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Human induced pluripotent stem cells (hiPSCs) hold promise for myocardial repair following injury, but preclinical studies in large animal models are required to determine optimal cell preparation and delivery strategies to maximize functional benefits and to evaluate safety. Here, we utilized a porcine model of acute myocardial infarction (MI) to investigate the functional impact of intramyocardial transplantation of hiPSC-derived cardiomyocytes, endothelial cells, and smooth muscle cells, in combination with a 3D fibrin patch loaded with insulin growth factor (IGF)-encapsulated microspheres. hiPSC-derived cardiomyocytes integrated into host myocardium and generated organized sarcomeric structures, and endothelial and smooth muscle cells contributed to host vasculature. Trilineage cell transplantation significantly improved left ventricular function, myocardial metabolism, and arteriole density, while reducing infarct size, ventricular wall stress, and apoptosis without inducing ventricular arrhythmias. These findings in a large animal MI model highlight the potential of utilizing hiPSC-derived cells for cardiac repair.

Occurrence of immediate post-transplant heart failure in the cardiac transplant recipient is typically attributed to elevated pulmonary vascular resistance, however other etiologies may play a role. At the completion of the transplant, free air, which has collected in the donor heart, is vented via an aortotomy. Free air may rise into the right coronary artery and obstruct reperfusion of the right ventricle. Cardiac perfusion MRI may offer a method of non-invasively determining the presence of air embolus. The objectives in the pilot study were to identify steps in the donor process where free air could enter into the aortic root causing obstruction of perfusion of the coronary arteries. A change in surgical technique could then be used to eliminate a portal of entry and cardiac perfusion MRI could validate the technique. Standard cardiectomy was compared to a variation in technique in two animals. Pulmonary vein ligation was completed in the experimental model before completion of cardiectomy. Both hearts were isolated and imaged using T1-weighted FLASH sequence and gadolinium contrast via the aortic root. Cardiac perfusion MRI imaging of the heart with the unligated pulmonary vein revealed evidence of air embolus and no perfusion of the right coronary artery compared to the ligated heart. Anatomically, the right coronary artery is anterior in the mediastinum compared to the left coronary artery. Air emboli preferentially rise into the right coronary and can obstruct flow into the right heart. Cardiac perfusion MRI offers an effective method to evaluate the isolated pre-transplant heart for perfusion defects. [ABSTRACT FROM AUTHOR]

Preclinical and clinical studies have demonstrated that stem cell transplantation can improve the left ventricular (LV) contractile performance, yet the underlying mechanisms remain unknown. We examined whether mesenchymal stem cell (MSC) transplantation-induced beneficial effects are secondary to paracrine-associated improvements in LV contractile performance, wall stress, and myocardial bioenergetics in hearts with postinfarction LV remodeling. Myocardial contractile function and bioenergetics were compared 4 wk after acute myocardial infarction in normal pigs (n = 6), untreated pigs with myocardial infarction (MI group; n = 6), and pigs receiving autologous MSC transplantation (Ml + MSC group; n = 5). A distal occlusion of the left anterior descending coronary artery instigated significant myocardial hypertrophy. Ejection fraction decreased from 55.3 ± 3.1% (normal) to 30.4 ± 2.3% (Ml group; P < 0.01) and to 45.4 ± 3.1% (MI + MSC group; P < 0.01 vs. Ml). Hearts in the Ml group developed severe contractile dyskinesis in the infarct zone and border zone (BZ). MSC transplantation significantly improved contractile performance from dyskinesis to active contraction (P < 0.01 vs. Ml). BZ systolic wall stress was severely increased in Ml hearts but significantly improved after MSC transplantation (P < 0.01 vs. Ml). The BZ demonstrated profound bioenergetic abnormalities in Ml pigs; this was significantly improved after MSC transplantation (P < 0.01 vs. Ml). Patchy spared myocytes were found in the infarct zone of hearts receiving MSC transplantation but not in control hearts. These data demonstrate that MSC transplantation into the BZ causes significant improvements in myocardial contractile performance and reduction in wall stress, which ultimately results in significant bioenergetic improvements. Low cell engraftment indicates that MSCs did not provide a structural contribution to the damaged heart and that the observed beneficial effects likely resulted from paracrine repair mechanisms. [ABSTRACT FROM AUTHOR]

