The role of diastolic pump flow in centrifugal blood pump hemodynamics.

We tried to verify the hypothesis that increases in pump flow during diastole are matched by decreases in left ventricular (LV) output during systole. A calf (80 kg) was implanted with an implantable centrifugal blood pump (EVAHEART, SunMedical Technology Research Corp., Nagano, Japan) with left ventricle to aorta (LV-Ao) bypass, and parameters were recorded at different pump speeds under general anesthesia. Pump inflow and outflow pressure, arterial pressure, systemic and pulmonary blood flow, and electrocardiogram (ECG) were recorded on the computer every 5 ms. All parameters were separated into systolic and diastolic components and analyzed. The pulmonary flow was the same as the systemic flow during the study (p > 0.1). Systemic flow consisted of pump flow and LV output through the aortic valve. The ratio of systolic pump flow to pulmonary flow (51.3%) did not change significantly at variable pump speeds (p > 0.1). The other portions of the systemic flow were shared by the left ventricular output and the pump flow during diastole. When pump flow increased during diastole, there was a corresponding decrease in the LV output (Y = -1.068X + 51.462; R(insert)(2) = 0.9501). These show that pump diastolic flow may regulate expansion of the left ventricle in diastole.

HeartMate III: pump design for a centrifugal LVAD with a magnetically levitated rotor.

A long-term, compact left ventricular assist device (LVAD), the HeartMate III, has been designed and fabricated, featuring a centrifugal pump with a magnetically levitated rotor. The pump has been optimized by in vitro testing to achieve a design point of 7 L/min against 135 mm Hg at high hydrodynamic efficiency (30%) and to be capable of up to 10 L/min under such a load. Furthermore, the pump has demonstrated no mechanical failures, low hemolysis (4-10 mg/dl plasma free Hb), and low thrombogenicity during six (40, 27, 59, 42, 27, and 49-day) in vivo bovine studies.

The Heartmate III: design and in vivo studies of a maglev centrifugal left ventricular assist device.

A compact implantable centrifugal left ventricular assist device (LVAD) (HeartMate III) featuring a magnetically levitated impeller is under development. The goal of our ongoing work is to demonstrate feasibility, low hemolysis, and low thrombogenicity of the titanium pump in chronic bovine in vivo studies. The LVAD is based on so-called bearingless motor technology and combines pump rotor, drive, and magnetic bearing functions in a single unit. The impeller is rotated (theta z) and levitated with both active (X, Y) and passive (Z, theta x, theta y) suspension. Six prototype systems have been built featuring an implantable titanium pump (69 mm diameter, 30 mm height) with textured blood contacting surfaces and extracorporeal electronics. The pumps were implanted in 9 calves (< or = 100 kg at implant) that were anticoagulated with Coumadin (2.5 < or = INR < or = 4.0) throughout the studies. Six studies were electively terminated (at 27-61 days), 1 study was terminated after the development of severe pneumonia and lung atelectasis (at 27 days) another study was terminated after cardiac arrest (at 2 days) while a final study is ongoing (at approximately 100 days). Mean pump flows ranged from 2 to 7 L/min, except for brief periods of exercise at 6 to 9 L/min. Plasma free hemoglobin ranged from 4 to 10 mg/dl. All measured biochemical indicators of end organ function remained within normal range. The pumps have met performance requirements in all 9 implants with acceptable hemolysis and no mechanical failures.

Relationship of blood pressure and pump flow in an implantable centrifugal blood pump during hypertension.

