Myofiber Orientation Research Articles

Reduced left ventricular dynamics modeling based on a cylindrical assumption.

Biomechanical modeling and simulation is expected to play a significant role in the development of the next generation tools in many fields of medicine. However, full-order finite element models of complex organs such as the heart can be computationally very expensive, thus limiting their practical usability. Therefore, reduced models are much valuable to be used, for example, for pre-calibration of full-order models, fast predictions, real-time applications, and so forth. In this work, focused on the left ventricle, we develop a reduced model by defining reduced geometry & kinematics while keeping general motion and behavior laws, allowing to derive a reduced model where all variables & parameters have a strong physical meaning. More specifically, we propose a reduced ventricular model based on cylindrical geometry & kinematics, which allows to describe the myofiber orientation through the ventricular wall and to represent contraction patterns such as ventricular twist, two important features of ventricular mechanics. Our model is based on the original cylindrical model of Guccione, McCulloch, & Waldman (1991); Guccione, Waldman, & McCulloch (1993), albeit with multiple differences: we propose a fully dynamical formulation, integrated into an open-loop lumped circulation model, and based on a material behavior that incorporates a fine description of contraction mechanisms; moreover, the issue of the cylinder closure has been completely reformulated; our numerical approach is novel aswell, with consistent spatial (finite element) and time discretizations. Finally, we analyze the sensitivity of the model response to various numerical and physical parameters, and study its physiological response.

PO-02-212 TWISTED FIBROTICALLY-INSULATED MYOBUNDLES CREATE THE COMMON TRACK AND PIVOTING POINTS FOR REENTRANT DRIVERS MAINTAINING PERSISTENT AF IN-VIVO

Localized reentrant drivers have been suggested to maintain persistent atrial fibrillation (perAF). However, their existence, electrophysiological mechanisms, and structural substrates in-vivo are controversial. Define AF driving mechanism in-vivo and ex-vivo in a perAF canine model. A clinically relevant canine model (n=5) of self-sustained (4 months) perAF was developed to study AF maintaining mechanisms both in-vivo and ex-vivo by integrated 3D imaging, including high-resolution near-infrared optical mapping (NIOM, 0.3-0.5mm resolution), contrast-enhanced magnetic resonance imaging (CE-MRI, 0.15mm resolution). Transmural fibrosis was validated with histology. CE-MRI was used to quantitatively analyze 3D atrial structure, including transmural fiber orientation and fibrosis content in the sub-epi, intramural, and sub-endocardial layers. In all canine models (n=5), sequential in-vivo to ex-vivo sub-surface NIOM and 3D CE-MRI revealed that the self-sustained perAF was primarily maintained by localized reentrant drivers with temporal stability 53±17% (n=10), which recurrent on the same arrhythmogenic hubs. The 3D structure of the arrhythmogenic driver hubs includes a common intramural reentry track and multiple return paths formed by fibrotically-insulated bundles. The in-vivo spatio-temporal activation patterns of reentrant drivers were preserved ex-vivo (Figure A), including driver AFCL (91±7 ms vs 96±12 ms, p>0.05) and temporal driver stability (51±8% vs 52±6%, n=4, p>0.05). Targeted ablation of the driver reentrant tracks terminated or converted AF to slower tachycardia. Quantitative CE-MRI analysis of 3D myofiber orientations revealed that the transmural fiber twist along the common reentry tracks and pivoting points is greater than in non-driver regions (Figure B-C). The driver reentrant tracks had increased interstitial fibrotic strands and significantly higher intramural fibrosis than the neighboring non-driver regions (Figure D-E). Our results provide in-vivo proof-of-concept that spatially stable reentrant drivers are a mechanism of perAF maintenance. Intramural fibrotic strands and fibrotically-insulated myobundles in regions with the highest transmural myofiber twist which may form the common tracks and pivoting points for return paths of the spatially-stable reentrant AF driver.

On the possibility of estimating myocardial fiber architecture from cardiac strains.

