Reconstruction of pulmonary artery with porcine small intestinal submucosa in a lamb surgical model: Viability and growth potential Lorenzo Boni, MD,

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Presentation transcript:

Reconstruction of pulmonary artery with porcine small intestinal submucosa in a lamb surgical model: Viability and growth potential Lorenzo Boni, MD, Fariba Chalajour, MD, Takashi Sasaki, MD, Radhika Lal Snyder, BA, Walter D. Boyd, MD, R. Kirk Riemer, PhD, V. Mohan Reddy, MD The Journal of Thoracic and Cardiovascular Surgery Volume 144, Issue 4, Pages 963-969.e1 (October 2012) DOI: 10.1016/j.jtcvs.2012.07.024 Copyright © 2012 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 A, In situ macroscopic appearance of patched area 1 month after implantation. Arrows show patched area externally. There were no visible adherences around the patch area. The external caliber of the left pulmonary artery (LPA) was free from stricture formation and distortions. B, Macroscopic view of the luminal surface of the explanted patch 1 month after implantation shows the patched area in the middle of the specimen (the left pulmonary artery was opened longitudinally). The intimal layer was found covering the luminal surface of the constructed left pulmonary artery, including the patched area. MPA, Main pulmonary artery; SIS, small intestine submucosal patch. The Journal of Thoracic and Cardiovascular Surgery 2012 144, 963-969.e1DOI: (10.1016/j.jtcvs.2012.07.024) Copyright © 2012 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Computed tomographic angiographic scan with 3-dimensional reconstruction 6 months after patch implantation. The left pulmonary artery (LPA) was patent, and there were no signs of stenosis or aneurysm formation. The left pulmonary artery and right pulmonary artery (RPA) sizes were normal and comparable with each other, as was the distal arborization after each pulmonary arterial branch. Ao, Aorta; MPA, main pulmonary artery. The Journal of Thoracic and Cardiovascular Surgery 2012 144, 963-969.e1DOI: (10.1016/j.jtcvs.2012.07.024) Copyright © 2012 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Scanning electron microscopy of the luminal surface of the small intestine submucosal patch 1 month after implantation. The patch area was covered with confluent cells with the morphologic characteristics of endothelial cells. Original magnification ×1600. The Journal of Thoracic and Cardiovascular Surgery 2012 144, 963-969.e1DOI: (10.1016/j.jtcvs.2012.07.024) Copyright © 2012 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 Russell-Movat pentachrome staining of small intestine submucosal (SIS) patch explants. The cross sections of small intestine submucosal patch tissue explants after 1 month (A) shows cellularization of intima and media (arrows; B). Formation of extracellular matrix is confirmed by production of collagen (yellow) in subintimal layer and proteoglycans (blue) in the tunica media; however, elastin fibers (black) are only detected in the wall of native pulmonary artery (C), not in the small intestine submucosal patch. Comparison of tunica adventitia in the small intestine submucosal explants (D) with native pulmonary artery (E) shows migration of inflammatory cells into the patch, thus increasing the thickness of patch. The cross section of small intestine submucosal patch tissue explants after 3 months shows increasing the size of pulmonary arterial lumen (F) and formation of blood vessels in the tunica media (arrows; G). Partial replacement of proteoglycans by collagen is evidence that matrix production in the small intestine submucosal patch tunica media is dynamic. Distribution of proteoglycans in small intestine submucosal patch follows a pattern similar to that in native pulmonary artery (H). No elastin fiber is observed in the small intestine submucosal patch after 3 months. Although the number of inflammatory cells in the tunica adventitia of the 3-month-old small intestine submucosal patch (I) is higher than that in the native pulmonary artery (J), it is dramatically decreased relative to a 1-month sample (D). The cross section of small intestine submucosal patch tissue explants after 6 months shows reduction of patch thickness (K) and formation of elastin fiber in the patch (L) with a pattern similar to that in native pulmonary artery (M). The similarity in the distribution of cells and extracellular matrix between small intestine submucosal patch tunica adventitia (N) and native pulmonary artery (O) indicates the reduction of inflammatory response after 6 months. Original magnifications ×16 (A, F, and K) and ×400 (B-E, G-J, and L-O). The Journal of Thoracic and Cardiovascular Surgery 2012 144, 963-969.e1DOI: (10.1016/j.jtcvs.2012.07.024) Copyright © 2012 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 Immunohistologic analysis of porcine small intestine submucosal patch explants. Formation of tunica intima was confirmed by expression of endothelial cell marker von Willebrand factor (vWF) in the cells covering the luminal side of grafted patch 1 month, 3 months, and 6 months after implantation (A, E, and I, respectively). Expression of smooth muscle actin (SMA) was observed in the medial layer of small intestine submucosal patch after 1 month (B), 3 months (F), and 6 months (J); however, the expression of elastin was observed in the medial layer of only 6-month small intestine submucosal patch (K), not in the 1-month (C) and 3-month (G) samples. The expression of stem cell marker CD117 (c-Kit) was identified in the small intestine submucosal patch 1 month (D), 3 months (H), and 6 months (L) after implantation; however, the number and distribution of CD117+ cells increased with time, indicating the remodeling potential of small intestine submucosal patch. Original magnification ×1000 (A-L). The Journal of Thoracic and Cardiovascular Surgery 2012 144, 963-969.e1DOI: (10.1016/j.jtcvs.2012.07.024) Copyright © 2012 The American Association for Thoracic Surgery Terms and Conditions

Figure E1 Activation of angiogenesis in small intestine submucosal patch. Vascularization of small intestine submucosal patch was confirmed by expression of endothelial marker von Willebrand factor (vWF) in the lumen of newly formed blood vessels in 1-month (A), 3-month (B), and 6-month (C) explants. The highest density of blood vessels was observed 3 months after patch implantation (B), demonstrating the proliferative phase of patch repair, whereas regression of blood vessels in the medial layer was identified in 6-month samples (C), indicating maturation and remodeling of grafted patch. Original magnification ×400 (A-C). The Journal of Thoracic and Cardiovascular Surgery 2012 144, 963-969.e1DOI: (10.1016/j.jtcvs.2012.07.024) Copyright © 2012 The American Association for Thoracic Surgery Terms and Conditions