Delivery of an engineered HGF fragment in an extracellular matrix-derived hydrogel prevents negative LV remodeling post-myocardial infarction.

Hepatocyte growth factor (HGF) has been shown to have anti-fibrotic, pro-angiogenic, and cardioprotective effects; however, it is highly unstable and expensive to manufacture, hindering its clinical translation. Recently, a HGF fragment (HGF-f), an alternative c-MET agonist, was engineered to possess increased stability and recombinant expression yields. In this study, we assessed the potential of HGF-f, delivered in an extracellular matrix (ECM)-derived hydrogel, as a potential treatment for myocardial infarction (MI). HGF-f protected cardiomyocytes from serum-starvation and induced down-regulation of fibrotic markers in whole cardiac cell isolate compared to the untreated control. The ECM hydrogel prolonged release of HGF-f compared to collagen gels, and in vivo delivery of HGF-f from ECM hydrogels mitigated negative left ventricular (LV) remodeling, improved fractional area change (FAC), and increased arteriole density in a rat myocardial infarction model. These results indicate that HGF-f may be a viable alternative to using recombinant HGF, and that an ECM hydrogel can be employed to increase growth factor retention and efficacy.

Safety and efficacy of an injectable extracellular matrix hydrogel for treating myocardial infarction.

New therapies are needed to prevent heart failure after myocardial infarction (MI). As experimental treatment strategies for MI approach translation, safety and efficacy must be established in relevant animal models that mimic the clinical situation. We have developed an injectable hydrogel derived from porcine myocardial extracellular matrix as a scaffold for cardiac repair after MI. We establish the safety and efficacy of this injectable biomaterial in large- and small-animal studies that simulate the clinical setting. Infarcted pigs were treated with percutaneous transendocardial injections of the myocardial matrix hydrogel 2 weeks after MI and evaluated after 3 months. Echocardiography indicated improvement in cardiac function, ventricular volumes, and global wall motion scores. Furthermore, a significantly larger zone of cardiac muscle was found at the endocardium in matrix-injected pigs compared to controls. In rats, we establish the safety of this biomaterial and explore the host response via direct injection into the left ventricular lumen and in an inflammation study, both of which support the biocompatibility of this material. Hemocompatibility studies with human blood indicate that exposure to the material at relevant concentrations does not affect clotting times or platelet activation. This work therefore provides a strong platform to move forward in clinical studies with this cardiac-specific biomaterial that can be delivered by catheter.

Development and validation of a noninvasive method to estimate cardiac output using cuff sphygmomanometry.

BACKGROUND: Obtaining cardiac output (CO) measurements noninvasively during routine blood pressure recording can improve hypertension management. A new method has been developed that estimates cardiac output using pulse-waveform analysis (PWA) from a brachial cuff sphygmomanometer. This study evaluates the ability of PWA to track changes in CO as derived by Doppler ultrasound during dobutamine stimulation. HYPOTHESIS: This study aims to validate the PWA CO estimation over a wide CO range as would be obtained by dobutamine stimulation during Doppler ultrasound evaluation. METHOD: A total of 48 patients undergoing standard dobutamine stress echocardiography testing for accepted clinical indications were enrolled. Among them, 44 patients (age 36-83, 18 females, 26 males) with good waveform data for analyses provided estimates of CO in this study. Noninvasive measurements of CO were performed using both Doppler ultrasound recordings and PWA techniques simultaneously at each stage of dobutamine infusion. RESULTS: A total of 207 simultaneous pulse-waveform analyses and Doppler measurements were taken during dobutamine stress on 44 cardiac patients. Linear regression analysis revealed good intra-patient correlation between pulse-waveform analysis and Doppler at different dobutamine-induced CO with coefficients from r = 0.69 to 0.98 (p < 0.05). Analysis of all patients yielded an overall correlation of r = 0.82 (p < 0.001, bias = 0.4 L/min, standard deviation = 1.8 L/min). CONCLUSION: The CO measured noninvasively from a sphygmomanometer using this PWA method correlates well with those of Doppler through a range of dobutamine-stimulated levels. The CO by PWA should be useful for monitoring hemodynamic changes in hypertensive and cardiac patients during routine blood pressure measurement.

Difference of optimal dose of contrast agent between gray-scale and power Doppler imaging in assessing graded coronary stenosis by myocardial contrast echocardiography.

RATIONALE AND OBJECTIVES: In myocardial contrast echocardiography (MCE), power Doppler imaging is more sensitive to contrast agent (microbubble) than gray-scale B-mode imaging; however, no data exist regarding the optimal contrast dose in power Doppler imaging. This study examined the optimal dose of contrast agent for power Doppler in assessing coronary stenosis. METHODS: Three grades of coronary stenosis were produced in 6 open-chest dogs. MCE was performed with gray-scale and power Doppler during continuous infusion of 0.2 mL/min FS-069. Thereafter, MCE was repeated with power Doppler during continuous infusion of 0.1 mL/min FS-069. RESULTS: Although the videointensity in the stenosed bed with power Doppler (214 +/- 14) was greater than gray scale (35 +/- 17) during 0.2 mL/min FS-069 infusion (P < 0.0001), power Doppler failed to identify milder coronary stenoses because videointensity in stenosed bed was quickly saturated with contrast agent. The videointensity in the stenosed bed with power Doppler (127 +/- 49) during 0.1 mL/min FS-069 infusion was greater than gray scale (35 +/- 17) during 0.2 mL/min FS-069 infusion (P < 0.0001), and all levels of stenosis were identified with power Doppler, even though the dose of contrast agent was half of that of gray scale imaging. The correlation between videointensity and myocardial blood flow was better in the case of power Doppler at 0.1 mL/min FS-069 infusion (r = 0.77, P < 0.0001) than in the case of gray scale imaging at 0.2 mL/min FS-069 infusion (r = 0.66, P < 0.01). CONCLUSIONS: These data support the need for a lower dose of contrast agent for power Doppler than for gray scale to detect milder coronary stenosis and avoid saturation of imaging fields.