The myocardium is composed of a complex network of contractile myofibers that are organized in such a way as to produce efficient contraction and relaxation of the heart. The myofiber architecture in the myocardium is a key determinant of cardiac motion and the global or organ-level function of the heart. Reports of architectural remodeling in cardiac diseases, such as pulmonary hypertension and myocardial infarction, potentially contributing to cardiac dysfunction call for the inclusion of an architectural marker for an improved assessment of cardiac function. However, the in-vivo quantification of three-dimensional myo-architecture has proven challenging. In this work, we examine the sensitivity of cardiac strains to varying myofiber orientation using a multiscale finite-element model of the LV. Additionally, we present an inverse modeling approach to predict the myocardium fiber structure from cardiac strains. Our results indicate a strong correlation between fiber orientation and LV kinematics, corroborating that the fiber structure is a principal determinant of LV contractile behavior. Our inverse model was capable of accurately predicting the myocardial fiber range and regional fiber angles from strain measures. A concrete understanding of the link between LV myofiber structure and motion, and the development of non-invasive and feasible means of characterizing the myocardium architecture is expected to lead to advanced LV functional metrics and improved prognostic assessment of structural heart disease.

Personalization of biomechanical simulations of the left ventricle by in-vivo cardiac DTI data: Impact of fiber interpolation methods.

Simulations of cardiac electrophysiology and mechanics have been reported to be sensitive to the microstructural anisotropy of the myocardium. Consequently, a personalized representation of cardiac microstructure is a crucial component of accurate, personalized cardiac biomechanical models. In-vivo cardiac Diffusion Tensor Imaging (cDTI) is a non-invasive magnetic resonance imaging technique capable of probing the heart's microstructure. Being a rather novel technique, issues such as low resolution, signal-to noise ratio, and spatial coverage are currently limiting factors. We outline four interpolation techniques with varying degrees of data fidelity, different amounts of smoothing strength, and varying representation error to bridge the gap between the sparse in-vivo data and the model, requiring a 3D representation of microstructure across the myocardium. We provide a workflow to incorporate in-vivo myofiber orientation into a left ventricular model and demonstrate that personalized modelling based on fiber orientations from in-vivo cDTI data is feasible. The interpolation error is correlated with a trend in personalized parameters and simulated physiological parameters, strains, and ventricular twist. This trend in simulation results is consistent across material parameter settings and therefore corresponds to a bias introduced by the interpolation method. This study suggests that using a tensor interpolation approach to personalize microstructure with in-vivo cDTI data, reduces the fiber uncertainty and thereby the bias in the simulation results.

Abstract 9829: Exercise-Induced CITED4 Expression is Necessary for Regional Remodeling of Cardiac Microstructural Tissue Helicity

Background: Exercise promotes cardioprotective benefits for the adult heart; however, the exercise-induced molecular and microstructural remodeling processes have yet to be fully characterized. This study sought to define the spatial interactions between molecular and microstructural remodeling that are induced by exercise and might result in cardiac protection by evaluating regional correlations of expressed cardiac CITED4, a transcriptional co-regulator previously shown to increase in hearts with exercise, and myocardial architecture mapping with diffusion tensor cardiac MRI (DT-CMR). Methods: In the first cohort, male wild-type C57BL/6J mice were housed with or without free access to a running wheel. In the second cohort, CITED4 cardiomyocyte specific knockout (KO) mice were generated along with control littermates (fl/fl mice) and the methods were repeated. After 8-weeks of voluntary wheel running, the mice were euthanized and their hearts were excised, scanned at 14.1 T MRI, and sectioned. RNA Fluorescent in situ hybridization (RNA-FISH) was used to detect CITED4 and all slides were counterstained with DAPI. All experiments were performed under approval of the IACUC of the Massachusetts General Hospital. Results: The exercised group of cohort 1 showed a significant increase in CITED4 expression and microstructural DT-CMR markers in the lateral wall of the left ventricle (p<0.05 for all). In particular, myofiber orientation, evaluated as helicity (helix angle transmurality), was significantly increased within the exercised group (p<0.05), indicating beneficial changes in the myocardial microstructure. Cohort 2 demonstrated that CITED4 was required for these changes to take place, as the CITED4 knock out exercise group did not replicate the same structural changes seen in cohort 1, while the fl/fl control mice reproduced the expected changes in tissue microstructure following exercise. Conclusion: Exercise induces a regional increase in CITED4 expression that coincides with microstructural remodeling. In addition, we have demonstrated that CITED4 is required for these beneficial exercise-induced microstructural changes to occur.

