Development of Decellularized Aortic Valvular Conduit Coated by Heparin–SDF-1α Multilayer Jingxin Zhou, PhD, Xiaofeng Ye, MD, PhD, Zhe Wang, MD, Jun.

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Development of Decellularized Aortic Valvular Conduit Coated by Heparin–SDF-1α Multilayer Jingxin Zhou, PhD, Xiaofeng Ye, MD, PhD, Zhe Wang, MD, Jun Liu, MD, Busheng Zhang, PhD, Jiapei Qiu, MD, Yanjun Sun, MD, Haiqing Li, MD, PhD, Qiang Zhao, MD The Annals of Thoracic Surgery Volume 99, Issue 2, Pages 612-618 (February 2015) DOI: 10.1016/j.athoracsur.2014.09.001 Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Morphology of decellularized rat aortic valvular conduits. H&E staining shows the cellular components of (A) the decellularized rat aortic conduits and (B) the aortic valve leaflets, which were completely removed. (C, D) Scanning electron micrographic (SEM) images of the decellularized graft show reserved collagen fibers with no cellular components remaining. Scale bars: (A) 100 μm; (B) 200 μm; (C) 50 μm; (D) 20 μm. The Annals of Thoracic Surgery 2015 99, 612-618DOI: (10.1016/j.athoracsur.2014.09.001) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 Quantification of lactate dehydrogenase activity. The number of adhered platelets on decellularized valvular conduits (DVCs) as determined by quantification of lactate dehydrogenase activity. ∗Corresponds to a p < 0.05 of polyelectrolyte multilayer films–treated DVCs (PEM-DVC) in comparison with untreated decellularized valvular conduits (UnDVC). The Annals of Thoracic Surgery 2015 99, 612-618DOI: (10.1016/j.athoracsur.2014.09.001) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 Attachment, proliferation, and migration of BMSCs. More attached cells on polyelectrolyte multilayer films–treated decellularized valvular conduits (PEM-DVC) at 6 hours, represented by OD value. (A) The cellular number at the 24-hour and 48-hour time points suggests better proliferation of BMSCs on PEM-DVC compared with untreated decellularized valvular conduits (UnDVC). (B) In comparison with UnDVC, PEM-DVC significantly increased migration of rat BMSCs to the lower surface of the membrane of Transwell chambers. ∗Corresponds to a p < 0.05 of PEM-DVC in comparison with Un-DVC. The Annals of Thoracic Surgery 2015 99, 612-618DOI: (10.1016/j.athoracsur.2014.09.001) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Functional evaluation after transplantation. (A) Color Doppler imaging shows the conduit in the abdominal cavity with pulsatile flow through the graft but no blood flow in the native aorta between the two anastomoses. (B) Three-dimensional reconstruction computed tomography (CT) reveals patency and no blood flow in the native aorta between the two anastomoses. (∗ = DVC graft, # = native aorta.) The Annals of Thoracic Surgery 2015 99, 612-618DOI: (10.1016/j.athoracsur.2014.09.001) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 H&E staining of polyelectrolyte multilayer films–treated decellularized valvular conduits (PEM-DVC) 4 weeks after implantation reveals (A) confluent luminal cell layer but (B) no endothelialization in untreated decellularized valvular conduits (UnDVC). The adventitia of both groups was covered by fibrous hyperplasia with no resident cells infiltrating into the media. ∗Luminal side of DVC. Scale bars: 100 μm. The Annals of Thoracic Surgery 2015 99, 612-618DOI: (10.1016/j.athoracsur.2014.09.001) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

Fig 6 Immunohistochemical analysis of explanted grafts 4 weeks after implantation. (A) Considerably more von Willebrand factor+ and (B) CD34+ cells could be found on the luminal side of polyelectrolyte multilayer films–treated decellularized valvular conduits (PEM-DVC) in contrast to (C, D) untreated decellularized valvular conduits (UnDVC). Scale bars: 50 μm. The Annals of Thoracic Surgery 2015 99, 612-618DOI: (10.1016/j.athoracsur.2014.09.001) Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions