Blood Rheology and Hemodynamics.

Seminars in Thrombosis and Hemostasis (STH) celebrates 50 years of publishing in 2024. To celebrate this landmark event, STH is republishing some archival material. This manuscript represents the most highly cited paper ever published in STH. The original abstract follows.Blood is a two-phase suspension of formed elements (i.e., red blood cells [RBCs], white blood cells [WBCs], platelets) suspended in an aqueous solution of organic molecules, proteins, and salts called plasma. The apparent viscosity of blood depends on the existing shear forces (i.e., blood behaves as a non-Newtonian fluid) and is determined by hematocrit, plasma viscosity, RBC aggregation, and the mechanical properties of RBCs. RBCs are highly deformable, and this physical property significantly contributes to aiding blood flow both under bulk flow conditions and in the microcirculation. The tendency of RBCs to undergo reversible aggregation is an important determinant of apparent viscosity because the size of RBC aggregates is inversely proportional to the magnitude of shear forces; the aggregates are dispersed with increasing shear forces, then reform under low-flow or static conditions. RBC aggregation also affects the in vivo fluidity of blood, especially in the low-shear regions of the circulatory system. Blood rheology has been reported to be altered in various physiopathological processes: (1) Alterations of hematocrit significantly contribute to hemorheological variations in diseases and in certain extreme physiological conditions; (2) RBC deformability is sensitive to local and general homeostasis, with RBC deformability affected by alterations of the properties and associations of membrane skeletal proteins, the ratio of RBC membrane surface area to cell volume, cell morphology, and cytoplasmic viscosity. Such alterations may result from genetic disorders or may be induced by such factors as abnormal local tissue metabolism, oxidant stress, and activated leukocytes; and (3) RBC aggregation is mainly determined by plasma protein composition and surface properties of RBCs, with increased plasma concentrations of acute phase reactants in inflammatory disorders a common cause of increased RBC aggregation. In addition, RBC aggregation tendency can be modified by alterations of RBC surface properties because of RBC in vivo aging, oxygen-free radicals, or proteolytic enzymes. Impairment of blood fluidity may significantly affect tissue perfusion and result in functional deteriorations, especially if disease processes also disturb vascular properties.

The functional significance of the rho/rho-kinase pathway in human erythrocytes.

OBJECTIVE: Erythrocyte deformability, which can be influenced by various intracellular signaling mechanisms, such as nitric oxide, cAMP, cGMP, and protein kinases, is the most important physiological factor providing the blood flow in microcirculation. However, the functional significance of the Rho/Rho-kinase pathway, which contributes cell shape changes and the reorganization of the actin cytoskeleton, has yet to be explored in erythrocytes. Therefore, we examined the influence of several activators and inhibitors of Rho/Rho-kinase signaling on human erythrocyte deformability. MATERIALS AND METHODS: RhoA and ROCK-2 proteins were studied by western blotting. Influences of 2 Rho-kinase inhibitors, fasudil and Y-27632 (both 10-7 to 10-4 M), on erythrocyte deformability was determined by ektacytometer at various shear stresses (0-30 Pa) in the presence or absence of a known Rho activator, lysophosphatidic acid (LPA, 10-5 to 5x10-5 M, 1-15 min). RESULTS: LPA incubation reduced deformability with concomitant RhoA-GTP inhibition. Y-27632 and fasudil also decreased deformability, but had no effect on LPA-induced reduction of deformability. Rho inhibitor C3 had no effect on RhoA activation. Reduction in RhoA activation was induced by sub-hemolytic mechanical stress. CONCLUSION: Our findings may indicate that the Rho/Rho-kinase pathway could contribute to the regulation of deformability of human erythrocytes.

Tissue oxygen demand in regulation of the behavior of the cells in the vasculature.

