*2.2. Preparation of Magnetoliposomes*

For magnetoliposomes preparation, the lipid 1,2-dipalmitoyl-*sn*-glycero-3-phosphatidylcholine (DPPC) (from Sigma-Aldrich, St. Louis, MO, USA), was used. The aqueous magnetoliposomes (AMLs) were developed through the ethanolic injection method [12,13]. Briefly, a 10 mM lipid solution in ethanol was injected, under vigorous agitation, to an aqueous dispersion of magnetic nanoparticles, above the melting transition temperature of DPPC (41 ◦C) [14]. The mixture was washed with water and purified by magnetic decantation to remove non-encapsulated nanoparticles, as previously reported [15].

Solid magnetoliposomes (SMLs) were developed by a reported method for manganese ferrite nanoparticles [12]. First, 10 μl of a nanoparticle solution (0.02 mg/mL), previously dispersed by sonication at 180 W for one minute, was added to 3 mL of chloroform. After brief sonication and under vigorous agitation, 150 μL of a DPPC 20 mM methanolic solution was added to form the first lipid layer. The first layer-coated nanoparticles were thoroughly washed with water to remove the lipids not attached to the nanoparticles' surfaces. The nanoparticles were dispersed in 3 mL of water and, under strong agitation, 150 μL of DPPC 20 mM methanolic solution was injected to form the second layer. The resulting solid magnetoliposomes were then washed and purified with ultrapure water by magnetic decantation [12,13].

Curcumin and Nile Red were loaded in AMLs through the co-injection method, while in SMLs they were incorporated through the injection of an ethanolic solution upon formation of the second lipid layer [12,13].
