Development of a three-dimensional bioprinter: construction of cell supporting structures using hydrogel and state-of-the-art inkjet technology.

We have developed a new technology for producing three-dimensional (3D) biological structures composed of living cells and hydrogel in vitro, via the direct and accurate printing of cells with an inkjet printing system. Various hydrogel structures were constructed with our custom-made inkjet printer, which we termed 3D bioprinter. In the present study, we used an alginate hydrogel that was obtained through the reaction of a sodium alginate solution with a calcium chloride solution. For the construction of the gel structure, sodium alginate solution was ejected from the inkjet nozzle (SEA-Jet, Seiko Epson Corp., Suwa, Japan) and was mixed with a substrate composed of a calcium chloride solution. In our 3D bioprinter, the nozzle head can be moved in three dimensions. Owing to the development of the 3D bioprinter, an innovative fabrication method that enables the gentle and precise fixation of 3D gel structures was established using living cells as a material. To date, several 3D structures that include living cells have been fabricated, including lines, planes, laminated structures, and tubes, and now, experiments to construct various hydrogel structures are being carried out in our laboratory.

Development of mechanical circulatory support devices at the University of Tokyo.

The development of mechanical circulatory support devices at the University of Tokyo has focused on developing a small total artificial heart (TAH) since achieving 532 days of survival of an animal with a paracorporial pneumatically driven TAH. The undulation pump was invented to meet this purpose. The undulation pump total artificial heart (UPTAH) is an implantable TAH that uses an undulation pump. To date, the UPTAH has been implanted in 71 goats weighting from 39 to 72 kg. The control methods are very important in animal experiments, and sucking control was developed to prevent atrial sucking. Rapid left-right balance control was performed by monitoring left atrial pressure to prevent acute lung edema caused by the rapid increase in both arterial pressure and venous return associated with the animal becoming agitated. Additionally, 1/R control was applied to stabilize the right atrial pressure. By applying these control methods, seven goats survived more than 1 month. The maximum survival period was 63 days. We are expecting to carry out longer term animal experiments with a recent model of TAH. In addition to the TAH, an undulation pump ventricular assist device (UPVAD), which is an implantable ventricular assist device (VAD), has been in development since 2002, based on the technology of the UPTAH. The UPVAD was implanted in six goats; three goats survived for more than 1 month. While further research and development is required to complete the the UPVAD system, the UPVAD has good potential to be realized as an implantable pulsatile-flow VAD.

Development of a novel intrafascicular nerve electrode.

Artificial organs could be controlled using autonomic neural signals, because they exhibit rapid responses to physical needs similar to those of natural organs. A nerve electrode must satisfy many requirements to measure autonomous neural signals such as a long lifetime, high signal-to-noise ratio, multichannel recording, simple installation into a nerve fascicle, and good manufacturing productivity. The purpose of our study is to propose and evaluate a novel nerve electrode that satisfies these conditions, which to date has not been developed. A novel intrafascicular nerve electrode was designed, fabricated, and evaluated on autonomic nerves. Conventional extrafascicular and intrafascicular nerve electrodes were fabricated and tested for comparison to our novel intrafascicular nerve electrode. The novel intrafascicular nerve electrode had a 3-week lifetime, whereas the conventional extrafascicular nerve electrode had a 2-week lifetime. The signal-to-noise ratio was improved from 1.6 to 2.0 compared with the conventional extrafascicular nerve electrode. The novel intrafascicular nerve electrode was easier to install into a nerve fascicle and had better manufacturing productivity than the conventional intrafascicular nerve electrode. We succeeded in demonstrating the feasibility of our novel intrafascicular nerve electrode.

Microcirculation of the bulbar conjunctiva in the goat implanted with a total artificial heart: effects of pulsatile and nonpulsatile flow.

A new system to observe the microcirculation on the bulbar conjunctiva was developed using a digital high definition microscope to investigate the influence of the flow patterns on the microcirculation in a goat with a total artificial heart (TAH). The undulation pump TAH was implanted into the goat. When the whole body condition became stable, the flow pattern was modulated between the pulsatile and the nonpulsatile mode, and the changes in the microcirculation were observed. When the flow pattern was changed from pulsatile to nonpulsatile mode, the erythrocyte velocity in capillaries dropped from 526+/-83 to 132+/-41 microm/s and remained at a low level. The number of perfused capillaries decreased as well. Then the nonpulsatile flow mode was maintained for 20 minutes. After the flow pattern was returned to the pulsatile mode again, the erythrocyte velocity recovered to the initial level (433+/-71 microm/s). In many cases, the flow of the nonperfused capillaries in the nonpulsatile mode recovered to the initial level after the flow pattern was changed to the pulsatile mode again. The perfused capillary density in the nonpulsatile mode (19.7+/-4.1 number of capillaries/mm) was significantly lower than that in the pulsatile mode (34.7+/-6.3 number of capillaries/mm). It is thought that the basal and flow stimulated endothelium derived nitric oxide release in the microvessels decreased because of the disappearance of pulsatility and that the nitric oxide induced the constriction of arterioles after the flow pattern was changed to the nonpulsatile mode. At the same time, the baroceptors might sense the decrease in the arterial peak pressure or dp/dt, and the sympathetic nerve increases activities and induce the constriction of arterioles. Then, the erythrocyte velocity in capillaries would decrease. Because of the flow pattern further in the chronic phase, it is important to follow the change in the microcirculation.