The control of arteriolar diameters in microvasculature has been in the focus of studies on mechanisms matching oxygen demand and supply at the tissue level. Functionally, important vascular elements include EC, VSMC, and RBC. Integration of these different cell types into functional units aimed at matching tissue oxygen supply with tissue oxygen demand is only achieved when all these cells can respond to the signals of tissue oxygen demand. Many vasoactive agents that serve as signals of tissue oxygen demand have their receptors on all these types of cells (VSMC, EC, and RBC) implying that there can be a coordinated regulation of their behavior by the tissue oxygen demand. Such functions of RBC as oxygen carrying by Hb, rheology, and release of vasoactive agents are considered. Several common extra- and intracellular signaling pathways that link tissue oxygen demand with control of VSMC contractility, EC permeability, and RBC functioning are discussed.

Iatrogenic hyperviscosity and thrombosis.

It is well known that hemostatic-thrombotic mechanisms are influenced by hemodynamic factors, such as shear forces affecting platelets or red blood cell aggregation, in turn affecting flow in stenotic regions. Endothelial cell function is also significantly influenced by shear forces acting on the vessel wall. Further, the distribution of shear forces in the vasculature is complex and closely associated with factors determining the flow properties of blood. Therefore, there is a link among alterations in the rheological properties of blood and its elements and the risk for thrombosis, with this linkage confirmed by numerous clinical studies. After discussing relevant rheological and hemodynamic concepts, this review focuses on selected drug-induced conditions that are known to be associated with both hyperviscosity conditions and increased thrombotic risk: oral contraceptives, diuretics, intravenous immunoglobulin, erythropoiesis-stimulating agents, chemotherapy, and radio-contrast media. Alterations of relationships between blood rheology and thrombotic risk related to artificial circulatory environments and physical exercise are also briefly discussed.

Shear stress activation of nitric oxide synthase and increased nitric oxide levels in human red blood cells.

Red blood cells (RBC) play an important role in the balance between generation and scavenging of nitric oxide (NO) and hence its local bioavailability and influence on vasomotor control. Previous studies have reported increased NO levels in RBC suspensions subsequent to exposure to shear forces; the present study was designed to further investigate changes in intracellular NO concentration and possible mechanisms involved for RBC exposed to well-controlled shear forces. Attached human RBC were subjected to shear stresses up to 0.1Pa in a parallel-plate flow channel; fluorescent methods were used to monitor changes in intracellular NO and calcium concentrations. Intracellular NO concentration, estimated by the fluorescence level of 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate (DAF-FM), increased sharply within 30s following the application of shear stress between 0.013 and 0.1Pa. This increase was only partially prevented by the absence of l-arginine and by the presence of l-N-acetyl-methyl-arginine (L-NAME), strongly suggesting that this response was in part related to the activation of NO-synthase (NOS) enzyme. The increase in intracellular NO concentration under shear stress was also inhibited by calcium chelation in the suspending medium, indicating the role of calcium entry for NOS activation. Increases of intracellular calcium concentrations under the same shearing conditions were demonstrated by monitoring Fluo-3/AM fluorescence in RBC exposed to shear stress. Serine 1177 phosphorylated NOS protein, the activated form of the enzyme determined by immunohistochemistry, was found to be significantly increased following the exposure of RBC to 0.1Pa shear stress for 1min. These data confirm that RBC possess a NOS enzyme that is actively synthesizing NO and activated by effective shear forces. The data also suggest that there may be additional (e.g., non-enzymatic) NO generating mechanisms in RBC that are also enhanced under shear stress.

Immune and hemorheological changes in chronic fatigue syndrome.

