A comparison of two quasi-static computational models for assessment of intra-myocardial injection as a therapeutic strategy for heart failure.

Myocardial infarction, or heart attack, is the leading cause of mortality globally. Although the treatment of myocardial infarct has improved significantly, scar tissue that persists can often lead to increased stress and adverse remodeling of surrounding tissue and ultimately to heart failure. Intra-myocardial injection of biomaterials represents a potential treatment to attenuate remodeling, mitigate degeneration, and reverse the disease process in the tissue. In vivo experiments on animal models have shown functional benefits of this therapeutic strategy. However, a poor understanding of the optimal injection pattern, volume, and material properties has acted as a barrier to its widespread clinical adoption. In this study, we developed two quasistatic finite element simulations of the left ventricle to investigate the mechanical effect of intra-myocardial injection. The first model employed an idealized left ventricular geometry with rule-based cardiomyocyte orientation. The second model employed a subject-specific left ventricular geometry with cardiomyocyte orientation from diffusion tensor magnetic resonance imaging. Both models predicted cardiac parameters including ejection fraction, systolic wall thickening, and ventricular twist that matched experimentally reported values. All injection simulations showed cardiomyocyte stress attenuation, offering an explanation for the mechanical reinforcement benefit associated with injection. The study also enabled a comparison of injection location and the corresponding effect on cardiac performance at different stages of the cardiac cycle. While the idealized model has lower fidelity, it predicts cardiac function and differentiates the effects of injection location. Both models represent versatile in silico tools to guide optimal strategy in terms of injection number, volume, site, and material properties.

Monte Carlo Simulations of Diffusion Weighted MRI in Myocardium: Validation and Sensitivity Analysis.

A model of cardiac microstructure and diffusion MRI is presented, and compared with experimental data from ex vivo rat hearts. The model includes a simplified representation of individual cells, with physiologically correct cell size and orientation, as well as intra- to extracellular volume ratio. Diffusion MRI is simulated using a Monte Carlo model and realistic MRI sequences. The results show good correspondence between the simulated and experimental MRI signals. Similar patterns are observed in the eigenvalues of the diffusion tensor, the mean diffusivity (MD), and the fractional anisotropy (FA). A sensitivity analysis shows that the diffusivity is the dominant influence on all three eigenvalues of the diffusion tensor, the MD, and the FA. The area and aspect ratio of the cell cross-section affect the secondary and tertiary eigenvalues, and hence the FA. Within biological norms, the cell length, volume fraction of cells, and rate of change of helix angle play a relatively small role in influencing tissue diffusion. Results suggest that the model could be used to improve understanding of the relationship between cardiac microstructure and diffusion MRI measurements, as well as in testing and refinement of cardiac diffusion MRI protocols.

Temporal accumulation and localization of isoflurane in the C57BL/6 mouse and assessment of its potential contamination in 19 F MRI with perfluoro-crown-ether-labeled cardiac progenitor cells at 9.4 Tesla.

PURPOSE: To assess the uptake, accumulation, temporal stability, and spatial localization of isoflurane (ISO) in the C57BL/6 mouse, and to identify its potential interference with the detection of labeled cardiac progenitor cells using 19 F MRI/MR spectroscopy (MRS). MATERIALS AND METHODS: Objectives are demonstrated using (a) in vitro ISO tests, (b) in vivo temporal accumulation/spatial localization C57BL/6 studies (n = 3), and (c) through injections of perfluoro-crown-ether (PFCE) labeled cardiac progenitor cells into femoral muscle areas of the murine hindlimb post-mortem (n = 1) using 1 H/19 F MRI/MRS at 9.4 Tesla. Data were acquired using double-gated spoiled gradient echo images and pulse-acquire spectra. For the in vivo study, the temporal stability of ISO resonances was quantified using coefficient of variability (CV) (5 min) estimates. RESULTS: Two ISO resonances were observed in vivo that correspond to the -CF3 and -OCHF2 moieties. CV values ranged between 3.2 and 6.4% (-CF3 ) and 6.4 and 11.2% (-OCHF2 ). Reductions of the ISO dose (2.0 to 1.7%) at 80 min postinduction had insignificant effects on ISO signals (P = 0.23; P = 0.71). PFCE-labeled cells exhibited a resonance at -16.25 ppm in vitro that did not overlap with the ISO resonances, a finding that is confirmed with MRS post-mortem using injected, labeled cells. Based on 19 F MRI, similar in vivo/post-mortem ISO compartmentalization was also confirmed in peripheral and thoracic skeletal muscles. CONCLUSION: Significant ISO accumulation was observed by 19 F MRS in vivo with temporally stable signals over 90 min postinduction. ISO effects on PFCE labels are anticipated to be minimal but may be more prominent for perfluoropolyether or perfluorooctyl bromide labels. LEVEL OF EVIDENCE: 1 Technical Efficacy: Stage 1 J. MAGN. RESON. IMAGING 2017;45:1659-1667.

