Dynamic Model for Characterizing Contractile Behaviors and Mechanical Properties of a Cardiomyocyte.

Studies on the contractile dynamics of heart cells have attracted broad attention for the development of both heart disease therapies and cardiomyocyte-actuated micro-robotics. In this study, a linear dynamic model of a single cardiomyocyte cell was proposed at the subcellular scale to characterize the contractile behaviors of heart cells, with system parameters representing the mechanical properties of the subcellular components of living cardiomyocytes. The system parameters of the dynamic model were identified with the cellular beating pattern measured by a scanning ion conductance microscope. The experiments were implemented with cardiomyocytes in one control group and two experimental groups with the drugs cytochalasin-D or nocodazole, to identify the system parameters of the model based on scanning ion conductance microscope measurements, measurement of the cellular Young's modulus with atomic force microscopy indentation, measurement of cellular contraction forces using the micro-pillar technique, and immunofluorescence staining and imaging of the cytoskeleton. The proposed mathematical model was both indirectly and qualitatively verified by the variation in cytoskeleton, beating amplitude, and contractility of cardiomyocytes among the control and the experimental groups, as well as directly and quantitatively validated by the simulation and the significant consistency of 90.5% in the comparison between the ratios of the Young's modulus and the equivalent comprehensive cellular elasticities of cells in the experimental groups to those in the control group. Apart from mechanical properties (mass, elasticity, and viscosity) of subcellular structures, other properties of cardiomyocytes have also been studied, such as the properties of the relative action potential pattern and cellular beating frequency. This work has potential implications for research on cytobiology, drug screening, mechanisms of the heart, and cardiomyocyte-based bio-syncretic robotics.

[1]  Gordana Vunjak-Novakovic,et al.  Scaffold stiffness affects the contractile function of three‐dimensional engineered cardiac constructs , 2010, Biotechnology progress.

[2]  R. Lal,et al.  Dynamic micromechanical properties of cultured rat atrial myocytes measured by atomic force microscopy. , 1995, The American journal of physiology.

[3]  W. Kraus,et al.  Endothelial, cardiac muscle and skeletal muscle exhibit different viscous and elastic properties as determined by atomic force microscopy. , 2001, Journal of biomechanics.

[4]  K. Pister,et al.  Surface micromachined polysilicon heart cell force transducer , 2000, Journal of Microelectromechanical Systems.

[5]  Jeong-Woo Choi,et al.  Phototactic guidance of a tissue-engineered soft-robotic ray , 2016, Science.

[6]  L. Tung.  An ultrasensitive transducer for measurement of isometric contractile force from single heart cells , 1986, Pflügers Archiv.

[7]  V. Markhasin,et al.  Cooperative mechanisms of thin filament activation and their contribution to the myocardial contractile function: Assessment in a mathematical model , 2009 .

[8]  V S Markhasin,et al.  Heart muscle: mathematical modelling of the mechanical activity and modelling of mechanochemical uncoupling. , 1990, General physiology and biophysics.

[9]  Małgorzata Lekka,et al.  Applicability of AFM in cancer detection. , 2009, Nature nanotechnology.

[10]  Christopher S. Chen,et al.  Cells lying on a bed of microneedles: An approach to isolate mechanical force , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Analysis of Cytoskeleton-Destabilizing Agents by Optimized Optical Navigation and AFM Force Measurements , 2010, Microscopy Today.

[12]  H. Kragh.  The Politics of Excellence: Behind the Nobel Prize in Science , 2002 .

[13]  Giovanni Dietler,et al.  Mechanical properties of biological specimens explored by atomic force microscopy , 2013 .

[14]  G. Whitesides,et al.  Muscular Thin Films for Building Actuators and Powering Devices , 2007, Science.

[15]  P. Mattila,et al.  Contractility-dependent actin dynamics in cardiomyocyte sarcomeres , 2009, Journal of Cell Science.

[16]  C. Luo,et al.  A model of the ventricular cardiac action potential. Depolarization, repolarization, and their interaction. , 1991, Circulation research.