BACKGROUND: Chronic Fatigue Syndrome (CFS) is a multifactorial disorder that affects various physiological systems including immune and neurological systems. The immune system has been substantially examined in CFS with equivocal results, however, little is known about the role of neutrophils and natural killer (NK) phenotypes in the pathomechanism of this disorder. Additionally the role of erythrocyte rheological characteristics in CFS has not been fully expounded. The objective of this present study was to determine deficiencies in lymphocyte function and erythrocyte rheology in CFS patients. METHODS: Flow cytometric measurements were performed for neutrophil function, lymphocyte numbers, NK phenotypes (CD56(dim)CD16(+) and CD56(bright)CD16(-)) and NK cytotoxic activity. Erythrocyte aggregation, deformability and fibrinogen levels were also assessed. RESULTS: CFS patients (n = 10) had significant decreases in neutrophil respiratory burst, NK cytotoxic activity and CD56(bright)CD16(-) NK phenotypes in comparison to healthy controls (n = 10). However, hemorheological characteristic, aggregation, deformability, fibrinogen, lymphocyte numbers and CD56(dim)CD16(+) NK cells were similar between the two groups. CONCLUSION: These results indicate immune dysfunction as potential contributors to the mechanism of CFS, as indicated by decreases in neutrophil respiratory burst, NK cell activity and NK phenotypes. Thus, immune cell function and phenotypes may be important diagnostic markers for CFS. The absence of rheological changes may indicate no abnormalities in erythrocytes of CFS patients.

Depletion of high molecular weight dextran from the red cell surface measured by particle electrophoresis.

The reversible aggregation of human red blood cells (RBC) by proteins or polymers continues to be of biological and biophysical interest, yet the mechanistic details governing the process are still being explored. A depletion model has been proposed for aggregation by the neutral polyglucose dextran and its applicability at high molecular weights has been recently documented. In the present study the depletion of high molecular weight dextrans on the red cell surface was measured as a function of polymer molecular mass (40 kDa-28 MDa), ionic strength (5 and 15 mM NaCl) and polymer concentration (< or =0.9 g/dL). The experimental data clearly indicate an increasing depletion effect with increasing molecular weight: the effects of medium viscosity on RBC mobility were markedly overestimated by the Helmholtz-Smoluchowski relation, with the difference increasing with dextran molecular mass. These results agree with the concept of polymer depletion near the RBC surface and lend strong support to a "depletion model" mechanism for dextran-mediated RBC aggregation. Our findings provide important new insight into polymer-RBC interactions and suggest the usefulness of this model for fundamental studies of cell-cell affinity and for the development of new agents to stabilize or destabilize specific bio-fluids.

Effect of exercise training on resistance arteries in rats with chronic NOS inhibition.

Regular exercise has blood pressure-lowering effects, as shown in different types of experimental hypertension models in rats, including the nitric oxide synthase (NOS) inhibition model. We aimed to investigate possible mechanisms implicated in the exercise effect by evaluating the vasoreactivity of resistance arteries. Exercise effects on agonist-induced vasodilatory responses and flow-mediated dilation were evaluated in vessel segments of the rat chronic NOS inhibition model. Normotensive and hypertensive rats were subjected to swimming exercise (1 h/day, 5 days/wk, 6 wk), while rats in other sedentary and hypertensive groups did not. Hypertension was induced by oral administration of the nonselective NOS inhibitor l-NAME (25 mg/kg day) for 6 wk. Systolic blood pressure, as measured by the tail-cuff method, was significantly decreased by the training protocol in exercising hypertensive rats. The vasoreactivity of resistance arteries was evaluated by both wire and pressure myography studies. An impaired nitric oxide-mediated relaxation pathway in untrained hypertensive rats led to decreased relaxation responses in vessels with intact endothelium. Exercise training significantly improved the responses to acetylcholine and flow-mediated dilation in exercise-trained hypertensive rats in parallel with a decrease in blood pressure. On the other hand contraction (norepinephrine and KCl) and relaxation (sodium nitroprusside) responses of vascular smooth muscle were not different between the groups. Vascular endothelial NOS protein expression was found to be increased in both exercising groups. In conclusion, these results revealed evidence of an increased role of the nitric oxide-dependent relaxation pathway in exercising hypertensive rats.

Role of hemoglobin oxygenation in the modulation of red blood cell mechanical properties by nitric oxide.

