Lipid distribution on ethnic hairs by Fourier transform infrared synchrotron spectroscopy.

BACKGROUND: A synchrotron-based Fourier transform infrared micro-spectrometer (µ-FTIR) allows the spatial determination of lipids across the different layers of ethnic hairs and differentiates between the lipid order arrangement and quantity. MATERIALS AND METHODS: The three ethnic fibers were delipidized, the lipid extracts were characterized, and the delipidized fibers were studied by dynamic vapor sorption experiments (DVS) and FTIR-synchrotron techniques. RESULTS: The average spectra from the different hair regions exhibited the most intense CH2 sym peaks on the medulla, followed by those from the cuticle and cortex for all hairs of different ethnicities. Differences in the lipid fraction of the three hair types have been observed, and they can explain some barrier properties. African virgin hair was demonstrated to have more lipids mainly in the medulla, which implies an important hydrophobicity with low hysteresis between absorption and desorption water vapor processes. In addition, these lipids are highly disordered, mainly in the cuticle, which can be related to its high water vapor diffusion. Asian and Caucasian virgin hairs presented a similar lipid order in all regions, with similar diffusion coefficients. Results indicate that the higher order of the lipid bilayer hinders water permeation kinetics in some way. CONCLUSION: The differences in the presence and organization of the lipids in the different regions of the African hair can account for its differentiation with regards to moisturization and swelling from the other types of fibers.

Lanolin-Based Synthetic Membranes for Transdermal Permeation and Penetration Drug Delivery Assays.

Due to the high similarity in composition and structure between lanolin and human SC lipids, we will work with two models from wool wax. Two types of lanolin were evaluated: one extracted with water and surfactants (WEL) and the other extracted with organic solvents (SEL). Skin permeation and skin penetration studies were performed with two active compounds to study the feasibility of the use of lanolin-based synthetic membranes as models of mammalian skin. Diclofenac sodium and lidocaine were selected as the active compounds considering that they have different chemical natures and different lipophilicities. In the permeation assay with SEL, a better correlation was obtained with the less permeable compound diclofenac sodium. This assay suggests the feasibility of using artificial membranes with SEL as a model for percutaneous absorption studies, even though the lipophilic barrier should be improved. Penetration profiles of the APIs through the SEL and WEL membranes indicated that the two membranes diminish penetration and can be considered good membrane surrogates for skin permeability studies. However, the WEL membranes, with a pH value similar to that of the skin surface, promoted a higher degree of diminution of the permeability of the two drugs, similar to those found for the skin.

Assessment of Finite and Infinite Dose In Vitro Experiments in Transdermal Drug Delivery.

Penetration, usually with finite dosing, provides data about the total active amount in the skin and permeation, being the most used methodology, usually with infinite dosing, leads to data about pharmacokinetic parameters. The main objective of this work is to assess if results from permeation, most of them at finite dose, may be equivalent to those from penetration usually at infinite dose. The transdermal behavior of four drugs with different physicochemical properties (diclofenac sodium, ibuprofen, lidocaine, and caffeine) was studied using penetration/finite and kinetic permeation/infinite dose systems using vertical Franz diffusion cells to determine the relationships between permeation and penetration profiles. Good correlation of these two in vitro assays is difficult to find; the influence of their dosage and the proportion of different ionized/unionized compounds due to the pH of the skin layers was demonstrated. Finite and infinite dose regimens have different applications in transdermal delivery. Each approach presents its own advantages and challenges. Pharmaceutical industries are not always clear about the method and the dose to use to determine transdermal drug delivery. Being aware that this study presents results for four actives with different physicochemical properties, it can be concluded that the permeation/infinite results could not be always extrapolated to those of penetration/finite. Differences in hydrophilicity and ionization of drugs can significantly influence the lack of equivalence between the two methodologies. Further investigations in this field are still needed to study the correlation of the two methodologies and the main properties of the drugs that should be taken into account.

Ethnic hair: Thermoanalytical and spectroscopic differences.

