Direct evaluation of myocardial viability and stem cell engraftment demonstrates salvage of the injured myocardium.

RATIONALE: The mechanism of functional restoration by stem cell therapy remains poorly understood. Novel manganese-enhanced MRI and bioluminescence reporter gene imaging were applied to follow myocardial viability and cell engraftment, respectively. Human-placenta-derived amniotic mesenchymal stem cells (AMCs) demonstrate unique immunoregulatory and precardiac properties. In this study, the restorative effects of 3 AMC-derived subpopulations were examined in a murine myocardial injury model: (1) unselected AMCs, (2) ckit(+)AMCs, and (3) AMC-derived induced pluripotent stem cells (MiPSCs). OBJECTIVE: To determine the differential restorative effects of the AMC-derived subpopulations in the murine myocardial injury model using multimodality imaging. METHODS AND RESULTS: SCID (severe combined immunodeficiency) mice underwent left anterior descending artery ligation and were divided into 4 treatment arms: (1) normal saline control (n=14), (2) unselected AMCs (n=10), (3) ckit(+)AMCs (n=13), and (4) MiPSCs (n=11). Cardiac MRI assessed myocardial viability and left ventricular function, whereas bioluminescence imaging assessed stem cell engraftment during a 4-week period. Immunohistological labeling and reverse transcriptase polymerase chain reaction of the explanted myocardium were performed. The unselected AMC and ckit(+)AMC-treated mice demonstrated transient left ventricular functional improvement. However, the MiPSCs exhibited a significantly greater increase in left ventricular function compared with all the other groups during the entire 4-week period. Left ventricular functional improvement correlated with increased myocardial viability and sustained stem cell engraftment. The MiPSC-treated animals lacked any evidence of de novo cardiac differentiation. CONCLUSION: The functional restoration seen in MiPSCs was characterized by increased myocardial viability and sustained engraftment without de novo cardiac differentiation, indicating salvage of the injured myocardium.

Telmisartan in the diabetic murine model of acute myocardial infarction: dual contrast manganese-enhanced and delayed enhancement MRI evaluation of the peri-infarct region.

BACKGROUND: A novel MRI technique, employing dual contrast manganese-enhanced MRI (MEMRI) and delayed enhancement MRI (DEMRI), can evaluate the physiologically unstable peri-infarct region. Dual contrast MEMRI-DEMRI enables comprehensive evaluation of telmisartan to salvage the peri-infarct injury to elucidate the underlying mechanism of restoring the ischemic cardiomyopathy in the diabetic mouse model. METHODS AND RESULTS: Dual contrast MEMRI-DEMRI was performed on weeks 1, 2, and 4 following initiation of telmisartan treatment in 24 left anterior descendent artery ligated diabetic mice. The MRI images were analyzed for core infarct, peri-infarct, left ventricular end-diastolic, end-systolic volumes, and the left ventricular ejection fraction (LVEF). Transmission electron microscopy (TEM) and real-time PCR were used for ex vivo analysis of the myocardium. Telmisartan vs. control groups demonstrated significantly improved LVEF at weeks 1, 2, and 4, respectively (33 ± 7 %*** vs. 19 ± 5 %, 29 ± 3 %*** vs. 22 ± 4 %, and 31 ± 2 %*** vs 18 ± 6 %, ***p < 0.001). The control group demonstrated significant differences in the scar volume measured by MEMRI and DEMRI, demonstrating peri-infarct injury. Telmisartan group significantly salvaged the peri-infarct injury. The myocardial effects were validated by TEM, which confirmed the presence of the injured but viable cardiomyocyte morphology in the peri-infarct region and by flow cytometry of venous blood, which demonstrated significantly increased circulating endothelial progenitor cells (EPCs). CONCLUSION: The improved cardiac function in ischemic cardiomyopathy of diabetic mice by telmisartan is attributed to the attenuation of the peri-infarct injury by the angiogenic effects of EPCs to salvage the injured cardiomyocytes. Dual-contrast MEMRI-DEMRI technique tracked the therapeutic effects of telmisartan on the injured myocardium longitudinally.

Manganese-Enhanced Magnetic Resonance Imaging Enables In Vivo Confirmation of Peri-Infarct Restoration Following Stem Cell Therapy in a Porcine Ischemia-Reperfusion Model.

