A novel vascularized patch enhances cell survival and modifies ventricular remodeling in a rat myocardial infarction model Qi Zhou, MD, PhD, Jian-Ye.

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A novel vascularized patch enhances cell survival and modifies ventricular remodeling in a rat myocardial infarction model Qi Zhou, MD, PhD, Jian-Ye Zhou, MS, Zhe Zheng, MD, PhD, Hao Zhang, MD, PhD, Sheng-Shou Hu, MD The Journal of Thoracic and Cardiovascular Surgery Volume 140, Issue 6, Pages 1388-1396.e3 (December 2010) DOI: 10.1016/j.jtcvs.2010.02.036 Copyright © 2010 Terms and Conditions

Figure 1 Angiogenesis assessed based on microvessel density and vascular endothelial growth factor (VEGF) protein evaluated by means of Western blotting. The microvessels in the myocardium were characterized with immunohistochemistry by using anti–von Willebrand factor antibody. The scarred myocardium in rats with myocardial infarction (MI) was excised at 1 (A, D, and G), 2 (B, E, and H), and 4 (C, F, and I) weeks after polylactic acid-co-glycolic acid–mesenchymal stem cell (PLGA–MSC) transplantation (A, B, and C) or transplantation of omentum alone (D, E, and F) or omentum-based PLGA–MSC-supporting patch transplantation (G, H, and I). J, Microvessel density was quantified as the numbers in blood vessel lumen per square millimeter (scale bar = 50 μm [original magnification 40×]). Although increased expression of VEGF was observed in rats with MI who received omentum or PLGA–MSCs compared with that seen in the control group, the VEGF expression was the highest in rats that received the omentum-based PLGA–MSC-supporting patch at 4 weeks after transplantation (K and L). ΔP < .05 versus group P, ∗P < .05 versus group O, and #P < .05 versus the control group. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 1388-1396.e3DOI: (10.1016/j.jtcvs.2010.02.036) Copyright © 2010 Terms and Conditions

Figure 2 Quantification of transplanted MSC apoptosis by means of terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling the (TUNEL) assay. TUNEL-positive cells were shown as green; the transplanted cells were shown as blue (DAPI). Merged images show that there were significantly less TUNEL-positive cells in rats with MI that received omentum-based polylactic acid-co-glycolic acid–mesenchymal stem cell (PLGA–MSC)–supporting patches compared with the PLGA+MSC group at 1 week after transplantation. At 2 or 4 weeks after transplantation, TUNEL-positive cells were comparable between the 2 groups. Tissue samples of the hearts were excised at 1 (A and B), 2 (C and D), and 4 (E and F) weeks after PLGA–MSC transplantation (A, C, and E) or omentum-based PLGA–MSC-supporting patch transplantation (B, D, and F). Scale bar = 20 μm (original magnification 40×) G, The percentage of apoptotic MSC nuclei both in group O+P and group; quantitative estimations of TUNEL-positive cells in each group are shown in the bar chart. ΔP < .05 versus group P. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 1388-1396.e3DOI: (10.1016/j.jtcvs.2010.02.036) Copyright © 2010 Terms and Conditions

Figure 3 Quantifying transplanted male mesenchymal stem cells (MSCs) in the female ischemic myocardium. A, Sry is expressed only in male rat cells. The polymerase chain reaction (PCR) product size is 213 bp. B, Dissociation curve of the real-time PCR. C, Time course of transplanted MSC survival. The survival of MSCs decreased from 1 to 4 weeks after transplantation in both groups. Viable MSCs were significantly higher in rats that received omentum-based polylactic acid-co-glycolic acid (PLGA)–MSC-supporting patch at each time point. ΔP < .05 versus group P. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 1388-1396.e3DOI: (10.1016/j.jtcvs.2010.02.036) Copyright © 2010 Terms and Conditions

Figure 4 Evaluation of ventricular remodeling and expression of stem cell factor (SCF) and connexin 43 (Cx43) after transplantation. A to H, Picrosirius red staining. I to L, Observed under polarized light. A to D, Gross view of the hearts. E to L, Sections from peri-infarction zone. The samples were excised 4 weeks after transplantation from rats with myocardial infarction (MI) that received no treatment (A, E, and I), rats with MI that received only omentum (B, F, and J), rats with MI treated with polylactic acid-co-glycolic acid–mesenchymal stem cells (PLGA–MSCs; C, G, and K), and rats with MI that received the omentum-based PLGA–MSC-supporting patch (D, H, and L). Collagen volume in the ventricle was reduced in rats treated with PLGA–MSCs than in the rats with MI that received no treatment. Rats with MI that received the omentum-based PLGA–MSC-supporting patch demonstrated further reduction in the collagen. M, The percentage of collagen content. Western blotting demonstrated that SCF expression was upregulated in rats that received MSCs compared with control rats or rats with MI that received either no treatment or omentum only (N and O). Western blotting demonstrated that there was a marked increase in Cx43 expression in the host myocardium after omentum-based PLGA–MSC-supporting patch transplantation compared with that seen in all other groups of rats (P and Q). Scale bar = 20 μm (original magnification 40×). ΔP < .05 versus group P, ∗P < .05 versus group O; ▴P < .05 versus group MI, and #P < .05 versus the control group. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 1388-1396.e3DOI: (10.1016/j.jtcvs.2010.02.036) Copyright © 2010 Terms and Conditions

Macroscopic characteristics of the rats with MI and PLGA scaffold Macroscopic characteristics of the rats with MI and PLGA scaffold. A, Photomicrographs of the PLGA patch. The PLGA patch was a dense layer with uniform pores. B, The punched 1-cm diameter PLGA patch is the size used to carry the MSCs for transplantation. C, Myocardial infarction was generated by means of LAD ligation. Eight weeks after ligation, the infarcted myocardium showed as a pale color, as indicated by the red arrow. D, Four weeks after transplantation, the macroscopic view of the animal revealed that the omentum supporting the cell patch was tightly connected with the host heart. The Journal of Thoracic and Cardiovascular Surgery 2010 140, 1388-1396.e3DOI: (10.1016/j.jtcvs.2010.02.036) Copyright © 2010 Terms and Conditions

Correlation between Sry gene signal and DAPI cell number Correlation between Sry gene signal and DAPI cell number. DAPI-positive cells were counted under fluorescence microscopy (original magnification 200×) in group P and group O+P before MSC patch transplantation and 1, 2, and 4 weeks after transplantation, respectively. Sry gene signal was evaluated at the same time points. The correlation of DAPI-positive cell numbers and Sry gene signal was analyzed and shown in Figure E2. The results showed that Sry gene signal was closely correlated with the delivered cells that were still present in the infarction area (r2 = .99). The Journal of Thoracic and Cardiovascular Surgery 2010 140, 1388-1396.e3DOI: (10.1016/j.jtcvs.2010.02.036) Copyright © 2010 Terms and Conditions

The histologic change of the PLGA polymer after transplantation: PLGA degradation in vivo. There is no obvious difference in PLGA fibers between the MSC–PLGA patch transplantation group (A) and the omentum-wrapped MSC–PLGA patch group (B) 1 week after transplantation. However, 4 weeks after transplantation, the remnants of PLGA in the omentum-wrapped MSC–PLGA patch group (D) were less than that in the MSC–PLGA patch group (C; PLGA fibers are indicated by a yellow arrow). The Journal of Thoracic and Cardiovascular Surgery 2010 140, 1388-1396.e3DOI: (10.1016/j.jtcvs.2010.02.036) Copyright © 2010 Terms and Conditions