Effect of microbubble fragility on transit rate measurement by contrast echography.

We sought to propose a simplified method to measure flow velocity based on ultrasonic microbubble destruction, and investigated the effect of microbubble shell fragility on such measurement. Acoustic density (AD) from the second harmonic short axis image of flow was obtained at variable velocities (2 to 73 mm/s) in an in vitro model during long (1000 ms) and short (33 ms) interval ultrasound (US) pulsing, allowing complete and partial microbubble replenishment between pulses, respectively. Microbubbles with shell elastic modulus of 0.4 MPa and 16 MPa were tested. By shortening pulsing interval, AD diminished gradually, rather than abruptly, to a plateau level for both microbubbles. The extent of AD decay was greater for the fragile than the strong microbubbles. A linear relationship existed between the magnitude of AD decay and flow velocity only in the higher and lower velocity range for the fragile and the strong microbubbles, respectively. Thus, difference in contrast intensities during long and short pulsing intervals, respectively, allowing complete and partial replenishment may provide for velocity measurement, in which choice of optimal microbubble fragility for the range of velocity to measure may increase the accuracy.

Contrast echocardiography can assess risk area and infarct size during coronary occlusion and reperfusion: experimental validation.

OBJECTIVES: We sought to validate the ability of real-time myocardial contrast echocardiography (MCE) measures of opacification defect and contrast refilling parameters to estimate risk area (RA) and infarct area (IA) during coronary occlusion and reperfusion. BACKGROUND: No data exist establishing the accuracy of MCE in determining RA and IA size. We hypothesized that in the setting of coronary occlusion, MCE should identify RA as a perfusion defect early after bubble destruction, collateral flow to viable myocardium as opacification late during refilling and IA as absent opacification. METHODS: Three hours of coronary occlusion and reperfusion were each produced in 11 dogs in which real-time MCE was performed during intravenous infusion of Sonovue (Bracco). Real-time contrast echocardiography was performed at baseline, during occlusion and reperfusion. Early (BEGIN) and end (END) images from a FLASH refilling sequence were acquired, as well as late refilling images (LATE) 1 min after FLASH. Real-time contrast echocardiography defect size and quantitative refilling parameters were compared with RA and IA determined by tissue staining. RESULTS: During occlusion, defect size varied with refilling time; defects from BEGIN images correlated best to RA and those from LATE images to IA. Refilling parameters, but not LATE peak intensity, did not predict the IA size during occlusion. During reperfusion, defects from BEGIN images were well correlated to RA and END images to IA, whereas peak plateau intensity and refilling slope parameters predicted IA size. CONCLUSIONS: Real-time contrast echocardiography defect size varies throughout microbubble refilling. Appropriately selected defect sizes and refilling parameters provide estimates of RA and IA during coronary occlusion and reperfusion.

Automatic Border Detection to Assess Right Ventricular Function Following Surgical Treatment of Thromboembolic Pulmonary Hypertension.

Automatic border detection (ABD) has been developed as a potentially useful means for evaluating ventricular function on line in an automatic fashion. Its success with tracking left ventricular function is established, but little is known about its ability to assess right ventricular (RV) function. Accordingly, 20 patients with severe pulmonary hypertension due to chronic thromboembolic disease underwent standard two-dimensional echocardiography and imaging with ABD before and after pulmonary thromboendarterectomy to correct pulmonary hypertension. ABD-derived results were compared to manually planimetered RV areas calculated from the apical four-chamber view. Doppler tricuspid regurgitant velocity fell significantly with surgery from 4.4 +/- 0.6 to 2.9 +/- 0.7 m/sec (P < 0.001). The mean values for RV areas derived by manual planimetry and ABD were similar, as was fractional area shortening, which improved significantly with surgery (manual 0.24 +/- 0.01 preoperative vs 0.31 +/- 0.11 postoperative, P < 0.05; and ABD 0.19 +/- 0.05 preoperative vs 0.32 +/- 0.15 postoperative, P < 0.001). There was, however, very little correlation between the individual values for ABD versus manually derived RV areas and fractional area shortening, with the best correlation being the RV end-diastolic areas after surgery (y = 0.684x + 7.9, r = 0.564, P = 0.01). These results demonstrate that both manually planimetered RV areas and those determined by ABD can adequately follow changes in ventricular function over time. However, variability within each technique may prevent direct comparison of the absolute values of the two techniques. (ECHOCARDIOGRAPHY, Volume 13, March 1996)