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Mechanical circulatory support device (MCS) design has improved over the years and yet blood damage (e.g., hemolysis) remains a problem. Accumulating evidence indicates a subhemolytic threshold for red blood cells (RBC)—a threshold at which RBC deformability is impaired prior to hemolysis. The current study aimed to assess the deformability of RBC exposed to supra-physiological shear stresses that are typical of MCS devices and assess whether a method used to estimate an individualized subhemolytic threshold, accurately demarcates whether future application of shear stress was damaging. Suspensions of RBC were “conditioned” with discrete magnitudes of shear stress (5–100 Pa) for specific durations (1–16 s). Cellular deformability was subsequently measured via ektacytometry and a mechanical sensitivity (MS) index was calculated to identify the subhemolytic threshold. Thereafter, fresh RBC suspensions were exposed to a magnitude of shear stress 10 Pa above, 10 Pa below, or matched to a donor’s previously estimated subhemolytic threshold for a given duration (1, 4, 16 s) to ascertain the sensitivity of the subhemolytic threshold. The MS index of RBC was significantly impaired following exposure to 10 Pa above the subhemolytic threshold (p

Right ventricular failure is a common postoperative com-plication following left ventricular assist device (LVAD) im-plantation. Left ventricular assist devices are adapted for right ventricular assist device (RVAD) support by reducing the right pump speed or restricting the diameter of the outflow graft by “banding.” We sought to conduct in vitro hemocompatibility testing in a pulmonary flow condition for current modification of an LVAD for RVAD support, with a specific aim to provide benchmark values for future RVAD development. Two Heart-Ware HVADs coupled to custom-built blood circulation loops, as RVADs, were tested using human blood (n = 6). The RVADs were either used in reduced speed (1,920 ± 50 RPM) or band-ing conditions (3,050 ± 50 RPM) to mimic healthy pulmonary circulation hemodynamics. Blood from the loop was sampled at 0, 15, 60, 150, and 300 min to investigate the level of he-molysis, red blood cell (RBC) deformability, and the activation and aggregation of platelets. The amount of hemolysis and RBC deformability were significantly increased with banding compared with reduced speed (p < 0.05). No significant dif-ferences were found between the two conditions for platelet activation and platelet aggregation. In conclusion, we have evaluated the hemocompatibility of the HVAD when used for RVAD support in the clinically used modes of reduced speed and outflow graft banding under a pulmonary flow conditions that are commonly used in the biventricular failure population. We anticipate the benchmark values in the current study will facilitate future RVAD development.

BACKGROUND Red blood cells (RBC) are exposed to varying shear stress while traversing the circulatory system; this shear initiates RBC-derived nitric oxide (NO) production. OBJECTIVE The current study investigated the effect of varying shear stress dose on RBC-derived NO production. METHODS Separated RBC were prepared with the molecular probe, diamino-fluoreoscein diacetate, for fluorometric detection of NO. Prepared RBC were exposed to discrete magnitudes of shear stress (1-100 Pa), and intracellular and extracellular fluorescence was quantified via fluorescence microscopy at baseline (0 min) and discrete time-points (1-30 min). RESULTS Intracellular RBC-derived NO fluorescence was significantly increased (p

Introduction:Accumulating evidence demonstrates that subhaemolytic mechanical stresses, typical of circulatory support, induce physical and biochemical changes to red blood cells. It remains unclear, however, whether cell age affects susceptibility to these mechanical forces. This study thus examined the sensitivity of density-fractionated red blood cells to sublethal mechanical stresses.Methods:Red blood cells were isolated and washed twice, with the least and most dense fractions being obtained following centrifugation (1500 g × 5 min). Red blood cell deformability was determined across an osmotic gradient and a range of shear stresses (0.3–50 Pa). Cell deformability was also quantified before and after 300 s exposure to shear stresses known to decrease (64 Pa) or increase (10 Pa) red blood cell deformability. The time course of accumulated sublethal damage that occurred during exposure to 64 Pa was also examined.Results:Dense red blood cells exhibited decreased capacity to deform when compared with less dense cells. Cellular response to mechanical stimuli was similar in trend for all red blood cells, independent of density; however, the magnitude of impairment in cell deformability was exacerbated in dense cells. Moreover, the rate of impairment in cellular deformability, induced by 64 Pa, was more rapid for dense cells. Relative improvement in red blood cell deformability, due to low-shear conditioning (10 Pa), was consistent for both cell populations.Conclusion:Red blood cell populations respond differently to mechanical stimuli: older (more dense) cells are highly susceptible to sublethal mechanical trauma, while cell age (density) does not appear to alter the magnitude of improved cell deformability following low-shear conditioning.