It has been previously demonstrated that both externally generated and internally synthesized nitric oxide (NO) can affect red blood cell (RBC) deformability. Further studies have shown that the RBC has active NO synthesizing mechanisms and that these mechanisms may play role in maintaining normal RBC mechanical properties. However, hemoglobin within the RBC is known to be a potent scavenger of NO; oxy-hemoglobin scavenges NO faster than deoxy-hemoglobin via the dioxygenation reaction to nitrate. The present study aimed at investigating the role of hemoglobin oxygenation in the modulation of RBC rheologic behavior by NO. Human blood was obtained from healthy volunteers, anticoagulated with sodium heparin (15 IU/mL), and the hematocrit was adjusted to 0.4 L/L by adding or removing autologous plasma. Several two mL aliquots of blood were equilibrated at room temperature (22+/-2 degrees C) with moisturized air or 100% nitrogen by a membrane gas exchanger, The NO donor sodium nitroprusside (SNP), at a concentration range of 10(-7)-10(-4)M, was added to the equilibrated aliquots which were maintained under the same conditions for an additional 60 min. The effect of the non-specific NOS inhibitor l-NAME was also tested at a concentration of 10(-3)M. RBC deformability was measured using an ektacytometer with an environment corresponding to that used for the prior incubation (i.e., oxygenated or deoxygenated). Our results indicate an improvement of RBC deformability with the NO donor SNP that was much more pronounced in the deoxygenated aliquots. SNP also had a more pronounced effect on RBC aggregation for deoxygenated RBC. Conversely, l-NAME had no effect on deoxygenated blood but resulted in impaired deformability, with no change in aggregation for oxygenated blood. These findings can be explained by a differential behavior of hemoglobin under oxygenated and deoxygenated conditions; the influence of oxygen partial pressure on NOS activity may also play a role. It is therefore critical to consider the oxygenation state of intracellular hemoglobin while studying the role of NO as a regulator of RBC mechanical properties.

Vascular dilation responses of rat small mesenteric arteries at high intravascular pressure in spontaneously hypertensive rats.

BACKGROUND: Hypertension is associated with remodeling and mechanical alterations of resistance arteries. Numerous studies have investigated the mechanical and morphometric properties of small arteries obtained from hypertensive animals and humans. However, the functional properties of resistance arteries from normotensive and hypertensive subjects have only been examined under normotensive conditions. The objective of the present study was to evaluate the dilation responses of small mesenteric arteries (SMA) from spontaneously hypertensive rats (SHR) at various levels of intraluminal pressure. METHODS AND RESULTS: SMA segments from Wistar Kyoto (WKY) rats and SHR were pressurized using pressure myography. Endothelium-dependent and -independent dilation responses of the SMA were examined under 3 different intravascular pressures (50, 80 and 120 mmHg). Endothelium-dependent dilation was evaluated by measuring vasodilator responses to increasing doses of acetylcholine or increases in intraluminal flow rate. Endothelium-independent vasodilator function was examined by using sodium nitroprusside. The results indicate that both endothelium-dependent and -independent dilation responses of SMA from WKY progressively decrease with increased intravascular pressure. In contrast, all dilatation responses of the SMA from SHR were enhanced at higher intraluminal pressures. CONCLUSIONS: These findings of differential sensitivity to luminal pressure should be considered during in vitro examination of vessels from normotensive and hypertensive subjects.

Parameterization of red blood cell elongation index--shear stress curves obtained by ektacytometry.

Measurement of red blood cell (RBC) deformability by ektacytometry yields a set of elongation indexes (EI) measured at various shear stresses (SS) presented as SS-EI curves, or tabulated data. These are useful for detailed analysis, but may not be appropriate when a simple comparison of a global parameter between groups is required. Based on the characteristic shape of SS-EI curves, two approaches have been proposed to calculate the maximal RBC elongation index (EI(max)) and the shear stress required for one-half of this maximal deformation (SS(1/2)): (i) linear Lineweaver-Burke (LB) model; (ii) Streekstra-Bronkhorst (SB) model. Both approaches have specific assumptions and thus may be subject to the measurement conditions. Using RBC treated with various concentrations of glutaraldehyde (GA) and data obtained by ektacytometry, the two approaches have been compared for nine different ranges of SS between 0.6-75 Pa. Our results indicate that: (i) the sensitivity of both models can be affected by the SS range and limits employed; (ii) over the entire range of SS-data, a non-linear curve fitting approach to the LB model gave more consistent results than a linear approach; (iii) the LB method is better for detecting SS(1/2) differences between RBC treated with 0.001-0.005% glutaraldehyde (GA) and for a 40% mixture of rigid cells but is equally sensitive to SB for 10% rigid cells; and (iv) the LB and SB methods for EI(max) are equivalent for 0.001% and 0.003% GA and 40% rigid, with the SB better for 0.005% GA and the LB better for 10% rigid.

