Stratum Corneum Lipid Matrix Research Articles

Sensing the Changes in Stratum Corneum Using Fourier Transform Infrared Microspectroscopy and Hyperspectral Data Processing.

The stratum corneum (SC) forms the outermost layer of the skin, playing a critical role in preventing water loss and protecting against external biological and chemical threats. Approximately 90% of the SC consists of large, flat corneocytes, yet its barrier function primarily relies on the intercellular lipid matrix that surrounds these cells. Traditional methods for characterizing these lipids, such as Fourier transform infrared spectroscopy (FTIR), typically involve macroscopic analysis using attenuated total reflection (ATR) techniques. In this study, we introduce a novel approach for investigating SC samples at a microscopic level to gain detailed chemical insights and assess sample heterogeneity. Special emphasis is placed on advanced hyperspectral data pre-processing to ensure the accuracy and reliability of the results. We also evaluate methods for filtering out spectral data that significantly deviate from the mean and analyze the extracted mean spectra, the intensities of specific infrared peaks, and their ratios. The novelty of this work lies in its microscopic approach to analyzing the SC lipid matrix, diverging from the traditional macroscopic FTIR-ATR methods. By focusing on hyperspectral imaging and developing robust pre-processing techniques, this study provides more localized, high-resolution chemical insights. This microscopic perspective opens up the possibility of detecting subtle heterogeneities within the skin's lipid matrix, offering deeper, previously unattainable understanding of the SC's barrier function. Additionally, the exploration of spectral filtering methods enhances the precision of the analysis, paving the way for more refined and reliable investigations of skin structure and behavior in future research.

Structure and function of skin barrier lipids: Effects of hydration and natural moisturizers in vitro

Lipid membranes play a crucial role in regulating the body’s water balance by adjusting their properties in response to hydration. The intercellular lipid matrix of the stratum corneum (SC), the outermost skin layer, serves as the body’s primary defense against environmental factors. Osmolytes, including urocanic acid (UCA) and glycerol, are key components of the natural moisturizing factor that help the SC resist osmotic stress from dry environments. This study examines the effects of UCA and glycerol (each at 5 mol %) on isolated human SC lipids. For this, different techniques were employed, offering complementary information of the system’s multiscale characteristics, including humidity-scanning quartz crystal microbalance with dissipation monitoring, infrared spectroscopy, x-ray diffraction, electrical impedance spectroscopy, and studies of water loss and permeability. Our results show that UCA increases water sorption and makes lipid films more liquid-like at high relative humidity, without significantly altering the lipid lamellar structure, chain order, or orthorhombic chain packing. Lipid films containing UCA exhibited higher water loss and significantly higher model drug permeability compared to lipid films without UCA. Further, incorporation of UCA resulted in kinetically faster changes in electrical properties upon contact with aqueous solution compared with control lipids. These observations suggest that UCA reduces lipid cohesion in regions other than the acyl chain-rich leaflets, which may impact SC desquamation. In contrast, glycerol did not influence the hydration or permeability of the SC lipid matrix. However, it increased the proportion of orthorhombic domains at high humidities and slowed the kinetics of the hydration process, as evidenced by slower changes in the dielectric properties of the lipid film. These findings suggest that glycerol enhances lipid cohesion rather than increasing water uptake, which is typically the expected function of humectants. Consequently, UCA and glycerol appear to have distinct roles in maintaining epidermal homeostasis.

Clinical efficacy of a multilamellar cream on skin physiology and microbiome in an epidermal stress model: A controlled double-blinded study.

