New engineering treatment of bovine pericardium confers outstanding resistance to calcification in mitral and pulmonary implantations in a juvenile sheep.

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New engineering treatment of bovine pericardium confers outstanding resistance to calcification in mitral and pulmonary implantations in a juvenile sheep model  Christian P. Brizard, MD, MS, Johann Brink, MBBS, Steven B. Horton, PhD, Glenn Anthony Edwards, BVSc, MANZCSc, John C. Galati, PhD, BSc, William M.L. Neethling, PhD, FACA  The Journal of Thoracic and Cardiovascular Surgery  Volume 148, Issue 6, Pages 3194-3201 (December 2014) DOI: 10.1016/j.jtcvs.2014.08.002 Copyright © 2014 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Epicardial echocardiography. A, Long axis study of the mitral valve. The white arrow points at the posterior leaflet bearing the hyperechogenic autologous pericardial patch. B, Long axis study of the mitral valve. The white arrow points at the posterior leaflet that does not display hyperechogenicity. C, Long axis of the pulmonary valve. The white arrow points at the unremarkable autologous patch. D, Short axis of the pulmonary valve. The white arrow points at the CardioCel patch, thin and flexible. The Journal of Thoracic and Cardiovascular Surgery 2014 148, 3194-3201DOI: (10.1016/j.jtcvs.2014.08.002) Copyright © 2014 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Macroscopy. A and B, both the autologous and the CardioCel patches have the same macroscopic aspect. C and D, Both the autologous and the CardioCel are flexible. The CardioCel patch after 8 months implantation is remarkably comparable to a fresh patch. The Journal of Thoracic and Cardiovascular Surgery 2014 148, 3194-3201DOI: (10.1016/j.jtcvs.2014.08.002) Copyright © 2014 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Hematoxylin and eosin. A, B, C, and D, All slides show a central layer made of the original patch and 2 outer layers of new collagen. An endothelial layer is continuous on the outer surface of the new collagen layers. Some mononucleated cells and lymphocytes can be identified in the autologous group mostly the mitral slide (A). The Journal of Thoracic and Cardiovascular Surgery 2014 148, 3194-3201DOI: (10.1016/j.jtcvs.2014.08.002) Copyright © 2014 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 Factor VIII immunohistochemistry. A, B, C, and D, The presence of a continuous endothelium in all groups on the outer margin of the neocollagen layers. The Journal of Thoracic and Cardiovascular Surgery 2014 148, 3194-3201DOI: (10.1016/j.jtcvs.2014.08.002) Copyright © 2014 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 Picrosirius red. In the control group ([A] mitral and [C] pulmonary) the outer layers are made of new collagen, predominantly collagen I, of the same thickness than the central layer (autologous pericardial patch). The outer layers are much less dense than the autologous patch and contain some immature collagen III in green. In the CardioCel (Admedus, Brisbane, Australia) group ([B] mitral and [D] pulmonary, the outer layers are very dense but thin and made exclusively of collagen I. The Journal of Thoracic and Cardiovascular Surgery 2014 148, 3194-3201DOI: (10.1016/j.jtcvs.2014.08.002) Copyright © 2014 The American Association for Thoracic Surgery Terms and Conditions

Figure 6 Trichrome staining with differential color adjustment demonstrating endothelial cells on the surface followed by a mixture of smooth muscle cells (confirmed by the smooth muscle cell α actin immunohistochemistry; photo not shown) and interstitial cells in the newly formed collagen. A and C, Control groups. B and D, CardioCel (Admedus, Brisbane, Australia). Note the difference in the distribution of the cells between groups. (High contrast light microscopy with; original magnification × 100). The Journal of Thoracic and Cardiovascular Surgery 2014 148, 3194-3201DOI: (10.1016/j.jtcvs.2014.08.002) Copyright © 2014 The American Association for Thoracic Surgery Terms and Conditions