[17]  Byungkyu Kim,et al.  Real-time measurement of the contractile forces of self-organized cardiomyocytes on hybrid biopolymer microcantilevers. , 2005, Analytical chemistry.

[18]  D E Ingber,et al.  Cell shape, cytoskeletal mechanics, and cell cycle control in angiogenesis. , 1995, Journal of biomechanics.

[19]  D. Atar,et al.  Relationship between intracellular calcium and contractile force in stunned myocardium. Direct evidence for decreased myofilament Ca2+ responsiveness and altered diastolic function in intact ventricular muscle. , 1995, Circulation research.

[20]  P Wadsworth,et al.  Nanomolar concentrations of nocodazole alter microtubule dynamic instability in vivo and in vitro. , 1997, Molecular biology of the cell.

[21]  H. Rothuizen,et al.  Translating biomolecular recognition into nanomechanics. , 2000, Science.

[22]  Vasanti Gharpuray,et al.  Gel stretch method: a new method to measure constitutive properties of cardiac muscle cells. , 1998, American journal of physiology. Heart and circulatory physiology.

[23]  A. Khademhosseini,et al.  Carbon-nanotube-embedded hydrogel sheets for engineering cardiac constructs and bioactuators. , 2013, ACS nano.

[24]  H. Asada,et al.  Utilization and control of bioactuators across multiple length scales. , 2014, Lab on a chip.

[25]  Jing Fang,et al.  Dynamical stress characterization and energy evaluation of single cardiac myocyte actuating on flexible substrate. , 2007, Biochemical and biophysical research communications.

[26]  K. Chien,et al.  Cardiac mechanotransduction and implications for heart disease , 2003, Journal of Molecular Medicine.

[27]  M. Günther,et al.  A macroscopic ansatz to deduce the Hill relation. , 2010, Journal of theoretical biology.

[28]  Junhyong Kim,et al.  Transcriptome transfer provides a model for understanding the phenotype of cardiomyocytes , 2011, Proceedings of the National Academy of Sciences.

[29]  J. Howard,et al.  Mechanics of Motor Proteins and the Cytoskeleton , 2001 .

[30]  P. Doevendans,et al.  Extracellular matrix formation after transplantation of human embryonic stem cell-derived cardiomyocytes , 2009, Cellular and Molecular Life Sciences.

[31]  Stefano Severi,et al.  Mathematical modelling of the action potential of human embryonic stem cell derived cardiomyocytes , 2012, BioMedical Engineering OnLine.

[32]  J. Cooper,et al.  Effects of cytochalasin and phalloidin on actin , 1987, The Journal of cell biology.

[33]  J. Cheung,et al.  Relaxation abnormalities in single cardiac myocytes from renovascular hypertensive rats. , 1992, The American journal of physiology.

[34]  George I. Zahalak,et al.  Modeling Muscle Mechanics (and Energetics) , 1990 .

[35]  Jinchao Xu,et al.  Measuring single cardiac myocyte contractile force via moving a magnetic bead. , 2005, Biophysical journal.

[36]  U. Keyser,et al.  Real-time deformability cytometry: on-the-fly cell mechanical phenotyping , 2015, Nature Methods.

[37]  Lianqing Liu,et al.  Nanoscale imaging and mechanical analysis of Fc receptor-mediated macrophage phagocytosis against cancer cells. , 2014, Langmuir : the ACS journal of surfaces and colloids.

[38]  D. Ingber.  Tensegrity: the architectural basis of cellular mechanotransduction. , 1997, Annual review of physiology.

[39]  D. Ingber,et al.  Mechanical behavior in living cells consistent with the tensegrity model , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[40]  S. Tideswell,et al.  Filament compliance and tension transients in muscle , 1996, Journal of Muscle Research & Cell Motility.

[41]  Takumi Washio,et al.  A three-dimensional simulation model of cardiomyocyte integrating excitation-contraction coupling and metabolism. , 2011, Biophysical journal.

[42]  L. Addadi,et al.  Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates , 2001, Nature Cell Biology.

[43]  E. Evans,et al.  Cortical shell-liquid core model for passive flow of liquid-like spherical cells into micropipets. , 1989, Biophysical journal.