The purpose of this study was to evaluate the real time relationship between pump flow and pump differential pressure (D-P) during experimentally induced hypertension (HT). Two calves (80 and 68 kg) were implanted with the EVA-HEART centrifugal blood pump (SunMedical Technology Research Corp., Nagano, Japan) under general anesthesia. Blood pressure (BP) in diastole was increased to 100 mm Hg by norepinephrine to simulate HT. Pump flow, D-P, ECG, and BP were measured at pump speeds of 1,800, 2,100, and 2,300 rpm. All data were separated into systole and diastole, and pump flow during HT was compared with normotensive (NT) conditions at respective pump speeds. Diastolic BP was increased to 99.3+/-4.1 mm Hg from 66.5+/-4.4 mm Hg (p<0.01). D-P in systole was under 40 mm Hg (range of change was 10 to 40 mm Hg) even during HT. During NT, the average systolic pump flow volume was 60% of the total pump flow. However, during HT, the average systolic pump flow was 100% of total pump flow volume, although the pump flow volume in systole during HT decreased (33.1+/-5.7 vs. 25.9+/-4.0 ml/systole, p<0.01). In diastole, the average flow volume through the pump was 19.6+/-6.9 ml/diastole during NT and -2.2+/-11.1 ml/diastole during HT (p<0.01). The change in pump flow volume due to HT, in diastole, was greater than the change in pump flow in systole at each pump speed (p<0.001). This study suggests that the decrease of mean pump flow during HT is mainly due to the decrease of the diastolic pump flow and, to a much lesser degree, systolic pump flow.

Continuously maintaining positive flow avoids endocardial suction of a rotary blood pump with left ventricular bypass.

This study showed the usefulness of maintaining positive pump flow to avoid endocardial suction and as an assist bypass. Three calves were implanted with centrifugal pumps. Hemodynamics and pump parameters were measured at varying pump speeds (from 1,100 to 2,300 rpm). In each test pump, speed was adjusted to create 3 hemodynamic states: both positive and negative flow (PNF), positive and zero flow (PZF), and continuously positive flow (CPF). The pump flow volume was determined during systole (Vs) and diastole (Vd). Vs in PNF was 29.6 ml and was not significantly different from Vs in PZF (p > 0.15). Vd in PNF was significantly different from Vd in PZF (p < 0.05). All bypass rates of PNF were over 30% of pulmonary flow. All PZF bypass rates were between the PNF rate and the CPF rate. These data showed that PZF satisfied the minimum requirement of assist flow and was under 100% bypass. Thus, PZF may avoid endocardial suction.

Rotary blood pump flow spontaneously increases during exercise under constant pump speed: results of a chronic study.

Many types of rotary blood pumps and pump control methods have recently been developed with the goal of clinical use. From experiments, we know that pump flow spontaneously increases during exercise without changing pump control parameters. The purpose of this study was to determine the hemodynamics associated with the long-term observation of calves implanted with centrifugal blood pumps (EVAHEART, Sun Medical Technology Research Corporation, Nagano, Japan). Two healthy female Jersey calves were implanted with devices in the left thoracic cavity. A total of 22 treadmill exercise tests were performed after the 50th postoperative day. During exercise, the following parameters were compared with conditions at rest: heart rate, blood pressure, central venous oxygen saturation (SvO2), pump speed, and pump flow. The pump flow in a cardiac cycle was analyzed by separating the systole and diastole. Compared to the base data, statistically significant differences were found in the following interrelated parameters: the heart rate (66.8 +/- 5.2 vs. 106 +/- 9.7 bpm), mean pump flow (4.8 +/- 0.2 vs. 7.0 +/- 0.3 L/min), and volume of pump flow in diastole (26.0 +/- 1.8 vs. 13.5 +/- 2.5 ml). During exercise, the volume of pump flow in systole was 3 times larger than that measured in diastole. Blood pressure, SvO2, and pump speed did not change significantly from rest to exercise. These results suggested that the mean pump flow depends on the systolic pump flow. Therefore, the increase in the mean pump flow during exercise under constant pump speed was caused by an increase in the heart rate.

Decrease in red blood cell deformability caused by hypothermia, hemodilution, and mechanical stress: factors related to cardiopulmonary bypass.

