Proposal of an in vitro thrombus-growth model for evaluating anticoagulants.

The general anticoagulant evaluation requires high expense equipment, reagents, and space. Therefore, not all laboratories can perform research related to anticoagulant. In this study, we propose a novel simple method "in vitro thrombus-growth model" that can evaluate anticoagulant ability by measuring weight. The in vitro thrombus-growth model is prepared by creating a "growth-clot" with citrate plasma, calcium chloride, and thrombin, and then pouring new citrate plasma onto it. The prepared growth-clots were increased in volume in citrated human plasma, including surpluses calcium chloride, which was released slowly, leading to clot coagulation around the plasma. As a result of evaluating the anticoagulant ability of direct thrombin inhibitor using this in vitro thrombus-growth model, it was confirmed that clot growth was suppressed in a concentration-dependent manner. Therefore, this thrombus-growth model is useful as a primary anticoagulant test that can to discover compounds with anticoagulant activity perform in any laboratory.

Novel temporary left ventricular assist system with hydrodynamically levitated bearing pump for bridge to decision: initial preclinical assessment in a goat model.

The management of heart failure patients presenting in a moribund state remains challenging, despite significant advances in the field of ventricular assist systems. Bridge to decision involves using temporary devices to stabilize the hemodynamic state of such patients while further assessment is performed and a decision can be made regarding patient management. We developed a new temporary left ventricular assist system employing a disposable centrifugal pump with a hydrodynamically levitated bearing. We used three adult goats (body weight, 58-68 kg) to investigate the 30-day performance and hemocompatibility of the newly developed left ventricular assist system, which included the pump, inflow and outflow cannulas, the extracorporeal circuit, and connectors. Hemodynamic, hematologic, and blood chemistry measurements were investigated as well as end-organ effect on necropsy. All goats survived for 30 days in good general condition. The blood pump was operated at a rotational speed of 3000-4500 rpm and a mean pump flow of 3.2 ± 0.6 L min. Excess hemolysis, observed in one goat, was due to the inadequate increase in pump rotational speed in response to drainage insufficiency caused by continuous contact of the inflow cannula tip with the left ventricular septal wall in the early days after surgery. At necropsy, no thrombus was noted in the pump, and no damage caused by mechanical contact was found on the bearing. The newly developed temporary left ventricular assist system using a disposable centrifugal pump with hydrodynamic bearing demonstrated consistent and satisfactory hemodynamic performance and hemocompatibility in the goat model.

Development of a stent-biovalve with round-shaped leaflets: in vitro hydrodynamic evaluation for transcatheter pulmonary valve implantation (TPVI).

This study evaluates a newly designed autologous heart valve-shaped tissue with a stent [stent-biovalve (SBV)] for transcatheter pulmonary valve implantation using the "in-body tissue architecture" technology. In the previously developed SBV with flat-shaped leaflets (FS-SBV), the valve could not close rapidly, because the leaflets were fixed in the open position, which induced regurgitant volume in the closing phase. Therefore, a novel mold to fabricate an SBV with round-shaped leaflets (RS-SBV) was developed, and its hydrodynamic performance with different valve diameters was evaluated in this study. A specially designed, self-expandable, stent-mounted, acrylic mold, which has 3 hemispheres, was placed in dorsal subcutaneous pouches of goats for 2 months. After extraction, the acrylic mold was removed from the implant, and a tubular tissue impregnated with the stent strut was obtained. Half of the tubular tissue with 3 hemispheres was completely folded in half inwards. The acrylic mold was designed, such that the folded half of the tubular tissue became the round-shaped leaflets. The 3 commissure parts were connected to form 3 leaflets, resulting in the preparation of the RS-SBV (internal diameter 25 mm). The RS-SBV closed more rapidly than the FS-SBV in a pulsatile mock circulation circuit under the pulmonary circulation conditions. The regurgitant fraction of the RS-SBV was approximately 6 %, which was lower than that of the FS-SBV. The appropriate pulmonary annulus diameter of the RS-SBV was from 24 to 25 mm based on the pressure difference and effective orifice area.

In vitro hydrodynamic evaluation of a biovalve with stent (tubular leaflet type) for transcatheter pulmonary valve implantation.