[44]  K. Walley,et al.  Novel regulatory mechanism of cardiomyocyte contractility involving ICAM-1 and the cytoskeleton. , 2004, American journal of physiology. Heart and circulatory physiology.

[45]  A. Hill.  The heat of shortening and the dynamic constants of muscle , 1938 .

[46]  Kevin D. Costa,et al.  Osteoblast Elastic Modulus Measured by Atomic Force Microscopy Is Substrate Dependent , 2005, Annals of Biomedical Engineering.

[47]  Sung-Jin Park,et al.  Instrumented cardiac microphysiological devices via multi-material 3D printing , 2016, Nature materials.

[48]  J. Ohayon,et al.  Theoretical analysis of the adaptive contractile behaviour of a single cardiomyocyte cultured on elastic substrates with varying stiffness. , 2008, Journal of theoretical biology.

[49]  Mark D. Huffman,et al.  Executive Summary: Heart Disease and Stroke Statistics—2015 Update A Report From the American Heart Association , 2011, Circulation.

[50]  Topi Korhonen,et al.  Mathematical Model of Mouse Embryonic Cardiomyocyte Excitation–Contraction Coupling , 2008, The Journal of general physiology.

[51]  W. Danforth,et al.  GLYCOGENOLYSIS DURING TETANIC CONTRACTION OF ISOLATED MOUSE MUSCLES IN THE PRESENCE AND ABSENCE OF PHOSPHORYLASE A. , 1964, The Journal of biological chemistry.

[52]  Bernard Nysten,et al.  Nanoscale mapping of the elasticity of microbial cells by atomic force microscopy , 2003 .

[53]  C. Luo,et al.  A dynamic model of the cardiac ventricular action potential. I. Simulations of ionic currents and concentration changes. , 1994, Circulation research.

[54]  B. Williams,et al.  A self-propelled biohybrid swimmer at low Reynolds number , 2014, Nature Communications.

[55]  Milica Radisic,et al.  Influence of substrate stiffness on the phenotype of heart cells , 2010, Biotechnology and bioengineering.

[56]  J. Xi,et al.  Self-assembled microdevices driven by muscle , 2005, Nature materials.

[57]  Megan L. McCain,et al.  A tissue-engineered jellyfish with biomimetic propulsion , 2012, Nature Biotechnology.

[58]  Lianqing Liu,et al.  Phase modulation mode of scanning ion conductance microscopy , 2014 .

[59]  D. Ingber,et al.  Mechanotransduction across the cell surface and through the cytoskeleton , 1993 .

[60]  R. Bashir,et al.  Development of Miniaturized Walking Biological Machines , 2012, Scientific Reports.

[61]  Yuechao Wang,et al.  Modeling and analysis of bio-syncretic micro-swimmers for cardiomyocyte-based actuation , 2016, Bioinspiration & biomimetics.

[62]  Daniel W. Repperger,et al.  Biomimetic model of skeletal muscle isometric contraction: I. an energetic-viscoelastic model for the skeletal muscle isometric force twitch , 2004, Comput. Biol. Medicine.

[63]  N. Gavara,et al.  Probing mechanical properties of living cells by atomic force microscopy with blunted pyramidal cantilever tips. , 2005, Physical review. E, Statistical, nonlinear, and soft matter physics.

[64]  P. Zorlutuna,et al.  Development and characterization of muscle-based actuators for self-stabilizing swimming biorobots. , 2016, Lab on a chip.

[65]  D. Kass,et al.  What Mechanisms Underlie Diastolic Dysfunction in Heart Failure? , 2004, Circulation research.

[66]  M. Olschewski,et al.  Diminished post-rest potentiation of contractile force in human dilated cardiomyopathy. Functional evidence for alterations in intracellular Ca2+ handling. , 1996, The Journal of clinical investigation.

[67]  S Sugiura,et al.  A novel method to study contraction characteristics of a single cardiac myocyte using carbon fibers. , 2001, American journal of physiology. Heart and circulatory physiology.

[68]  G. Stoney.  The Tension of Metallic Films Deposited by Electrolysis , 1909 .