Naoyuki Yokoyama

Mechanical Damage of Red Blood Cells by Rotary Blood Pumps: Selective Destruction of Aged Red Blood Cells and Subhemolytic Trauma

In this study, mean cell volume (MCV), mean cell hemoglobin concentration (MCHC), and mean cell hemoglobin (MCH) were measured to quantify RBC damage by rotary blood pumps. Six-hour hemolysis tests were conducted with a Bio-pump BPX-80, a Sarns 15200 roller pump, and a prototype mag-lev centrifugal pump (MedTech Heart) using fresh porcine blood circulated at 5 L/min against a 100 mm Hg head pressure. The temperature of the test and noncirculated control blood was maintained at 37 degrees C. The normalized index of hemolysis (NIH) of each pump was determined by measuring the plasma-free hemoglobin level. The MCV was measured with a Coulter counter, and MCHC was derived from total hemoglobin and hematocrit. MCH was derived from MCV and MCHC. A multivariance statistical analysis (ANOVA) revealed statistically significant differences (n = 15, P < 0.05) in MCV, MCHC, and MCH between the blood sheared by the rotary blood pumps and the nonsheared control blood. Normalized to the control blood, the Bio-pump BPX-80 showed an MCV of 1.04 +/- 0.03, an MCHC of 0.95 +/- 0.04, and an MCH of 0.98 +/- 0.02; the mag-lev MedTech Heart had an MCV of 1.02 +/- 0.02, an MCHC of 0.97 +/- 0.02, and an MCH of 0.99 +/- 0.01; and the roller pump exhibited an MCV of 1.03 +/- 0.03, an MCHC of 0.96 +/- 0.03, and an MCH of 0.99 +/- 0.01. Per 0.01 increase in NIH, the BPX-80 showed a normalized MCV change of +10.1% and a normalized MCHC change of -14.0%; the MedTech Heart demonstrated a +6.9% MCV and -9.5% MCHC change; and the roller pump had a +0.5% MCV and -0.6% MCHC change. Due to shear in the pump circuits, the RBC increased while the MCHC decreased. The likely mechanism is that older RBCs with smaller size and higher hemoglobin concentration were destroyed fast by the shear, leaving younger RBCs with larger size and lower hemoglobin concentration. Subhemolytic trauma caused the intracellular hemoglobin to decrease due to gradual hemoglobin leakage through the micropores formed in the thinned membrane. In conclusion, the rate of change in MCV and MCHC with respect to NIH change provides useful information relating to selective destruction of RBCs, while the MCH level reflects subhemolytic damage.

In vivo evaluation of the "TinyPump" as a pediatric left ventricular assist device.

Pediatric patients with end-stage heart failure require mechanical circulatory support (MCS) just as adults do. In order to meet the special requirements for neonates and infants MCS, pediatric circulatory support devices must be compact with low priming volume, easily controllable with low flow, less traumatic for blood cells and tissues, and biocompatible with minimum anticoagulation. We have designed and developed a miniature rotary centrifugal blood pump, TinyPump, with a priming volume of 5 mL, which has already demonstrated its controllable performance for low flow and durability in vitro. To evaluate the feasibility of the TinyPump as a left ventricular assist device (LVAD) suitable for neonates and infants, we have examined the biocompatibility and hemodynamic performance of the TinyPump in a pediatric animal model using Shiba goats. The TinyPump is a miniaturized centrifugal pump weighing 150 g comprising a disposable pump head with a 30-mm diameter impeller having six straight-vanes and a reusable motor driver. The impeller in the pump head is supported by a hydrodynamic bearing at its center and is driven by radial magnetic force coupled to the motor driver. TinyPump implantations were performed in 22 Shiba goats (17 female and 5 male), with body weights ranging from 8.4 to 27.2 kg. Under gas anesthesia, via left lateral thoracotomy, a 22 Fr inflow cannula was inserted through the left ventricular apex, while a 6-mm outflow graft was anastomosed to the descending aorta, which were then connected to a TinyPump mounted on the animals back. Postoperative hemodynamic monitoring included heart rate, arterial and central venous pressure, pump flow, and rotation speed. Target pump flow in all animals was maintained at 0.9 ± 0.1 L/min, which is approximately half the normal pulmonary artery flow measured in control animals. Blood samples were collected to evaluate peripheral organ functions, hemolysis, and thrombosis. Goats were divided into three groups-acute phase (6 h; n = 4), subchronic phase (6 h 2 postoperative days [POD]; n = 11), and chronic phase (3 POD-16 POD; n = 8)-based on their survival duration. In the early experiments, hemolysis and thrombi formation at the impeller bearing resulted in termination of the study. Subsequent modifications of the bearing design, pump housing design, and magnetic coupling force helped to minimize the hemolysis and thrombi formation, prolonging the survival duration of the Shiba goats to 2 weeks with minimum adverse effects on the blood components and organ functions. With further experiments and improvements in pump durability and hemocompatibility, the TinyPump can serve as a suitable circulatory support device for neonates and infants bridging to heart transplantation as well as to heart recovery.