Abstract 15525: Resolving Human Sinoatrial Node Leading Pacemaker vs Atrial Early Activation Sites by 3D High Resolution Imaging to Guide Ablation Treatment of Atrial Arrhythmia

Introduction: Surgical/ablation treatment for atrial arrhythmias such as anatomic sinoatrial node (SAN) tachycardia and atrial fibrillation either target or preserve the SAN. However, defining SAN is challenging because clinical electrophysiological (EP) mapping can visualize only exits of SAN activation as early activation sites (EAS) and not the leading pacemaker (LP) within the 3D intramural SAN. Methods: High resolution (300-900μm 2 ) including epi/endocardial (Epi/Endo) dual sided near infrared optical mapping (NIOM) was conducted on coronary perfused explanted human atria (n=26, 19-69 y.o.). SAN was defined by optical action potential morphologies as the region of slow diastolic depolarization (slow upstroke) preceding atrial excitation. Serial histology and contrast enhanced MRI (CE-MRI, 100μm 3 resolution) were used to define the 3D fibrotic structure of the SAN pacemaker complex. Results: During sinus rhythm (SR) (83±22 bpm), the LP was primarily located in the SAN center. Electrical impulses exit from SAN to atria through 1-2 sinoatrial conduction pathways (SACP), leading to discrete EAS along crista terminalis (CT), preferentially from lateral superior/middle SACPs. The distance between SAN LP and EAS varied from 3.5-23 mm. Heterogeneous atrial wall thickness, fibrosis content and myofiber orientation along the SAN and CT (5-15mm thick) regions led to complex intramural conduction from SAN LP to EAS and substantial Epi-Endo activation dyssynchrony. Conclusions: EAS visualized on both Epi/Endo mapping mainly distributed along thick CT, but not on the surface projection of SAN, due to intramural fiber orientation of preferential SACPs. The higher fibrotic content in the human SAN than CT detected by CE-MRI, integrated with EP mapping can be helpful to accurately define SAN structure and pathways for reentrant SAN arrhythmias ablations.

Refraction-corrected backscatter tensor imaging of excised porcine ventricular myocardium.

Backscatter tensor imaging (BTI) is performed on excised porcine right- and left-ventricular myocardium to estimate the transmural myofiber orientation. Calculation of the backscatter spatial coherence employs measured sound speeds of the myocardium and the fluid that separates the tissue from the imaging array to account for effects of refraction during the delay-and-sum beamforming calculation. Compared to the assumption of a homogeneous sound speed in the imaging region, accounting for refraction yields significantly increased average spatial coherence as well as contrast of spatial coherence between the along- and across-fiber directions, thus improving sensitivity of BTI for myofiber orientation estimation.

A groupwise registration and tractography framework for cardiac myofiber architecture description by diffusion MRI: An application to the ventricular junctions.

BackgroundKnowledge of the normal myocardial–myocyte orientation could theoretically allow the definition of relevant quantitative biomarkers in clinical routine to diagnose heart pathologies. A whole heart diffusion tensor template representative of the global myofiber organization over species is therefore crucial for comparisons across populations. In this study, we developed a groupwise registration and tractography framework to resolve the global myofiber arrangement of large mammalian sheep hearts. To demonstrate the potential application of the proposed method, a novel description of sub-regions in the intraventricular septum is presented.MethodsThree explanted sheep (ovine) hearts (size ~12×8×6 cm3, heart weight ~ 150 g) were perfused with contrast agent and fixative and imaged in a 9.4T magnet. A group-wise registration of high-resolution anatomical and diffusion-weighted images were performed to generate anatomical and diffusion tensor templates. Diffusion tensor metrics (eigenvalues, eigenvectors, fractional anisotropy …) were computed to provide a quantitative and spatially-resolved analysis of cardiac microstructure. Then tractography was performed using deterministic and probabilistic algorithms and used for different purposes: i) Visualization of myofiber architecture, ii) Segmentation of sub-area depicting the same fiber organization, iii) Seeding and Tract Editing. Finally, dissection was performed to confirm the existence of macroscopic structures identified in the diffusion tensor template.ResultsThe template creation takes advantage of high-resolution anatomical and diffusion-weighted images obtained at an isotropic resolution of 150 μm and 600 μm respectively, covering ventricles and atria and providing information on the normal myocardial architecture. The diffusion metric distributions from the template were found close to the one of the individual samples validating the registration procedure. Small new sub-regions exhibiting spatially sharp variations in fiber orientation close to the junctions of the septum and ventricles were identified. Each substructure was defined and represented using streamlines. The existence of a fiber-bundles in the posterior junction was validated by anatomical dissection. A complex structural organization of the anterior junction in comparison to the posterior junction was evidenced by the high-resolution acquisition.ConclusionsA new framework combining cardiac template generation and tractography was applied on the whole sheep heart. The framework can be used for anatomical investigation, characterization of microstructure and visualization of myofiber orientation across samples. Finally, a novel description of the ventricular junction in large mammalian sheep hearts was proposed.