We have been developing an autologous heart valve-shaped tissue with a stent (stent-biovalve) for transcatheter pulmonary valve implantation (TPVI) using "in-body tissue architecture" technology. In this study, the hydrodynamic performance of a stent-biovalve with tubular leaflets was evaluated by changing its leaflet height in an in vitro test in order to determine the appropriate stent-biovalve form for the pulmonary valve. A specially designed, self-expandable, stent-mounted, cylindrical acrylic mold was placed in a dorsal subcutaneous pouch of goat, and the implant was extracted 2 months later. Only the cylindrical acrylic mold was removed from the implant, and a tubular hollow structure of membranous connective tissue impregnated with the stent strut was obtained. Half of tubular tissue was completely folded in half inwards, and 3 commissure parts were connected to form 3 leaflets, resulting in the preparation of a stent-biovalve with tubular leaflets (25-mm ID). The stent-biovalve with adjusting leaflet height (13, 14, 15, 17, 20, and 25 mm) was fixed to a specially designed pulsatile mock circulation circuit under pulmonary valve conditions using 37 °C saline. The mean pressure difference and effective orifice area were better than those of the biological valve. The lowest and highest leaflet heights had a high regurgitation rate due to lack of coaptation or prevention of leaflet movement, respectively. The lowest regurgitation (ca. 11%) was observed at a height of 15 mm. The leaflet height was found to significantly affect the hydrodynamics of stent-biovalves, and the existence of an appropriate leaflet height became clear.

Development of a flow rate monitoring method for the wearable ventricular assist device driver.

Our research institute has been working on the development of a compact wearable drive unit for an extracorporeal ventricular assist device (VAD) with a pneumatically driven pump. A method for checking the pump blood flow on the side of the drive unit without modifying the existing blood pump and impairing the portability of it will be useful. In this study, to calculate the pump flow rate indirectly from measuring the flow rate of the driving air of the VAD air chamber, we conducted experiments using a mock circuit to investigate the correlation between the air flow rate and the pump flow rate as well as its accuracy and error factors. The pump flow rate was measured using an ultrasonic flow meter at the inflow and outflow tube, and the air flow was measured using a thermal mass flow meter at the driveline. Similarity in the instantaneous waveform was confirmed between the air flow rate in the driveline and the pump flow rate. Some limitations of this technique were indicated by consideration of the error factors. A significant correlation was found between the average pump flow rate in the ejecting direction and the average air flow rate in the ejecting direction (R2 = 0.704-0.856), and the air flow rate in the filling direction (R2 = 0.947-0.971). It was demonstrated that the average pump flow rate was estimated exactly in a wide range of drive conditions using the air flow of the filling phase.

In vitro evaluation of a novel autologous aortic valve (biovalve) with a pulsatile circulation circuit.

We have used in-body tissue architecture technology to develop an autologous valved conduit with intact sinuses of Valsalva (biovalve). In this study, we fabricated three different forms of biovalves and evaluated their function in vitro using a mock circulation model to determine the optimal biovalve form for aortic valve replacement. A cylindrical mold for biovalve organization was placed in a dorsal subcutaneous pouch of a goat, and the implant that was encapsulated with connective tissue was extracted 2 months later. The cylindrical mold was removed to obtain the biovalve (16 mm inside diameter) that consisted of pure connective tissue. The biovalve was connected to a pulsatile mock circulation system in the aortic valve position. The function of the three biovalves (biovalve A: normal leaflets with the sinuses of Valsalva; biovalve B: extended leaflets with the sinuses of Valsalva; biovalve C: extended leaflets without the sinuses of Valsalva) was examined under pulsatile flow conditions using saline. In addition, the mock circuit was operated continuously for 40 days to evaluate the durability of biovalve C. The regurgitation rate (expressed as a percent of the mean aortic flow rate during diastole) was 46% for biovalve A but only 3% for biovalves B and C. The durability test demonstrated that even after biovalve C pulsated more than four million times (heart rate, 70 bpm; mean flow rate, 5.0 L/min; mean aortic pressure, 92 mm Hg), stable continuous operation was possible without excessive reduction of the flow rate or bursting. The developed biovalve demonstrated good function and durability in this initial in vitro study.

Preparation of an autologous heart valve with a stent (stent-biovalve) using the stent eversion method.

We designed a novel method for constructing an autologous heart valve with a stent, called a stent-biovalve. In constructing completely autologous heart valves, named biovalves, which used in-body tissue architecture technology, tissues for leaflets were formed via ingrowths into narrow apertures in the preparation molds, frequently leading to delayed or incomplete biovalve preparation. In this technique, self-expandable nitinol stents after everting were mounted on an acrylic column-shaped part and partially covered with an acrylic cylinder-shaped part with three slits. This assembled mold was placed into subcutaneous abdominal pouches in beagles or goats for 4 weeks. Upon removing the acrylic parts after harvesting and trimming of capsulated tissues, a tubular hollow structure with three pocket-flaps of membranous tissue rigidly fixed to the stent's outer surface was obtained. Then, the stent was turned inside out to the original form, thus moving the pocket-flaps from outside to the inside. Stent-biovalves with a sufficient coaptation area were thus obtained with little tissue damage in all cases. The valve opened smoothly, and high aperture ratio was noted. This novel technique was thus highly effective in constructing a robust, completely autologous stent-biovalve with adequate valve function.