During extracorporeal circulation in cardiopulmonary bypass (CPB) surgery, blood is exposed to anomalous mechanical and environmental factors, such as high shear stress, turbulence, decreased oncotic pressure caused by dilution of plasma, and moderate and especially deep hypothermia widely applied during CPB in infants. These factors cause damage to the red blood cells (RBCs), which is manifest by immediate and delayed hemolysis and by changes in the mechanical properties of RBCs. These changes include, in particular, decrease in RBC deformability impeding the passage of RBCs through the microvessels and may contribute to the complications associated with CPB surgery. We investigated in vitro the independent and combined effects of hypothermia, plasma dilution, and mechanical stress on deformability of bovine RBCs. Our studies showed each of these factors to cause a significant decrease in the deformability of RBCs, especially acting synergistically. The impairment of RBC deformability caused by hypothermia was found to be more pronounced for RBCs suspended in phosphate buffered saline (PBS) than for RBCs suspended in plasma. The decrease in RBC deformability caused by mechanical stress was significantly exacerbated by dilution of plasma with PBS. In summary, results of our in vitro study strongly point to a possible detrimental consequence of conventional CPB arising from increased RBC rigidity, which may lead to impaired microcirculation and tissue oxygen supply.

Chronic animal health assessment during axial ventricular assistance: importance of hemorheologic parameters.

Chronic testing of the Nimbus/UOP Axial Flow Pump was performed on 22 calves for periods of implantation ranging from 27 to 226 days (average, 74 days). The following parameters were measured: plasma free hemoglobin, blood and plasma viscosity, erythrocyte deformability and mechanical fragility, oxygen delivery index (ODI), blood cell counts, hematocrit, hemoglobin, blood urea nitrogen, creatinine, bilirubin, total protein, fibrinogen, and plasma osmolality. Most of the above parameters were stable during the full course of support. Compared with baseline, statistically significant differences during the entire period of implantation were only found in: hematocrit (p<0.001), hemoglobin (p<0.005), red blood cell (RBC) count (p<0.001), and whole blood viscosity (p<0.01). Plasma viscosity and ODI were mostly stable during the period of implantation. In some animals, an acute increase in fibrinogen concentration, plasma and blood viscosity, and a decrease in ODI were found to be early signs of the onset of infection. A small (10%) decrease in deformability of RBCs was found during the first 2 weeks after implantation. This alteration in RBC deformability was highly correlated (r = 0.793) with changes in total plasma protein concentration that fell more than 15% (p<0.001) during the same period. Mechanical fragility of RBCs was found to be slightly increased after implantation. Plasma free hemoglobin remained close to baseline level (p>0.2). After the first 2 weeks of the postoperative period, pump performing parameters for all animals were consistent and stable. In general, the Nimbus/UOP Axial Flow Pump demonstrated basic reliability and biocompatibility and did not produce significant alterations in the mechanical properties of blood or animal health status. The pump provided adequate hemodynamics and was well tolerated by the experimental animal for periods as long as 7.5 months. Monitoring rheologic parameters of blood is very helpful for evaluation of health during heart-assist device application.

Gender difference in rheologic properties of blood and risk of cardiovascular diseases.

According to official statistical data there is a significant difference between pre-menopausal women and age-matched men in morbidity and mortality from cardiac diseases and especially from myocardial infarction. There are several speculations regarding the nature of this phenomenon which have both supporting and refuting evidence. Our hypothesis was that due to regular physiologic bleeding, rheological properties of blood of pre-menopausal women are superior to those of men, and place such women at a lower risk of cardiovascular diseases than men in any age group. We believe that this difference in hemorheological properties is due to the reduced concentration of red blood cells (RBCs) and due to greater population of younger and less population of older RBCs in female blood. We studied mechanical properties of blood from 47 pre-menopausal women and 50 age-matched men. Compared to female blood, male blood had higher viscosity and RBC aggregation and lower RBC deformability. Oxygen Delivery Index, calculated as a ratio of hematocrit to blood viscosity, was found to be significantly lower in male blood. Decreased oxygen delivery along with increased RBC aggregation and decreased RBC deformability may contribute to the higher risk for the development of cardiovascular diseases. Regular blood donation may reduce hematocrit and blood viscosity, improve rheological properties of blood, and increase oxygen delivery in men.