Stratum corneum (SC) is essential for skin barrier function, mitigating water loss and shielding against potentially harmful substances and allergens. The SC's lipid matrix, arranged in a lamellar structure, is integral to its protective role. Our study explores the restoration effects of a multilamellar cream with an acidic pH compared to a basic placebo cream on skin physiology and its interaction with the skin microbiome after stress induction via tape stripping (TS). In this double-blind study, 14 healthy participants aged 21-58 years were assessed pre- and post-tape stripping, followed by a 14 days application of a multilamellar test cream and a placebo cream with evaluations on days 7, 14 and 17 for sustained effects. Skin physiology was analysed in terms of epidermal barrier function, SC hydration and surface pH. The microbiome was analysed by 16S rRNA amplicon sequencing the 16S rRNA gene using Illumina MiSeq, with subsequent species identification. Our study showed significant improvements in skin barrier repair and SC hydration with verum, particularly after 14 days of application, while both creams initially enhanced stratum corneum hydration. No significant changes in surface-pH were detected. The skin microbiome analysis revealed that TS slightly decreased alpha diversity, a trend that verum significantly reversed, enhancing diversity beyond baseline levels after 14 days. Overall, while both creams contributed to a broader microbial phyla diversity over time, no significant changes in the abundance of specific genera or species were noted between treatments. Our study delineates the efficacy of a pH-optimized multilamellar cream in enhancing epidermal barrier recovery and SC hydration post-sequential TS, in contrast to an unstructured basic placebo. Verum cream significantly improved skin barrier function and SC hydration at day 14, with sustained effects evident beyond the treatment period. Furthermore, the multilamellar formulation facilitated the restitution of cutaneous microbiome diversity, a key indicator of healthy skin ecology, underscoring the symbiotic relationship between barrier integrity and microbial composition. These findings underscore the importance of multilamellar emollient structures in dermatological therapeutics, with potential implications for the design of advanced skincare interventions that holistically support cutaneous resilience and homeostasis.

Palmitoylation enhances short polar peptide permeation across stratum corneum lipid bilayer: A molecular dynamics study

Designing chemical molecules that target the skin for non-invasive transdermal drug delivery is of significant interest for both wound healing and skincare applications. These skin-targeting molecules must permeate the outermost protective layer of the skin, the stratum corneum (SC), which consists of dead corneocytes embedded in a lipid matrix, to fulfill their biological functions. Adsorption onto and diffusion through the lipid matrix in the SC represent two key steps for the successful permeation of a skin-targeting molecule across the SC into the underlying skin layers. Here we compare the effects of cyclization and palmitoylation on the adsorption and diffusion of a short polar peptide across a model SC lipid bilayer using molecular dynamics simulations. The cyclized peptide showed slightly better binding to the SC lipid bilayer and similar interaction energies with SC lipids compared to the unmodified peptide. In contrast, the palmitoylated peptide exhibited much stronger interaction with SC lipids via insertion of its attached fatty acid tail into the SC lipid bilayer. The average diffusivity of the cyclized peptide across the SC lipid bilayer was approximately twice that of the unmodified peptide, whereas the palmitoylated peptide’s diffusivity was about 2.7 times higher. Thus, palmitoylation appears to be a promising strategy for enhancing the binding and permeability of short polar peptides across the SC lipid matrix.

Guselkumab treatment normalizes the stratum corneum ceramide profile and alleviates barrier dysfunction in psoriasis: results of a randomized controlled trial

The epidermal inflammation associated with psoriasis drives skin barrier perturbations. The skin barrier is primarily located in stratum corneum (SC). Its function depends on the SC lipid matrix of which ceramides constitute important components. Changes in the ceramide profile directly correlate to barrier function. In this study, we characterized the dynamics of the barrier function and ceramide profile of psoriatic skin during anti-Interleukin-23 therapy with guselkumab. We conducted a double-blind, randomized controlled trial in which 26 mild-to-severe plaque psoriasis patients were randomization 3:1–100 mg guselkumab or placebo for 16 weeks and barrier dynamics monitored throughout. Barrier function was measured by trans-epidermal water loss measurements. Untargeted ceramide profiling was performed using liquid chromatography-mass spectrometry after SC was harvested using tape-stripping. The barrier function and ceramide profile of lesional skin normalized to that of controls during treatment with guselkumab, but not placebo. This resulted in significant differences compared to placebo at the end of the treatment. Changes in the lesional ceramide profile during treatment correlated with barrier function and target lesion severity. Nonlesional skin remained similar throughout treatment. Guselkumab therapy restored the skin barrier in psoriasis. Concomitant correlations between skin barrier function, the ceramide profile, and disease severity demonstrate their interdependency.

