Lyotropic Liquid Crystalline Phases for the Formulation of Future Functional Foods

The especial features and advanced characteristics of Lyotropic Liquid Crystalline (LLC) phases as potent nano materials for encapsulation and the development of novel delivery systems for nutraceuticals and other bioactive compounds are reviewed. Exemplary, a focus is set on the health benefits of flavonoids and their current restrictions in bioavailability. Accordingly, our visions for application of LLC phases in the engineering of enhanced flavonoid-based food supplements and correlated challenges to over come are highlighted

Hybrid Vesicle Stability under Sterilisation and Preservation Processes Used in the Manufacture of Medicinal Formulations

Sterilisation and preservation of vesicle formulations are important considerations for their viable manufacture for industry applications, particular those intended for medicinal use. Here, we undertake an initial investigation of the stability of hybrid lipid-block copolymer vesicles to common sterilisation and preservation processes, with particular interest in how the block copolymer component might tune vesicle stability. We investigate two sizes of polybutadiene-block-poly(ethylene oxide) polymers (PBd12-PEO11 and PBd22-PEO14) mixed with the phospholipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) considering the encapsulation stability of a fluorescent cargo and the colloidal stability of vesicle size distributions. We find that autoclaving and lyophilisation cause complete loss of encapsulation stability under the conditions studied here. Filtering through 200 nm pores appears to be viable for sterilisation for all vesicle compositions with comparatively low release of encapsulated cargo, even for vesicle size distributions which extend beyond the 200 nm filter pore size. Freeze-thaw of vesicles also shows promise for the preservation of hybrid vesicles with high block copolymer content. We discuss the process stability of hybrid vesicles in terms of the complex mechanical interplay between bending resistance, stretching elasticity and lysis strain of these membranes and propose strategies for future work to further enhance the process stability of these vesicle formulations

Experimental Modeling of Flavonoid-Biomembrane Interactions

Nonspecific interactions of flavonoids with lipids can alter the membrane's features (e.g., thickness and fluctuations) as well as influence their therapeutic potentials. However, relatively little is known about the details of how flavonoids interact with lipid components. Structure-dependent interactions of a variety of flavonoids with phospholipid monolayers on a mercury (Hg) film electrode were established by rapid cyclic voltammetry (RCV). The data revealed that flavonoids adopting a planar configuration altered the membrane properties more significantly than nonplanar flavonoids. Quercetin, rutin, and tiliroside were selected for follow-up experiments with Langmuir monolayers, Brewster angle microscopy (BAM), and small-angle X-ray scattering (SAXS). Relaxation phenomena in DOPC monolayers and visualization of the surface with BAM revealed a pronounced monolayer stabilization effect with both quercetin and tiliroside, whereas rutin disrupted the monolayer structure rendering the surface entirely smooth. SAXS showed a monotonous membrane thinning for all compounds studied associated with an increase in the mean fluctuations of the membrane. Rutin, quercetin, and tiliroside decreased the bilayer thickness of DOPC by ∼0.45, 0.8, and 1.1 Å at 6 mol %, respectively. In addition to the novelty of using lipid monolayers to systematically characterize the structure-activity relationship (SAR) of a variety of flavonoids, this is the first report investigating the effect of tiliroside with biomimetic membrane models. All the flavonoids studied are believed to be localized in the lipid/water interface region. Both this localization and the membrane perturbations have implications for their therapeutic activity

Discontinuous unbinding of lipid multibilayers

We have observed a discontinuous unbinding transition of lipid bilayer stacks composed of phosphatidylethanolamine and phosphatidylglycerol using X-ray diffraction. The unbinding is reversible and coincides with the main (Lβ→Lα) transition of the lipid mixture. Interbilayer interaction potentials deduced from the diffraction data reveal that the bilayers in the Lβ phase are only weakly bound. The unbinding transition appears to be driven by an abrupt increase in steric repulsion resulting from increased thermal undulations of the bilayers upon entering the fluid Lαphase

