Physical Stability of Sonicated Arsonoliposomes: Effect of Calcium Ions

The physical stability of sonicated arsonoliposomes in the absence and presence of Ca2+ ions is evaluated. Cholesterol‐containing arsonoliposomes composed of arsonolipids [having different acyl chains (C12-C18)], or mixtures of arsonolipids with phospholipids (phosphatidylcholine or distearoyl‐phosphatidylcholine) were prepared, and physical stability was evaluated in the absence and presence of CaCl2, by vesicle dispersions turbidity measurements and cryo‐electron microscopy morphological assessment. In some cases, vesicle ζ‐potential was measured, under identical conditions. Results demonstrate that self‐aggregation of the vesicles studied is low and influenced by the acyl chain length of the arsonolipid used, whereas calcium‐induced aggregation is higher, correlating well with the decreased values of vesicle ζ‐potential in the presence of Ca2+ ions (weaker electrostatic repulsion). Acyl chain length of arsonolipids used has a significant quantitative effect on Ca2+‐induced vesicle aggregation mainly for arsonoliposomes that contain phospholipids (mixed), compared with the vesicles that consist of plain arsonolipids (significant effect only for initial aggregation at time 0). Another difference between plain and mixed arsonoliposomes is that for mixed arsonoliposomes Ca2+‐induced increases in turbidity are irreversible by ethylenediaminotetraacetic acid, suggesting that vesicle fusion is taking place. This was confirmed by cryo‐electron microscopy observations. Finally, when phosphatidylcholine is replaced by distearoyl‐phosphatidylcholine, arsonoliposomes are more stable in terms of self‐aggregation, but in the presence of calcium, the turbidity and morphology results are similar.