Protein-stabilized emulsions and nanoemulsions

This presentation will summarize briefly our current understanding of the scientific and technological basis for the formation of emulsions containing micrometer and nanometer sized droplets, with applications in food, cosmetic and pharma industries [1-3]. The focus will be on protein-stabilized emulsions. Their specific features will be summarized in comparison to the surfactant- and particle-stabilized emulsions [3]. The subtle relations between the material characteristics of the emulsions (oil type, emulsifier, pH, etc.) and the optimal hydrodynamic conditions for emulsification will be discussed [3] in the context of obtaining emulsions with desired properties. Examples of appropriate mixtures of proteins with other emulsifiers (lipids, lysolipids, natural gums) will be given. The differences between oil-in-water and water-in-oil emulsions will be briefly discussed [4,5]. Special focus will be given on the recent advance in production of nanoemulsions using high pressure homogenizers, high viscosity of the continuous phase, and/or high oil volume fraction during emulsification [6,7]. Some new methods for self-emulsification will be briefly reviewed [8,9]. The basic physicochemical and hydrodynamic concepts will be illustrated by multiple examples with real systems. References: David Julian McClements, Food Emulsions: Principles, Practices, and Techniques, 3rd Edition, CRC Press, 2015. Andreas Håkansson, Emulsion Formation by Homogenization: Current Understanding and Future Perspectives, Annu. Rev. Food Sci. Technol. 10 (2019) 239–258 (review article). S. Tcholakova, N. D. Denkov, and A. Lips, Comparison of Solid Particles, Globular Proteins and Surfactants as Emulsifiers. Phys. Chem. Chem. Phys. 10 (2008) 1608-1627 (review article). N. Politova, S. Tcholakova, N. D. Denkov, Factors Affecting the Stability of Water-oil-water Emulsion Films. Colloids Surfaces A 522 (2017) 608–620. N. Politova, S. Tcholakova, S. Tsibranska, N. D. Denkov, K. Muelheims, Coalescence Stability of Water-in-Oil drops: Effects of Drop Size and Surfactant Concentration. Colloids Surfaces A 531 (2017) 32–39. S. Tcholakova, I. Lesov, K. Golemanov, N. Denkov, S. Judat, R. Engel, T. Daner, Efficient Emulsification of Viscous Oils at High Drop Volume Fraction. Langmuir 27 (2011) 14783-14796. D. Gazolu-Rusanova, I. Lesov, S. Tcholakova, N. Denkov, B. Ahtchi, Food grade nanoemulsion preparation by rotor-stator homogenization. Food Hydrocolloids (2019) under review. S. Tcholakova, Z. Valkova, D. Cholakova, Z. Vinarov, I. Lesov, N. D. Denkov, K. Smoukov, Efficient Self-Emulsification via Cooling-Heating Cycles. Nature Comm. 8 (2017) 15012. Zh. Valkova, D. Cholakova, S. Tcholakova, N. Denkov, S. K. Smoukov, Mechanisms and Control of Self-Emulsification upon Freezing and Melting of Dispersed Alkane Drops. Langmuir 33 (2017) 12155−12170

Self-shaping of oil droplets via the formation of intermediate rotator phases upon cooling.

Revealing the chemical and physical mechanisms underlying symmetry breaking and shape transformations is key to understanding morphogenesis. If we are to synthesize artificial structures with similar control and complexity to biological systems, we need energy- and material-efficient bottom-up processes to create building blocks of various shapes that can further assemble into hierarchical structures. Lithographic top-down processing allows a high level of structural control in microparticle production but at the expense of limited productivity. Conversely, bottom-up particle syntheses have higher material and energy efficiency, but are more limited in the shapes achievable. Linear hydrocarbons are known to pass through a series of metastable plastic rotator phases before freezing. Here we show that by using appropriate cooling protocols, we can harness these phase transitions to control the deformation of liquid hydrocarbon droplets and then freeze them into solid particles, permanently preserving their shape. Upon cooling, the droplets spontaneously break their shape symmetry several times, morphing through a series of complex regular shapes owing to the internal phase-transition processes. In this way we produce particles including micrometre-sized octahedra, various polygonal platelets, O-shapes, and fibres of submicrometre diameter, which can be selectively frozen into the corresponding solid particles. This mechanism offers insights into achieving complex morphogenesis from a system with a minimal number of molecular components.European Research Council (Grant ID: EMATTER 280078), European networks COST MP 1106 and 1305 and the capacity building project BeyondEverest of the European Commission (Grant ID: 286205)This is the author accepted manuscript. The final version is available from Nature Publishing Group via http://dx.doi.org/10.1038/nature1618

