Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 © 2011 S. Karger AG, Basel

Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 Fig. 1. Dacron and C-ECM patches. a Macroscopic photo of the patches. Dacron is on the left and C-ECM is on the right. b Approximate placement of a 6-mm patch in the RVOT shown on excised RV free wall. SEM of Dacron: ×30 (c) and ×1,000 (d). SEM of C-ECM with endocardium on the right side: ×30 (e) and ×1,000 (f). © 2011 S. Karger AG, Basel

Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 Fig. 2. Macroscopic images of the rat heart with the patched area at 2, 4, 8, and 16 weeks postsurgery are enclosed in red ovals. The C-ECM patch decreased in thickness as it remodeled. Fibrous adhesions can be seen at 2 and 4 weeks on the Dacron-patched hearts. The adhesions were removed from the hearts at the later time points so that the patch could be better visualized. C-ECM at 2 (a), 4 (b), 8 (c), and 16 weeks (d) after fixation. Dacron at 2 (e), 4 (f), 8 (g), and 16 weeks (h). © 2011 S. Karger AG, Basel

Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 Fig. 3. Masson’s trichrome C-ECM patch at 2, 4, 8, and 16 weeks postsurgery. The C-ECM patch recellularizes and remodels over 16 weeks whereas there are few changes in the Dacron-patched hearts over the 16 weeks. C-ECM at 2 (a), 4 (b), 8 (c), and 16 weeks (d). e C-ECM from the inset in d at 16 weeks. Dacron at 2 (f), 4 (g), 8 (h), and 16 weeks (i). j Dacron from the inset in i at 16 weeks. Scale bar = 500 µm. RV and LV intracavitary spaces are labeled where appropriate. © 2011 S. Karger AG, Basel

Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 Fig. 4. Masson’s trichrome and IF staining of the Dacron-patched area at 16 weeks showing little cellular infiltrate of the tissue surrounding the patch and no cellular infiltrate into the Dacron weave. There is an intact endothelium on the endocardial surface but no cardiomyocytes surrounding patch. a ×2, Masson’s trichrome. b ×10 magnification with transmission light capture of the inset in a. c ×60 magnification of the inset in b. vWF = Green; α-actinin = red; DRAQ5 = blue. Scale bar = 100 µm. © 2011 S. Karger AG, Basel

Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 Fig. 5. IF staining of the remodeled C-ECM patch showing vWF-positive endothelium and vasculature and SMMHCII-positive smooth muscle throughout the middle of the patch at 16 weeks. a ×10. b ×60 magnification of inset B in a. c ×60 magnification of inset C in a. d ×60 magnification of inset D in a. vWF = Green; SMMHCII = red; DRAQ5 = blue. Scale bar = 100 µm. © 2011 S. Karger AG, Basel

Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 Fig. 6. IF staining of the remodeled C-ECM patch showing α-actinin-positive cardiomyocytes in the middle of the patch on the epicardial surface at 16 weeks. α-Actinin = Red; nuclei = blue. a ×10. b ×40 magnification of the inset in a. Scale bar = 100 µm. © 2011 S. Karger AG, Basel

Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 Fig. 7. IF staining of the remodeled C-ECM patch showing α-actinin and cardiac troponin T-positive cardiomyocytes with connexin 43-labeled gap junctions, vWF-labeled endothelium, and SMMHCII-labeled smooth muscle in the middle of the patched area on the endocardial surface at 16 weeks. a ×10. vWF = Green; α-actinin = red; DAPI = blue. Scale bar = 200 µm. b–e Magnifications of the inset area on sequential slides. Scale bar = 50 µm. b ×63. α-Actinin = Green; cardiac troponin T = red; DRAQ5 = blue. c ×60. vWF = Green; α-actinin = red; DAPI = blue. d ×63. Connexin 43 = Green; α-actinin = red; DRAQ5 = blue. e ×60. SMMHCII = Green; α-actinin = red; DAPI = blue. © 2011 S. Karger AG, Basel

Right Ventricular Outflow Tract Repair with a Cardiac Biologic Scaffold Cells Tissues Organs 2012;195:159–170 - DOI:10.1159/000331400 Fig. 8. Echocardiographic analysis at presurgery and 4 and 16 weeks postsurgery with one standard deviation. a RV minimum diameter. b RV maximum diameter. c LV minimum diameter. d LV maximum diameter. e LV fractional area shortening diameter. For the C-ECM patch, none of the measures were different from presurgery values at either 4 or 16 weeks. For the Dacron patch the RV minimum and maximum diameters had dilated by 16 weeks. * p < 0.05 compared to presurgery; ** p < 0.05 compared to presurgery and 4 weeks postsurgery. © 2011 S. Karger AG, Basel