BACKGROUND: The aim of this study is to characterize and detect the possible differences among the hair of three different ethnicities: African, Asiatic and Caucasian. MATERIALS AND METHODS: The differences in water adsorption/desorption behaviour of hairs were studied using a thermogravimetric balance and compared with the analysis of the lipid distribution and order using synchrotron-based Fourier transform infrared microspectroscopy. Besides, the thermal thermogravimetry (TG) and differential scanning calorimetry (DSC) analyses on human hair were executed. RESULTS: Differences in the diffusion coefficients were evidenced. African hair exhibited increased permeability. Caucasian hair displayed a higher water absorption capability with increasing humidity but with a slow diffusion rate. The Asian fibre appeared to be more resistant to hydration changes. The spectroscopic analysis showed notable differences in the cuticle lipids. The African cuticle exhibited more lipids with a lower order bilayer. The outmost layer of Caucasian fibres contained more ordered lipids, and the Asian fibres show a very low level of lipids on the cuticle region. The DSC results indicate no difference in the thermal stability and TG showed higher water content in the Caucasian fibre and a possible lower cysteine disulphide bond content in the African hair matrix. CONCLUSION: The triple approach demonstrated the permeability differences among the ethnic fibres and their correlation with the properties of their cuticle lipids. These differences could have particular relevance to the hair care cosmetic market.

Effect of propylene glycol on the skin penetration of drugs.

Propylene glycol (PG) has been used in formulations as a co-solvent and/or to enhance drug permeation through the skin from topical preparations. Two skin in vitro permeation approaches are used to determine the effect of PG on drug penetration. The in vitro Skin-PAMPA was performed using 24 actives applied in aqueous buffer or PG. PG modulates permeability by increasing or diminishing it in the compounds with poor or high permeability, respectively. Percutaneous absorption using pigskin on Franz diffusion cells was performed on seven actives and their commercial formulations. The commercial formulations evaluated tend to have a lower permeability than their corresponding PG solutions but maintain the compound distribution in the different strata: stratum corneum, epidermis and dermis. The results indicate the enhancer properties of PG for all compounds, especially for the hydrophilic ones. Additionally, the Synchrotron-Based Fourier Transform Infrared microspectroscopy technique is applied to study the penetration of PG and the molecular changes that the vehicle may promote in the different skin layers. Results showed an increase of the areas under the curve indicating the higher amount of lipids in the deeper layers and altering the lipidic order of the bilayer structure to a more disordered lipid structure.

Selective modification of skin barrier lipids.

The molecular organization of stratum corneum (SC) lipids is important for maintaining the barrier properties of the skin. The majority of intercellular lipids are in a solid state at normal humidity (RH) and ambient temperature; however, several studies indicate that a small fraction exist in a fluid state. In a previous work, a preferential solubilization of fluid skin lipids by acetone (Ac) was envisaged. A different change in the skin permeability related to the different lipid structures of the extracted lipids was suggested. To increase the knowledge of the specific role of different lipids on skin structure, a selective lipid modification is proposed. This study assess the effect of Ac on skin barrier lipids in-depth. Synchrotron based Fourier-transform infrared microspectroscopy (FTIR), which is used to study SC lipid organization, revealed a more ordered lipid organization after Ac treatment. In vitro experiments using Franz cells, which were selected to follow the SC barrier function capability, demonstrated that Ac-treated skin retained caffeine and ibuprofen on the SC with very low permeation of both compounds into the deeper skin layers. In vitro transepidermal water loss (TEWL) measurements revealed the ability of Ac to induce a less water permeated skin. Although an important lipid fraction has been removed, Ac skin treatment brings to a skin where the remaining lipids promote an improved barrier function. These results could lead to a better understanding of the role of different lipid components in skin structure.

Lipid loses and barrier function modifications of the brown-to-white hair transition.

BACKGROUND: The main objective of this study was to determine the lipid profile of brown and white Caucasian hair fibres and the effects of lipids on the properties of fibres. MATERIALS AND METHODS: To determine the structures of white and brown hair lipid bilayers, cross sections of fibres of both hair types were examined using synchrotron-based µ-FTIR mapping. Dynamic vapour sorption (DVS) analyses were also performed to determine the differences in the barrier function of both fibres. RESULTS: Spatial identification of lipids showed that a great amount of lipids was present in the medulla of fibres of both hair types, but important differences were also observed between cuticles of the different fibres. The cuticle of a white hair fibre showed a significant decrease in its lipid content, but did not show differences in the lateral packing order with respect to the cuticle of a brown hair fibre. The cortex and medulla of the white hair fibre also exhibited a significant decrease in its lipid content but with a higher lateral packing order than brown hair. Using DVS analysis, it was found that the water dynamics of white hair fibres differed from those of brown hair fibres, showing a decrease in their total capacity to absorb water and an increase in the velocity of the exchange of water with the environment. CONCLUSION: The results of both techniques demonstrated a high correlation between the characteristics of the lipids located in the cuticle and the water dynamics of the fibres.