BACKGROUND: The exact mechanism of stem cell therapy in augmenting the function of ischemic cardiomyopathy is unclear. In this study, we hypothesized that increased viability of the peri-infarct region (PIR) produces restorative benefits after stem cell engraftment. A novel multimodality imaging approach simultaneously assessed myocardial viability (manganese-enhanced magnetic resonance imaging [MEMRI]), myocardial scar (delayed gadolinium enhancement MRI), and transplanted stem cell engraftment (positron emission tomography reporter gene) in the injured porcine hearts. METHODS AND RESULTS: Twelve adult swine underwent ischemia-reperfusion injury. Digital subtraction of MEMRI-negative myocardium (intrainfarct region) from delayed gadolinium enhancement MRI-positive myocardium (PIR and intrainfarct region) clearly delineated the PIR in which the MEMRI-positive signal reflected PIR viability. Human amniotic mesenchymal stem cells (hAMSCs) represent a unique population of immunomodulatory mesodermal stem cells that restored the murine PIR. Immediately following hAMSC delivery, MEMRI demonstrated an increased PIR viability signal compared with control. Direct PIR viability remained higher in hAMSC-treated hearts for >6 weeks. Increased PIR viability correlated with improved regional contractility, left ventricular ejection fraction, infarct size, and hAMSC engraftment, as confirmed by immunocytochemistry. Increased MEMRI and positron emission tomography reporter gene signal in the intrainfarct region and the PIR correlated with sustained functional augmentation (global and regional) within the hAMSC group (mean change, left ventricular ejection fraction: hAMSC 85±60%, control 8±10%; P<0.05) and reduced chamber dilatation (left ventricular end-diastole volume increase: hAMSC 24±8%, control 110±30%; P<0.05). CONCLUSIONS: The positron emission tomography reporter gene signal of hAMSC engraftment correlates with the improved MEMRI signal in the PIR. The increased MEMRI signal represents PIR viability and the restorative potential of the injured heart. This in vivo multimodality imaging platform represents a novel, real-time method of tracking PIR viability and stem cell engraftment while providing a mechanistic explanation of the therapeutic efficacy of cardiovascular stem cells.

Preliminary evaluation of EVP 1001-1: a new cardiac-specific magnetic resonance contrast agent with kinetics suitable for steady-state imaging of the ischemic heart.

RATIONALE AND OBJECTIVES: To investigate the potential of a novel manganese-based magnetic resonance (MR) contrast agent, EVP 1001-1 for the evaluation of myocardial ischemia. METHODS: MR imaging with EVP 1001-1 was performed on 6 Yorkshire pigs, and T1 relaxation times were calculated. One animal served as a control, 2 were subjected to an acute coronary artery occlusion and 3 provided a model of chronic ischemia. RESULTS: Administration of the agent in the control and acute coronary occlusion model demonstrated a short plasma half-life (approximately 1.5 minutes) and rapid myocardial uptake in nonoccluded regions, with long retention times in the myocardium (>1 hour) and no evidence of redistribution. In the chronic ischemia model, differential enhancement was observed between normal and ischemic tissue, particularly under dobutamine-induced stress. CONCLUSIONS: These properties suggest the use of EVP 1001-1 for steady-state imaging of myocardial perfusion. Contrast administration could be performed under stress conditions outside the scanner, with high-resolution MR images reflecting the stress condition acquired after the stress has subsided.

Dual manganese-enhanced and delayed gadolinium-enhanced MRI detects myocardial border zone injury in a pig ischemia-reperfusion model.