Regions of myocardial infarct (MI) are surrounded by a border zone (BZ) of normally perfused but dysfunctional myocardium. Although systolic dysfunction has been attributed to elevated wall stress in this region, there is evidence that intrinsic abnormalities of contractile performance exist in BZ myocardium. This study examined whether decreases of high-energy phosphates (HEP) and mitochondrial F1F0-ATPase (mtATPase) subunits typical of failing myocardium exist in BZ myocardium of compensated postinfarct remodeled hearts. Eight pigs were studied 6 wk after MI was produced by ligation of the left anterior descending coronary artery (LAD) distal to the second diagonal. Animals developed compensated LV remodeling with a decrease of ejection fraction from 54.6 ± 5.4% to 31 ± 2.1% (MRI) 5 wk after LAD occlusion. The remote zone (RZ) myocardium demonstrated modest decreases of ATP and mtATPase components. In contrast, BZ myocardium demonstrated profound abnormalities with ATP levels decreased to 42% of normal, and phosphocreatine-to-ATP ratio (31P-magnetic resonance spectroscopy) decreased from 2.06 ± 0.19 in normal hearts to 1.07 ± 0.10, with decreases in α-, β-, OSCP, and IF1 subunits of mtATPase, especially in the subendocardium. The reduction of myocardial creatine kinase isoform protein expression was also more severe in the BZ relative to the RZ myocardium. These abnormalities were independent of a change in mitochondrial content because the mitochondrial citrate synthase protein level was not different between the BZ and RZ. This regional heterogeneity of ATP content and expression of key enzymes in ATP production suggests that energetic insufficiency in the peri-infarct region may contribute to the transition from compensated LV remodeling to congestive heart failure. [ABSTRACT FROM AUTHOR]

The donor heart undergoes degradation during hypothermic storage. An assessment of donor heart preservation is typically done with histological or biochemical methods that are not feasible in the clinical setting. We describe a method to study the donor heart using cardiac perfusion MRI that is potentially feasible for clinical use. Standard cardiectomy was performed in the pig model and the hearts were stored in normal saline at 5 degrees C. Imaging was performed by using a rapid gradient-echo sequence (FLASH) with saturation-recovery preparation for T1-weighting in the short axis and horizontal long axis views. Approximately 80 serial images were acquired at a rate of 1/s during administration of 0.006 mmol/ml Gd-DTPA (500 ml, 1 l/min). Signal intensity vs. time curves were generated for each heart and slice imaged and compared to a 0.006 mmol/ml Gd-DTPA reference. H&E stained biopsies of the LV, RV, and septum were also obtained. The mean duration of heart storage (N=10) was 8.8 h (range 4.2-19.2 h). Histologically, no differences were seen in H&E stained biopsies among hearts at different storage times. However cardiac MRI revealed a decrease in perfusion units in each subsequent heart tested after 4.2 h. (R=0.49). Average peak up-slope was used as a surrogate measure for flow capacity through the microvasculature and peak contrast enhancement was used as a measurement of viable microvasculature. The 4 h heart had 83% peak contrast enhancement of the reference standard, as compared to 44% for the 19.2 h heart. The decrease in peak enhancement is directly related to the duration of storage time. No correlation of peak up-slope of the intensity curve to storage time was found. This new application of cardiac MRI in the donor heart is applicable to: (1) assessing marginal hearts, (2) evaluating donor heart preservation techniques, and (3) correlating pre- to post-transplant viability. [ABSTRACT FROM AUTHOR]

Introduction: Studies of hibernating myocardium suggest that functional recovery following CABG may be incomplete. Application of stem cells at the time of CABG may have promise for improving function, possibly by enhanced expression of mitochondrial antioxidant proteins. Hypothesis: The application of MSCs on a vicryl-based patch that is sutured onto the epicardium at the time of CABG enhances the expression of mitochondrial antioxidant proteins. Methods: Anesthetized swine underwent thoracotomy with placement of a constrictor around the LAD. At 12 weeks, pigs underwent off-pump revascularization with a LIMA graft and a vicryl patch seeded with (N=4) or without (N=4) MSCs (4 million cells, CD90+, CD105+ & CD45-) was sutured to the surface. At 4 weeks, regional function with MRI and expression of mitochondria proteins by TMT proteomics were determined. Results: At 4 weeks following CABG, swine that received the stem cell patch showed improvement in regional wall thickening in response to inotropic stimulation, indicating improved recovery of function as compared to revascularization alone (63±22% versus 47±2%; P<0,05). As shown in the Figure, expression of PGC1α as well as mitochondrial antioxidant proteins from the chronically ischemic territories were higher in animals with cell-based patch compared to the pigs that received CABG without a cell-based therapy. Conclusions: In summary, placement of an epicardial patch loaded with MSCs at the time of CABG improves contractile reserve within the revascularized territory, potentially by enhanced expression of mitochondrial antioxidant proteins, potentially by a PGC1α mechanism. These data support the utility of a cell-based approach for hibernating myocardial regions at the time of CABG, potentially by enhanced antioxidant pathways. [ABSTRACT FROM AUTHOR]