Assessment of bovine platelet life span with biotinylation and flow cytometry.

Reduced platelet life span is associated with the implantation of a variety of cardiovascular devices and may be used as a gauge of device biocompatibility. In the bovine model, platelet life span has previously been assessed with radioisotope labeling of removed platelets followed by reinjection and periodic gamma counting of blood samples. We report here the use of protein-reactive biotin (sulfo-N-hydroxysuccinimido [NHS]-biotin) as an alternative to radioisotope techniques whereby reinjected biotinylated platelets are subsequently detected in blood samples using phycoerythrin-streptavidin and flow cytometric techniques. Platelet life span was quantified in a normal calf (4.9 days) and in a calf prior to (6.1 days) and following (3.1 days) implantation of a Nimbus Axial Flow Pump ventricular assist device. The assessment of bovine platelet life span with biotinylation and flow cytometry avoids the technical, regulatory, and safety considerations associated with radioisotope usage and appears readily amenable to application in cardiovascular device testing.

Red blood cell aging and risk of cardiovascular diseases.

We hypothesized that due to monthly bloodloss, the mechanical properties of blood of premenopausal women are superior to men, and place them at less risk for cardiovascular diseases than men in any age group. Rheological properties of blood of premenopausal women and age-matched men were compared. It was found that male blood possesses an increased viscosity, RBC aggregability and RBC rigidity. Additionally, male RBCs were found to have higher mechanical fragility. Since women in reproductive age have almost half as many old RBCs and almost twice as many young RBCs as men, we investigated the effect of in vivo aging of RBCs on their mechanical properties. Old RBCs were shown to have an increased mechanical fragility and aggregability, and decreased deformability as compared to young RBCs. Decreased deformability and increased aggregability of RBCs cause an increase in blood viscosity and are known as risk factors of cardiovascular diseases. Since men possess a higher number of old RBCs with suboptimum mechanical properties than premenopausal women, who due to monthly bloodloss have a higher number of young cells and a lower number of old RBCs than their male counterparts, our results suggest that an elevated hemorheological risk for males is associated with the age distribution of RBCs. This, in addition to significantly higher hematocrit, may be the reason for the increased risk of morbidity and mortality from cardiovascular diseases of men as compared to women of reproductive age.

Flow cytometric assays to detect platelet activation and aggregation in device-implanted calves.

Cardiovascular device development often relies upon large-animal models to assess blood biocompatibility prior to initiating clinical trials. Unfortunately, the amount of information gleaned from such trials is limited by simple assays that do not take full advantage of immunotechnological advances that increasingly are applied in clinical studies. Thus we have developed and tested new flow cytometric techniques for measuring circulating activated bovine platelets and platelet microaggregates. Monoclonal antibodies (MAbs) raised against both activated and quiescent bovine platelets were incubated with control and PMA-or ADP-stimulated whole blood. Selected MAbs detected activated bovine platelets and platelet microaggregates in vitro with flow cytometry. Five calves implanted with one of two designs of nonpulsatile ventricular-assist devices (VADs) were followed with these assays prior to and during VAD implantation. Circulating activated bovine platelets and microaggregates increased after implantation in all animals and, alternatively, remained elevated or returned toward preimplant levels. Platelet activation percentages as detected temporally by three MAbs were correlated with one another, and platelet activation was correlated with microaggregate formation. In summary, these new methods for the sensitive measurement of circulating activated bovine platelets and microaggregates may provide valuable information for the development and assessment of future cardiovascular device designs.

Plasma protective effect on red blood cells exposed to mechanical stress.