Toward Hierarchical Assembly of Aligned Cell Sheets into a Conical Cardiac Ventricle Using Microfabricated Elastomers.

Despite current efforts in organ-on-chip engineering to construct miniature cardiac models, they often lack some physiological aspects of the heart, including fiber orientation. This motivates the development of bioartificial left ventricle models that mimic the myofiber orientation of the native ventricle. Herein, an approach relying on microfabricated elastomers that enables hierarchical assembly of 2D aligned cell sheets into a functional conical cardiac ventricle is described. Soft lithography and injection molding techniques are used to fabricate micro-grooves on an elastomeric polymer scaffold with three different orientations ranging from -60° to +60°, each on a separate trapezoidal construct. The width of the micro-grooves is optimized to direct the majority of cells along the groove direction and while periodic breaks are used to promote cell-cell contact. The scaffold is wrapped around a central mandrel to obtain a conical-shaped left ventricle model inspired by the size of a human left ventricle 19 weeks post-gestation. Rectangular micro-scale holes are incorporated to alleviate oxygen diffusional limitations within the 3D scaffold. Cardiomyocytes within the 3D left ventricle constructs showed high viability in all layers after 7 days of cultivation. The hierarchically assembled left ventricle also provided functional readouts such as calcium transients and ejection fraction.

The distribution of neuromuscular junctions depends on muscle pennation, when botulinum neurotoxin receptors and SNAREs expression are uniform in the rat

Botulinum neurotoxins (BoNTs) are used to treat spastic disorders. Depending on muscle size, one or multiple injections are recommended according to labels to target neuromuscular junctions (NMJ). However, information about NMJ distribution and number in muscles, as well as expression of receptors and molecular targets of toxins is scarce in human and animal models.Seven muscles from adult rats were used to identify expression of BoNT receptors and SNAREs using immunohistochemistry (IHC), and fluorescent α-Bungarotoxin combined to light-sheet microscopy used to determine their distribution.The location, number, and density of NMJ were muscle specific and mostly dependent on the type of pennation (myofiber orientation). In the Flexor Digitorum Brevis (a very small muscle) NMJ were as numerous as in the Gastrocnemius lateralis. A strong expression of SV2C, Synaptotagmin 2, SNAP25 and VAMP1 were observed in all muscles, and SV2A, Synaptotagmin 1 and VAMP2 were never detected.This work highlights the specific distribution of NMJ in muscles which seems to depend on the type of pennation. Detailed observation of myofibers organization might help clinicians to better evaluate the location of NMJ in humans; the molecular phenotyping of NMJ will contribute to better integrate the rat model into research of BoNT therapeutics.

Contraction Control of Aligned Myofiber Sheet Tissue by Parallel Oriented Induced Pluripotent Stem Cell-Derived Neurons.

