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Ann Biomed Eng. 2005 Oct;33(10):1375-86.

De-activation of neutrophils in suspension by fluid shear stress: a requirement for erythrocytes.

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Department of Bioengineering, The Whitaker Institute for Biomedical Engineering, University of California-San Diego, La Jolla, CA 92093-0412, USA.


Leukocyte de-activation in response to a mechanical stimulus may be an important mechanism to reduce inflammation in the circulation and cardiovascular complications. We examine here a specific form of leukocyte activation in the form of pseudopod projection, a process that is important during cell spreading and migration, but if it occurs in circulating leukocytes, may also lead to their entrapment in the microvascular network. Fresh neutrophils were activated with fMLP, suspended without adhesion to endothelium, and sheared in a cone-and-plate device while both shear stress and shear rate were measured. A fraction of the activated neutrophils retracted their pseudopods under the influence of fluid shear and returned to round shape. Pseudopod retraction was observed only in the presence of erythrocytes (at shear stresses up to approximately 25 dyn/cm(2)). At a constant hematocrit and increasing plasma viscosities with addition of macromolecules, the number of de-activated neutrophils scaled with shear stress and less so with shear rate. We examined a biochemical and rheological role of erythrocytes during shear de-activation of neutrophils. Addition of superoxide dismutase (SOD) in phosphate buffer served to enhance neutrophil de-activation by fluid shear. Replacement of erythrocytes by solid microspheres (5.4 mum) to simulate the particle properties of the erythrocytes, did not serve to enhance neutrophil de-activation unless in the presence of SOD. At higher shear stresses without erythrocytes (38-77 dyn/cm(2)), we also observed neutrophil de-activation but only in the presence of SOD. These results suggest that erythrocytes play an important role in neutrophil de-activation by reducing the superoxide level in plasma. Shear stress, rather than shear rate, is the key determinant that regulates neutrophil de-activation.

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