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Journal of Lipid Research, Vol 37, 606-630, Copyright © 1996 by Lipid Research, Inc.
DQ Wang and MC Carey
Using complementary physical-chemical techniques we defined five different
crystallization pathways as functions of time (30 days) and increasing
lecithin (egg yolk) content in pathophysiologically relevant model biles
super-saturated (cholesterol saturation indices, 1.2 - 2.7) by dilution of
approximately equal to 29 g/dl bile salt-lecithin- cholesterol micellar
solutions. As evidenced by quasi-elastic light- scattering spectroscopy,
supersaturation was heralded by the appearance of unilamellar vesicles.
With the lowest lecithin contents, arc-like crystals with habit and density
(d 1.030 g/mL) consistent with anhydrous cholesterol appeared first and
evolved via helical and tubular crystals to form plate-like cholesterol
monohydrate crystals (d 1.045 g/mL). With higher lecithin fractions,
cholesterol monohydrate crystals appeared earlier than arc and other
transitional crystals. With typical physiological lecithin contents, early
liquid crystals (d 1.020 g/mL) were followed by cholesterol monohydrate
crystals and subsequent appearances of arc and other intermediate crystals.
With higher lecithin contents, liquid crystals were followed by cholesterol
monohydrate crystals only, and at the highest lecithin mole fractions,
liquid crystals appeared that did not generate solid crystals. Added
calcium increased solid crystal number in proportion to its concentration
(5 - 20 mM) but did not influence appearance times, crystallization
pathways, or micellar cholesterol solubilities. Decreases in temperature
(37 degrees --> 4 degrees C), total lipid concentration (7.3 --> 2.4
g/dL), and bile salt hydrophobicity (3 alpha, 12 alpha --> 3 alpha, 7
alpha, 12 alpha --> 3 alpha, 7 beta hydroxylated taurine conjugates)
progressively shifted all crystallization pathways to lower lecithin
contents, retarded crystallization, and decreased micellar cholesterol
solubilities. The lecithin content of mother biles decreased markedly
during crystallization especially where liquid crystals were a coexisting
phase at equilibrium. This systematic study provides a framework for
understanding cholesterol crystallization in human and animal biles and for
examining factors that influence the kinetics of phase separation.
ARTICLES
Complete mapping of crystallization pathways during cholesterol precipitation from model bile: influence of physical-chemical variables of pathophysiologic relevance and identification of a stable liquid crystalline state in cold, dilute and hydrophilic bile salt-containing systems
Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
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