Preservation of Dry Liposomes Does not Require Retention of Residual Water

Certain sugars, particularly trehalose, dramatically alter physical properties of dry phospholipids in ways that mimic the presence of water. As a result, these sugars are capable of preserving the integrity of dry liposomes and membranes. Since these effects could conceivably be due to the presence of small amounts of water in the dry preparations of sugar and lipid, we have done careful measurements of the residual water contents in the dry samples and report the results here. Lyophilized liposomes composed of palmitoyloleoylphosphatidylcholine and phosphatidylserine (9:1) contain at most 0.2 mol of H2O per mol of lipid. When the trehalose concentration in the dry mixtures is increased, there is no increase in the apparent water content of the samples over a wide range of sugar concentrations. Over the same range of trehalose contents the maximal effect of trehalose on physical properties of the lipids and on stabilization of liposomes is achieved. We conclude that the stabilization does not require retention of residual amounts of water in the dry trehalose-phospholipid preparations. Similar studies with other sugars show a relationship between the amount of sugar interacting with the lipid and the ability of the same sugar to stabilize dry liposomes.