Relationships between hemodynamic, hemorheological and metabolic responses during exercise.

Aerobic performance is dependent on both cardio-respiratory and peripheral factors with hemodynamic parameters playing a major role. However, whether blood rheology might affect aerobic performance through an effect on hemodynamic factors is not known. The aim of the present study was to assess the relationships between hemodynamic, hemorheological and metabolic parameters in response to a sub-maximal cycling exercise protocol consisting of three successive levels of nine min duration (50, 100 and 150 W). Ten young sportsmen participated in the present study. Mean arterial pressure (MAP) was measured manually, with thoracic impedance used to monitor cardiac output (Qc): systemic vascular resistance (SVR) was then calculated. Whole blood viscosity (etab) was measured and used to calculate systemic vascular hindrance. Hematocrit (Hct) was determined by micro-centrifugation and red blood cell (RBC) deformability (EI) was determined by ecktacytometry. A breath-by-breath gas analyzer was used to measure oxygen uptake (VO2); the Fick equation was used to calculate arterio-venous oxygen difference [(a-v)O(2)] from VO(2) and Qc. All measurements were performed at rest, during exercise and during recovery. Compared to baseline, Qc, MAP, Hct, EI, VO(2), and (a-v)O(2) increased during exercise. etab increased above baseline only at 150 W and remained elevated during recovery; the increase in etab during the last level of exercise was associated with a decrease of SVR and systemic vascular hindrance. There was a significant negative correlation between EI and SVR (r=-0.35, p<0.01) and a significant positive relationship between EI and (a-v)O(2) (r=0.35, p<0.01) and between EI and VO(2) (r=0.37, p<0.01) across all exercise workloads, thus suggesting a potential role for RBC deformability as a factor affecting aerobic performance via oxygen delivery to tissues. These data lend support to the concept that hemorheological parameters may contribute to hemodynamic and cardio-respiratory adaptations in response to exercise in moderately trained sportsmen.

Effect of hemoglobin oxygenation level on red blood cell deformability and aggregation parameters.

Measurements of red blood cell (RBC) deformability and aggregation can be subject to influence by pre-analytical handling procedures, with the degree of hemoglobin oxygenation having the potential to affect the results. To examine such effects, RBC deformability and aggregation were studied before and after oxygenation or deoxygenation of human blood samples. RBC deformability was assessed using a laser-diffraction ektacytometer having Couette geometry. RBC aggregation was assessed using the same system by monitoring light backscattering after a sudden cessation of high shear; aggregation was also measured by monitoring light transmittance through RBC suspensions. RBC deformability was found to be significantly increased after equilibrating RBC with ambient air (pO2: 142.0+/-3.1 mmHg) compared to the non-oxygenated sample (pO2: 42.4+/-1.8 mmHg). In contrast, equilibration with 100% nitrogen resulted in significant impairment in RBC deformability. RBC aggregation parameters were also affected by oxygenation if measured based on light backscattering, but not if measured using light transmittance. It is thus recommended that blood samples be oxygenated by repeated exposure to ambient air prior to the measurement of hemorheological parameters.

Effects of storage duration and temperature of human blood on red cell deformability and aggregation.

