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

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Presentation transcript:

Vascular smooth muscle enhances functionality of tissue-engineered blood vessels in vivo Lucas P. Neff, MD, Bryan W. Tillman, MD, PhD, Saami K. Yazdani, PhD, Masood A. Machingal, MS, James J. Yoo, MD, PhD, Shay Soker, PhD, Brian W. Bernish, BS, Randolph L. Geary, MD, George J. Christ, PhD Journal of Vascular Surgery Volume 53, Issue 2, Pages 426-434 (February 2011) DOI: 10.1016/j.jvs.2010.07.054 Copyright © 2011 Society for Vascular Surgery Terms and Conditions

Fig 1 A, Hematoxylin and eosin (H&E) histology (original magnification ×100) of a preimplant dual-seeded tissue-engineered blood vessel (TEBV) reveals an endothelial monolayer (bottom) and multilayered smooth muscle cells (SMCs) on the extraluminal surface (top). Scale bar = 400 μm. Inset H&E histologic images of the (B) SMC layers (arrowhead) and (C) the endothelial monolayer (arrow) (original magnification ×400; scale bar = 100 μm). D, Operative photograph of TEBV implantation as an interposition graft of the common carotid artery, denoted as A on the top and bottom of the panel. Anastomotic suture lines are noted (arrows). E, A representative anterior-posterior subtraction angiography view of an implanted TEBV at 4 months after implantation. Examination of the proximal and distal anastomoses (arrows) demonstrates a slight narrowing due to the small luminal size discrepancy between the native vessel and TEBV. Journal of Vascular Surgery 2011 53, 426-434DOI: (10.1016/j.jvs.2010.07.054) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

Fig 2 Representative images (original magnification ×100) of (A, B, C, D) native artery control, the (E, F, G, H) dual-seeded tissue-engineered blood vessel (dsTEBV), and (I, J, K, L) endothelial cell-seeded TEBV (ecTEBV) after 4 months in vivo stained with (A, E, I) Movat, (B, F, J) hematoxylin and eosin (H&E), (C, G, K) α-actin, and (D, H, L) smooth muscle cell (SMC) myosin heavy chain (MHC). Representative images demonstrate (E, I) elastin fibers that define the internal and external elastic lamellae of the TEBV scaffold and increased overall cellularity of the (F) dsTEBV compared with the (J) ecTEBV on H&E histology as well as increased immunohistochemical staining positive for (G vs K) α-actin and (H vs L) MHC. Scale bar = 400 μm. Journal of Vascular Surgery 2011 53, 426-434DOI: (10.1016/j.jvs.2010.07.054) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

Fig 3 Representative images (original magnification ×400) of (A, B, C) native artery control, the (D, E, F) dual-seeded tissue-engineered blood vessel (TEBV), and (G, H, I) endothelial cell (EC)-seeded TEBV after 4 months in vivo stained with (A, D, G) hematoxylin and eosin (H&E), (B, E, H) α-actin, and (C, F, I) myosin heavy chain (MHC). Representative images demonstrate increased overall cellularity of (D) the dual-seeded vessel compared with (G) the EC-seeded vessel on H&E and eosin histology as well as increased immunohistochemical staining that is positive for (E vs H) α-actin and (F vs I) MHC. Scale bar = 100 μm. Journal of Vascular Surgery 2011 53, 426-434DOI: (10.1016/j.jvs.2010.07.054) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

Fig 4 Graphic comparison and statistical summary of percentage of cells with positive immunohistochemical staining in the media of the dual-seeded tissue-engineered blood vessel (dsTEBV) grafts and the endothelial cell-seeded TEBV (ecTEBV) grafts. As illustrated, the dsTEBVs revealed a greater percentage of cells in the TEBV wall that stained positive for α-actin and myosin heavy chain (MHC) compared with the ecTEBVs. The error bars show the standard error of the mean. Journal of Vascular Surgery 2011 53, 426-434DOI: (10.1016/j.jvs.2010.07.054) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

Fig 5 Representative tracings of pharmacologic response of (A, C) dual-seeded tissue-engineered blood vessels (dsTEBV) and (B, D) endothelial cell-seeded TEBV in response to (A, B) 10 μM 5-hydroxytryptamine and (C, D) 10 μM phenylephrine. The arrows indicate the application of agonists. Journal of Vascular Surgery 2011 53, 426-434DOI: (10.1016/j.jvs.2010.07.054) Copyright © 2011 Society for Vascular Surgery Terms and Conditions

Fig 6 Summary graph of the increased steady-state maximal contractile response that was observed in the dual-seeded tissue-engineered blood vessel (dsTEBV) relative to the endothelial cell-seeded TEBV (ecTEBV) in response to stimulation of the α1-adrenergic receptor subtype with phenylephrine (10 μM; P = .01) and the 5-hyroxytryptamine 2 (5-HT) receptor subtype with 5-HT (10 μM; P = .01). No significant difference was observed in the mean maximal contractile response to potassium chloride (KCl) between the two TEBV treatment groups (n = a mean of 3 rings per TEBV implanted). Error bars show the standard error of the mean. Journal of Vascular Surgery 2011 53, 426-434DOI: (10.1016/j.jvs.2010.07.054) Copyright © 2011 Society for Vascular Surgery Terms and Conditions