The Unique Crystallization Behavior of Buffalo Milk Fat

A full comprehension of milk fat crystallization is important for the structural development of dairy products such as butter, ice cream, and cheese. The influence of triacylglycerol (TAG) composition on the dynamic of milk fat crystallization and the nanostructure of the formed crystals was investigated using two chemically different types of milk fat, namely buffalo and cow milk fats (BMF; CMF). The TAG composition was determined using liquid chromatography and mass spectrophotometry (LCMS), whereas differential scanning calorimetry (DSC), small- and wide-angle X-ray scattering (SAXS and WAXS), and polarized light microscopy (PLM) were used to characterize the crystallization behavior of the two milk fats. A total of 37 TAG species were identified in both BMF and CMF, but in different proportions. In particular, BMF was found to have a higher amount of low-molecular-weight TAGs in comparison to CMF. This difference in chemical composition explains the different kinetics of polymorphic transformation in the two samples. Specifically, it clarifies the delay in the nucleation of the β′-polymorph in BMF in comparison to CMF. BMF also showed a higher nucleation rate due to its higher proportion of saturated TAGs and higher melting range. Finally, this work presents a novel interpretation of the mechanism of formation of the β-polymorph (53 Å), which has recently become the subject of a vivid debate in milk fat crystallization studies

From angular to round: in depth interfacial analysis of binary phosphatidylethanolamine mixtures in the inverse hexagonal phase

Packing stress in the lipidic inverse hexagonal HII phase arises from the necessity of the ideally cylinder-shaped micelles to fill out the hexagonally-shaped Wigner–Seitz unit cell. Thus, hydrocarbon chains stretch towards the corners and compress in the direction of the flat side of the hexagonal unit cell. Additionally, the lipid/water interface deviates from being perfectly circular. To study this packing frustration in greater detail, we have doped 1-palmitoyl-2-oleoyl-sn-phosphatidylethanolamine (POPE) with increasing molar concentrations of 1,2-palmitoyl-sn-phosphatidylethanolamine (DPPE: 0 to 15 mol%). Due to its effectively longer hydrophobic tails, DPPE tends to aggregate in the corner regions of the unit cell, and thus, increases the circularity of the lipid/water interface. From small angle X-ray diffraction (SAXD) we determined electron density maps. Using those, we analysed the size, shape and homogeneity of the lipid/water interface as well as that of the methyl trough region. At 6 and 9 mol% DPPE the nanotubular water core most closely resembles a circle; further to this, in comparison to its neighbouring concentrations, the 9 mol% DPPE sample has the smallest water core area and smallest number of lipids per circumference, best alleviating the packing stress. Finally, a three-water layer model was applied, discerning headgroup, perturbed and free water, demonstrating that the hexagonal phase is most stable in the direction of the flat faces (compression zones) and least stable towards the vertices of the unit cell (decompression zones)

Effect of β-sitosterol on the curcumin-loaded liposomes: Vesicle characteristics, physicochemical stability, in vitro release and bioavailability

In this work, the effect of β-sitosterol (Sito) on vesicle characteristics, physicochemical stability as well as the in vitro release and bioavailability of curcumin-loaded liposomes (Cur-LP) was studied. When 20–33 mol% of Sito was incorporated, encapsulation efficiency of curcumin was improved due to the high amount of liquid-ordered domains in membranes. At 50 mol% Sito a lower encapsulation efficiency was observed possibly due to membrane defects. The physical, thermal and photo stability of curcumin in liposomes were markedly improved with increasing the amount of Sito. First-order kinetics fitted best the curcumin release dynamics of Sito containing liposomes, clearly showing that sustained release improved with increasing amounts of Sito in liposomes. Simulated digestion studies suggested that Sito concentration of about 20–33 mol% improved the bioavailability of curcumin in liposomes. These study shows that Sito is an applicable and potential route in forming healthier cholesterol-free curcumin-loaded liposomes for functional supplements

