Bruce H. Smaill

Cardiac Microstructure: Implications for Electrical Propagation and Defibrillation in the Heart

Abstract— Our understanding of the electrophysiological properties of the heart is incomplete. We have investigated two issues that are fundamental to advancing that understanding. First, there has been widespread debate over the mechanisms by which an externally applied shock can influence a sufficient volume of heart tissue to terminate cardiac fibrillation. Second, it has been uncertain whether cardiac tissue should be viewed as an electrically orthotropic structure, or whether its electrical properties are, in fact, isotropic in the plane orthogonal to myofiber direction. In the present study, a computer model that incorporates a detailed three-dimensional representation of cardiac muscular architecture is used to investigate these issues. We describe a bidomain model of electrical propagation solved in a discontinuous domain that accurately represents the microstructure of a transmural block of rat left ventricle. From analysis of the model results, we conclude that (1) the laminar organization of myocytes determines unique electrical properties in three microstructurally defined directions at any point in the ventricular wall of the heart, and (2) interlaminar clefts between layers of cardiomyocytes provide a substrate for bulk activation of the ventricles during defibrillation.

Three-dimensional transmural organization of perimysial collagen in the heart

There is strong support for the view that the ventricular myocardium has a laminar organization in which myocytes are grouped into branching layers separated by cleavage planes. However, understanding of the extent and functional implications of this architecture has been limited by the lack of a systematic three-dimensional description of the organization of myocytes and associated perimysial collagen. We imaged myocytes and collagen across the left ventricular wall at high resolution in seven normal rat hearts using extended volume confocal microscopy. We developed novel reconstruction and segmentation techniques necessary for the quantitative analysis of three-dimensional myocyte and perimysial collagen organization. The results confirm that perimysial collagen has an ordered arrangement and that it defines a laminar organization. Perimysial collagen is composed of three distinct forms: extensive meshwork on laminar surfaces, convoluted fibers connecting adjacent layers, and longitudinal cords. While myolaminae are the principal form of structural organization throughout most of the wall, they are not seen in the subepicardium, where perimysial collagen is present only as longitudinal cords.

Laminar arrangement of ventricular myocytes influences electrical behavior of the heart.

The response of the heart to electrical shock, electrical propagation in sinus rhythm, and the spatiotemporal dynamics of ventricular fibrillation all depend critically on the electrical anisotropy of cardiac tissue. A long-held view of cardiac electrical anisotropy is that electrical conductivity is greatest along the myocyte axis allowing most rapid propagation of electrical activation in this direction, and that conductivity is isotropic transverse to the myocyte axis supporting a slower uniform spread of activation in this plane. In this context, knowledge of conductivity in two directions, parallel and transverse to the myofiber axis, is sufficient to characterize the electrical action of the heart. Here we present new experimental data that challenge this view. We have used a novel combination of intramural electrical mapping, and experiment-specific computer modeling, to demonstrate that left ventricular myocardium has unique bulk conductivities associated with three microstructurally-defined axes. We show that voltage fields induced by intramural current injection are influenced by not only myofiber direction, but also the transmural arrangement of muscle layers or myolaminae. Computer models of these experiments, in which measured 3D tissue structure was reconstructed in-silico, best matched recorded voltages with conductivities in the myofiber direction, and parallel and normal to myolaminae, set in the ratio 4:2:1, respectively. These findings redefine cardiac tissue as an electrically orthotropic substrate and enhance our understanding of how external shocks may act to successfully reset the fibrillating heart into a uniform electrical state. More generally, the mechanisms governing the destabilization of coordinated electrical propagation into ventricular arrhythmia need to be evaluated in the light of this discovery.

The architecture of the heart: a data–based model

An appropriate mathematical representation of heart structure is central to advancing an integrative approach to cardiac function. The Auckland heart model provides a realistic representation of important aspects of ventricular structure. The finite–element model is based on extensive anatomical data and incorporates detailed information on ventricular geometry and myocyte organization. It includes preliminary descriptions of the Purkinje fibre network, coronary vessels and collagen organization. Comprehensive extension of these data is required to exploit the full potential of computer modelling. In particular, we need to quantify the morphology of the atria, cardiac conduction system and the intramyocardial coronary vessels. This information must span an appropriate range of species and disease states.

Three Distinct Directions of Intramural Activation Reveal Nonuniform Side-to-Side Electrical Coupling of Ventricular Myocytes

Background—The anisotropy of cardiac tissue is a key determinant of 3D electric propagation and the stability of activation wave fronts in the heart. The electric properties of ventricular myocardium are widely assumed to be axially anisotropic, with activation propagating most rapidly in the myofiber direction and at uniform velocity transverse to this. We present new experimental evidence that contradicts this view. Methods and Results—For the first time, high-density intramural electric mapping (325 electrodes at ≈4×4×1-mm spacing) from pig left ventricular tissue was used to reconstruct 3D paced activation surfaces projected directly onto 3D tissue structure imaged throughout the same left ventricular volume. These data from 5 hearts demonstrate that ventricular tissue is electrically orthotropic with 3 distinct propagation directions that coincide with local microstructural axes defined by the laminar arrangement of ventricular myocytes. The maximum conduction velocity of 0.67±0.019 ms−1 was aligned with the myofiber axis. However, transverse to this, the maximum conduction velocity was 0.30±0.010 ms−1, parallel to the myocyte layers and 0.17±0.004 ms−1 normal to them. These orthotropic conduction velocities give rise to preferential activation pathways across the left ventricular free wall that are not captured by structurally detailed computer models, which incorporate axially anisotropic electric properties. Conclusions—Our findings suggest that current views on uniform side-to-side electric coupling in the heart need to be revised. In particular, nonuniform laminar myocardial architecture and associated electric orthotropy should be included in future models of initiation and maintenance of ventricular arrhythmia.

