1,366 research outputs found

    Image Processing Techniques for Assessing Contractility in Isolated Neonatal Cardiac Myocytes

    Get PDF
    We describe a computational framework for the quantitative assessment of contractile responses of isolated neonatal cardiac myocytes. To the best of our knowledge, this is the first report on a practical and accessible method for the assessment of contractility in neonatal cardiocytes. The proposed methodology is comprised of digital video recording of the contracting cell, signal preparation, representation by polar Fourier descriptors, and contractility assessment. The different processing stages are variants of mathematically sound and computationally robust algorithms very well established in the scientific community. The described computational approach provides a comprehensive assessment of the neonatal cardiac myocyte contraction without the need of elaborate instrumentation. The versatility of the methodology allows it to be employed in determining myocyte contractility almost simultaneously with the acquisition of the Ca2+ transient and other correlates of cell contraction. The proposed methodology can be utilized to evaluate changes in contractile behavior resulting from drug intervention, disease models, transgeneity, or other common applications of neonatal cardiocytes

    Introduction of non-linear elasticity models for characterization of shape and deformation statistics: application to contractility assessment of isolated adult cardiocytes

    Get PDF
    <p>Abstract</p> <p>Background</p> <p>We are exploring the viability of a novel approach to cardiocyte contractility assessment based on biomechanical properties of the cardiac cells, energy conservation principles, and information content measures. We define our measure of cell contraction as being the distance between the shapes of the contracting cell, assessed by the minimum total energy of the domain deformation (warping) of one cell shape into another. To guarantee a meaningful <it>vis-à-vis </it>correspondence between the two shapes, we employ both a data fidelity term and a regularization term. The data fidelity term is based on nonlinear features of the shapes while the regularization term enforces the compatibility between the shape deformations and that of a hyper-elastic material.</p> <p>Results</p> <p>We tested the proposed approach by assessing the contractile responses in isolated adult rat cardiocytes and contrasted these measurements against two different methods for contractility assessment in the literature. Our results show good qualitative and quantitative agreements with these methods as far as frequency, pacing, and overall behavior of the contractions are concerned.</p> <p>Conclusions</p> <p>We hypothesize that the proposed methodology, once appropriately developed and customized, can provide a framework for computational cardiac cell biomechanics that can be used to integrate both theory and experiment. For example, besides giving a good assessment of contractile response of the cardiocyte, since the excitation process of the cell is a closed system, this methodology can be employed in an attempt to infer statistically significant model parameters for the constitutive equations of the cardiocytes.</p

    Image Processing Techniques for Assessing Contractility in Isolated Adult Cardiac Myocytes

    Get PDF
    We describe a computational framework for the comprehensive assessment of contractile responses of enzymatically dissociated adult cardiac myocytes. The proposed methodology comprises the following stages: digital video recording of the contracting cell, edge preserving total variation-based image smoothing, segmentation of the smoothed images, contour extraction from the segmented images, shape representation by Fourier descriptors, and contractility assessment. The different stages are variants of mathematically sound and computationally robust algorithms very well established in the image processing community. The physiologic application of the methodology is evaluated by assessing overall contraction in enzymatically dissociated adult rat cardiocytes. Our results demonstrate the effectiveness of the proposed approach in characterizing the true, two-dimensional, “shortening” in the contraction process of adult cardiocytes. We compare the performance of the proposed method to that of a popular edge detection system in the literature. The proposed method not only provides a more comprehensive assessment of the myocyte contraction process but also can potentially eliminate historical concerns and sources of errors caused by myocyte rotation or translation during contraction. Furthermore, the versatility of the image processing techniques makes the method suitable for determining myocyte shortening in cells that usually bend or move during contraction. The proposed method can be utilized to evaluate changes in contractile behavior resulting from drug intervention, disease modeling, transgeneity, or other common applications to mammalian cardiocytes

    Cell Shortening and Calcium Homeostasis Analysis in Adult Cardiomyocytes via a New Software Tool

    Get PDF
    Intracellular calcium (Ca2+) is the central regulator of heart contractility. Indeed, it couples the electrical signal, which pervades the myocardium, with cardiomyocytes contraction. Moreover, alterations in calcium management are the main factors contributing to the mechanical and electrical dysfunction observed in failing hearts. So, simultaneous analysis of the contractile function and intracellular Ca2+ is indispensable to evaluate cardiomyocytes activity. Intracellular Ca2+ variations and fraction shortening are commonly studied with fluorescent Ca2+ indicator dyes associated with microscopy techniques. However, tracking and dealing with multiple files manually is time-consuming and error-prone and often requires expensive apparatus and software. Here, we announce a new, user-friendly image processing and analysis tool, based on ImageJ-Fiji/MATLAB® software, to evaluate the major cardiomyocyte functional parameters. We succeeded in analyzing fractional cell shortening, Ca2+ transient amplitude, and the kinematics/dynamics parameters of mouse isolated adult cardiomyocytes. The proposed method can be applied to evaluate changes in the Ca2+ cycle and contractile behavior in genetically or pharmacologically induced disease models, in drug screening and other common applications to assess mammalian cardiomyocyte functions

    Image-based evaluation of contraction–relaxation kinetics of human-induced pluripotent stem cell-derived cardiomyocytes: Correlation and complementarity with extracellular electrophysiology

