A New High‐Speed Biaxial Testing System for Rodent Myocardium Viscoelastic Measurement under Dynamic Loadings
Authors: Roth Kellan , Liu Wenqiang , LeBar Kristen , Ahern Matt , Wang Zhijie 
Published: May 2022

IntroductionMyocardium stiffening is well known in heart failure (HF) development, and it contributes to cardiac dysfunction. Myocardium is viscoelastic, which means that both elastic and viscous resistance exist under dynamic loading. There are limited studies of myocardium viscoelasticity, particularly for the right ventricle (RV). Because viscoelasticity is a time‐ and strain‐dependent mechanical behavior, the measurement at physiological rates & non‐linear deformation is critical. This study aims to develop a high‐speed biaxial tester that induces physiological deformation to characterize RV viscoelasticity in rats.MethodsTo enable the viscoelasticity measurement at physiological conditions, we built a biaxial tester that induces non‐linear, cyclic stretch of myocardium at rodent heart rates. Biaxial sinusoidal stretches were induced by actuators (1400mm/s) (Zaber), forces were recorded by load cells (Honeywell) at 1 kHz, and strains were obtained by a high‐speed camera (Baumer) at up to 200 fps. The LabVIEW and MATLAB codes were developed for performing biaxial tests at various frequencies with synchronized force and image data acquisition. To validate the biaxial tester, isotropic polydimethylsiloxane (PDMS) was used due to its similar viscoelasticity to cardiovascular tissues. PDMS sheets were prepared (Sylgard 184) for equibiaxial tests. Then, consistency between different axes’ measurements, synchronization of the axes’ movement, and accurate viscoelastic measurement were evaluated. Next, we performed equibiaxial cyclic sinusoidal tests on healthy rat RVs. After euthanasia, RV free wall was dissected and mounted. The outflow track direction was defined as the longitudinal direction. The RV was placed in a relaxant solution prior to testing, then preloaded by ~0.1 N and preconditioned by 15 cycles. Stress‐strain loops were derived for viscoelastic analysis.ResultsFirst, we examined if the biaxial tester could achieve planar biaxial tests at desired frequencies. The equibiaxial data obtained from PDMS sheets showed that the tester induced cyclic sinusoidal stretch at various frequencies up to 8 Hz (Fig. 1a), and the viscoelastic behavior measured at two axes were very close (Fig. 1b), thus confirming the tester’s accurate biaxial measurement of the isotropic material. The PDMS clearly showed frequency‐dependent changes in the hysteresis loops (Fig. 1c), with trends of increasing elasticity (slope of loop) and viscosity (area of loop) at higher frequencies. We further derived the viscosity by measuring Tan(δ) (Fig. 1d). The frequency‐dependent elasticity and viscosity of the PDMS sheet were consistent with literature data. Next, we obtained preliminary viscoelasticity data of healthy rat RVs (at 0.1‐2 Hz). Our data showed strain‐rate dependent changes in the hysteresis loops (Fig. 2). The dissipated energy (loop area) as well as the stiffness (loop slope) of the RV tended to increase with increasing frequency. The RV tissue showed similar viscoelastic behavior in both directions at all frequencies.ConclusionOur data showed that this novel testing system enables biaxial viscoelastic measurement of myocardium under physiological conditions (0.1‐8Hz) across small and large animal species. The establishment of this system allows further exploration of RV tissues at higher frequencies (5‐8 Hz) and in diseased states, which will improve the understanding of cardiac biomechanics in HF progression.