Action of surfactants on the mammal epidermal skin barrier.

BACKGROUND: Daily skin washing routines can promote undesirable effects on skin barrier function. The stratum corneum (SC) lipid matrix is crucial for skin barrier function. Skin cleansing products are mostly composed of surfactants: surface-active molecules that interact with skin lipids in several ways. The main aim of this work was to investigate the effect produced by surfactants on skin barrier permeability. Porcine skin is a well-accepted and readily available model of the human skin barrier. The effect of two cleansing formulations (based on different surfactant mixtures) on the barrier properties of mammalian skin were evaluated. METHODS: Water sorption/desorption (DVS) experiments were used to measure skin permeability. Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy and confocal Raman were useful to study SC lipid organization. RESULTS: The results showed that while anionic surfactants (SLS) had a negative impact on the skin barrier, with a clear increase of alkyl chain disorder; cosurfactants present in the shampoo formulation diminished the detrimental effect of their primary ionic surfactant, inducing less modification on lipid intramolecular chain disorder. CONCLUSIONS: The obtained results confirmed that the mild cleansing formulations studied had gentle interaction with skin. The capacity to discriminate between detergent systems was clearly established with both DVS and spectroscopy techniques.

Soybean-fragmented proteoglycans against skin aging.

OBJECTIVE: The majority of age-dependent skin changes happen in the dermis layer inducing changes in skin collagen and in the proteoglycans. The main aim of this work is to study the efficacy of a Proteum serum, containing soybean-fragmented proteoglycans, against skin aging. MATERIALS AND METHODS: In vitro tests were performed to evaluate the Proteum serum ability on activating the production of collagen and proteoglycans. An in vivo long-term study was performed to determine the efficacy of the Proteum serum when applied on skin. Protection of healthy skin against detergent-induced dermatitis and the antioxidant properties of the applied Proteum serum were also studied. RESULTS AND DISCUSSION: The in vitro tests demonstrated that the Proteum serum was able to elevate the production of molecules which are essential for supporting the dermal extracellular matrix organization. These results were correlated by the in vivo measurements where a clear trend on improving the measured skin parameters due to the Proteum serum application was found. CONCLUSIONS: A beneficial effect of the Proteum serum was demonstrated with an improvement in the skin roughness and a reinforcement of the skin barrier function. Moreover, a significant protector effect on human stratum corneum against lipids peroxides (LPO) was demonstrated.

Skin barrier modification with organic solvents.

The primary barrier to body water loss and influx of exogenous substances resides in the stratum corneum (SC). The barrier function of the SC is provided by patterned lipid lamellae localized to the extracellular spaces between corneocytes. SC lipids are intimately involved in maintaining the barrier function. It is generally accepted that solvents induce cutaneous barrier disruption. The main aim of this work is the evaluation of the different capability of two solvent systems on inducing changes in the SC barrier function. SC lipid modifications will be evaluated by lipid analysis, water sorption/desorption experiments, confocal-Raman visualization and FSTEM images. The amount of SC lipids extracted by chloroform/methanol was significantly higher than those extracted by acetone. DSC results indicate that acetone extract has lower temperature phase transitions than chloroform/methanol extract. The evaluation of the kinetics of the moisture uptake and loss demonstrated that when SC is treated with chloroform/methanol the resultant sample reach equilibrium in shorter times indicating a deterioration of the SC tissue with higher permeability. Instead, acetone treatment led to a SC sample with a decreased permeability thus with an improved SC barrier function. Confocal-Raman and FSTEM images demonstrated the absence of the lipids on SC previously treated with chloroform/methanol. However, they were still present when the SC was treated with acetone. Results obtained with all the different techniques used were consistent. The results obtained increases the knowledge of the interaction lipid-solvent, being this useful for understanding the mechanism of reparation of damaged skin.

Skin delivery of antioxidant surfactants based on gallic acid and hydroxytyrosol.

