Niosomes are vesicular carriers formed by a bilayer shell, which is composed of non-ionic surfactants with the addition of a structural supporting agent. Cholesterol is typically used as an additive to increase the stability or drug entrapment efficiency of niosomes. Although increasing the amount of cholesterol is reported to improve niosomal properties, an excessive amount of cholesterol may not be accommodated in the bilayer shell, and thus remain in the crystalline form in the niosomal solution. The presence of a crystalline phase is a potential risk for further medical application. Therefore, Tween 80-based niosomes were prepared using a well-established thin-film hydration method and organic phase injection method, followed by their thorough characterization in order to estimate the cholesterol incorporation into the niosomal shell. To detect the crystalline phase in the niosomal suspensions, a novel approach based on depolarized dynamic light scattering combined with cryo-transmission electron microscopy, X-ray diffraction and optical microscopy is used to confirm the presence of cholesterol crystals. This method is fast, quantitative, and allows the sample analysis in a natural liquid environment, thus eliminating biased results influenced by sample drying.
Keywords: Crystalline cholesterol; Depolarization; Niosomes; Static and dynamic light scattering; X-ray diffraction; cryo-TEM.
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