Three-dimensional histology: tools and application to quantitative assessment of cell-type distribution in rabbit heart.

AIMS: Cardiac histo-anatomical organization is a major determinant of function. Changes in tissue structure are a relevant factor in normal and disease development, and form targets of therapeutic interventions. The purpose of this study was to test tools aimed to allow quantitative assessment of cell-type distribution from large histology and magnetic resonance imaging- (MRI) based datasets. METHODS AND RESULTS: Rabbit heart fixation during cardioplegic arrest and MRI were followed by serial sectioning of the whole heart and light-microscopic imaging of trichrome-stained tissue. Segmentation techniques developed specifically for this project were applied to segment myocardial tissue in the MRI and histology datasets. In addition, histology slices were segmented into myocytes, connective tissue, and undefined. A bounding surface, containing the whole heart, was established for both MRI and histology. Volumes contained in the bounding surface (called 'anatomical volume'), as well as that identified as containing any of the above tissue categories (called 'morphological volume'), were calculated. The anatomical volume was 7.8 cm(3) in MRI, and this reduced to 4.9 cm(3) after histological processing, representing an 'anatomical' shrinkage by 37.2%. The morphological volume decreased by 48% between MRI and histology, highlighting the presence of additional tissue-level shrinkage (e.g. an increase in interstitial cleft space). The ratio of pixels classified as containing myocytes to pixels identified as non-myocytes was roughly 6:1 (61.6 vs. 9.8%; the remaining fraction of 28.6% was 'undefined'). CONCLUSION: Qualitative and quantitative differentiation between myocytes and connective tissue, using state-of-the-art high-resolution serial histology techniques, allows identification of cell-type distribution in whole-heart datasets. Comparison with MRI illustrates a pronounced reduction in anatomical and morphological volumes during histology processing.

Accelerating global left-ventricular function assessment in mice using reduced slice acquisition and three-dimensional guide-point modelling.

BACKGROUND: To investigate the utility of three-dimensional guide-point modeling (GPM) to reduce the time required for CMR evaluation of global cardiac function in mice, by reducing the number of image slices required for accurate quantification of left-ventricular (LV) mass and volumes. METHODS: Five female C57Bl/6 mice 8 weeks post myocardial infarction induced by permanent occlusion of the left coronary artery, and six male control (un-operated) C57Bl/6 mice, were subject to CMR examination under isoflurane anaesthesia. Contiguous short axis (SAX) slices (1 mm thick 7-9 slices) were obtained together with two long axis (LAX) slices in two chamber and four chamber orientations. Using a mathematical model of the heart to interpolate information between the available slices, GPM LV mass and volumes were determined using full slice (all SAX and two LAX), six slice (four SAX and two LAX) and four slice (two SAX and two LAX) analysis protocols. All results were compared with standard manual volumetric analysis using all SAX slices. RESULTS: Infarct size was 39.1±5.1% of LV myocardium. No significant differences were found in left ventricular mass and volumes between the standard and GPM full and six slice protocols in infarcted mice (113±10, 116±11, and 117±11 mg respectively for mass), or between the standard and GPM full, six and four slice protocols in control mice, (105±14, 106±10, 104±12, and 105±7 mg respectively for mass). Significant differences were found in LV mass (135±18 mg) and EF using the GPM four slice protocol in infarcted mice (p<0.05). CONCLUSION: GPM enables accurate analysis of LV function in mice with relatively large infarcts using a reduced six slice acquisition protocol, and in mice with normal/symmetrical left-ventricular topology using a four slice protocol.

Assessment of global cardiac function.