Losartan’s affinity to fluid bilayers modulates lipid–cholesterol interactions

Losartan is an angiotensin II receptor antagonist mainly used for the regulation of high blood pressure. Since it was anticipated that losartan reaches the receptor site via membrane diffusion, the impact of losartan on model membranes has been investigated by small angle X-ray scattering. For this purpose 2–20 mol% losartan was incorporated into dimyristoylphosphatidylcholine (DMPC) and palmitoyl-oleoyl-phosphatidylcholine (POPC) bilayers and into their binary mixtures with cholesterol in the concentration range of 0 to 40 mol%. Effects of losartan on single component bilayers are alike. Partitioning of losartan into the membranes confers a negative charge to the lipid bilayers that causes the formation of unilamellar vesicles and a reduction of the bilayer thickness by 3–4%. Analysis of the structural data resulted in an estimate for the partial area of losartan, ALos E 40 A˚ 2 . In the presence of cholesterol, differences between the effects of losartan on POPC and DMPC are striking. Membrane condensation by cholesterol is retarded by losartan in POPC. This contrasts with DMPC, where an increase of the cholesterol content shifts the partitioning equilibrium of losartan towards the aqueous phase, such that losartan gets depleted from the bilayers from 20 mol% cholesterol onwards. This indicates (i) a chain-saturation dependent competition of losartan with lipid–cholesterol interactions, and (ii) the insolubility of losartan in the liquid ordered phase of PCs. Consequently, losartan’s action is more likely to take place in fluid plasma membrane patches rather than in domains rich in cholesterol and saturated lipid species such as in membrane raft

Stability and release performance of curcumin-loaded liposomes with varying content of hydrogenated phospholipids

The aim of this study was to substitute part of soybean phospholipid (SPC) with hydrogenated soybean phospholipid (HSPC) in curcumin-loaded liposomes (Cur-LP), in order to further enhance stability and release performances of curcumin. When the SPC/HSPC mass ratio changed from 10:0 to 5:5, vesicle size, encapsulation efficiency and alkali resistance of curcumin increased, although a small decrease in centrifugal stability was observed. Salt stability became worse as more HSPC was used (3:7 and 0:10). Owing storage at 4 °C and 25 °C, Cur-LP at a SPC/HSPC mass ratio of 5:5 performed well considering vesicle size, lipid oxidation and curcumin retention. These vesicles displayed also the best sustained-release performance in simulated digestion, attributed to the tighter lipid packing in membranes as indicated by fluorescence probes, DSC and FTIR. This study can guide the development of a Cur-LP product with improved shelf-life stability by using HSPC

Membrane mixing and dynamics in hybrid POPC/Poly(1,2-butadiene-block-ethylene oxide) (PBd-b-PEO) lipid/block co-polymer giant vesicles

Lipids and block copolymers can individually self-assemble into vesicles, each with their own particular benefits and limitations. Combining polymers with lipids allows for further optimisation of the vesicle membranes for bionanotechnology applications. Here, POPC lipid is mixed with poly(1,2-butadiene-block-ethylene oxide) of two different molecular weights (PBd22–PEO14, Mr = 1800 g mol−1 and PBd12–PEO11, Mr = 1150 g mol−1) in order to investigate how increasing the polymer fraction affects membrane mixing, hydration and fluidity. Intensity contributions of fluorescently labelled lipid and polymer within mixed GUV membranes confirm membrane homogeneity within the hybrids. General polarisation measurements of Laurdan in GUVs showed little change in membrane hydration as polymer fraction is increased, which suggests good structural compatibility between lipids and polymers that gives rise to well-mixed vesicles. Membrane fluidity in hybrid GUVs was found to decrease non-linearly with increasing polymer fraction. However, the diffusion coefficients for the fluorescent polymer in hybrid membranes did not change significantly with increasing polymer content. While increasing the polymer fraction does reduce the movement of lipids through a polymer-rich matrix, insignificant difference in diffusion coefficients of the polymer suggests that its diffusion is minimally affected by increasing lipid composition in the range studied. These results lay further foundations for the wider development of hybrid vesicles with controlled properties for advanced biotechnologies