Grafting an Acellular 3-Dimensional Collagen Scaffold Onto a Non-transmural Infarcted Myocardium Induces Neo-angiogenesis and Reduces Cardiac Remodeling

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Grafting an Acellular 3-Dimensional Collagen Scaffold Onto a Non-transmural Infarcted Myocardium Induces Neo-angiogenesis and Reduces Cardiac Remodeling Mohamed A. Gaballa, PhD, Julia N.E. Sunkomat, MD, Hoang Thai, MD, Eugene Morkin, MD, Gordon Ewy, MD, Steven Goldman, MD The Journal of Heart and Lung Transplantation Volume 25, Issue 8, Pages 946-954 (August 2006) DOI: 10.1016/j.healun.2006.04.008 Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions

Figure 1 Pressure–volume curves for the four groups. Solid line: treatment group (infarcted rats with collagen scaffold); dashed line: untreated sham and sham treated with collagen scaffold; dotted line: untreated infarcted groups (cryoinjury). The curve for sham with collagen scaffold is superimposed upon the untreated sham curve. Note that, 6 weeks after transplantation, the P-V curve is shifted to the left. N = 4 for each group. *p < 0.05. The Journal of Heart and Lung Transplantation 2006 25, 946-954DOI: (10.1016/j.healun.2006.04.008) Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions

Figure 2 (A) High magnification (original magnification ×40) of the H&E stain of the engrafted scaffold showing large vessel (arrows). These vessels are thick-walled and have multiple cell layers. (B) Same magnification as (A) of the H&E staining of infarcted myocardium without the scaffold. Note that the ischemia-induced vascularization is characterized by thin-walled vessels. The Journal of Heart and Lung Transplantation 2006 25, 946-954DOI: (10.1016/j.healun.2006.04.008) Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions

Figure 3 (A) A typical Factor VII staining for endothelial cells in cryoinjured heart with scaffold shows neo-vascularization in the scaffold (top) at 6 weeks after grating onto native myocardium (bottom). (B) A different field from the same section. The Journal of Heart and Lung Transplantation 2006 25, 946-954DOI: (10.1016/j.healun.2006.04.008) Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions

Figure 4 Vascular α-smooth-muscle actin staining. (A) Native (non-injured) myocardium (control). (B) Scaffold. Brown staining within the neo-vessels indicates the presence of mural cells 6 weeks after grafting. The Journal of Heart and Lung Transplantation 2006 25, 946-954DOI: (10.1016/j.healun.2006.04.008) Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions

Figure 5 Effects of scaffold grafting, with and without G-CSF administration, on vascular density. Vascular density increased by 8-fold with the scaffold alone and by 40% with scaffold and G-CSF treatment. *p < 0.05 infarcted + scaffold compared with infarcted alone. **p < 0.05 infarcted + scaffold + G-CSF compared with infarcted + scaffold. N = 5 for each group. The Journal of Heart and Lung Transplantation 2006 25, 946-954DOI: (10.1016/j.healun.2006.04.008) Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions

Figure 6 Coronary artery perfusion of isolated hearts with Evans blue. (A) Infarcted heart without scaffold (control). (B) Infarcted heart with scaffold. Note the neo-vasculature within the scaffold that perfuses blue, indicating a connection to the coronary arteries. The Journal of Heart and Lung Transplantation 2006 25, 946-954DOI: (10.1016/j.healun.2006.04.008) Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions

Figure 7 Micrographs showing Evans blue within myocardial vessels (white arrows, red color). H&E myocardial sections examined under fluorescence microscopy (original magnification ×40). (A) Non-infarcted myocardium. (B) 3D scaffold. The Journal of Heart and Lung Transplantation 2006 25, 946-954DOI: (10.1016/j.healun.2006.04.008) Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions

Figure 8 (A) Detection of cardiac myofibril bundle within the scaffold (left) by MF20 (A) and cardiac troponin T (B) immunohistochemical staining (brown, arrows) in the infarcted + scaffold groups. Native myocardium is shown on the right. (C) Control staining for sham rats (uninjured myocardium). The Journal of Heart and Lung Transplantation 2006 25, 946-954DOI: (10.1016/j.healun.2006.04.008) Copyright © 2006 International Society for Heart and Lung Transplantation Terms and Conditions