Biorheology

Since the sweet ketohexose L-sorbose causes overt hemolysis in dogs but not in man, we examined the possibility that L-sorbose induces a “prehemolytic state” of human red cells, manifesting itself as impairment of rheological red cell properties. After 2 hours incubation at 37°C relative viscosity of red cell suspensions measured by radial spreading in filter paper and packing ability of red cells were normal. Incubation for 24 and 48 hours of red cells in media containing L-sorbose, glucose or no sugar showed that relative viscosity was best maintained in glucose. Relative viscosity and packing ability of red cells in L-sorbose containing suspensions decreased less than in suspensions without sugar. This difference was independent of the glucose metabolism, red cell ATP, osmolality and pH of the suspending media, but appeared to be related to different degrees of spheroechinocytic red cell shape transformation observed in different suspending media. It is possible that L-sorbose has some antiechinocytic properties and/or that it induces an alteration of red cell membrane flexibility. There is no indication of an I.-sorbose induced “prehemolytic state” in human red cells.

The aggregation behaviour of normal and heat treated (48.4°C, 48.8°C, 49.5°C) red blood cells (RBCs) suspended in dextran-saline solutions (Dx70, Dx173) was investigated by a laser light reflectometric method over a wide range of bridging energies. The characteristic times of rouleau formation were found to be increased after RBC heat treatment. The disaggregation shear stress is not significantly different between normal RBCs and heat treated RBCs. The loss of cell deformability is nevertheless shown to improve slightly the dissociation efficiency of the flowing liquid in a shear flow resulting in a small reduction of the disaggregation shear rate after heat treatment. Heat treatment is also shown to alter the structure of RBC network at equilibrium. These results indicate that heat induced alterations of erythrocytes only affects the mechanical properties of the cell membrane without significant changes in the macromolecular bridging energy.

In order to attempt in vitro evaluation of antithrombogenecity for materials of artificial blood vessel tube, a new type of rheometer was developed. The rheometer originally consists of a cylindrical tube suspended from a torsion wire and filled with blood. The tube is excited in torsional oscillation and subsequent damped oscillation is observed. The apparatus can sensitively follow the change of fluidity during coagulation of blood. The damped oscillation curves during coagulation for fibrinogen-thrombin solution and blood put in a cylindrical tube made of the artificial material were measured. For fibrinogen-thrombin solution with lower fibrinogen and thrombin concentrations, the values of logarithmic damping factor (LDF) during coagulation increased and then decreased through a maximum. For blood and fibrinogen-thrombin solution with the higher concentrations of fibrinogen and thrombin, LDF monotonously decreased with the progress of coagulation. With a glass tube, the decrease of LDF for whole blood taken without anticoagulant rapidly occurred within about 15 min after sampling, while, with expanded poly tetrafluoroethylene (EPTFE ; Goar tex^®) and polydimethylsiloxane (Silastic^®) tubes, the decrease of LDF proceeds over 40–60 min. The present method is probably available for in vitro evaluation of anticoagulability or antithrombogenecity of artificial materials.

The viscoelastic behaviour of hardened or aggregated red blood cells is compared with the flow pattern of native red blood cells, all suspended in buffer solution at a hematocrit of 45%. The rheological properties are investigated under oscillatory shear at the constant frequency of 2Hz. Variation of the amplitude covers a range of shear-rates from 0.5/s to 200/s. It can be seen that rigidification of the red cells by treatment with glutardialdehyde leads to changes of the flow properties in the range of shear-rates above 10/s, whereas aggregate formation due to addition of dextran distinctly alters the flow properties in the range of shear-rates below 10/s.

The fluorescence polarization of probe molecules gives information on the “fluidity” of probe environment. Although the data cannot be related with absolute values of microviscosity, the method is largely used for probing the “fluidity” of lipid regions of biological membranes. Therefore, fluorescence polarization is of interest in clinical research, for membrane alterations are associated with either pathological processes of red cells, platelets, leukocytes or important cell functions.