M. Cócera

Conformational Changes in Stratum Corneum Lipids by Effect of Bicellar Systems

Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy was applied to study the effects of the bicelles formed by dimyristoyl-glycero-phosphocholine (DMPC) and dihexanoyl-glycero-phosphocholine (DHPC) in porcine stratum corneum (SC) in vitro. A comparison of skin samples treated and untreated with bicelles at different temperatures was carried out. The analysis of variations after treatment in the position of the symmetric CH2 stretching, CH2 scissoring, and CH2 rocking vibrations reported important information about the effect of bicelles on the skin. Bicellar systems caused a phase transition from the gel or solid state to the liquid crystalline state in the lipid conformation of SC, reflecting the major order-disorder transition from hexagonally packed to disordered chains. Grazing incidence small and wide X-ray scattering (GISAXS and GIWAXS) techniques confirmed this effect of bicelles on the SC. These results are probably related to with the permeabilizing effect previously described for the DMPC/DHPC bicelles.

Permeability investigations of phospholipid liposomes by adding cholesterol

Abstract The sublytic changes caused by sodium dodecyl sulfate (SDS) in egg phosphatidylcholine (PC) liposomes with rising concentrations of cholesterol (CH) (PC:CH mole ratios from 9:1 to 7:3) were investigated. The effective molar ratios Re of membrane-bound surfactant to lipid and the bilayer/aqueous phase partition coefficients (K) were determined. A linear increase and decrease in Re and K, respectively, take place as the CH proportion increase. Given that the surfactant capacity to interact with liposomes is inversely related to the Re value, the rise of CH reduces linearly both the SDS activity against vesicles (increase in Re) and its affinity with these structures (decrease in K). These linear variations may be due to the linear increase of hydrophobicity in the interfacial region of bilayers and to their reduced translational fluidity by changes in the bilayer packing and ordering. The increase in Re always results in two opposite effects on K. At low Re, K firstly increases because only the outer vesicle leaflet is available for interaction with SDS. The following abrupt fall in K is due to the fact that the binding of additional SDS molecules to bilayer is hampered. The K peaks obtained are correlated in all cases with the saturation of the outer vesicle leaflet by SDS. Increasing Re values, lead to an increased rate of flip–flop of the surfactant molecules (or permeabilization of the bilayers to SDS), thus also making the inner monolayer available for interaction with added SDS. The free SDS concentrations are always lower than its critical micelle concentration indicating that the SDS–liposome interaction is mainly ruled by the action of surfactant monomers regardless of the CH concentration in bilayers.

Bicelles: Lipid Nanostructured Platforms with Potential Dermal Applications

Bicelles emerge as promising membrane models, and because of their attractive combination of lipid composition, small size and morphological versatility, they become new targets in skin research. Bicelles are able to modify skin biophysical parameters and modulate the skins barrier function, acting to enhance drug penetration. Because of their nanostructured assemblies, bicelles have the ability to penetrate through the narrow intercellular spaces of the stratum corneum of the skin to reinforce its lipid lamellae. The bicelle structure also allows for the incorporation of different molecules that can be carried through the skin layers. All of these characteristics can be modulated by varying the lipid composition and experimental conditions. The remarkable versatility of bicelles is their most important characteristic, which makes their use possible in various fields. This system represents a platform for dermal applications. In this review, an overview of the main properties of bicelles and their effects on the skin are presented.

Application of Bicellar Systems on Skin: Diffusion and Molecular Organization Effects

The effect of bicelles formed by dipalmitoylphosphatidylcholine (DPPC)/dihexanoylphosphatidylcholine (DHPC) on stratum corneum (SC) lipids was studied by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy at different temperatures. Analysis of the lipid organization in terms of chain conformational order and lateral packing shows that the use of bicelles hampers the fluidification of SC lipids with temperature and leads to a lateral packing corresponding to a stable hexagonal phase. Grazing incidence small- and wide-angle X-ray scattering (GISAXS and GIWAXS) techniques confirm these results and give evidence of higher lamellar order after treatment with these bicelles. Additionally, the effects of DPPC/DHPC and dimyristoylphosphatidylcholine (DMPC)/DHPC bicelles at different SC depths were compared. The combination of ATR-FTIR spectroscopy and the tape-stripping method was very useful for this purpose.