Hemodilution with plasma expanders is a widely applied practice during extracorporeal circulation and hemodialysis. Despite the immediate beneficial effects of hemodilution, such as reduction of blood viscosity and red blood cell (RBC) aggregation, elevation of blood flow in the microcirculation, etc., the dilution of plasma may cause some unfavorable effects on RBCs, amplifying the mechanical damage caused by circulatory assist devices. The authors investigated the effect of partial and total replacement of plasma on susceptibility of human and bovine RBCs to mechanical stress in vitro. Hemolysis was measured after the exposure of RBCs suspended in different media to similar mechanical stress. Experiments were performed at room temperature with control of osmolality and viscosity of the suspension media. The lowest hemolysis was obtained for RBCs suspended in serum, plasma, and albumin solutions. Hemolysis in PBS and Dextran suspensions was more than three times higher than that in plasma (p < 0.001). The protective effect depended upon protein concentration. Human RBCs were found to be significantly more sensitive to mechanical stress than bovine RBCs in all investigated suspension media (p < 0.005). Human RBCs from men suspended in plasma were found to be significantly (p < 0.05) more fragile than RBCs from women. The presence of even small amounts of plasma (such as 25%) in the suspension media significantly (p < 0.001) decreased hemolysis. However, a 30% replacement of plasma with PBS or Dextran solutions caused a statistically significant (p < 0.001) increase in mechanical hemolysis. This suggests that a decrease in the concentration of plasma proteins due to hemodilution may elevate blood damage during extracorporeal circulation and hemodialysis.

The vasoregulatory role of endothelium derived nitric oxide during pulsatile cardiopulmonary bypass.

The role of pulsatile flow as a physiologic stimulus for endothelium mediated vasoregulation is poorly understood. Furthermore, non pulsatile flow, which is associated with increased vascular resistance and end-organ failure, has been demonstrated to lead to a decrease in nitric oxide (NO) production in vitro. Anesthetized pigs (23.4 +/- 0.3 kg) were placed on cardiopulmonary bypass using either non pulsatile or pulsatile perfusion for 60 min. In both groups, animals were maintained with a constant mean aortic flow (1.0-1.3 L/min). Serum samples obtained during bypass were assayed for the stable end-products of NO (nitrate [NO3-] and nitrite [NO2-]) by a method based on the Greiss reaction. Systemic vascular resistance was higher after 60 min in the non pulsatile (3712.5 +/- 481.2 dyne sec cm(-5)) vs the pulsatile group (2672.6 +/- 427.0 dyne sec cm(-5)), but not statistically significant (p > .05). However, NO production was decreased in the non pulsatile flow group (27 +/- 6%) vs the pulsatile flow group (14 +/- 5%) at a statistically significant level (p < .005). The results suggest that non pulsatile flow is associated with diminished endothelial shear stress and a reduction in endothelial nitric oxide production. This may contribute to the detrimental physiologic effects observed in prolonged non pulsatile flow states.

Mechanisms of red blood cell trauma in assisted circulation. Rheologic similarities of red blood cell transformations due to natural aging and mechanical stress.

Clinical experience with circulatory support devices has typically shown alteration of patient blood rheology exhibited through increasing blood viscosity and decreasing erythrocyte deformability. Our hemorheologic studies have additionally shown a remarkable increase in red blood cell (RBC) aggregation in the blood of artificial heart patients as compared to healthy donors. These hemorheologic changes may be caused by mechanical trauma to RBCs. The authors hypothesize that the mechanical trauma process, from a rheologic point of view, could be analogous to an "accelerated" RBC aging process. The hypothesis was examined through in vivo and in vitro experiments on RBCs, age-separated on the basis of density, specifically to identify the rheologic similarities between aged and mechanically traumatized RBCs. Older RBCs demonstrated an increased mechanical fragility, a decreased deformability, and a increased ability to aggregate as compared to younger RBCs. RBCs exposed to mechanical stress demonstrated similar alterations in the same rheologic parameters. Our experiments have also shown that mechanical stress decreases the negative surface charge of RBCs as is known to occur in aged RBCs. Similarities found between the processes of RBC mechanical trauma and senescence enhance our understanding of mechanisms of subhemolytic trauma incurred in assisted circulation. This may improve the design and evaluation of future heart assist devices through minimizing shear induced blood trauma.