Fabrication and application of engineered complex tissues composed of different types of cells is a crucial milestone in the next phase of tissue engineering. The delicate organization structure of each tissue component and its physiological connections enable all the functions in the human body. In this study, cell sheet-based engineering allowed us to fabricate a complex myofiber sheet tissue using motor neurons derived from human-induced pluripotent stem cells. In contrast with previous studies of other groups, a myofiber sheet with a biomimetic aligned structure was produced from human myoblasts using a striped-patterned thermoresponsive dish, which enabled manipulation of the sheet tissue by simply lowering the culture temperature. The myofiber sheet was transferred onto a gel that promotes functional maturation of human myofibers, resulting in production of contractile human muscle tissue. Just by seeding motor neurons onto the sheet tissue, all the neurons physically contacted to the aligned myofibers, and autonomously elongated in parallel to the myofiber orientation. In addition, the neurite outgrowth was enlarged by coculturing on the myofiber sheet. The presence of the neurons enhanced clustering of myofiber acetylcholine receptors (AChRs), typically found at the neuromuscular junctions (NMJs). Consequently, contraction behaviors of the myofiber sheet were regulated by neuronal signal transduction through NMJs. Muscle contraction was induced when the motor neurons were stimulated by glutamic acid, and effectively blocked by administration of d-tubocurarine as an antagonistic inhibitor for the AChR. The fibrin-based gel was useful as a culture environment for tissue maturation and as a favorable substrate for unobstructed contractions. Our neuron-muscle sheet tissue will be scalable by simply enlarging the micropatterned substrate and manipulable three dimensionally; fabrication of a thick tissue and a bundle-like structured tissue will be possible just by layering multiple sheets or rolling up the sheet. Simplified control over self-orientation of neurite elongation will be advantageous for fabrication of such a large and complex tissue. Therefore, our methodology, established in this study, will be instrumental in future applications of regenerative medicine for locomotion apparatus. Impact Statement A complex tissue containing skeletal myofibers and induced pluripotent stem cell-derived motor neurons was fabricated from human cells based on the cell sheet engineering technology. A micropatterned thermoresponsive culture dish and a fibrin-based gel substrate enabled production of aligned, and functionally matured myofiber sheet tissue. The motor neurons were autonomously oriented simply by seeding on the aligned myofiber sheet tissue. Induction and inhibition of the muscle contraction were effectively controlled by neuronal signal transduction. Considering the potential scalability and manipulability of the neuron-muscle sheet tissue, our methodology will contribute to future applications of regenerative medicine for locomotion apparatus.

Abstract 14303: Structural Remodeling in the Left Ventricular Myocardium Underlies Systolic Dysfunction in Myocardial Infarction

Introduction: Myocardial infarction (MI) leads to myofiber death and the formation of a fibrotic scar in the left ventricular free wall (LVFW) myocardium. Acute MI induces remodeling events in the LV consisting of alterations in the architecture and biomechanical properties of the LVFW, collectively leading to LV systolic dysfunction. We implemented a multi-modality pipeline bridging architectural remodeling, LVFW passive behavior adaptation, and ventricular dysfunction in MI. We hypothesize that biomechanical remodeling of the LV serves as an important predictor of clinical outcomes in MI. Methods: MI was induced in rats via the ligation of the LAD artery, and hearts were harvested at five time points: pre-MI, and 1-wk to 4-wk post-MI (n=6 for each time point). Echocardiography and terminal LV catheterization were performed. LVFW specimens were harvested and subjected to biaxial testing, followed by picrosirius red staining. Histological slides were analyzed to quantify fiber architecture. Results: Comparisons for 3-wk MI versus control groups are reported (animal-specific results for representative rats are shown in Fig. 1). Echocardiography imaging showed a decrease in EF (84±5 vs 73±10%; p=0.053) and a decrease in ESV (0.122±0.045 vs 0.31±0.195ml; p=0.013). LV catheterization revealed a decrease in SV (50±0.86 vs 10±0.38μL; p<0.001) and an increase in peak systolic pressure (36.62±0.87 vs 44.91±0.99 mmHg; p<0.001), indicating systolic dysfunction. Biaxial testing revealed longitudinal stiffening of the LVFW at 3-wk post-MI. Scarring and a shift in myofiber orientation towards the longitudinal direction corroborated changes in the LVFW mechanical behavior in the MI group. Conclusions: LV systolic dysfunction in MI is significantly influenced by disruption in myofiber architecture and fiber orientation remodeling in the LVFW. Assessment of LVFW remodeling events provides important incremental information over traditional organ-level indices.