Mechanisms of Drug Solubilization by Polar Lipids in Biorelevant Media

Despite the widespread use of lipid excipients in both academic research and oral formulation development, rational selection guidelines are still missing. In the current study, we aimed to establish a link between the molecular structure of commonly used polar lipids and drug solubilization in biorelevant media. We studied the effect of 26 polar lipids of the fatty acid, phospholipid or monoglyceride type on the solubilization of fenofibrate in a two-stage in vitro GI tract model. The main trends were checked also with progesterone and danazol.Based on their fenofibrate solubilization efficiency, the polar lipids can be grouped in 3 main classes. Class 1 substances (n = 5) provide biggest enhancement of drug solubilization (>10-fold) and are composed only by unsaturated compounds. Class 2 materials (n = 10) have an intermediate effect (3-10 fold increase) and are composed primarily (80 %) of saturated compounds. Class 3 materials (n = 11) have very low or no effect on drug solubilization and are entirely composed of saturated compounds.The observed behaviour of the polar lipids was rationalized by using two classical physicochemical parameters: the acyl chain phase transition temperature (Tm) and the critical micellar concentration (CMC). Hence, the superior performance of class 1 polar lipids was explained by the double bonds in their acyl chains, which: (1) significantly decrease Tm, allowing these C18 lipids to form colloidal aggregates and (2) prevent tight packing of the molecules in the aggregates, resulting in bigger volume available for drug solubilization. Long-chain (C18) saturated polar lipids had no significant effect on drug solubilization because their Tm was much higher than the temperature of the experiment (T = 37 C) and, therefore, their association in colloidal aggregates was limited. On the other end of the spectrum, the short chain octanoic acid manifested a high CMC (50 mM), which had to be exceeded in order to enhance drug solubilization. When these two parameters were satisfied (C > CMC, Tm Texp), the increase of the polar lipid chain length increased the drug solubilization capacity (similarly to classical surfactants), due to the decreased CMC and bigger volume available for solubilization.The hydrophilic head group also has a dramatic impact on the drug solubilization enhancement, with polar lipids performance decreasing in the order: choline phospholipids > monoglycerides > fatty acids.As both the acyl chain length and the head group type are structural features of the polar lipids, and not of the solubilized drugs, the impact of Tm and CMC on solubilization by polar lipids should hold true for a wide variety of hydrophobic molecules. The obtained mechanistic insights can guide rational drug formulation development and thus support modern drug discovery pipelines.<br /

Structure and Undulations of Escin Adsorption Layer at Water Surface Studied by Molecular Dynamics

The saponin escin, extracted from horse chestnut seeds, forms adsorption layers with high viscoelasticity and low gas permeability. Upon deformation, escin adsorption layers often feature surface wrinkles with characteristic wavelength. In previous studies, we investigated the origin of this behavior and found that the substantial surface elasticity of escin layers may be related to a specific combination of short-, medium-, and long-range attractive forces, leading to tight molecular packing in the layers. In the current study, we performed atomistic molecular dynamics simulations of 441 escin molecules in a dense adsorption layer with an area per molecule of 0.49 nm2. We found that the surfactant molecules are less submerged in water and adopt a more upright position when compared to the characteristics determined in our previous simulations with much smaller molecular models. The number of neighbouring molecules and their local orientation, however, remain similar in the different-size models. To maintain their preferred mutual orientation, the escin molecules segregate into well-ordered domains and spontaneously form wrinkled layers. The same specific interactions (H-bonds, dipole–dipole attraction, and intermediate strong attraction) define the complex internal structure and the undulations of the layers. The analysis of the layer properties reveals a characteristic wrinkle wavelength related to the surface lateral dimensions, in qualitative agreement with the phenomenological description of thin elastic sheets