Mechanical properties of bovine aortic endothelial cells in suspension studied by ultrasonic interferometry.

OBJECTIVE: Cell adhesion phenomenon has been extensively studied in the last decade and was shown to be mediated by specialized molecules and driven by physical forces. Cohesion of the vessel wall cells is also dependent on adhesion molecules but less is known about the physical forces involved. To investigate endothelial cell/endothelial cell interaction from a mechanical point of view, we have used an ultrasonic interferometry device, named EchoCell, which has been previously designed to study red blood cell-red bood cell (RBC-RBC) interaction. METHODS: Bovine aortic endothelial (BAE) cells were cultured, detached, then suspended in buffer and their mechanical and geometrical properties studied with the EchoCell system. The ultrasonic apparatus measures both the accumulation rate of cells in suspension on a solid plate and the acoustical impedances of the suspension and the sediment. RESULTS: In suspension, BAE exhibited, in our experimental conditions (3x10(6) cells per ml), a spherical size evaluated by calculation at a mean radius of 7+/-2 microm. Moreover, no BAE aggregation occurred at the concentrations used. The acoustical impedance of the BAE suspensions calculated from all the samples studied, in the cell concentration range from 1.5x10(6) to 6x10(6) cells per ml, was 1.52x10(6) Rayl (kg m(-2) s(-1)). Furthermore, the acoustical impedance of the cell sediment was found to be independent on the initial cell suspension concentration and equal to 1.63x10(6) Rayl (kg m(-2) s(-1)). Estimation of the volume fraction of BAE inside the sediment allows to evaluate the ultrasonic velocity and the elastic bulk modulus of cells. CONCLUSION: The ultrasonic interferometry method appears particularly interesting to study geometrical and mechanical (acoustical impedance, sound velocity, elastic bulk modulus) properties of BAE cells.

Covalent binding of polyethylene glycol to the surface of red blood cells as detected and followed up by cell electrophoresis and rheological methods.

Cyanuric chloride activated polyethylene glycol (PEG)-5000 was covalently coupled to murine and human red blood cells (pegylated RBC). Our purpose was to camouflage RBC receptors, which is necessary for parasite invasion, a process essential to sustain parasitemia. Cell electrophoretic mobility analysis (CEM) of pegylated RBC distinguished a new population of cells bearing characteristic CEM. Pegylation of RBC also modified their rheological properties, which were documented by evaluation of cell deformability (based on cell transit time through calibrated micropores) and cell aggregation (as measured by ultrasonic interferometry). Homologous transfusion of pegylated RBC into murine malaria-infected mice had no significant effect on the cerebral malaria death rate in Plasmodium berghei-infected mice, but it reduced the peripheral blood parasitemia by a factor 2 while in Plasmodium yoelii infected mice, the parasitemia was dramatically reduced by a factor of 4. These experiments demonstrate that transfusion of pegylated RBC may inhibit peripheral parasitemia. Cell electrophoresis appears to be a useful tool to allow in vivo detection and to investigate the fate of transfused pegylated RBC.

Gender, menstrual cycle, oral contraceptives and red blood cell deformability in healthy adult subjects.

Gender, menstrual cycle and oral contraceptives may have influence on mechanical properties of Red Blood Cell (RBC) and particularly on RBC deformability. So cell transit parameters have been assessed by filtration with the Cell Transit Analyser (CTA) for a large healthy adult population (seventy-nine males and one-hundred-fifteen females). The CTA provides the distribution of cell transit times of 5000 red blood cells, the mean transit time of the population and different percentiles such as p50, p75, p90 and p95. No effect of oral contraceptives was found. Nevertheless, influence of sex and menstrual cycle were demonstrated. A significant increase of the filtration parameters measured in the female population with respect to the male population and during menstruation, preovulation and post-ovulation periods was observed. During ovulation, the CTA parameters are comparable to the same parameters found in males.

Casein peptide release and passage to the blood in humans during digestion of milk or yogurt.

In adult humans, after milk or yogurt ingestion, many peptides derived from alpha s1-, beta- or kappa-caseins were detected in stomach, including the kappa-caseinoglycopeptide, an inhibitor of platelet aggregation. Smaller peptides derived from casein and lactoferrin were recovered from duodenum. Two long peptides, the kappa-caseinoglycopeptide and the N-terminal peptide of alpha s1-casein, were absorbed and detected in plasma. These results support the concept that food-born peptides could have physiological activities in man.

Measurement of platelet aggregation peptide inhibitors by ultrasonic interferometry.

Several peptide inhibitors of thrombin- or collagen-induced platelet aggregation and of the interaction between glycoprotein Ib and von Willebrand factor were studied by a new method--ultrasonic interferometry (Echo Cell). Inhibition of aggregate formation in a concentration-dependent manner was observed. The sensitivity of the method was 3 to 40 times higher than that of classical turbidimetry.

Measurement of human platelet microaggregates by a new method: ultrasonic interferometry.

We have adapted the ultrasonic interferometry technique (Echo-Cell), which was initially designed to study red blood cell aggregation and agglutination, to the detection of human platelet microaggregates. The experimental parameter chosen was the slope of the signal over the first 5 minutes of sedimentation. We compared our new method with the conventional aggregometry for the measurement of aggregates after thrombin-, collagen-, and epinephrine-induced platelet activation. Under these conditions we demonstrated the particular sensibility of the present method in detecting small platelet aggregates induced in the first phase of aggregation and formed by low concentrations of agonists. Furthermore, as an illustration of this method, we showed an inhibition of the formation of thrombin-induced platelet aggregates in a concentration-dependent manner by the well known antagonist arginine-glycine-aspartic acid-serine with a median inhibitory concentration of 0.4 micromol/L, which is 30 times lower than the median inhibitory concentration found by aggregometry.