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GB Nash, CS Johnson and HJ Meiselman
Department of Physiology and Biophysics, University of Southern California
School of Medicine, Los Angeles 90033.
The transformation of less-dense, discoidal homozygous sickle cells (HbSS)
RBCs into dehydrated, rheologically impaired cells is believed to be an
important factor in the pathophysiology of sickle cell disease. We
investigated this process by subjecting the less-dense fraction of HbSS
RBCs, which contains a low percentage of irreversibly sickled cells (ISCs),
to cyclic deoxygenation-reoxygenation for 15 hours at 37 degrees C. This
incubation procedure caused cell shrinkage, shifts in intracellular Na and
K content, and formation of ISCs that closely resembled endogenous ISCs
found in sickle blood. The viscoelasticity of the treated cells was tested
using micropipette techniques to measure the membrane shear elastic modulus
(mu) and the time constant for extensional shape recovery (tc); mu
represents the "static rigidity" of the cells, and the product mu.tc was
taken as a measure of their "dynamic rigidity". Density separation of the
incubated cells showed that their rheologic impairment (ie, elevation of
both static and dynamic rigidities) paralleled cellular dehydration and
that the newly formed dense cells had viscoelastic characteristics very
similar to those of endogenous dense cells. Rehydration by osmotic swelling
tended to normalize the dynamic rigidities of dense cells but had no
significant effect on their static rigidities. Thus, cellular dehydration
contributes to the observed changes of viscoelasticity, although an
irreversible alteration of membrane structure also appears to be involved.
Dense ISCs could be formed without added calcium, implying that entry of
external calcium is not an essential requirement for cellular dehydration;
ISCs formed without calcium tended to be less rigid (ie, to have lower
static and dynamic rigidities) than those formed with calcium. Our results
indicate that the cyclic incubation procedure closely mimics RBC rheologic
deterioration in vivo and thus suggest its usefulness as a model for
investigating this phenomenon.
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| Copyright © 1988 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||