Charged hybrid block copolymer-lipid-cholesterol vesicles: pH, ionic environment, and composition dependence of phase transitions

Abstract

The impacts of pH, salt concentration (expressed as Debye length), and composition on the phase behavior of hybrid block copolymer-lipid-cholesterol bilayers incorporating carboxyl-terminated poly(butadiene)-block-poly(ethylene oxide) copolymer (PBdPEO1800^(-)) or/and non-carboxyl-terminated PBdPEO (PBdPEO1800 or/and PBdPEO950), egg sphingomyelin (egg SM), and cholesterol were examined using fluorescence spectroscopy of laurdan. Laurdan emission spectra were decomposed into three lognormal curves as functions of energy. The ratio of the area of the mid-energy peak to the sum of the areas of all three peaks was evaluated as vesicles were cooled, yielding temperature breakpoint values (T_break) expected to be within the range of the phase transition temperature. T_break values displayed dependence on pH, Debye length, and vesicle composition consistent with an electrostatic repulsion contribution to vesicle phase behavior. Increased pH and Debye length, for which a greater dissociated fraction of PBdPEO1800^(-) and a greater energy of electrostatic repulsion would be expected, resulted in T_break values as much as 10 °C less than at low pH or short Debye lengths. Additionally, at Debye lengths comparable to those at physiologically relevant ionic strength, T_break at pH 5.9 was observed to be slightly higher than at pH 7.0 for vesicles containing 50 mol% PBdPEO1800^(-). Electrostatic effects observed for hybrid vesicles incorporating significant amounts of carboxyl-terminated polymer may have the ability to drive phase separation in response to pH drops-such as those observed after endocytosis-in physiologically relevant conditions, suggesting the utility of such materials for drug delivery.

Keywords: Copolymer; Electrostatics; Fluorescence spectroscopy; Hybrid vesicles; Liquid-ordered; Membrane phase behavior.