High-resolution magnetic resonance cine imaging (cine-MRI) allows for a non-invasive assessment of ventricular function and mass in normal mice and in genetically and surgically modified mouse models of cardiac disease. The assessment of myocardial mass and function by cine-MRI does not rely on geometric assumptions, as the hearts are covered from the base to the apex, typically by a stack of two-dimensional images. The MR data acquisition is then followed by image segmentation of specific cine frames in each slice to obtain geometric and functional parameters, such as end-diastolic volume (EDV), end-systolic volume (ESV) or ejection fraction (EF). This technique has been well established in clinical routine application and it is now also becoming the reference method in experimental cardiovascular MRI. The cine images are typically acquired in short- and long-axis orientations of the heart to facilitate an accurate assessment of cardiac functional parameters. These views can be difficult to identify, particularly in animals with diseased hearts. Furthermore, data analysis can be the source of a systematic error, mainly for myocardial mass measurement. We have established protocols that allow for a quick and reproducible way of obtaining the relevant cardiac views for cine-MRI, and for accurate image analysis.

Development of an anatomically detailed MRI-derived rabbit ventricular model and assessment of its impact on simulations of electrophysiological function.

Recent advances in magnetic resonance (MR) imaging technology have unveiled a wealth of information regarding cardiac histoanatomical complexity. However, methods to faithfully translate this level of fine-scale structural detail into computational whole ventricular models are still in their infancy, and, thus, the relevance of this additional complexity for simulations of cardiac function has yet to be elucidated. Here, we describe the development of a highly detailed finite-element computational model (resolution: approximately 125 microm) of rabbit ventricles constructed from high-resolution MR data (raw data resolution: 43 x 43 x 36 microm), including the processes of segmentation (using a combination of level-set approaches), identification of relevant anatomical features, mesh generation, and myocyte orientation representation (using a rule-based approach). Full access is provided to the completed model and MR data. Simulation results were compared with those from a simplified model built from the same images but excluding finer anatomical features (vessels/endocardial structures). Initial simulations showed that the presence of trabeculations can provide shortcut paths for excitation, causing regional differences in activation after pacing between models. Endocardial structures gave rise to small-scale virtual electrodes upon the application of external field stimulation, which appeared to protect parts of the endocardium in the complex model from strong polarizations, whereas intramural virtual electrodes caused by blood vessels and extracellular cleft spaces appeared to reduce polarization of the epicardium. Postshock, these differences resulted in the genesis of new excitation wavefronts that were not observed in more simplified models. Furthermore, global differences in the stimulus recovery rates of apex/base regions were observed, causing differences in the ensuing arrhythmogenic episodes. In conclusion, structurally simplified models are well suited for a large range of cardiac modeling applications. However, important differences are seen when behavior at microscales is relevant, particularly when examining the effects of external electrical stimulation on tissue electrophysiology and arrhythmia induction. This highlights the utility of histoanatomically detailed models for investigations of cardiac function, in particular for future patient-specific modeling.

Fast left ventricular mass and volume assessment in mice with three-dimensional guide-point modeling.

PURPOSE: To investigate the accuracy (vs. standard manual analysis) and precision (scan-rescan reproducibility) of three-dimensional guide-point modeling (GPM) for the assessment of left ventricular (LV) function in mice. METHODS: Six male wildtype C57/Bl6 mice (weight 26.2 +/- 1.1 g) were scanned twice, 3 days apart. Each scan was performed twice, at 0.2 mm/pixel with one average and at 0.1 mm/pixel with two averages. The 24 studies were anonymized and analyzed in blinded fashion using GPM and standard manual slice summation. RESULTS: The average error between GPM and standard analysis was 2.3 +/- 5.8 mg in mass, 1.7 +/- 3.2 microL in end-diastolic volume, 2.3 +/- 3.1 microL in end-systolic volume, -2.7 +/- 4.3% in ejection fraction, -0.6 +/- 3.3 microL in stroke volume, and -0.31 +/- 1.56 ml . min(-1) in cardiac output (mean difference +/- SD of differences, n = 24). The average time taken was 8.0 +/- 2.5 minutes for 3D GPM and 48.5 +/- 8.9 minutes for standard analysis (n = 24). Scan-rescan reproducibility results were similar to the standard analysis. No significant differences were found using linear mixed effects modeling in either accuracy or precision between scan resolutions or analysis method. CONCLUSION: 3D GPM enables fast analysis of mouse LV function, with similar accuracy and reproducibility to standard analysis. An image resolution of 0.2 mm/pixel with one average is adequate for LV function studies.