Lipid Nanostructures: Self-Assembly and Effect on Skin Properties

This work evaluates the relation between the composition and the self-assembly of some lipid aggregates with their effects on the skin. To this end, liposomes, bicelles and micelles formed by dipalmitoylphosphatidylcholine (DPPC), dimyristoylphosphatidylcholine (DMPC) and dihexanoylphosphatidylcholine (DHPC) were characterized by electron microscopy and dynamic light scattering techniques, and applied on the skin. The results revealed that nanostructures with similar assembly but different composition caused different effects on the skin parameters. In general, samples containing DMPC affected the barrier function to a greater extent than systems containing DPPC. Additionally, our results showed that samples with the same lipid composition but different assembly exerted different effects on the skin. Liposomes decreased or did not modify the transepidermal water loss (TEWL), while bicelles and micelles increased this parameter. Hydration of the skin diminished especially after the application of micellar and bicellar samples. In vitro experiments showed structures like vesicles inside cutaneous SC (stratum corneum) incubated with DPPC/DHPC bicelles. These structures were not detected in SC samples incubated with DMPC/DHPC bicelles probably due to the different thermotropic behavior of DMPC and DPPC at physiological temperatures. Results reported in this work should be considered in terms of design of more efficient and specific skin delivery systems.

Influence of the alkyl chain length of alkyl glucosides on their ability to solubilize phosphatidylcholine liposomes

Abstract The solubilizing alterations caused by a series of alkyl glucosides (alkyl chain length ranging from C8 to C12) in neutral and electrically charged phosphatidylcholine (PC) liposomes were investigated. The surfactant to phospholipid molar ratios (Re) and the bilayer/aqueous phase partition coefficients (K) were determined by monitoring the changes in the static light scattering (SLS) of the system during solubilization. Liposomes were formed by PC, to which phosphatidic acid (PA) or stearylamine (SA) was added when required to increase the negative or positive surface charge. The fact that at the two interaction levels investigated (100 and 0% of SLS of the surfactant/PC systems), the free surfactant concentration for each surfactant was always comparable to its critical micelle concentration (CMC) indicates that the liposome solubilization was mainly ruled by the formation of mixed micelles. The rise in the surfactant CMC (decrease in its alkyl chain length) led to an increase in the surfactant ability to saturate or solubilize liposomes and inversely in an abrupt decrease in its bilayer affinity, regardless of the electrical charge of liposomes. The overall balance of these opposite tendencies shows that the octyl glucoside showed the highest ability to saturate and solubilize liposomes (lowest Re values), whereas the dodecyl glucoside exhibited the highest degree of partitioning into liposomes or affinity with these bilayer structures (highest K values). The use of C9-Glu reduced approximately 2.5 times the concentration needed to saturate and solubilize 1.0 mM PC liposomes with respect to that needed for C8-Glu, regardless of the type of electrical charge present in bilayers.

Different stratum corneum lipid liposomes as models to evaluate the effect of the sodium dodecyl sulfate

The stability of stratum corneum (SC) liposomes against the action of surfactants has been revised. To this end, two types of vesicles were used; vesicles formed with the lipid and protein material extracted from SC, and lipid mixtures approximating the SC composition. In this case, the proportion of ceramides (Cer) and cholesteryl sulfate (Chol-sulf) was varied and the relative proportion of the other lipids remained constant. The increasing presence of these two lipids increased the resistance of liposomes against the action of the anionic surfactant sodium dodecyl sulfate (SDS). The rise in the cell-to-cell cohesion that occurred in recessive X-linked ichthyosis due to the accumulation of Chol-sulf could be associated in part to the enhanced stability of (Chol-sulf)-enriched bilayers. It is noteworthy that the surfactant partitioning between bilayers and the aqueous phase increased and decreased, respectively, as the proportion of Cer and Chol-sulf increased. This effect may be attributed to the variations in both the electrostatic interactions lipid-surfactant (electrostatic repulsion between the sulfate groups of both Chol-sulf and SDS), and the hydrophilic lipophilic balance of the lipid mixtures, in which Cer is replaced by the major polar lipid of the mixture (Chol-sulf). The fact that the free surfactant concentration was always smaller than its critical micelle concentration indicates that the permeability alterations were mainly ruled by the action of surfactant monomers, in agreement with the results reported for sublytic interactions of this surfactant with PC liposomes.

