doi:10.1016/j.bbamem.2006.01.002
Copyright © 2006 Elsevier B.V. All rights reserved.
Effects of trehalose on the phase behavior of DPPC–cholesterol unilamellar vesicles
References and further reading may be available for this article. To view references and further reading you must
purchase this article.
Satoshi Ohtakea, Carolina Scheborb, 1 and Juan J. de Pabloa,
, 
aChemical and Biological Engineering Department, University of Wisconsin, 1415 Engineering Drive, Madison, WI 53706, USA
bDepartamento de Industrias, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, (1428) Ciudad Autónoma de Buenos Aires, Argentina
Received 12 August 2005;
revised 6 January 2006;
accepted 6 January 2006.
Available online 27 January 2006.
Abstract
A systematic study is presented of the effects of trehalose on the physical properties of extruded DPPC–cholesterol unilamellar vesicles. Particular emphasis is placed on examining how the interactions present in the hydrated state translate into those in the dehydrated state. Observations from HSDSC and DSC are used to examine the phase behavior of hydrated and dehydrated vesicles, respectively. The concentration of trehalose inside and outside the vesicles is manipulated, and is shown to affect the relative stability of the membranes. Our results show for the first time that a combination of high inner and low outer trehalose concentration is able to decrease the gel-to-liquid crystalline phase temperature (Tm), while any other combination will not. Upon dehydration, the Tm of all lipid mixtures increases. The extent of the increase depends on the trehalose distribution across the bilayer. The Tm changes in the same direction with trehalose concentration for both freeze-dried and fully hydrated samples, suggesting that the trehalose distribution across the vesicle membrane, as well as the trehalose–phospholipid interaction, is maintained upon lyophilization. The results presented in this work may aid in the formulation of systems to be used in the lyophilization of liposomes for drug delivery applications.
Keywords: Trehalose; DPPC; Cholesterol; Liposome
Fig. 1. Thermograms for DPPC liposomes (diameter = 100 nm) in trehalose solutions. Trehalose concentration (mM) is indicated for each curve.
Fig. 2. Thermograms for (a) 100 nm and (b) 50 nm diameter DPPC liposomes containing asymmetric trehalose concentrations (values indicated in the figure denote inner/outer mM trehalose concentrations) across the vesicle bilayer.
Fig. 3. Thermograms for 100 nm diameter DPPC–cholesterol liposomes containing (a) 9 mol% and (b) 17 mol% cholesterol. The values indicate the concentration (mM) of trehalose inside/outside the vesicle. The thermograms for DPPC–cholesterol liposomes without trehalose are identical to those obtained at 11/11 mM trehalose (data not shown).
Fig. 4. Thermograms obtained from the first (a) and second (b) heating scans for 100 nm diameter DPPC vesicles freeze-dried in the presence of various trehalose concentrations (values indicated in the figure). The samples contain less than 1% (wt) water.
Fig. 5. Thermograms obtained from the first (a) and second (b) heating scans for various 100 nm diameter DPPC–cholesterol (Ch) vesicles (values indicated in the figure) freeze-dried in the presence of 146/11 mM trehalose. The samples contain less than 1% (wt) water.
Table 1.
The various trehalose concentrations considered in this work

Table 2.
The enthalpy of the phase transition (ΔH), the vesicle diameter (d), and the size of the cooperative unit of the transition (CU) for 100 nm DPPC liposomes containing the indicated trehalose concentrations

Standard deviation data are indicated for each ΔH, d and CU value.
Table 3.
Average diameters (nm) of DPPC–cholesterol vesicles containing various trehalose concentrations

Standard deviation data are indicated for each size value.
Table 4.
Size of the cooperative unit of the phase transition (CU) for DPPC and DPPC–cholesterol vesicles at 9 and 17 mol% cholesterol

Standard deviation data are indicated for each CU value.
Table 5.
Tm of DPPC–cholesterol vesicles obtained during the first and second heating scans

Standard deviation data are indicated for each Tm value.

Corresponding author. Tel.: +1 608 262 7727; fax: +1 608 262 5434.
1 Member of CONICET, Argentina.