Ultra-fast and accurate assessment of cardiac function in rats using accelerated MRI at 9.4 Tesla.

MRI can accurately and reproducibly assess cardiac function in rodents but requires relatively long imaging times. Therefore, parallel imaging techniques using a 4-element RF-coil array and MR sequences for cardiac MRI in rats were implemented at ultra-high magnetic fields (9.4 Tesla [T]). The hypothesis that these developments would result in a major reduction in imaging time without loss of accuracy was tested on female Wistar rats under isoflurane anesthesia. High-resolution, contiguous short-axis slices (thickness 1.5 mm) were acquired covering the entire heart. Two interleaved data sets (i) with the volume coil (eight averages) and (ii) with the four-element coil array (one average) were obtained. In addition, two-, three-, and fourfold accelerated data sets were generated through postprocessing of the coil array data, followed by a TGRAPPA reconstruction, resulting in five data sets per rat (in-plane voxel size 100 x 100 microm). Using a single blinded operator, excellent agreement was obtained between volume coil (acquisition time: 88 min) and the fourfold accelerated (<3 min) data sets (e.g., LV mass 436 +/- 21 mg vs 433 +/- 19 mg; ejection fraction 74 +/- 5% vs 75 +/- 4%). This finding demonstrates that it is possible to complete a rat cine-MRI study under 3 min with low variability and without losing temporal or spatial resolution, making high throughput screening programs feasible.

How to perform an accurate assessment of cardiac function in mice using high-resolution magnetic resonance imaging.

High-resolution magnetic resonance cine imaging (cine-MRI) is a method that allows for a non-invasive assessment of left ventricular function and mass. To perform this quantitation, hearts are imaged from the base to the apex by a stack of two-dimensional images. Thus, analysis of myocardial mass and function by cine-MRI does not rely on geometric assumptions. Geometric and functional parameters, such as end-diastolic volume (EDV), end-systolic volume (ESV) or ejection fraction (EF), are obtained by subsequent image segmentation of the respective cine frames in each slice. While this technique has been well established in clinical practice, it is now rapidly becoming the reference method in experimental cardiovascular MRI for accurate quantification of cardiac parameters, thereby aiding the phenotyping of the increasing number of transgenic and surgical mouse models. However, accurate measurement of cardiac functional parameters requires the images to be acquired in short-axis orientation of the heart, which can be difficult to define, particularly in animals with diseased hearts. Furthermore, data analysis can be the source of a systematic error, mainly for myocardial mass measurement. Here, we describe a protocol that allows for a quick and reproducible approach of obtaining the relevant cardiac views for cine-MRI, and we explain how an accurate experimental image analysis can be performed.

Assessment of motion gating strategies for mouse magnetic resonance at high magnetic fields.

PURPOSE: To assess the performance of motion gating strategies for mouse cardiac magnetic resonance (MR) at high magnetic fields by quantifying the levels of motion artifact observed in images and spectra in vivo. MATERIALS AND METHODS: MR imaging (MRI) of the heart, diaphragm, and liver; MR angiography of the aortic arch; and slice-selective 1H-spectroscopy of the heart were performed on anesthetized C57Bl/6 mice at 11.75 T. Gating signals were derived using a custom-built physiological motion gating device, and the gating strategies considered were no gating, cardiac gating, conventional gating (i.e., blanking during respiration), automatic gating, and user-defined gating. Both automatic and user-defined modes used cardiac and respiratory gating with steady-state maintenance during respiration. Gating performance was assessed by quantifying the levels of motion artifact observed in images and the degree of amplitude and phase stability in spectra. RESULTS: User-defined gating with steady-state maintenance during respiration gave the best performance for mouse cardiac imaging, angiography, and spectroscopy, with a threefold increase in signal intensity and a sixfold reduction in noise intensity compared to cardiac gating only. CONCLUSION: Physiological gating with steady-state maintenance during respiration is essential for mouse cardiac MR at high magnetic fields.