Application of a search algorithm using stochastic behaviors to autonomous control of a ventricular assist device.

A ventricular assist device (VAD) is a device with mechanical pumps implanted adjacent to the patient's native heart to support the blood flow. Mechanical circulatory support using VADs has been an essential therapeutic tool for patients with severe heart failure waiting for a heart transplant in clinical site. Adaptive control of VADs that automatically adjust the pump output with changes in a patient state is one of the important approaches for enhanced therapeutic efficacy, prevention of complications and quality of life improvement. However adaptively controlling a VAD in the realistic situation would be difficult because it is necessary to model the whole including the VAD and the cardiovascular dynamics. To solve this problem, we propose an application of attractor selection algorithm using stochastic behavior to a VAD control system. In this study, we sought to investigate whether this proposed method can be used to adaptively control of a VAD in the simple case of a continuous flow VAD. The flow rate control algorithm was constructed on the basis of a stochastically searching algorithm as one example of application. The validity of the constructed control algorithm was examined in a mock circuit. As a result, in response to a low-flow state with the different causes, the flow rate of the pump reached a target value with self adaptive behavior without designing the detailed control rule based on the experience or the model of the control target.

Development and evaluation of endurance test system for ventricular assist devices.

We developed a novel endurance test system that can arbitrarily set various circulatory conditions and has durability and stability for long-term continuous evaluation of ventricular assist devices (VADs), and we evaluated its fundamental performance and prolonged durability and stability. The circulation circuit of the present endurance test system consisted of a pulsatile pump with a small closed chamber (SCC), a closed chamber, a reservoir and an electromagnetic proportional valve. Two duckbill valves were mounted in the inlet and outlet of the pulsatile pump. The features of the circulation circuit are as follows: (1) the components of the circulation circuit consist of optimized industrial devices, giving durability; (2) the pulsatile pump can change the heart rate and stroke length (SL), as well as its compliance using the SCC. Therefore, the endurance test system can quantitatively reproduce various circulatory conditions. The range of reproducible circulatory conditions in the endurance test circuit was examined in terms of fundamental performance. Additionally, continuous operation for 6 months was performed in order to evaluate the durability and stability. The circulation circuit was able to set up a wide range of pressure and total flow conditions using the SCC and adjusting the pulsatile pump SL. The long-term continuous operation test demonstrated that stable, continuous operation for 6 months was possible without leakage or industrial device failure. The newly developed endurance test system demonstrated a wide range of reproducible circulatory conditions, durability and stability, and is a promising approach for evaluating the basic characteristics of VADs.

Development and hydrodynamic evaluation of a novel inflow cannula in a mechanical circulatory support system for bridge to decision.

Recent progress in the development of implantable rotary blood pumps realized long-term mechanical circulatory support (MCS) for bridge to transplant, bridge to recovery, or a destination therapy. Meanwhile, a short-term MCS system is becoming necessary for bridge to decision. We developed a novel inflow cannula for the short-term MCS system, which gives sufficient bypass flow with minimal invasion at insertion, and evaluated its hydrodynamic characteristics. The novel inflow cannula, named the Lantern cannula, is made of elastic silicone reinforced with metal wires. The cannula tip has six slits on the side. This cannula tip can be extended to the axial direction by using an introducer and can be reduced in diameter, and the Lantern cannula enables easy insertion into the left ventricle apex with minimal invasion. The sufficient bypass flow rate can be obtained due to low pressure loss. Moreover, this Lantern shape also resists suction complication around the cannula tip. The pressure loss through the Lantern cannula was measured using a mock circulation and compared with two commercially available venous cannulae (Sarns4882, Terumo, Tokyo, Japan and Stockert V122-28, Sorin Group, Tokyo, Japan), which have almost same diameter as the Lantern cannula. Moreover, the flow patterns around the cannula tip were numerically analyzed by computational fluid dynamics (CFD). Acute animal experiment was also performed to confirm the practical effectiveness of the Lantern cannula. The pressure loss of the Lantern cannula was the lowest compared with those of the commercially available venous cannulae in in vitro experiment. CFD analysis results demonstrated that the Lantern cannula has low pressure loss because of wide inflow orifice area and a bell mouth, which were formed via Lantern shape. The highest bypass flow was obtained in the Lantern cannula because of the low pressure loss under pulsatile condition in in vivo experiments. The Lantern cannula demonstrated superior hydrodynamic characteristics as the inflow cannula in terms of pressure loss due to its specially designed Lantern shape.