OBJECTIVES: The aim of this study has been to investigate the dermal absorption profile of the antioxidant compounds gallic acid and hydroxytyrosol as well as their derivatives, hexanoate (hexyl gallate and hydroxytyrosol hexanoate) and octanoate (octyl gallate and octanoate derivative) alkyl esters (antioxidant surfactants). Previously, the scavenging capacity of these compounds, expressed as efficient dose ED50, has also determined. METHODS: The percutaneous absorption of these compounds was obtained by an in vitro methodology using porcine skin biopsies on Franz static diffusion cells. The antiradical activity of compounds was determined using the 1,1-diphenyl-2-picrylhydrazyl free radical method. KEY FINDINGS: The percutaneous penetration results show the presence of antioxidants in all layers of the skin. The content of the cutaneously absorbed compound is higher for the antioxidant surfactants (ester derivatives). This particular behaviour could be due to the higher hydrophobicity of these compounds and the presence of surface activity in the antioxidant surfactants. CONCLUSIONS: These new antioxidant surfactants display optimum properties, which may be useful in the preparation of emulsified systems in cosmetic and pharmaceutical formulations because of their suitable surface activity and because they can protect the skin from oxidative damage.

Antioxidative effects and percutaneous absorption of five polyphenols.

A new strategy was designed to evaluate the antioxidant effectiveness of five topically applied polyphenols following skin penetration profiles. The antioxidants were the following polyphenol derivatives: epicatechin, resveratrol, rutin, quercetin, and trolox, which was used as the reference antioxidant. The hydrophilic/lipophilic character of these compounds was evaluated, and their antioxidant activity was measured by the DPPH method. The percutaneous absorption of these polyphenols was obtained by an in vitro methodology using porcine skin biopsies. This methodology involves the quantification of the antioxidants present in each specific skin layer to evaluate antioxidant effectiveness. The antioxidant activity in each skin layer was also determined by the DPPH method. The results indicated that lipophilic antioxidants (epicatechin, resveratrol, quercetin, and trolox) penetrated deeper into the skin layers, whereas a more hydrophilic compound, rutin, remained on the skin surface. The antioxidant evaluation of each skin compartment suggested that resveratrol and rutin were the most effective topically applied compounds in view of their antioxidant activity and their skin penetration profile.

Characterization of new DOPC/DHPC platform for dermal applications.

Systems formed by mixtures of the phospholipids dioleoylphosphatidylcholine (DOPC) and dihexanoylphosphatidylcholine (DHPC) were characterized by use of differential scanning calorimetry, small angle X-ray scattering and two electron-microscopy techniques, freeze fracture electron microscopy and cryogenic transmission electron microscopy. These techniques allowed for the determination of the size, morphology, structural topology, self-assembly and thermotropic behavior of the nanostructures present in the mixtures. The interaction between the two phospholipids provides curvatures, irregularities and the increase of thickness and flexibility in the membrane. These effects led to the formation of different aggregates with a differential distribution of both phospholipids. The effect of these systems on the skin in vivo was evaluated by measurement of the biophysical skin parameters. Our results show that the DOPC/DHPC application induces a decrease in the permeability and in the hydration of the tissue. These effects in vivo are related to different microstructural changes promoted by these systems in the skin in vitro, published in a recent work. The fundamental biophysical analyses of DOPC/DHPC systems contribute to our understanding of the mechanisms that govern their interaction with the skin.

Efficacy of antioxidants in human hair.

Hair is exposed every day to a range of harmful effects such as sunlight, pollution, cosmetic treatments, grooming practices and cleansing. The UV components of sunlight damage human hair, causing fibre degradation. UV-B attacks the melanin pigments and the protein fractions (keratin) of hair and UV-A produces free radical/reactive oxygen species (ROS) through the interaction of endogenous photosensitizers. Hair was dyed and the efficacy of two antioxidant formulations was demonstrated after UV exposure by evaluating, surface morphology, protein and amino acid degradation, lipidic peroxidation, colour and shine changes and strength/relaxation properties. UV treatment resulted in an increase in protein and lipid degradation, changes in colour and shine and in adverse consequences for the mechanical properties. Natural antioxidants obtained from artichoke and rice applied to pretreated hair improved mechanical properties and preserved colour and shine of fibres, coating them and protecting them against UV. Furthermore, the lipidic peroxidation of the protein degradation caused by UV was reduced for some treated fibres, suggesting an improvement in fibre integrity. This was more marked in the case of the fibres treated using the artichoke extract, whereas the rice extract was better preserving shine and colour of hair fibres.

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 skin's 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.

Photodamage determination of human hair.