A sheep model for the study of hemorheology with assisted circulation. Effect of an axial flow blood pump.

Hemorheologic investigations were performed on nine sheep during the in vivo evaluation of a new axial flow ventricular assist device, the Nimbus AxiPump (Nimbus, Inc., Rancho Cordova, CA). Blood hematocrit, plasma and whole blood viscosity, red blood cell (RBC) deformability and aggregation, plasma fibrinogen, and free hemoglobin (hemolysis) levels were measured. Changes in the main rheologic parameters of sheep blood relative to the pre-implant values were minor and transient. The exception was RBC aggregation, which appeared on the third day of implantation. (Sheep blood does not normally demonstrate RBC aggregation.) Sheep RBCs started to form classic rouleaux typically on the third post-operative day simultaneously with increasing fibrinogen level. To investigate the relative effects of mechanical stress and elevated fibrinogen levels on RBC aggregability, in vitro studies were conducted with blood from control sheep. These studies indicated that neither mechanical trauma nor elevated fibrinogen alone caused RBC aggregation as seen in vivo. However, combined mechanical stress and elevated fibrinogen did cause this unusual effect for sheep blood.

Effect of perfluorochemical emulsion on blood trauma and hemorheology.

One of the major problems of development and improvement of heart assist devices is the reduction of blood cell damage. The extremely high levels of shear stress, turbulence, prolonged contact between blood and foreign surfaces, and other abnormal hydrodynamic circumstances have been shown to cause hemolysis, activation of platelets, and changes in mechanical properties of red blood cells. Hemolysis, in turn, can drastically increase red blood cell aggregation at low shear conditions. A new pharmacologic approach to reduce blood trauma and improve rheologic properties of blood subjected to mechanical stress was investigated. These experiments showed that the replacement of 20% of the plasma volume with Fluosol (Alpha Therapeutic Corp., Los Angeles, CA), a perfluorochemical that transports oxygen, reduced mechanical fragility of human (P < 0.01) and ovine (P = 0.054) red blood cells by approximately 30%. The same replacement of plasma with Fluosol reduced hemolysis (plasma free Hb) by approximately 40% compared to control (P < 0.05) during in vitro pumping of blood with a centrifugal pump. A 20% replacement of plasma volume with Fluosol remarkably reduced low shear blood viscosity (from 31.9 +/- 6.1 to 18.2 +/- 4.8 cP, for shear rate gamma = 0.277 sec-1) and erythrocyte sedimentation rate (from 16.7 +/- 9.2 to 3.1 +/- 3.1) mm/hr) in human blood. Decrease of these parameters indicates the reduction of red blood cell aggregation. Results of this study demonstrate the potential feasibility of Fluosol to improve mechanical properties of blood in patients with heart assist devices.

Heparin effect on red blood cell aggregation.

We studied the effect of relatively high dosages of heparin (25-150 U/ml) on some rheological parameters of donor blood. It was found that heparin increased the erythrocyte sedimentation rate (ESR) and low shear blood viscosity at all concentrations studied. The increase of these parameters indicates an elevation in RBC aggregation. In all samples with a heparin concentration of 100 U/ml, the average ESR was increased by approximately 75% (p < 0.001); whole blood viscosity increased under shear rate, gamma = 0.277 sec-1, by approximately 30% (p < 0.001). Since bolus injection of heparin may result in transient high local concentration, these results suggest a possible additional adverse effect to the well known heparin effect on platelet aggregation.