Rotary blood pumps (RBPs) are used for mechanical circulatory support in heart failure patients but exhibit a reduced response to preload changes, which can lead to ventricular suction events. A passive control system, in the form of a compliant inflow cannula (IC), has been developed to mitigate suction, although this device may cause significant hemolysis. This study compared the incidence of mechanically induced hemolysis of two compliant IC designs (strutted and nonstrutted) with a rigid IC (control) in a blood circulation loop over 90 min. The nonstrutted compliant IC introduced high frequency and high amplitude oscillations in RBP inlet pressure and RBP IC resistance. These oscillations were correlated with a significant increase in plasma-free hemoglobin (pfHb) and hemolysis: pfHb increased to 2.005 ± 0.665 g/L, while normalized index of hemolysis (NIH) and modified index of hemolysis (MIH) increased to 0.04945 ± 0.01276 g/100 L and 4.0505 ± 0.6589 after 90 min (P

The supra-physiological shear stress that blood is exposed to while traversing mechanical circulatory assist devices affects the physical properties of red blood cells (RBCs), impairs RBC deformability, and may induce hemolysis. Previous studies exploring RBC damage following exposure to supra-physiological shear stress have employed durations exceeding clinical instrumentation, thus we explored changes in RBC deformability following exposure to shear stress below the reported "hemolytic threshold" using shear exposure durations per minute (i.e., duty-cycles) reflective of that employed by circulatory assist devices. Blood collected from 20 male donors, aged 18-38 years, was suspended in a viscous medium and exposed to an intermittent shear stress protocol of 1 s at 100 Pa, every 60 s for 60 duty-cycles. During the remaining 59 s/min, the cells were left at stasis until the subsequent duty-cycle commenced. At discrete time points (15/30/45/60 duty-cycles), an ektacytometer measured RBC deformability immediately after shear exposure at 100 Pa. Plasma-free hemoglobin, a measurement of hemolysis, was quantified via spectrophotometry. Supra-physiological shear stress impaired RBC properties, as indicated by: (1) decreased maximal elongation of RBCs at infinite shear stress following 15 duty-cycles (P

Red blood cells (RBC) express a nitric oxide synthase isoform (RBC-NOS) that appears dependent on shear stress for Serine1177 phosphorylation. Whether this protein is equally activated by varied shears in the physiological range is less described. Here, we explored RBC-NOS Serine1177 phosphorylation in response to shear stress levels reflective of in vivo conditions. Whole blood samples were exposed to specific magnitudes of shear stress (0.5, 1.5, 4.5, 13.5 Pa) for discrete exposure times (1, 10, 30 min). Thereafter, RBC-NOS Serine1177 phosphorylation was measured utilising immunofluorescence labelling. Shear stress exposure at 0.5, 1.5, and 13.5 Pa significantly increased RBC-NOS Serine1177 phosphorylation following 1 min (p < 0.0001); exposure to 4.5 Pa had no effect after 1 min. RBC-NOS Serine1177 phosphorylation was significantly increased following 10 min at each magnitude of shear stress (0.5, 1.5, 13.5 Pa, p < 0.0001; 4.5 Pa, p = 0.0042). Shear stress exposure for 30 min significantly increased RBC-NOS Serine1177 phosphorylation at 0.5 Pa and 13.5 Pa (p < 0.0001). We found that RBC-NOS phosphorylation via shear stress is non-linear and differs for a given magnitude and duration of exposure. This study provides a new understanding of the discrete relation between RBC-NOS and shear stress.

Management of organ failure has improved in recent years in parallel with advancements in interventions, including organ transplant, although the shortage of donor organs remains the rate-limiting step. The advent of mechanical alternatives to biological organs is a burgeoning area available to clinicians in a variety of scenarios, including short-term procedures (e.g., cardiopulmonary bypass), longer and acute management (e.g., extracorporeal membrane oxygenation), and semi-to-permanent therapies (e.g., ventricular assist devices). A paradigm shift has recently effected a transition from “bridge” therapies toward destination therapies, with a resultant increase in clinical utilization. It is clear, however, that while mechanical circulatory and respiratory support devices can sustain life, damage to blood and its constituents, and/or activation of cellular processes, can negatively impact recovery and health. These adverse effects may be broadly related to blood exposure to high shear stress and/or interactions between biological and artificial materials. Only through advances in mechanical circulatory and respiratory support to minimize blood damage will complications be overcome and mechanical devices attain their true potential.