Sunlight on human hair causes photo-degradation. This results in bleaching due to melanin oxidation through free radicals, and induces keratin impairment. Protein degradation, tryptophan degradation, lipidic peroxidation and electron paramagnetic resonance can be used to evaluate proteic and lipidic photodecomposition and free radical formation in hair fibres subjected to antioxidant action and different UV intensities. All these methodologies have been optimised to determine protein, lipid and melanin degradation in hair subjected to different UV intensities.

An ex vivo methodology to assess the lipid peroxidation in stratum corneum.

Environmental risks, particularly UV radiation, provide a challenge to the function of the skin barrier. Protective measures such as the use of antioxidant products represent a possible method of providing protection to the skin. This paper reports the development of a non-invasive ex vivo method using tape strips of the outermost layers of stratum corneum (SC) from human volunteers in order to determine the effectiveness of an antioxidant emulsion topically applied to prevent lipid peroxidation (LPO) in the horny layer after an UV irradiation exposure. Two different formulations were used: formulation (A), containing Vitamin A, E and C, and formulation (B) containing fish extract. Both formulations were topically applied in vivo on volunteer forearms; then, a tape stripping of the SC of each volunteer was carried out. The lipid peroxidation was measured ex vivo after an UV irradiation of the SC samples. The amount of SC stripped to evaluate differences in lipid peroxidation, the UV irradiation intensity to form lipid peroxides and the accuracy of lipid peroxide analysis were optimized in this methodology using formulation (A). After an exposure application of seven days, a group of three strips of the outermost layers of SC of volunteers was irradiated with an intensity of 182.7 J/cm(2) to quantify the LPO inhibition. The percentage of LPO inhibition obtained after topical application of both formulations was in the range of 40-58% demonstrating the effectiveness of the formulations topically applied against lipid peroxidation on human SC. This methodology may be used as a quality control tool to determine ex vivo the percentage of the LPO inhibition on human SC for a variety of antioxidants topically applied.

Application of internal wool lipids to hair.

PURPOSE: Hair lipids can contribute to physicochemical phenomena such as diffusion, cell cohesion and mechanical strength, although lipids occur at much lower levels (1-6% dry weight) than proteins (>90%).Hair lipids can be diminished by hair washing and submission to chemical treatments. Studies have shown that internal wool lipids (IWL) resemble those membranes of other keratinic tissues such as human hair or stratum corneum. RESULTS: In this work, the IWL, in the form of liposomes or from an emulsion system, were applied to untreated hair fibres and also treated hair fibres. The results showed that application of IWL to pretreated hair samples lead to an improvement in mechanical strength properties of the fibres when the IWL were applied structured as liposomes. Differential scanning calorimetry studies demonstrated that the application of IWL to pretreated hair sample led to a slight increase in the crystalline material of the fibres. Moreover, scanning electron microscopy studies showed that application of IWL liposomes to damaged hair fibres lead to an improvement of the cuticle scale, demonstrating the importance of the use of a delivery vehicle with a bilayer structure similar to the one present in the hair fibre to restore the natural properties of the fibre.

Thermotropic behavior of ceramides and their isolation from wool.

The composition of internal wool lipids (IWL) resembles that of lipids present in other keratinic tissues such as human hair or the stratum corneum. Advances in the isolation of ceramides from wool and in the characterization of their thermotropic properties could facilitate their application in human skin care treatments. IWL are solvents extracted from wool fibers. Ceramide isolation is carried out by medium-pressure liquid chromatography. The different fractions obtained were analyzed quantitatively by thin layer chromatography coupled to an automated flame ionization detector and by high-performance thin layer chromatography using a densitometric detector. Two important fractions were isolated: one was a mixture of different ceramides and the other was exclusively made of ceramide 2 (nonhydroxy acid sphingosine [NS]). The thermotropic behavior of IWL and their isolated fractions were studied by thermogravimetric analysis, differential scanning calorimetry (DSC), and by attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR) methodologies. The transition temperature (Tm) obtained was compared with the results of the IWL extract, stratum corneum lipids, and the values found in the bibliography for isolated ceramides. The Tm obtained for IWL (48 degrees C) was lower than that achieved for SCL (65 degrees C). This discrepancy could be due to the different ceramide pattern and to the larger amounts of free fatty acids present in the IWL extract. Although the isolated ceramides had higher Tm values, they resembled the values reported in the bibliography. The suitability of the fraction composed exclusively of ceramide 2 [NS] for skin applications was confirmed by ATR-FTIR and DSC.