The Sphingosine and Phytosphingosine Ceramide Ratio in Lipid Models Forming the Short Periodicity Phase: An Experimental and Molecular Simulation Study.

The lipids located in the outermost layer of the skin, the stratum corneum (SC), play a crucial role in maintaining the skin barrier function. The primary components of the SC lipid matrix are ceramides (CERs), cholesterol (CHOL), and free fatty acids (FFAs). They form two crystalline lamellar phases: the long periodicity phase (LPP) and the short periodicity phase (SPP). In inflammatory skin conditions like atopic dermatitis and psoriasis, there are changes in the SC CER composition, such as an increased concentration of a sphingosine-based CER (CER NS) and a reduced concentration of a phytosphingosine-based CER (CER NP). In the present study, a lipid model was created exclusively forming the SPP, to examine whether alterations in the CER NS:CER NP molar ratio would affect the lipid organization. Experimental data were combined with molecular dynamics simulations of lipid models containing CER NS:CER NP at ratios of 1:2 (mimicking a healthy SC ratio) and 2:1 (observed in inflammatory skin diseases), mixed with CHOL and lignoceric acid as the FFA. The experimental findings show that the acyl chains of CER NS and CER NP and the FFA are in close proximity within the SPP unit cell, indicating that CER NS and CER NP adopt a linear conformation, similarly as observed for the LPP. Both the experiments and simulations indicate that the lamellar organization is the same for the two CER NS:CER NP ratios while the SPP NS:NP 1:2 model had a slightly denser hydrogen bonding network than the SPP NS:NP 2:1 model. The simulations show that this might be attributed to intermolecular hydrogen bonding with the additional hydroxide group on the headgroup of CER NP compared with CER NS.

Lipid Biomimetic Models as Simple Yet Complex Tools to Predict Skin Permeation and Drug-Membrane Biophysical Interactions.

The barrier function of the skin is primarily determined by its outermost layer, the Stratum Corneum (SC). The SC consists of corneocytes embedded in a lipid matrix composed mainly of ceramides, cholesterol, and free fatty acids in equimolar proportions and is organised in a complex lamellar structure with different periodicities and lateral packings. This matrix provides a diffusion pathway across the SC for bioactive compounds that are administered to the skin. In this regard, and as the skin administration route has grown in popularity, there has been an increase in the use of lipid mixtures that closely resemble the SC lipid matrix, either for a deeper biophysical understanding or for pharmaceutical and cosmetic purposes. This review focuses on a systematic analysis of the main outcomes of using lipid mixtures as SC lipid matrix models for pharmaceutical and cosmetic purposes. Thus, a methodical evaluation of the main outcomes based on the SC structure is performed, as well as the main recent developments in finding suitable new in vitro tools for permeation testing based on lipid models.

Molecular Dynamics Investigation into CerENP's Effect on the Lipid Matrix of Stratum Corneum.

The extracellular lipid matrix in the stratum corneum (SC) plays a critical role in skin barrier functionality, comprising three primary components: ceramides, cholesterol, and free fatty acids. The diverse ceramides, differentiated by molecular structures such as hydroxylations and varying chain lengths, are essential for the lipid matrix's structural integrity. Recently, a new subclass of ceramide, 1-O-acylceramide NP (CerENP), has been identified; however, its precise role in the lipid matrix of the SC is still elusive. Herein, we investigate the role of CerENP on the structure and permeability of the SC using molecular dynamics simulations. Our findings indicate that CerENP contributes to a compact lipid matrix in the lateral dimension of our SC model with a repeat distance of about 13 nm. Additionally, ethanol permeability assessments show that CerENP effectively reduces molecular penetration through the lipid matrix. This study provides an insight into the role of a new subclass of ceramide in the SC, enhancing our understanding of skin structure and the mechanisms behind barrier dysfunction in skin diseases.