A temporal and spatial analysis of cavitation on mechanical heart valves by observing faint light emission.

Cavitation on mechanical heart valves (MHVs) could cause the mechanical failure of the occluder. A simple and reliable in vitro test method to evaluate cavitation potential must be developed. The bubble implosion damages the MHV material; thus, observing the behavior of the bubble implosion is essential. According to sonoluminescence, the collapsing cavity emits faint light. Therefore, in this study, the bubble collapse was analyzed both temporally and spatially by observing faint light emission. A photon counting system has been developed using a photomultiplier tube (H7360-01, Hamamatsu Photonics, Japan). The highest time resolution of this system is 5 microsec. A quartz optical fiber bundle of 2 mm diameter can be connected to this photomultiplier tube and traversed two-dimensionally over the MHV. The closure of the MHV triggers the photon counter, and the photons through 500 beats are recorded and integrated. A 20 mm Björk-Shiley valve was submerged in a water tank containing 10 L deionized water, and the pressure difference of 120 mm Hg was exerted on the valve at a rate of 60 bpm with a pulse duplicator. Approximately 700 microsec after the valve closure, light emission was detected along the edge of the occluder on the inflow side in the major orifice. Then, approximately 1,000 microsec after the closure, light along the occluder's edge in the minor orifice was recorded as well. Compared with the analysis, using a stroboscope and a high-speed camera, faint light was emitted from the collapsing cavities. In conclusion, sonoluminescnece was successfully observed around the MHV, and the photon counting technique and the traversing mechanism of the optical fiber bundle revealed the temporal and spatial distribution of the cavity collapse on the MHV.

Preliminary study of a new type of energy transmission system for artificial hearts.

A transcutaneous energy transmission (TET) system is the most common way to power artificial hearts and ventricular assist devices. However, an external battery used with a TET system poses several problems, such as its heavy mass, small charge capacity, and long recharging time. The battery is indispensable when patients want to be ambulatory. This article proposes a new type of TET system that does not require an external battery because electrical energy is supplied remotely by using electromagnetic waves. For this system to operate, multiple transmitting antennas have to be mounted in a room or facility that has been shielded from electromagnetic waves, and a receiving antenna is attached to the patient. Electromagnetic waves transmit electrical power from the transmitting antennas to the receiving antenna. The received electrical power is sent to an implanted device through the TET system. The total power efficiency was plotted against the transmitter-receiver distance by measuring the power that was input to the transmitting antennas, and the final direct current (DC) power that was received by the receiving antenna. A 430-MHz frequency was applied in the experiments. The obtained efficiency was around 10% within a transmitter-receiver distance of 1 m when Yagi-Uda antennas were used for the transmitting antennas and two other types of antenna were used for the receiving antennas: a folded dipole with a reflector and a single loop with a reflector. The results suggested that the proposed system is worth considering. The proposed system would go a long way toward enhancing the patient's quality of life compared with the currently used conventional TET system.

Evaluation of pulsatile and nonpulsatile flow in microvessels of the bulbar conjunctiva in the goat with an undulation pump artificial heart.

This study has three purposes, as follows. The first is to develop a microscopic system to observe the microcirculation of animals implanted with an artificial heart. The second is to investigate the influence of flow pattern change from pulsatile to nonpulsatile on the microcirculation. The third is to study the effects of pulsatility in blood flow on endothelium-derived nitric oxide release in the microvasculature. When the flow pattern was changed from pulsatile to nonpulsatile, the velocity of erythrocytes in many capillaries dropped and remained at a low level, and the number of perfused capillaries decreased. After the flow pattern was returned to pulsatile, the velocity of erythrocytes recovered to the initial level. In many cases, the flow of nonperfused capillaries recovered to the initial level as well. Also, the pulsatile flow enhances the basal and flow-stimulated endothelium-derived nitric oxide release in microvessels.

Pathophysiologic study of goats with undulation pump total artificial heart: those that survived for more than 1 month.

