Shear stress-facilitated calcium ion transport across lipid bilayers

Abstract

Small unilamellar liposomes were used in this study of shear stress effects on the trans-bilayer flux of calcium ions (Ca 2+). Liposome suspensions were prepared from 99% egg phosphatidylcholine by a microporous filter extrusion technique. The inner aqueous phase of the unilamellar liposomes contained indo-1 5−, a fluorescent indicator of free Ca 2+. The external aqueous phase was composed of Hepes-buffered saline containing normal physiological levels of common ionic species. Calcium ion levels were set at 100 nM and 1 mM in the inner and outer aqueous phases, respectively. Liposome suspensions were exposed to graded levels of uniform shear stress in an optically modified rotational viscometer. Intraliposome Ca 2+ concentration was estimated from continuous measurement of indo-1 5− fluorescence. Electronically measured particle size distribution was used to determine liposome surface area for estimation of trans-bilayer Ca 2+ flux. Trans-bilayer Ca 2+ flux increased linearly with applied shear rate from 27 s −1 to 2700 s −1. Diffusional resistance of the lipid bilayer, not the convective resistance of the surrounding fluid, was the limiting step in the transport of Ca 2+. Liposome permeability to Ca 2+ increased by nearly two orders of magnitude over the physiologically relevant shear rate range studied. Solute transport in injectable liposome preparations may be dramatically influenced by cardiovascular fluid stress. Solute delivery rates determined in liposomes exposed to static conditions may not accurately predict in vivo, cardiovascular solute transport.