Efficacy of a miniature centrifugal rotary pump (TinyPump) for transfusion-free cardiopulmonary bypass in neonatal piglets.

We have developed a miniaturized semiclosed cardiopulmonary bypass (CPB) circuit incorporating a centrifugal blood pump (TinyPump) with a volume of 5 ml. The current study was undertaken to evaluate the hemolytic performance of the TinyPump in comparison with the BioPump and to investigate the impact of different CPB circuit volumes on hemodilution, coagulation, and the inflammatory response. Twelve 1-week-old piglets (3.4 ± 0.2 kg) were used. The circuit comprised a centrifugal pump, a membrane oxygenator, and a cardiotomy reservoir. Cardiopulmonary bypass was conducted with mild hypothermia at 150 ml/kg/min for 3 hours. Transfusion was not performed. Priming volume was 68 ml for the circuit with the TinyPump and 111 ml for the circuit with the BioPump. Although the TinyPump required higher speed, plasma free hemoglobin levels after CPB were not different between the groups. After CPB, the TinyPump group had a significantly higher hematocrit (27% ± 3% vs. 23% ± 3%) and lower platelet reduction rate, lower thrombin-antithrombin complex levels, and lower interleukin-6 levels. Better lung compliance with less water content was observed in the TinyPump group. The TinyPump maintained CPB with acceptable hemolysis and lower inflammatory responses. This miniaturized CPB circuit may make transfusion-free open heart surgery feasible in neonates and would help to prevent postoperative organ dysfunction.

Feasibility of a TinyPump system for pediatric CPB, ECMO, and circulatory assistance: hydrodynamic performances of the modified pump housing for implantable TinyPump.

The TinyPump is a miniature centrifugal blood pump with an extremely small priming volume of 5 ml, allowing blood transfusion free cardiopulmonary bypass as well as extracorporeal membrane oxygenation in pediatric patients. In this study, a new pump housing with the angled inlet port (25 degrees toward impeller center with respect to the flow axis) was designed to optimize the pump displaced volume and to extend the application of the TinyPump to implantable support The fluid dynamic performance analysis revealed that the head pressure losses increased from 3 to 17 mm Hg in comparison with straight port design as the pump rotational speed increased from 2,000 to 4,000 rpm. This was probably caused by perturbed flow patterns at the site of the inlet bent port area and streamline hitting the off-center of the impeller. No significant effect on pumping efficiency was observed because of modification in inlet port design. Modification in the inflow and outflow port designs together with the drive mechanism reduces the height of the pump system, including the motor, to 27 mm yielding the displaced volume of 68 ml in comparison with 40 mm of the paracorporeal system with the displaced volume of 105 ml. Further analysis in terms of hemolytic as well as antithrombogenic performance will be carried out to finalize the housing design for the implantable version of the TinyPump.

Alterations in Red Blood Cell Volume and Hemoglobin Concentration, Viscoelastic Properties, and Mechanical Fragility Caused by Continuous Flow Pumping in Calves

In this study, we have analyzed the changes in mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and the dynamic deformability and mechanical fragility of red blood cells (RBCs) in five male Holstein calves (body weight: 95.6 ± 10.8 kg) whose circulation was partially supported with a novel magnetically levitated extracorporeal centrifugal blood pump MedTech Dispo. One hour after the pumping has started, the MCV increased and the MCHC decreased by 1.064 ± 0.006 and 0.906 ± 0.050 times, respectively, as compared with those of the prepumped blood (P < 0.05). The deformability index L/W, where L and W are the long and short axes of the two-dimensional RBC images, respectively, sheared by a cyclically reversing shear flow increased indicating that the RBCs pumped for 1 h exhibited more elastic characteristics (P < 0.05). In addition, when the pumped blood cells were sheared for 30 min with a uniform shear stress of 25.38 Pa, the hemolysis level decreased dramatically as compared with the control blood, as more fragile RBCs were destroyed by pumping, leaving behind less fragile RBCs. All these characteristics of the RBCs exposed to continuous flow resemble those of young RBCs having larger MCV, lower MCHC, higher elasticity, and lower fragility. In conclusion, during continuous flow pumping, the RBCs having relatively lower threshold for hemolysis to mechanical shear stress generated by continuous flow blood pump (CFBP) are destroyed first and removed from circulation in the early stage of application of CFBP, thus leaving behind less fragile and stronger RBCs.