Cardiac mesh morphing method for finite element modeling of heart failure with preserved ejection fraction

Numerical modeling of heart biomechanics can realistically capture morphological variations in diseases and has been helpful in advancing our understanding of the physiology. Subject-specific models require anatomic representation of medical images, and it is desirable to have a consistently repeatable models for any given morphology. In this study, we propose a novel and easily adaptable cardiac reconstruction algorithm by morphing an existing discretized mesh of an advanced finite element (FE) model, to match anatomies acquired from porcine cardiac magnetic resonance imaging (cMRI) scans. The morphing algorithm involves iterative FE simulations with visco-hyperelastic material properties. The living heart porcine model (LHPM) was chosen as the input baseline FE mesh, in order to preserve detailed anatomical features that cannot be captured in routine scans such as myofiber orientations and conduction pathways. The algorithm was demonstrated for the recreation of porcine hearts of a healthy subject and of a subject induced with heart failure with preserved ejection fraction (HFpEF) conditions, where there were substantial hypertrophy and anatomical alterations. We further used the morphed meshes for FE modeling of cardiac contraction and relaxation, thus demonstrating the applicability of the proposed algorithm in producing viable meshes. The results show that our algorithm can recreate the characteristic anatomical changes of cardiac remodeling, including heart muscle thickening, as well as replicate the reduction in ventricular volume. This algorithm allows for the creation of subject-specific models with the same mesh connectivity, thus enabling spatial comparison and analysis of pathologic progress.

Abstract P138: Long-term Effects Of Severe Caloric Restriction To The Heart Function

Severe caloric restriction (sCR) increases the risk for acute cardiovascular disease. Less understood are the long-term effects on cardiovascular disease risk after the sCR period has ended. We investigated the effects of sCR on heart structure and function months after refeeding (sCR-Refed). Female Fischer rats (3-months-old) were maintained on (CT) ad libitum or a 60% caloric restricted diet for 2 weeks. Thereafter, all rats received ad libitum chow for 3 months and they were analyzed by precision ultrasound to assess their heart function. After imaging, the animals were sacrificed and the hearts were subjected to ischemia-reperfusion (I/R) using a Langendorff preparation. After 2 weeks of sCR, rats lost 15% of their initial body weight (BW) [% (100*(Final-Initial/Initial)): CT, 1.5±0.8 vs sCR, -15.4±1.1; p<0.001;n=8]. After 3 months of refeeding, there was no detectable difference in BW between CT and sFR-Refed groups. Isolated hearts from the sCR-Refed rats exhibited worse myocardial pathology after I/R compared to CT rats. The parallel orientation of myofibers and striations normally present in cardiomyocytes was lost in sCR-Refed rats. Further analysis revealed uneven blood-filling of the microcirculatory vessels and prominent interstitial edema of the myocardium. Hearts from sCR-Refed rats had more atrophied cardiomyocytes than CT [Atrophied/Total (%): CT, 0.2±0.1 vs sCR-Refed, 50.6±1.1; p<0.001; n=5]. The number of arrhythmic events during a 30 min ischemic interval in isolated hearts doubled after 2 weeks on the sCR diet ( data not shown ) and remained doubled 3 months later [Arrhythmias (% of time): CT, 34±8 vs sCR-Refed, 68±9; p=0.02; n=8]. Ultrasound imaging showed no difference in stroke volume, coronary perfusion pressure and left ventricular mass. However, the thickness of the left ventricular posterior wall was significantly reduced in sCR-Refed rats [(mm): CT, 2.55 ±0.03 vs sCR-Refed, 2.10±0.04; p=0.002; n=4]. These findings indicate heart structure and function remained damaged months after the sCR period ended and BW was restored. These studies have adverse cardiovascular risk implications for who are subjected either voluntarily (crash diets) or involuntarily (very low food security) to periods of inadequate caloric intake.

Ultrasound measurement of transmural fiber orientation and 3D tissue strain in excised myocardium under uniaxial loading

The noninvasive measurement with ultrasound of myofiber orientation and 3D tissue strain through the tissue thickness during the application of uniaxial tension is demonstrated on porcine myocardium. Complex tissue deformation, including rotation of myofiber sheets, during the application of stretch is observed.