Permeation of actives in the stratum corneum lipid matrix

Permeation of actives in the stratum corneum lipid matrix

Improvement of Human Epidermal Barrier Structure and Lipid Profile in Xerotic- and Atopic-Prone Skin via Application of a Plant-Oil and Urea Containing pH 4.5 Emulsion

Epidermal barrier dysfunction can lead to xerotic skin and promote skin disorders like atopic dermatitis. Atopic skin is characterized by reduced water-retaining compounds, altered lipid composition and elevated skin pH. Against this background, a study was conducted to investigate the impact of a specific skin care product on epidermal barrier function in dry and atopic-prone skin. A marketed pH 4.5 cosmetic formulation containing 10% urea and specific plant oils was evaluated on 25 subjects with dry and atopic-prone skin. Measurements of skin hydration, pH, and barrier function were performed before and after 3 weeks of product usage. Additionally, visual scoring and stratum corneum lipid analysis using electron microscopy were conducted to investigate lipid composition. An improved skin hydration compared to the untreated area and a tendency to decrease the baseline elevated skin surface pH were observed. The visual scoring showed reduced dryness, roughness, and tension through the application. Furthermore, the stratum corneum lipid matrix was improved in terms of lipid content and organization. The combination of an acidic product’s pH, a relevant urea content and effective plant oils is shown to be beneficial in terms of improving the skin barrier function, structure and appearance and is recommended for dry and atopic-prone skin.

Effect of sphingosine and phytosphingosine ceramide ratio on lipid arrangement and barrier function in skin lipid models

The lipids in the uppermost layer of the skin, the stratum corneum (SC), play an important role in the skin barrier function. The three main subclasses in the SC lipid matrix are ceramides (CER), cholesterol, and free fatty acids. In inflammatory skin diseases, such as atopic dermatitis and psoriasis, the SC lipid composition is modulated compared to the composition in healthy SC. One of the main alterations is the molar ratio between the concentration of CER N-(tetracosanoyl)-sphingosine (CER NS) and CER N-(tetracosanoyl)-phytosphingosine (CER NP), which correlated with an impaired skin barrier function. In the present study, we investigated the impact of varying the CER NS:CER NP ratios on the lipid organization, lipid arrangement, and barrier functionality in SC lipid model systems. The results indicate that a higher CER NS:CER NP ratio as observed in diseased skin did not alter the lipid organization or lipid arrangement in the long periodicity phase encountered in SC. The trans-epidermal water loss, an indication of the barrier functionality, was significantly higher for the CER NS:CER NP 2:1 model (mimicking the ratio in inflammatory skin diseases) compared to the CER NS:CER NP 1:2 ratio (in healthy skin). These findings provide a more detailed insight into the lipid organization in both healthy and diseased skin and suggest that in vivo the molar ratio between CER NS:CER NP contributes to barrier impairment as well but might not be the main factor.

Encapsulation of Vitamin C by Glycerol-Derived Dendrimers, Their Interaction with Biomimetic Models of Stratum corneum and Their Cytotoxicity.

Vitamin C is one of the most sensitive cosmetic active ingredients. To avoid its degradation, its encapsulation into biobased carriers such as dendrimers is one alternative of interest. In this work, we wanted to evaluate the potential of two biobased glycerodendrimer families (GlyceroDendrimers-Poly(AmidoAmine) (GD-PAMAMs) or GlyceroDendrimers-Poly(Propylene Imine) (GD-PPIs)) as a vitamin C carrier for topical application. The higher encapsulation capacity of GD-PAMAM-3 compared to commercial PAMAM-3 and different GD-PPIs, and its absence of cytotoxicity towards dermal cells, make it a good candidate. Investigation of its mechanism of action was done by using two kinds of biomimetic models of stratum corneum (SC), lipid monolayers and liposomes. GD-PAMAM-3 and VitC@GD-PAMAM-3 (GD-PAMAM-3 with encapsulated vitamin C) can both interact with the lipid representatives of the SC lipid matrix, whichever pH is considered. However, only pH 5.0 is suggested to be favorable to release vitamin C into the SC matrix. Their binding to SC-biomimetic liposomes revealed only a slight effect on membrane permeability in accordance with the absence of cytotoxicity but an increase in membrane rigidity, suggesting a reinforcement of the SC barrier property. Globally, our results suggest that the dendrimer GD-PAMAM-3 could be an efficient carrier for cosmetic applications.

