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The degree of hemolysis as a function of surface roughness value and roughened area under shear flow conditions was investigated using a rotational shear stressor. The shearing portion of the stressor is cone shaped in its upper and lower positions, with a cylindrical central section. Surface roughness was applied to the cylindrical section. Bovine blood was sheared for 30 min over a set of roughened surfaces of between arithmetic mean roughness (Ra) 0.1 and 0.8 mm covering 10% of the surface area of the cylindrical section (equivalent to 1.8% of the whole blood contact area) at a shear rate of 3750/s. The threshold value thus obtained for rapid increase in hemolysis was between Ra 0.6 and 0.8 mm. When sheared with a roughened surface of Ra 0.8 mm applied to the cylindrical surface at areas between 0 and 100% (equivalent to between 0 and 18% of the whole blood-contacting area), the hemolysis level did not increase from 10 to 100%, but a significant difference was obtained between 0 and 10%. This suggests that red blood cells were destroyed not by fatigue failure caused by rolling on the roughened surface, but due to the high shear stress generated by surface roughness. Moreover, it appears that the shear stress was generated over the entire cylindrical section, regardless of the area of surface roughness.