Bile Salts Caught in the Act: From Emulsification to Nanostructural Reorganization of Lipid Self-Assemblies

Bile salts (BSs) are important for the digestion and absorption of fats and fat-soluble vitamins in the small intestine. In this work, we scrutinized, with small-angle X-ray scattering (SAXS), the crucial functions of bile salts beyond their capacity for the interfacial stabilization of submicrometer-sized lipid particles. By studying a wide compositional range of BS-lipid dispersions using two widely applied lipids for drug-delivery systems (one a monoglyceride being stabilizer-sensitive and the other an aliphatic alcohol being relatively stabilizer-insensitive), we identified the necessary BS to lipid ratios to guarantee full emulsification. A novel ad hoc developed global small-angle-X-ray scattering analysis method revealed that the addition of BS hardly changes the bilayer thicknesses in bicontinuous phases, while significant membrane thinning is observed in the coexisting fluid lamellar phase. Furthermore, we show that a BS strongly decreases the average critical packing parameter. At increasing BS concentration, the order of phases formed is (i) the bicontinuous diamond cubic ( Pn3 m), (ii) the bicontinuous primitive cubic ( Im3 m), and (iii) the fluid lamellar phase ( L_α). These distinctive findings on BS-driven "emulsification" and "membrane curvature reduction" provide new molecular-scale insights for the understanding of the interfacial action of bile salts on lipid assemblies.