BACKGROUND: Gadolinium (Gd)-based delayed-enhancement MRI (DEMRI) identifies nonviable myocardium but is nonspecific and may overestimate nonviable territory. Manganese (Mn(2+))-enhanced MRI (MEMRI) denotes specific Mn(2+) uptake into viable cardiomyocytes. We performed a dual-contrast myocardial assessment in a porcine ischemia-reperfusion (IR) model to test the hypothesis that combined DEMRI and MEMRI identifies viable infarct border zone (BZ) myocardium in vivo. METHODS AND RESULTS: Sixty-minute left anterior descending coronary artery IR injury was induced in 13 adult swine. Twenty-one days post-IR, 3-T cardiac MRI was performed. MEMRI was obtained after injection of 0.7 mL/kg Mn(2+) contrast agent. DEMRI was then acquired after injection of 0.2 mmol/kg Gd. Left ventricular (LV) mass, infarct, and function were analyzed. Subtraction of MEMRI defect from DEMRI signal identified injured BZ myocardium. Explanted hearts were analyzed by 2,3,5-triphenyltetrazolium chloride stain and tissue electron microscopy to compare infarct, BZ, and remote myocardium. Average LV ejection fraction was reduced (30±7%). MEMRI and DEMRI infarct volumes correlated with 2,3,5-triphenyltetrazolium chloride stain analysis (MEMRI, r=0.78; DEMRI, r=0.75; P<0.004). MEMRI infarct volume percentage was significantly lower than that of DEMRI (14±4% versus 23±4%; P<0.05). BZ MEMRI signal-to-noise ratio (SNR) was intermediate to remote and core infarct SNR (7.5±2.8 versus 13.2±3.4 and 2.9±1.6; P<0.0001), and DEMRI BZ SNR tended to be intermediate to remote and core infarct SNR (8.4±5.4 versus 3.3±0.6 and 14.3±6.6; P>0.05). Tissue electron microscopy analysis exhibited preserved cell structure in BZ cardiomyocytes despite transmural DEMRI enhancement. CONCLUSIONS: The dual-contrast MEMRI-DEMRI detects BZ viability within DEMRI infarct zones. This approach may identify injured, at-risk myocardium in ischemic cardiomyopathy.

EVP-ABD-enhanced MRI to evaluate diffuse liver disease in a rat model.

PURPOSE: To evaluate the feasibility of using manganese-based MR imaging contrast agent EVP-ABD to detect diffuse liver disease in an established rat hepatitis model. MATERIALS AND METHODS: Hepatitis was induced by administration of CCl(4) in corn oil vehicle to rats intraperitoneally. MR images were acquired on a 3T scanner using a volume coil approximately 36 hours after the administration of CCl(4). EVP-ABD was administered via a tail vein at a dose of 10 mumol/kg. Multi-TI turboflash images were acquired to evaluate liver R1 (=1/T1) values before and after the EVP-ABD administration. Eighteen rats received various doses of CCl(4) and completed pre- and postcontrast MRI scans and liver histologic evaluation. RESULTS: The liver R1 after the EVP-ABD administration and the change of the liver R1 before and after the administration, DeltaR1, show significant correlations with the CCl(4) dose. A significant correlation was also found between the histologic scores and the CCl(4) doses despite known variability in the relationship of CCl(4) dose to histology. A significant correlation was found between the histologic score and DeltaR1. CONCLUSION: Our results indicate that EVP-ABD-enhanced MRI can detect diffuse liver disease generated by CCl(4) based on the significant correlation between proton R1 in liver following EVP-ABD and the CCl(4) doses as well as the histologic scores.

Magnetic resonance imaging of myocardial infarction using a manganese-based contrast agent (EVP 1001-1): preliminary results in a dog model.

PURPOSE: To investigate the MRI characteristics of an intracellular manganese-based contrast agent, EVP 1001-1 (Eagle Vision Pharmaceutical Corp.), in a canine model of myocardial infarction. MATERIALS AND METHODS: Three dogs were imaged 14-37 days following permanent ligation of the left anterior descending coronary artery (LAD). Measurements of the longitudinal relaxation rate R(1) were made prior to EVP 1001-1 administration (20 micromol/kg i.v.) and for one hour thereafter. Triphenyl tetrazolium chloride (TTC) staining was used to document infarction. RESULTS: In normal myocardium, EVP 1001-1 produced a substantial increase in the longitudinal relaxation rate, which remained fairly constant over the postcontrast imaging period (DeltaR1= 1.47 +/- 0.58 sec(-1) (mean +/- SD) at 35 minutes, P < 0.05). In the infarct, the response to EVP 1001-1 was small or negligible (DeltaR1= 0.27 +/- 0.28 sec(-1)). This resulted in a significant postcontrast difference in relaxation rate between normal and infarcted tissue (R1(normal) - R1(infarct) = 1.08 +/- 0.26 sec(-1), P < 0.05). The infarct remained clearly delineated in all animals throughout the steady-state imaging period, and qualitatively matched TTC results. CONCLUSION: The persistent enhancement pattern revealed by MRI following EVP 1001-1 administration may be beneficial for identifying and characterizing myocardial infarction.