Rotary blood pumps (RBPs) are used for mechanical circulatory support in heart failure patients but exhibit a reduced response to preload changes, which can lead to ventricular suction events. A passive control system, in the form of a compliant inflow cannula (IC), has been developed to mitigate suction, although this device may cause significant hemolysis. This study compared the incidence of mechanically induced hemolysis of two compliant IC designs (strutted and nonstrutted) with a rigid IC (control) in a blood circulation loop over 90 min. The nonstrutted compliant IC introduced high frequency and high amplitude oscillations in RBP inlet pressure and RBP IC resistance. These oscillations were correlated with a significant increase in plasma-free hemoglobin (pfHb) and hemolysis: pfHb increased to 2.005 ± 0.665 g/L, while normalized index of hemolysis (NIH) and modified index of hemolysis (MIH) increased to 0.04945 ± 0.01276 g/100 L and 4.0505 ± 0.6589 after 90 min (P 0.05). In contrast, the strutted compliant IC performed similar to the clinically utilized rigid IC and did not increase pfHb (0.300 ± 0.090 and 0.320 ± 0.171 g/L, respectively) and rate of hemolysis (NIH 0.00435 ± 0.00155 and 0.00543 ± 0.00371 g/100 L; MIH 0.3896 ± 0.1749 and 0.4261 ± 0.2792, respectively) within the RBP circuit. These data indicated that strutted, compliant ICs meet the hemocompatibility of clinically used rigid ICs while also offering a potential solution to prevent ventricular suction events.

Classic features of polycystic ovary syndrome (PCOS) include derangement of metabolic and cardiovascular health, and vascular dysfunction is commonly reported. These comorbidities indicate impaired blood flow; however, other than limited reports of increased plasma viscosity, surprisingly little is known regarding the physical properties of blood in PCOS. We aimed to investigate whether haemorheology was impaired in women with PCOS. We thus measured a comprehensive haemorheological profile, in a case-control design, of lean women with PCOS and age-matched healthy controls. A clinical examination determined similar cardiovascular risk for the two groups. Whole blood and plasma viscosity was measured using a cone-plate viscometer. The magnitude and rate of red blood cell (RBC) aggregation was determined using a light-transmission aggregometer, and the degree of RBC deformability was measured via laser-diffraction ektacytometry. Plasma viscosity was significantly increased in women with PCOS. Blood viscosity was also increased for PCOS at lower-to-moderate shear rates in both native and standardised haematocrit samples. The magnitude of RBC aggregation-a primary determinant of low-shear blood viscosity-was significantly increased in PCOS at native and 0.4 L·L-1 haematocrit. No difference was detected between PCOS and CON groups for RBC deformability measurements. A novel measure indicating the effectiveness of oxygen transport by RBC (i.e., the haematocrit-to-viscosity ratio; HVR) was decreased at all shear rates in women with PCOS. In a group of young and lean women with PCOS with an unremarkable cardiovascular risk profile based on clinical data, significant haemorheological impairment was observed. The degree of haemorheological derangement observed in the present study reflects that of overt chronic disease, and provides an avenue for future therapeutic intervention in PCOS.

Introduction Given the severity of the current imbalance between blood donor supply and recipient demand, discarded blood drawn from the routine venesections of haemochromatosis (HFE-HH) patients may serve as a valuable alternative source for blood banks and transfusion. We investigated whether functional or biochemical differences existed between HFE-HH and control blood samples, with particular focus upon the haemorheological properties, to investigate the viability of venesected blood being subsequently harvested for blood products. Methods Blood samples were collected from HFE-HH patients undergoing venesection treatment (n = 19) and healthy volunteers (n = 8). Moreover, a second experiment investigated the effects of a dose-response of iron (0, 40, 80, 320 mM FeCl3) on haemorheology in healthy blood samples (n = 7). Dependent variables included basic haematology, iron status, haematocrit, red blood cell (RBC) aggregation (native and standardised haematocrit) and “aggregability” (RBC tendency to aggregate in a standard aggregating medium; 0.4 L/L haematocrit in a Dx70), and RBC deformability. Results Indices of RBC deformability were significantly decreased for HFE-HH when compared with healthy controls: RBC deformability was significantly decreased at 1–7 Pa (p < 0.05), and the shear stress required for half maximal deformability was significantly increased (p < 0.05) for HFE-HH. RBC aggregation in plasma was significantly increased (p < 0.001) for HFE-HH, although when RBC were suspended in plasma-free Dx70 no differences were detected. No differences in RBC deformability or RBC aggregation/aggregability were detected when healthy RBC were incubated with varying dose of FeCl3. Conclusion HFE-HH impairs the haemorheological properties of blood; however, RBC aggregability was similar between HFE-HH and controls when cells were suspended in a plasma-free medium, indicating that plasma factor(s) may explain the altered haemorheology in HFE-HH patients. Acute exposure to elevated iron levels does not appear (in isolation) to account for these differences. Further consideration is required prior to utilising routine venesection blood for harvesting RBC concentrates due to the potential risk of microvascular disorders arising from impaired haemorheology.