Blood samples used in hemorheological studies may be stored for a period of time, the effects of storage have yet to be fully explored. This study evaluated the effects of storage temperature (i.e., 4 degrees C or 25 degrees C) and duration on RBC deformability and aggregation for blood from healthy controls and from septic patients. Our results indicate that for normal blood, RBC deformability over 0.3-50 Pa is stable up to six hours regardless of storage temperature; at eight hours there were no significant differences in EI but SS1/2 calculated via a Lineweaver-Burk method indicated impaired deformability. Storage temperature affected the stable period for RBC aggregation: the safe time was shorter at 25 degrees C whereas at 4 degrees C aggregation was stable up to 12 hours. Interestingly, blood samples from septic patients were less affected by storage. Blood can thus be stored at 25 degrees C for up to six hours for deformability studies, but should be limited to four hours for RBC aggregation; storage at 4 degrees C may prolong the storage period up to 12 hours for aggregation but not deformability measurements. Therefore, the time period between sampling and measurement should be as short as possible and reported together with results.

Sampling time after tourniquet removal affects erythrocyte deformability and aggregation measurements.

Venipuncture procedures are widely thought to influence biochemical, hematological or hemorheological measurements. In line with the preparation of the new Guidelines for the standardization of hemorheological measurement, we compared various blood rheological parameters (i.e., red blood cell deformability and aggregation indices) assessed in blood samples obtained after 5, 30, 60 and 90 s following the tourniquet removal and a blood sample obtained without applying a tourniquet (control sample). A slight but significant improvement in red blood cell (RBC) deformability after the removal of tourniquet compared to the control sample was observed. RBC deformability was maximal in the samples obtained 30 s after tourniquet removal and remained slightly higher than the control in the following samples (at 60 and 90 s after tourniquet removal). The aggregation index (AI) decreased with time after tourniquet removal reaching significantly lower values than the control at 90 s after tourniquet removal. This finding was supported by a greater half time for RBC aggregation in the samples obtained 60 and 90 s after tourniquet removal. In conclusion, this study revealed that RBC deformability and aggregation might be significantly altered in the samples obtained after the application and removal of a tourniquet, as a part of the blood sampling procedure. Recommendation "remove the tourniquet at least 5 s prior to the start of blood sampling" may need to be revised.

RBC aggregation: more important than RBC adhesion to endothelial cells as a determinant of in vivo blood flow in health and disease.

Although the shear-dependent and reversible phenomenon of red blood cell (RBC) aggregation has been studied for decades, its role as a determinant of in vivo blood flow in both health and disease has not yet been fully documented. In this brief review, we present compelling arguments, supported by literature evidence, that in vivo flow dynamics are more affected by RBC aggregation than by RBC adhesion to endothelial cells (ECs). A companion article (i.e., a "counter-point") published in this issue of the journal argues that in disease states, RBC-EC adhesion is the more important determinant.

In vivo correlates of altered blood rheology.

It is has been known for more than 80 years that compared to in vitro determinations, blood behaves as a less viscous fluid under in vivo flow conditions. The experiments of Whittaker and Winton were among the first dealing with the in vivo effects of altered blood rheology, and experimental studies during the second half of 20th century have provided additional evidence for the complexity of in vivo hemodynamics-hemorheology relationships. Careful studies indicate that the impact of a given blood rheology alteration is determined by the properties of the experimental model (e.g., organ or tissue under investigation), experimental approach (e.g., intravital microscopy, whole organ perfusion) and method used to modify blood rheology. In addition, vascular control mechanisms may play a major role in the resulting hemodynamic effects of a hemorheological alteration: (1) a response simply related to metabolic autoregulation in which there is a compensatory vasodilation due to altered in vivo blood flow and organ/tissue hypoxia; (2) modulation of endothelial function (e.g., NO production) via altering wall shear stress, thereby leading to changes of vascular hindrance. The in vivo effects of altered red blood cell (RBC) aggregation have been investigated in various experimental models. A novel technique for modifying RBC aggregability (i.e., intrinsic tendency of RBC to aggregate) by covalent attachment of specific co-polymers has been used in some studies, and has provided data reflecting the specific effects of RBC aggregation without the influence of altered suspending phase properties. These data indicate that both the magnitude of the hemodynamic effect and the direction of the alteration depend on the intensity of RBC aggregation. Using the same novel technique, RBC aggregation has been shown to be an important determinant of endothelial function through its effects on RBC axial distribution and wall shear stress. These somewhat diverse findings can be explained by considering the contribution of various in vivo hemorheological mechanisms that have opposite effects on in vivo flow resistance.