Structural modifications in the stratum corneum by effect of different solubilizing agents: a study based on high-resolution low-temperature scanning electron microscopy.

The action of different solubilizing agents (chloroform-methanol mixtures and the nonionic surfactant octyl glucoside, OG) on the structural organization of the stratum corneum (SC) was investigated by means of the double-layer high-resolution low-temperature scanning electron microscopy technique. Chloroform-methanol mixtures were able to remove mainly the lipid without a significant loss of cohesion in the SC tissue. However, OG treatment caused a partial disaggregation of the corneocytes and their envelopes and, at a macroscopic level, a loss of cohesion in the whole SC tissue. As for lipid domains, after OG treatment the formation of rough structures was detected, probably associated with the disorder in the lipid lamellae. The formation of these new structures could be attributable to the interaction of the lipids with the proteins liberated from the corneocytes. Hence, a direct correlation may be established between the preservation of the structure of the corneocytes and the corneocyte envelope and the cohesion of the whole SC tissue.

Bicellar systems as modifiers of skin lipid structure.

The characterization of different bicellar aggregates and the effects of these systems on the stratum corneum (SC) microstructure have been studied. Dynamic light scattering (DLS) and freeze fracture electron microscopy (FFEM) techniques showed that both of the systems studied, dimyristoyl-phosphatidylcholine/dihexanoyl-phosphocholine (DMPC/DHPC) and dipalmitoyl-phosphocholine (DPPC)/DHPC, were formed by small discoidal aggregates at room temperature (20°C). Treating skin with DMPC/DHPC bicelles does not affect the SC lipid microstructure, whereas bicellar systems formed by DPPC and DHPC can promote the formation of new structures in the SC lipid domains. This indicates the passage of lipids from bicelles through the SC layers and also a possible interaction of these lipids with the SC lipids. Given the absence of surfactant in the bicellar composition and the small size of these structures, the use of these smart nano-systems offers great advantages over other lipid systems for dermatological purposes. Bicelles could be promising applications as drug carriers through the skin. This contribution, based on the new biological use of bicelles, may be useful to scientists engaged in colloid science and offers a new tool for different applications in skin and cosmetic research.

Alterations in stratum corneum lipid liposomes due to the action of triton X-100. Influence of the level of ceramides on this process.

The role played by the ceramides (Cer) in the interaction of Triton X-100 (T(X-100)) with liposomes modeling the stratum corneum (SC) lipid composition was studied. The surfactant/lipid molar ratios (Re) and the bilayer/aqueous phase partition coefficients (K) were determined at sublytic level by monitoring the changes in the fluorescence intensity of liposomes due to the 5(6)-carboxyfluorescein (CF) released from the interior of vesicles. Higher and lower Cer proportions than those existing in the SC (40%) led respectively to a fall and to a rise in the surfactant ability to alter these liposomes. However, the surfactant partitioning between bilayers and water (bilayer affinity with vesicles) exhibited a maximum for 40% Cer. Thus, at low Cer proportions the ability of T(X-100) molecules to alter these bilayers was maximum despite their reduced partitioning into bilayers, in line with the reported interaction of the anionic surfactant model sodium dodecyl sulfate with these vesicles. These findings underline the fragility of these bilayers as an effective barrier and could explain in part the reported dependencies of low level of Cer in skin lipids and function barrier abnormalities. The fact that the free surfactant concentration needed to achieve the two interaction levels investigated was lower than the surfactant CMC indicates that permeability alterations were mainly ruled by the action of surfactant monomers, regardless of Cer proportion in bilayers.