A triaxial-measurement shear-test device for soft biological tissues.

A novel shear-test device for soft biological tissue, capable of applying simple shear deformations simultaneously in two orthogonal directions while measuring the resulting forces generated in three axes, is described. We validated the device using a synthetic gel, the properties of which were ascertained from independent tensile and rotational shear tests. Material parameters for the gel were fitted using neo-Hookean analytical solutions to the independent test data, and these matched the results from the device. Preliminary results obtained with rat septal myocardium are also presented to demonstrate the feasibility of the apparatus in determining the shear characteristics of living tissue.

Application of Micro-Computed Tomography With Iodine Staining to Cardiac Imaging, Segmentation, and Computational Model Development

Micro-computed tomography (micro-CT) has been widely used to generate high-resolution 3-D tissue images from small animals nondestructively, especially for mineralized skeletal tissues. However, its application to the analysis of soft cardiovascular tissues has been limited by poor inter-tissue contrast. Recent ex vivo studies have shown that contrast between muscular and connective tissue in micro-CT images can be enhanced by staining with iodine. In the present study, we apply this novel technique for imaging of cardiovascular structures in canine hearts. We optimize the method to obtain high-resolution X-ray micro-CT images of the canine atria and its distinctive regions-including the Bachmanns bundle, atrioventricular node, pulmonary arteries and veins-with clear inter-tissue contrast. The imaging results are used to reconstruct and segment the detailed 3-D geometry of the atria. Structure tensor analysis shows that the arrangement of atrial fibers can also be characterized using the enhanced micro-CT images, as iodine preferentially accumulates within the muscular fibers rather than in connective tissues. This novel technique can be particularly useful in nondestructive imaging of 3-D cardiac architectures from large animals and humans, due to the combination of relatively high speed ( ~ 1 h/per scan of the large canine heart) and high voxel resolution (36 μm) provided. In summary, contrast micro-CT facilitates fast and nondestructive imaging and segmenting of detailed 3-D cardiovascular geometries, as well as measuring fiber orientation, which are crucial in constructing biophysically detailed computational cardiac models.

Regional left ventricular epicardial deformation in the passive dog heart.

Epicardial wall motion was measured on the left ventricular free wall in six isolated potassium-arrested dog hearts using a biplane video technique. Significant regional variations in epicardial deformations were recorded during static ventricular filling. Epicardial stretches varied linearly with cavity volume, sometimes exceeding 20% at physiological left ventricular end-diastolic pressures. The maximum component of epicardial stretch and the derived wall thinning increased substantially from the base to the apex on both the anterior and the posterior free walls of the left ventricle. In five hearts, the direction of greatest epicardial stretch at moderate and high filling pressures coincided closely with the local epicardial fiber direction, suggesting that the left-handed epicardial fiber helices stretch preferentially during passive filling to maximize end-diastolic fiber lengths. Epicardial rotation was always counterclockwise, consistent with a reduction in the pitch of the fiber helix during filling. These results suggest that, on the epicardial surface, the passive myocardium is anisotropic with respect to the local fiber direction. We suggest that the resulting torsional shear acts to minimize transmural gradients of fiber stretch.

A finite volume method for modeling discontinuous electrical activation in cardiac tissue

This paper describes a finite volume method for modeling electrical activation in a sample of cardiac tissue using the bidomain equations. Microstructural features to the level of cleavage planes between sheets of myocardial fibers in the tissue are explicitly represented. The key features of this implementation compared to previous modeling are that it represents physical discontinuities without the implicit removal of intracellular volume and it generates linear systems of equations that are computationally efficient to construct and solve. Results obtained using this method highlight how the understanding of discontinuous activation in cardiac tissue can form a basis for better understanding defibrillation processes and experimental recordings.

Biaxial Testing of Membrane Biomaterials: Testing Equipment and Procedures

A testing facility for measuring the biaxial mechanical properties of highly deformable membranes is described. Forces are applied, via strain-gauge force transducers, to four points on each side of an initially square 12 to 25 mm membrane sample to produce biaxial extensions of up to 80 percent of undeformed length. Strain is estimated from the displacement of markers bounding a 1 to 2 mm central square. The accuracy of stress and strain field measurements has been assessed by finite element analysis of a biaxially-loaded isotropic elastic membrane. Major advantages of the present system over those previously described in the literature are that 1) sample mounting procedures are simplified, 2) there is provision for independent adjustment of stress field uniformity and measurement of the applied point forces and 3) faster strain rates can be imposed on the relatively small samples tested.