Histologic evaluation of therapeutic responses in ischemic myocardium elicited by dual growth factor delivery from composite glycosaminoglycan hydrogels

In this project, the ability of dual growth factor-preloaded, silk-reinforced, composite hyaluronic acid-based hydrogels to elicit advantageous histologic responses when secured to ischemic myocardium was evaluated in vivo. Reinforced hydrogels containing both Vascular Endothelial Growth Factor (VEGF) and Platelet-derived Growth Factor (PDGF) were prepared by crosslinking chemically modified hyaluronic acid and heparin with poly(ethylene glycol)-diacrylate around a reinforcing silk mesh. Composite patches were sutured to the ventricular surface of ischemic myocardium in Sprague-Dawley rats, and the resulting angiogenic response was followed for 28 days. The gross appearance of treated hearts showed significantly reduced ischemic area and fibrous deposition compared to untreated control hearts. Histologic evaluation showed growth factor delivery to restore myofiber orientation to pre-surgical levels and to significantly increase elicited microvessel density and maturity by day 28 in infarcted myocardial tissue (p < 0.05). In addition, growth factor delivery reduced cell apoptosis and decreased the density of elicited mast cells and both CD68+ and anti-inflammatory CD163+ macrophages. These findings suggest that HA-based, dual growth factor-loaded hydrogels can successfully induce a series of beneficial responses in ischemic myocardium, and offer the potential for therapeutic improvement of ischemic myocardial remodeling.

Myofiber strain in healthy humans using DENSE and cDTI.

Myofiber strain, Eff , is a mechanistically relevant metric of cardiac cell shortening and is expected to be spatially uniform in healthy populations, making it a prime candidate for the evaluation of local cardiomyocyte contractility. In this study, a new, efficient pipeline was proposed to combine microstructural cDTI and functional DENSE data in order to estimate Eff in vivo. Thirty healthy volunteers were scanned with three long-axis (LA) and three short-axis (SA) DENSE slices using 2D displacement encoding and one SA slice of cDTI. The total acquisition time was 11 minutes ± 3 minutes across volunteers. The pipeline first generates 3D SA displacements from all DENSE slices which are then combined with cDTI data to generate a cine of myofiber orientations and compute Eff . The precision of the post-processing pipeline was assessed using a computational phantom study. Transmural myofiber strain was compared to circumferential strain, Ecc , in healthy volunteers using a Wilcoxon sign rank test. In vivo, computed Eff was found uniform transmurally compared to Ecc (-0.14[-0.15, -0.12] vs -0.18 [-0.20, -0.16], P < .001, -0.14 [-0.16, -0.12] vs -0.16 [-0.17, -0.13], P < .001 and -0.14 [-0.16, -0.12] vs Ecc_C = -0.14 [-0.15, -0.11], P = .002, Eff_C vs Ecc_C in the endo, mid, and epi layers, respectively). We demonstrate that it is possible to measure in vivo myofiber strain in a healthy human population in 10 minutes per subject. Myofiber strain was observed to be spatially uniform in healthy volunteers making it a potential biomarker for the evaluation of local cardiomyocyte contractility in assessing cardiovascular dysfunction.

Smart ultrasound device for real-time myocardial stiffness quantification of the human heart

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): ERC Introduction Myocardial stiffness (MS) is crucial to understand cardiac biomechanics and evaluate cardiac function. We recently demonstrated that shear wave imaging using acoustic radiation force can provide quantitative end-diastolic MS in human patients [1] . However, the dependence of shear wave velocity with myofiber orientation remained a limitation and required to perform Shear Wave Velocity (SWV) estimations from different probe orientations which is challenging in clinical practice. We propose a new approach to provide real-time quantitative assessment of MS without dependence of the probe orientation based on a dedicated smart ultrasound (US) device. Methods A new US probe was designed and manufactured to generate acoustic radiation force along the central axis and track the SWV simultaneously along three different orientations to obtain an elliptic profile of SWS. The probe was connected to dedicated electronics and software to provide real-time end-diastolic MS with ECG gating. Validation was performed on 4 in-vitro calibrated phantoms (0.92 – 1.49 – 2.58 – 3.49 m/s) and on ex vivo porcine hearts. MS along and across the fibers were compared to the values measured by conventional shear wave imaging with a linear probe mounted on a rotation motor (angular step of 10°) (Aixplorer, Supersonic imaging). Finally, the in vivo feasibility and reproducibility of measuring MS of the antero-septal wall and of the right ventricular (RV) wall was assessed transthoracically on four human volunteer . Results In vitro results on phantoms showed a good agreement with calibrated value (r2 = 0.98, std = 4.8%). Elliptic profiles on ex-vivo porcine heart showed good agreement with Aixplorer measurements acquired at different angles, with a relative difference along the long axis (LA) of: Δ=7.0%, Δ=7.1%, Δ=9% respectively for left ventricle (LV), right ventricle (RV) and septum. Finally, myocardial SWV assessment in human volunteers was obtained successfully on the RV and on the septum in late diastole. The mean MS was 1.79+/- 0.15 m/s along the fiber direction, the fractional anisotropy (FA) was 0.25 +/- 0.06 on septal wall in good agreement with previous results [1] and 1.06 +/- 0.11 m/s along fibers orientation and a FA of 0.27 +/- 0.08 on RV. Finally the beat to beat reproducibility of MS measurement was estimated to be 8.22%. Conclusion The new smart US device allowed non-invasive quantification of anisotropic myocardial tissues in real time. Results showed the accuracy of the methods. This approach could offer a new clinical tool for the evaluation of the myocardium in cardiomyopathies and in heart failure patients. Abstract Figure. SWV on myocardium human volonteer