Transfusion-Free Neonatal Cardiopulmonary Bypass Using a TinyPump

BACKGROUNDnWe devised a miniaturized circuit incorporating a TinyPump in the venous line to amplify the venous return. We compared this system to the conventional blood-primed circuit and investigated whether this circuit could maintain hematocrit levels without blood transfusion and reduce coagulation and inflammatory cascades.nnnMETHODSnThirteen 1-week-old piglets (3.7 ± 0.2 kg) were divided into group M (miniaturized circuits with TinyPump-assisted venous drainage without blood, n = 7) and group C (conventional circuits with blood priming, n = 6). Cardiopulmonary bypass (CPB) was performed at 150 to 180 mL·kg(-1)·min(-1) for 2 hours, including 60 minutes of cardioplegic cardiac arrest. Modified ultrafiltration (MUF) was subsequently performed. Data were acquired before CPB and after the end of MUF.nnnRESULTSnThe priming volume including the hemofilter circuit of the main circuit required 152 mL in group M and 300 mL in group C. The mean hematocrit values in group M and group C were not significantly different during CPB (21.5% ± 2.0% versus 23.2% ± 1.3%) or after MUF (30.7% ± 2.1% versus 32.9% ± 4.0%). After MUF, group M had lower thrombin-antithrombin complex levels (16.7 ± 5.0 ng/mL versus 28.4 ± 8.4 ng/mL, p < 0.01) and interleukin-8 levels (2,867 ± 758 pg/mL versus 13,730 ± 5,220 pg/mL, p < 0.01) than group C. The pulmonary vascular resistance index was lower in group M after MUF (4,105 ± 862 dynes·cm(-5)·kg(-1) versus 6,304 ± 1,477 dynes·cm(-5)·kg(-1), p < 0.01). The lung water content was also better in group M (83.7% ± 0.5% versus 84.9% ± 0.5%, p < 0.01).nnnCONCLUSIONSnThe minicircuit with TinyPump-assisted venous drainage successfully maintained acceptable hematocrit levels and the cardiopulmonary function in neonatal piglets. Employing this technique may attenuate blood requirements and inflammatory responses, thereby improving the clinical outcomes of neonatal open-heart surgery.

Glucose Depletion Enhances Sensitivity to Shear Stress-induced Mechanical Damage in Red Blood Cells by Rotary Blood Pumps

The metabolic process in red blood cells (RBCs) is anaerobic. The life-dependent adenosine triphosphate (ATP) for survival of cells is produced through glycolytic process. The aim of the study was to evaluate the effects of the glucose level on the mean corpuscular volume, mean corpuscular hemoglobin concentration, and hemolysis rate during hemolysis study by rotary blood pumps. The shear stress generated by rotary blood pumps may enhance glucose utilization by RBCs with depletion of glucose affecting ATP production and, consequently, cell size, shape, and morphology. The shear stress increases metabolism of RBCs consuming more energy ATP which is produced anaerobically from glycolytic process. Hence, in the closed circuit testing of rotary blood pumps, depletion of glucose might occur after prolonged pumping, which in turn affects metabolic process of RBCs by changing their size, shape, and morphology. It is thus suggested to monitor and control the glucose level of the fluid that suspends RBCs depending on the study duration.

Time-resolved optical spectroscopic quantification of red blood cell damage caused by cardiovascular devices

Cardiovascular devices such as heart-lung machine generate un-physiological level of shear stress to damage red blood cells, leading to hemolysis. The diagnostic techniques of cell damages, however, have not yet been established. In this study, the time-resolved optical spectroscopy was applied to quantify red blood cell (RBC) damages caused by the extracorporeal circulation system. Experimentally, the fresh porcine blood was subjected to varying degrees of shear stress in the rotary blood pump, followed with measurement of the time-resolved transmission characteristics using the pico-second pulses at 651 nm. The propagated optical energy through the blood specimen was detected using a streak camera. The data were analyzed in terms of the mean cell volume (MCV) and mean cell hemoglobin concentration (MCHC) measured separately versus the energy and propagation time of the light pulses. The results showed that as the circulation time increased, the MCV increased with decrease in MCHC. It was speculated that the older RBCs with smaller size and fragile membrane properties had been selectively destroyed by the shear stress. The time-resolved optical spectroscopy is a useful technique in quantifying the RBCs damages by measuring the energy and propagation time of the ultra-short light pulses through the blood.