The undulation pump total artificial heart is an implantable total artificial heart that is being developed at the University of Tokyo. Many advances in our system have been made by the adoption of hardware and software solutions that enabled the animals with the undulation pump total artificial hearts to survive more than 1 month. Pathophysiologic observations were performed for these goats that survived. In this article, the pathophysiologic findings are described in detail for three goats that survived for 46 (goat 9916), 54 (0030), and 63 (0107) days. The microscopic findings indicated that in goat 0107 the histologic changes in key organs occurred at the termination of the experiments. Therefore, the pathophysiologic changes in goats 9916 and 0030 were mainly investigated in this study to evaluate the chronic effect caused by our total artificial heart system. The signs of chronic ischemia and cell dystrophy were observed in both the liver and kidney. Until now, 13 goats had survived more than 1 week and 6 goats survived more than 1 month, including one that survived for 63 days, which is the longest in our experimental series. The pathophysiologic results of goats 9916 and 0030 showed that the undulation pump total artificial heart might still cause some damage to the liver and the kidney. To accomplish long-term survival with the undulation pump total artificial heart, further pathophysiologic studies are required, and the necessary modifications to the total artificial heart system will need to be made.

Functions for detecting malposition of transcutaneous energy transmission coils.

A transcutaneous energy transmission system (TETS) for artificial hearts and ventricular assist devices uses electrical coupling of power between external and implanted coils. If the position of coils changes relative to each other, the TETS cannot feed the required power of the implanted device. During activity or sleep, the coils may move accidentally. TETS users and the people around them have to pay attention to this because the range of the position where the required power can be fed efficiently is not wide. Therefore, we added functions for the position changes of the coils to the TETS. Regular, cautious, and irregular positions were introduced, and the ranges of them were decided upon in our experiments. The cautious position was determined by the area where the change of the relative position of the coils was relatively small. When the coils were in the cautious position, the circuit was tuned by way of changing the resonant point. This modulation could give good power efficiency in the cautious position. When the coils were in the irregular position, an alarm switch was turned on. These functions ease the restriction of the coil position and give better quality of life (QOL) than do the conventional TETS.

A new approach to detection of the cavitation on mechanical heart valves.

The cavitation on the mechanical heart valves (MHVs) is thought to be a cause of the mechanical failure of the occluder; also, the free radicals that would be generated when the cavitation bubbles implode might affect the patients chemically. These cavitation effects are attributed to the bubble collapse. Therefore, it is important to detect the bubble implosion behavior to analyze the cavitation on MHVs. The cavitation bubbles induce the generation of free radicals at their implosion, and the excited hydroxyl radicals emit the faint light. Based on this fact, we have tried to observe the faint light emission from a MHV to specifically capture the implosion of the cavitation bubbles. A highly sensitive CCD (charge coupled device) camera (C2400-35 VIM camera, Hamamatsu Photonics, Hamamatsu, Japan) was adopted in this study. This camera can observe low light down to the single photon counting range, and it gives two-dimensional mapping of the light. A 20 mm Björk-Shiley valve was submerged in the water tank of 10 L deionized water with luminol as a light enhancer, and then the pressure difference of 150 mm Hg was exerted on the valve at a rate of 60 bpm with a pulse duplicator. The camera and the water tank were settled in the lightproof configuration. After 2 hours of exposure, faint light images have been obtained successfully. The light emits mostly from the edge of the occluder on the inflow side in the major orifice of the valve. Therefore the results suggest that the bubbles would implode around this region and that free radicals caused by cavitation might be produced on MHV, which has coincided with our preliminary result by an electron spin resonance spectrometry.

Progress in the control system of the undulation pump total artificial heart.

The undulation pump total artificial heart (UPTAH) is a small implantable total artificial heart. As the UPTAH generates outflow and inflow at the same time, control of the UPTAH is very difficult. Therefore suitable control methods specifically for the UPTAH should be established. Various motor control, left-right flow balance control, and physiological control methods were examined and tried for the UPTAH control in this study. The control system is divided into seven categories. It has a hierarchical structure and all control modes work at the same time. The UPTAH with the newly developed control method has been implanted into the chest cavities of 48 goats. Until now, six goats survived for more than one month, including 63 days in the longest case. The good condition of the UPTAH implanted animal could be maintained with the newly developed control scheme, consisting of the 1/R control and several other additional controls.

A study on an energy supply method for a transcutaneous energy transmission system.

This study proposes a new type of a transcutaneous energy transmission system (TETS) that can supply electrical power for an implanted device without an external battery. In this system, the power is supplied from the floor to the shoes of the patients through coils that are set beneath the floor and the bottom of the shoes. If the patients wear the special shoes, they will be able to move freely on the specially designed floor without an external battery. Direct current (DC)-DC power efficiency was measured in the experiments, and the results showed that it varies with relative positions between the shoe and the floor coils. The results suggested that three-layered floor coils would enable the system to meet the demand for providing the required power anywhere on the floor without intermission. DC-DC power efficiency could be kept over 60% under the practical condition. It can then be concluded that the proposed system has a potential to provide better quality of life for the patients using a TETS.