Effect of lanthanum on red blood cell deformability.

Prior reports describing the effects of lanthanum (La(3+)) on red blood cells (RBC) have focused on the effects of this lanthanide on cell fusion or on membrane characteristics (e.g., ion movement across membrane, membrane protein aggregation); the present study explores its rheological and biophysical effects. Normal human RBC were exposed to La(3+) levels up to 200 microM then tested for: (1) cellular deformability using a laser-based ektacytometer and an optical-based rheoscope; (2) membrane viscoelastic behavior via micropipettes; (3) surface charge via micro electrophoresis. La(3+) concentrations of 12.5 to 200 microM caused dose-dependent decreases of deformability that were greatest at low stresses: these rheological changes were completely reversible upon removing La(3+) from the media either by washing with La(3+)-free buffer or by suspending La(3+)-exposed cells in La(3+)-free media (i.e., viscous dextran solution). Both membrane shear elastic modulus and membrane surface viscosity were increased by 25-30% at 100 or 200 microM. As expected, La(3+) decreased RBC electrophoretic mobility (EPM), with EPM inversely but not linearly associated with deformability; changes of EPM were also completely reversible. These results thus indicate novel aspects of RBC cellular and membrane rheological behavior yet raise questions regarding specific mechanisms responsible for La(3+)-induced alterations.

Effect of decreased plasma cholesterol by atorvastatin treatment on erythrocyte mechanical properties.

3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are the most commonly used cholesterol-lowering drugs, with recent clinical trends usually aimed at achieving the lowest possible plasma cholesterol levels. Although the effects of increased plasma cholesterol have been previously reported, it is not obvious how very low plasma cholesterol levels would affect membrane composition and the deformability of red blood cells (RBC). The present study investigated the effects of hypocholesterolemia achieved by atorvastatin therapy on RBC membrane and mechanical properties in guinea pigs fed a normal diet. Two groups of animals were used (atorvastatin-treated, n=12; control n=12), and atorvastatin given orally in isotonic phosphate-buffered saline (PBS) at a dose of 20 mg/kg/day for a 21-day period. Our results indicate that the atorvastatin-treated group had significantly lower plasma total cholesterol (17.42+/-1.70 mg/dl), low-density lipoprotein cholesterol (5.25+/-2.22 mg/dl) and triglycerides (42.60+/-3.78 mg/dl) than the control group (34.08+/-1.72, 21.17+/-1.41 and 60.64+/-2.43 mg/dl, respectively). In addition, membrane cholesterol content was lower (p<0.0001) and phospholipid content higher (p<0.0001) in the atorvastatin-treated group, thus decreasing the cholesterol to phospholipid ratio; a significant enhancement in sodium-potassium-ATPase activity also occurred. However, in spite the marked changes of plasma and RBC membrane composition, there was no change of RBC deformability. Note that although our results indicate no adverse rheological alterations, extension of our findings to humans requires caution.

Hemodynamic effects of red blood cell aggregation.

The influence of red blood cell (RBC) aggregation on blood flow in vivo has been under debate since early 1900's, yet a full understanding has still has not been reached. Enhanced RBC aggregation is well known to increase blood viscosity measured in rotational viscometers. However, it has been demonstrated that RBC aggregation may decrease flow resistance in cylindrical tubes, due to the formation of a cell-poor zone near the tube wall which results from the enhanced central accumulation of RBC. There is also extensive discussion regarding the effects of RBC aggregation on in vivo blood flow resistance. Several groups have reported increased microcirculatory flow resistance with enhanced RBC aggregation in experiments that utilized intravital microscopy. Alternatively, whole organ studies revealed that flow resistance may be significantly decreased if RBC aggregation is enhanced. Recently, new techniques have been developed to achieve well-controlled, graded alterations in RBC aggregation without influencing suspending phase properties. Studies using this technique revealed that the effects of RBC aggregation are determined by the degree of aggregation changes, and that this relationship can be explained by different hemodynamic mechanisms.