Biophysical characterization of asolectin-squalene liposomes

Liposomes are shell nanoparticles able to embed hydrophobic molecules into their lipid layers to be released to cells. In pharmaceutical sciences, liposomes remain the delivery system with the highest biocompatibility, stability, loading characteristics, tunable physicochemical properties. Squalene is a natural, water insoluble, lipid, abundant in olive oil and shark liver. Studies in vitro and in animal models suggest protective and inhibitory effects of squalene against cancer. To study its effect on cells, and to overcome its insolubility in water, we have designed and produced large unilamellar liposomes containing different quantities of this terpene (0%, 2.8%, 5% w/w). Liposomes have been characterized by different biophysical techniques. Size-exclusion and affinity chromatography showed a unimodal size distribution and confirmed the squalene loaded dose. Laurdan fluorescence evidenced the changes in the hydration of the external layer of liposomes as a function of squalene concentration. Dynamic light scattering and small angle X-ray scattering revealed squalene induced structural differences in the hydrodynamic radius distribution and in the bilayer thickness respectively. Finally, preliminary experiments on the effects of liposome-delivered squalene on tumor and non-tumor cell lines showed time- and dose-dependent cytotoxic effects on LAN5 tumor cells and no effect on NIH-3T3 normal cells.