Nobuyuki Magome

Electrospun nanofibers as a tool for architecture control in engineered cardiac tissue

This paper presents an in vitro system for cardiac tissue engineering based on cardiomyocytes cultured on electrospun polymethylglutarimide (PMGI) nanofibrous meshes either imprinted on solid substrate or suspended in space. Special care was taken over the ability to control the tissue architecture. The electrospinning process allowed nano-scale diameter PMGI fibers with different positioning density to be collected in a random or in an aligned way that defines the general configuration of the mesh. Micro-imprinted on solid substrate nanofibers guarantee aligned cell growth, when the distance between them is 30 μm or less. Suspended in 3D space, nanofibers define the overall architecture of the tissue, depending on orientation and positioning density of the nanofibers. As a result, cardiac cells proliferated into contractile tissue filaments, open-worked tissue meshes and continuous anisotropic cell sheets. Alignment of the cells was characterized by elongation of the cell shape and orientation of the α-actin filaments supported by the FFT data. The advantage of this method is its ability to maintain both three-dimensionality and structural anisotropy.

Finding the optimal path with the aid of chemical wave

Abstract It is shown that the optimal path in a two-dimensional vector field is deduced by the use of chemical wave in the Belousov-Zhabotinsky reaction (BZ reaction). We also reproduced our experimental result in a numerical simulation based on a two-variable reaction-diffusion equation. The present result provides a simple model for the future application of excitable media to parallel computing, i.e., an excitable medium serves as a self-organized parallel processor.

Convective and periodic motion driven by a chemical wave

The generation of convective flow by a chemical wave was studied experimentally on a mm-sized droplet of Belousov–Zhabotinsky (BZ) reaction medium. A propagating chemical wave causes a transient increase in interfacial tension, and this local change in interfacial tension induces convection. The observed flow profile was reproduced with a numerical simulation by introducing the transient increase in interfacial tension to a modified Navier–Stokes equation coupled with a chemical kinetic equation; a modified Oregonator. We also observed the periodic motion of a BZ droplet floating on an oil phase. Such periodic motion is attributed to the rhythmic change in interfacial tension. The observed periodic convective motion coupled with a chemical reaction is discussed in relation to chemo-mechanical energy transduction under isothermal conditions.

Spontaneous mode-selection in the self-propelled motion of a solid/liquid composite driven by interfacial instability.

Spontaneous motion of a solid/liquid composite induced by a chemical Marangoni effect, where an oil droplet attached to a solid soap is placed on a water phase, was investigated. The composite exhibits various characteristic motions, such as revolution (orbital motion) and translational motion. The results showed that the mode of this spontaneous motion switches with a change in the size of the solid scrap. The essential features of this mode-switching were reproduced by ordinary differential equations by considering nonlinear friction with proper symmetry.

Liquid/liquid dynamic phase separation induced by a focused laser

We found that a focused laser can generate microscopic phase separation in an oil/water system. An oil droplet emerges and grows at the focus of the laser in a water-rich homogeneous medium. In contrast, in an oil-rich homogeneous phase, water droplets spring out in a successive manner from the focus of the laser, move away, and disappear in the surroundings, forming a flower-like pattern. The mechanism of this dynamic phase separation is discussed under the framework of the mean field theory.

Curvature-dependent excitation propagation in cultured cardiac tissue

The geometry of excitation wave front may play an important role on the propagation block and spiral wave formation. The wave front which is bent over the critical value due to interaction with the obstacles may partially cease to propagate and appearing wave breaks evolve into rotating waves or reentry. This scenario may explain how reentry spontaneously originates in a heart. We studied highly curved excitation wave fronts in the cardiac tissue culture and found that in the conditions of normal, non-inhibited excitability the curvature effects do not play essential role in the propagation. Neither narrow isthmuses nor sharp corners of the obstacles, being classical objects for production of extremely curved wave front, affect non-inhibited wave propagation. The curvature-related phenomena of the propagation block and wave detachment from the obstacle boundary were observed only after partial suppression of the sodium channels with Lidocaine. Computer simulations confirmed the experimental observations. The explanation of the observed phenomena refers to the fact that the heart tissue is made of finite size cells so that curvature radii smaller than the cardiomyocyte size loses sense, and in non-inhibited tissue the single cell is capable to transmit excitation to its neighbors.

Plastic bottle oscillator: Rhythmicity and mode bifurcation of fluid flow

The oscillatory flow of water draining from an upside-down plastic bottle with a thin pipe attached to its head is studied as an example of a dissipative structure generated under far-from-equilibrium conditions. Mode bifurcation was observed in the water/air flow: no flow, oscillatory flow, and counter flow were found when the inner diameter of the thin pipe was changed. The modes are stable against perturbations. A coupled two-bottle system exhibits either in-phase or anti-phase self-synchronization. These characteristic behaviors imply that the essential features of the oscillatory flow in a single bottle system can be described as a limit-cycle oscillation.

An Oil Droplet That Spontaneously Climbs up Stairs

It has been reported that an oil droplet on a glass surface moves spontaneously in an oil-water system. This motion of an oil droplet can be understood as the spreading of a reactive droplet, which is induced by the interfacial tension gradient at the glass surface. In this paper, we focus on the spontaneous motion of an oil droplet climbing up stairs. We found that an oil droplet tends to move up the stairs rather than to step down. We describe some of the mechanisms of this unique behavior.

Photo-Control of Excitation Waves in Cardiomyocyte Tissue Culture

Azobenzene photoswitches were recently reported to control the activity of neural cells and heart beat in leeches. Here, we report photocontrol of excitation of cultured cardiomyocytes that have been made light sensitive by using the addition of azobenzene trimethylammonium bromide (AzoTAB). The trans-isomer of AzoTAB reversibly suppresses spontaneous activity and propagation of excitation waves, whereas the cis-isomer has no detectable effect on the electrical properties of cardiomyocytes. Photoisomerization of AzoTAB was achieved by switching the illumination wavelength, λ, from ~440 nm (trans-isomer) to ~350 nm (cis-isomer). Simultaneous irradiation at two wavelengths with properly chosen intensities allowed for dynamic control of the cis-isomer/trans-isomer ratio and the level of excitability from normal to fully unexcitable. Experiments were conducted by using AzoTAB-treated confluent monolayers of neonatal rat cardiomyocytes. Excitation waves were monitored by using the Ca2+-sensitive fluorescent dye Fluo-4. By projecting two-wavelength illumination patterns onto otherwise uniform cell layers, we were able to create excitable networks with the desired topology, dimensions, and functional properties. The present article discusses potential applications of this technique for the analysis of complex patterns of electrical excitation and cardiac arrhythmias.

Digital photocontrol of the network of live excitable cells

Recent development of tissue engineering techniques allows creating and maintaining almost indefinitely networks of excitable cells with desired architecture. We coupled the network of live excitable cardiac cells with a common computer by sensitizing them to light, projecting a light pattern on the layer of cells, and monitoring excitation with the aid of fluorescent probes (optical mapping). As a sensitizing substance we used azobenzene trimethylammonium bromide (AzoTAB). This substance undergoes cis-trans-photoisomerization and trans-isomer of AzoTAB inhibits excitation in the cardiac cells, while cis-isomer does not. AzoTAB-mediated sensitization allows, thus, reversible and dynamic control of the excitation waves through the entire cardiomyocyte network either uniformly, or in a preferred spatial pattern. Technically, it was achieved by coupling a common digital projector with a macroview microscope and using computer graphic software for creating the projected pattern of conducting pathways. This approach allows real time interactive photocontrol of the heart tissue.