Direct physical factors and PGI2 and TXA2 secretions by a human endothelial cell line: in vitro investigation of pressure and shear stress applied independently or in synergy.

The direct effect of two types of mechanical stress was measured through the prostacyclin (PGI2) and thromboxane A2 (TXA2) secretions by a confluent monolayer of cells from the EA.hy926 line. Eight values of constant pressure were applied in the gas phase above the culture medium, around atmospheric pressure taken as a control (0 mm Hg), from -500 to +760 mm Hg. Three amplitudes of sinewave modulated pressure (+/- 40; +/- 80; +/- 160 mm Hg) were explored at a frequency of 1 Hz. Modulated pressure (+/- 40 mm Hg) was also applied synergetically to a shear stress generated under steady state conditions by a rectilinear laminar motion of the medium. The cells remained adherent and exhibited unchanged morphology and viability. Constant pressure or depressure increased both PGI2 and TXA2 release but to an extent depending on the pressure value. Under pressure, the PGI2/TXA2 ratio was unchanged, but was higher under depressure, compared to the control. Pressure modulation strongly stimulated the secretion of PGI2 but had no effect on TXA2. Modulation strongly increased the PGI2/TXA2 ratio to a similar extent for the three amplitudes. Pressure-shear synergy enhanced secretion of PGI2 markedly more than shear stress alone, but the level reached was similar to the one induced by pressure modulation. No cumulative effect on the secretion of PGI2 was observed, whereas TXA2 synthesis undergoes a more than cumulative effect. The PGI2/TXA2 ratio remained unchanged under shear alone or under combined shear-pressure modulation but was higher with the modulated pressure alone. These results demonstrate that pressure has an outstanding effect on secretion that may be origin to local disturbances of the vascular system, thus inducing pathologies such as thrombosis or atherosclerosis.

Influence of a constant or a modulated pressure on insulin secretion by intact murine islets of Langerhans: existence of a threshold effect.

Pressure effect on cultured islets was investigated to simulate the behavior of the endocrine pancreas when subjected to an anatomical change in its environment. Insulin secretion from islets seeded on glass was determined by radioimmunoassays after 12-h exposure to constant or sine-wave pressure perturbation, at 37 degrees C and pH 7.4, under well-defined conditions applied through the culture medium. A specially devised setup allowed control of the constant component Po, the amplitude A, and the frequency f of the pressure perturbation. Under constant conditions, three Po values (760, 760 + 110, 760 + 250 mm Hg) were explored. Under sine-wave conditions, two frequencies (f = 0.2 and 4.33 Hz) were investigated for Po = 760 + 110 mm Hg and A = 10 mm Hg, as well as three amplitude values (A = +/-10, +/-50, and +/-110 mm Hg) for Po = 760 mm Hg and f = 0.2 Hz. Constant pressure inhibits secretion, by a factor of up to 60%, as well as low frequency (also by approximately 60%). Low amplitudes enhance secretion (by 41 and 31%), whereas a higher amplitude has a marked inhibiting effect (by 70%). In all cases, no linearity was observed, showing the existence of a threshold pressure effect. These results suggest that pressure may be an important parameter in certain forms of endocrine pancreatic dysfunctions.

Adsorption of bovine serum albumin onto glassy carbon in a Couette flow. Effect of shear rate on the adsorption kinetics and on the structure of the adsorbed proteic layer.

Adsorption of bovine serum albumin onto glassy carbon is investigated by analysing the time-variation of the double-layer capacitance recorded during the adsorption process. The effect of shear rate is investigated under laminar conditions in a Couette flow. Stationary and sinusoidally modulated values of the shear rate are imposed over the (0-200 s-1) range. The flow conditions are shown to play an essential role by markedly modifying the rate of all the steps (three at most) involved in the adsorption mechanism. Moreover, the structure of the adsorbed layers in the intermediate and final states are also strongly modified, an increase of the shear rate increasing the interaction between the protein and the electrode. Piezoelectric properties of albumin are invoked to account for the experimental results.