Estimating cardiomyofiber strain in vivo by solving a computational model.

Since heart contraction results from the electrically activated contraction of millions of cardiomyocytes, a measure of cardiomyocyte shortening mechanistically underlies cardiac contraction. In this work we aim to measure preferential aggregate cardiomyocyte ("myofiber") strains based on Magnetic Resonance Imaging (MRI) data acquired to measure both voxel-wise displacements through systole and myofiber orientation. In order to reduce the effect of experimental noise on the computed myofiber strains, we recast the strains calculation as the solution of a boundary value problem (BVP). This approach does not require a calibrated material model, and consequently is independent of specific myocardial material properties. The solution to this auxiliary BVP is the displacement field corresponding to assigned values of myofiber strains. The actual myofiber strains are then determined by minimizing the difference between computed and measured displacements. The approach is validated using an analytical phantom, for which the ground-truth solution is known. The method is applied to compute myofiber strains using in vivo displacement and myofiber MRI data acquired in a mid-ventricular left ventricle section in N=8 swine subjects. The proposed method shows a more physiological distribution of myofiber strains compared to standard approaches that directly differentiate the displacement field.

Analysis of Differential Gene Expression in Response to Anisotropic Stretch using a Systems Model of Cardiac Myocyte Mechanotransduction

Cardiomyocytes transduce various mechanical signals into transcriptional responses associated with ventricular hypertrophy and remodeling. The organization of the sarcomere and cytoskeleton suggest that myocytes may respond differentially to mechanical loading applied either parallel or transverse to the long axis of the myofiber. The regulatory networks underlying these differential responses, however, remain unknown. Micropatterned mouse neonatal cardiomyocytes were stretched biaxially, the major axis of strain being either parallel or transverse to the myofiber orientation, to examine the differential effects of anisotropic stretch on gene expression. At 30 minutes and 4 hours of stretch, RNA sequencing identified 843 differentially expressed genes of which 830 were regulated by longitudinal stretch and 82 regulated by transverse stretch. An ordinary differential equation model of cardiac myocyte mechanosignaling predicted the expression of 784 genes with an accuracy of 72%. By simulating inhibition of mechanoreceptors, the model predicted the greatest inhibition of genes induced by transverse stretch when angiotensin II (AngII) and endothelin-1 (ET-1) receptors were blocked. These predictions were confirmed by experiments which showed a significantly greater inhibition (48%) in the transverse-stretch induced expression of Ctgf, Fosl2, Mafk and Nuak1 by combined inhibition of AngII and ET1 receptors with 1uM Losartan and 100uM/10uM BQ123/BQ788, respectively, compared with combined inhibition of the L-type Ca channel (5uM Nifedipine), sodium hydrogen exchanger (10uM HOE642) and TRP channels (1uM Ruthenium Red) which resulted in 18% inhibition. A graph node centrality analysis showed that 22 signaling molecules primarily in MAPK pathways are the dominant regulators of the expression of 20 genes in response to transverse stretch. Thus, we conclude that transverse stretch induces differential gene responses by stimulating AngII and ET1 receptors and activating 9 transcriptional regulators via the MAPK pathways.