adventitia. n = 3 vein segments per condition. Micrographs (×400) correspond to one representative image of three experiments performed. (* denotes P < .05 for comparisons). Journal of Vascular Surgery 2010 52, 1608-1615DOI: (10.1016/j.jvs.2010.06.169) Copyright © 2010 Society for Vascular Surgery Terms and Conditions

Fig 4 Endothelial specific nitric oxide synthase (eNOS) knockdown is achieved using all three transfection methods and is increased using distending pressure transfection. (A) Immunohistochemistry demonstrates greatest endothelial eNOS knockdown after distending pressure (DP) transfection with eNOS siRNA (panel b) as compared to vein segments treated with control siRNA (panel a). Protein knockdown is specific to eNOS as CD31 levels (panels c,d) are preserved. (B-E) Quantitation demonstrates greater eNOS knockdown using distending pressure as compared to hyperbaric (HBP) and non-pressurized (soak) transfection. n = 3 vein segments per condition. Micrographs (×400) correspond to one representative image of three experiments performed. (* denotes P < .05 for comparisons). Journal of Vascular Surgery 2010 52, 1608-1615DOI: (10.1016/j.jvs.2010.06.169) Copyright © 2010 Society for Vascular Surgery Terms and Conditions