    Get PDF
    AbstractIn this study, we used high-speed video microscopy with motion vector analysis to investigate the contractile characteristics of hiPS-CM monolayer, in addition to further characterizing the motion with extracellular field potential (FP), traction force and the Ca2+ transient. Results of our traction force microscopy demonstrated that the force development of hiPS-CMs correlated well with the cellular deformation detected by the video microscopy with motion vector analysis. In the presence of verapamil and isoproterenol, contractile motion of hiPS-CMs showed alteration in accordance with the changes in fluorescence peak of the Ca2+ transient, i.e., upstroke, decay, amplitude and full-width at half-maximum. Simultaneously recorded hiPS-CM motion and FP showed that there was a linear correlation between changes in the motion and field potential duration in response to verapamil (30–150nM), isoproterenol (0.1–10μM) and E-4031 (10–50nM). In addition, tetrodotoxin (3–30μM)-induced delay of sodium current was corresponded with the delay of the contraction onset of hiPS-CMs. These results indicate that the electrophysiological and functional behaviors of hiPS-CMs are quantitatively reflected in the contractile motion detected by this image-based technique. In the presence of 100nM E-4031, the occurrence of early after-depolarization-like negative deflection in FP was also detected in the hiPS-CM motion as a characteristic two-step relaxation pattern. These findings offer insights into the interpretation of the motion kinetics of the hiPS-CMs, and are relevant for understanding electrical and mechanical relationship in hiPS-CMs

    Cell-based gene therapy for mending infarcted hearts

    Get PDF
    The goal of this study was to analyse the efficiency of a combinatorial cell/growth factor therapy to improve function of infarcted murine hearts. The Insulin-like Growth Factor-1 (IGF-1) isoform, IGF-1Ea, has been shown to reduce scar formation and decrease cell death after MI. The present study utilized P19Cl6-derived, IGF-1Ea over-expressing cardiomyocytes to achieve its goal. The P19Cl6 cells were stably transduced with IGF-1Ea using a lentiviral vector and investigated first in vitro for their feasibility for in vivo cell therapy. The engineered pluripotent cells over-expressing IGF-1Ea survived better to hypoxia-induced injury than the control cells. The cells maintained their pluripotency and efficient differentiation capacity towards ventricular cardiomyocyte lineage, generating large quantities of cardiomyocytes optimal for the transplantation study. The generated cardiomyocytes were functionally active and exhibited a mature phenotype. Transplantation of the cardiomyocytes into allogeneic wild type murine infarcted hearts conferred a tendency for maintenance of function at short-term time point. At long-term however, this effect was lost, returning to the level of the control infarcted hearts. Cell tracing assessment revealed engraftment of both IGF-1Ea- and empty-cells, although the cells failed to couple with the recipient tissue. Scar size and capillary density analyses revealed no significant difference between the cells transplanted compared to the saline treated hearts, corroborating with the long-term functional data. Interestingly, the IGF- 1Ea-cell transplanted hearts expressed significantly higher amount of VEGFa compared to the controls, albeit no change in capillary density. Further investigation revealed that the enhanced VEGFa expression in IGF-1Ea-cells transplanted hearts was associated with reduced hypertrophy, marked by reduced cell cross-sectional area at the border-zone, aSK and bMHC expression compared to the control hearts. Nonetheless, modulation of hypertrophic response and transplantation of IGF-1Ea-cells were not able to confer lasting functional preservation, possibly due to lack of sufficient engraftment and coupling of the transplanted cells

    Regional Mechanical Function Changes Remain after Ventricular Pacing Cessation: Evidence of Mechanical Cardiac Memory

    Get PDF
    Every year 400,000 - 600,000 people in the United States die from sudden cardiac death. Sudden cardiac death is often caused by irregular electrical impulses, or arrhythmias, in the heart. Arrhythmias can be corrected through pharmacological therapies, device therapies, or both. One type of device therapy, pacemakers, are inserted in the heart to correct arrhythmias. After a period of ventricular pacing, cardiac memory is defined by changes in the T-wave that are persistent upon return to normal activation pathways. During ventricular pacing, regional stroke work in areas closest to the pacing electrode is significantly decreased. We hypothesize that the mechanical function in the region around the pacing site will continue to have altered mechanical function after cession of pacing, in effect showing a mechanical cardiac memory. To test the hypothesis, nine canine models were implanted with pacing electrodes in both the atrium and ventricle. After a forty- minute stabilization period, baseline data were obtained during atrial pacing. Cardiac memory was induced in five canine models through a two-hour period of ventricular pacing followed immediately by atrial pacing. The remaining canine models served as controls, undergoing atrial pacing for two hours. High- density mapper (HDM) was used to determine mechanical function in a region centered approximately 1 cm away from the pacing electrode. No differences in global function (tau, developed pressure, dP/dtmax, dP/dtmin) were found after two hours of ventricular pacing upon return to normal activation pathways. There was a significant decrease in regional stroke work in an area close to the electrode between baseline (5.7 ± 2.6 %), during ventricular pacing (-3.8 ± 0.9 %)(p\u3c0.05) and after two hours of ventricular pacing upon return to normal activation pathways (2.4 ± 1.6 %)(p\u3c0.05). Further, systolic area contraction was also significantly different between baseline (5.0 ± 6.6 %) and after two hours of ventricular pacing upon return to normal activation pathways (0.2 ± 7.4 %)(p\u3c0.05). Diastolic twist and diastolic twist rates showed no significant differences. Finally, contractile principal strain increased by inducing cardiac memory (-2.6 ± 0.3 %) as compared to baseline (-1.1 ± 0.5 %)(p\u3c0.05). These findings suggest there is a mechanical correlation to electrical cardiac memory
    corecore