Evaluating Cardiac Function in Animal Models

To assess the cardiac function of the experimental animals, echocardiography was performed in all of the experimental animals at different time points (prior to LAD ligation, 3 days post LAD ligation, 2 and 4 weeks after aFRLM treatment) under controlled anesthesia using a Toshiba Aplio XG Echocardiography with a PLT-1202 S linear array transducer [5 (link), 36 ]. In each experimental subject, the M-mode tracing and 2-dimensional echocardiography of at least ten consecutive cardiac cycles were recorded from the parasternal long- and short-axis views. The left ventricle systolic function was assessed by calculating the EF and FS with the Modified-Simpson method using conventional echocardiography [37 (link)]. The left ventricular end systolic diameter (LVESD) and the left ventricular end diastolic diameter (LVEDD), as well as the systolic and diastolic wall thickness, were measured from the M-mode tracings using the leading-edge convention of the American Society of Echocardiography [5 (link), 11 (link), 36 ]. All of the data were analyzed offline with software that was installed in the ultrasound system. All of the measurements were averaged from ten consecutive cardiac cycles per experiment and were analyzed by an experienced investigator.