Phytosphingosine ceramide mainly localizes in the central layer of the unique lamellar phase of skin lipid model systems

Understanding the lipid arrangement within the skin’s outermost layer, the stratum corneum (SC), is important for advancing knowledge on the skin barrier function. The SC lipid matrix consists of ceramides (CERs), cholesterol, and free fatty acids, which form unique crystalline lamellar phases, referred to as the long periodicity phase (LPP) and short periodicity phases. As the SC lipid composition is complex, lipid model systems that mimic the properties of native SC are used to study the SC lipid organization and molecular arrangement. In previous studies, such lipid models were used to determine the molecular organization in the trilayer structure of the LPP unit cell. The aim of this study was to examine the location of CER N-(tetracosanoyl)-phytosphingosine (CER NP) in the unit cell of this lamellar phase and compare its position with CER N-(tetracosanoyl)-sphingosine (CER NS). We selected CER NP as it is the most prevalent CER subclass in the human SC, and its location in the LPP is not known. Our neutron diffraction results demonstrate that the acyl chain of CER NP was positioned in the central part of the trilayer structure, with a fraction also present in the outer layers, the same location as determined for the acyl chain of CER NS. In addition, our Fourier transformed infrared spectroscopy results are in agreement with this molecular arrangement, suggesting a linear arrangement for the CER NS and CER NP. These findings provide more detailed insight into the lipid organization in the SC lipid matrix.

Investigating the nanostructure of a CER[NP]/CER[AP]-based stratum corneum lipid matrix model: A combined neutron diffraction & molecular dynamics simulations approach

The human skin provides a physiochemical and biological protective barrier due to the unique structure of its outermost layer known as the Stratum corneum. This layer consists of corneocytes and a multi-lamellar lipid matrix forming a composite, which is a major determining factor for the barrier function of the Stratum corneum. A substantiated understanding of this barrier is necessary, as controlled breaching or modulation of the same is also essential for various health and personal care applications such as topical drug delivery and cosmetics to a name few.In this study, we discuss the state-of-the-art of neutron diffraction techniques, using specifically deuterated lipids, combined with the information obtained from molecular models using molecular dynamics simulations, to understand the structure and barrier function of the Stratum corneum lipid matrix.As an example, the effect of ceramide concentration on a lipid lamella system consisting of CER[NP]/CER[AP]/Cholesterol/free fatty acid (deprotonated) is studied. This study demonstrates the usefulness of the combined approach of neutron diffraction and molecular dynamics simulations for effective analysis of the model systems created for the Stratum corneum lipid matrix. The optimization of force fields by comparison with experimental data is furthermore an important step in the direction of providing a predictive quality.

The importance of ceramide headgroup for lipid localisation in skin lipid models

The stratum corneum's lipid matrix is a critical for the skin's barrier function and is primarily composed of ceramides (CERs), cholesterol (CHOL) and free fatty acids (FFAs). The lipids form a long periodicity phase (LPP), a unique trilayer unit cell structure. An enzyme driven pathway is implemented to synthesize these key lipids. If these enzymes are down- or upregulated as in inflammatory diseases, the final lipid composition is affected often altering the barrier function. In this study, we mimicked down regulation of enzymes involved in the synthesis of the sphingosine and CER amide bond. In a LPP lipid model, we substituted CER N-(tetracosanoyl)-sphingosine (CER NS) with either i) FFA C24 and free sphingosine, to simulate the loss of the CER amide bond, or ii) with FFA C24 and C18 to simulate the loss of the sphingosine headgroup. Our study shows the lipids in the LPP would not phase separate until at least 25% of the CER NS is substituted keeping the lateral packing and conformational ordering unaltered. Neutron diffraction studies showed that free sphingosine chains localized at the outer layers of the unit cell, while the remaining CER NS head group was concentrated in the inner headgroup layers. However, when FFA C18 was inserted, CER NS was dispersed throughout the LPP, resulting in an even distribution between the inner and outer water layers. The presented results highlight the importance of the CER NS headgroup structure and its interaction in combination with the carbon chain invariability for optimal lipid arrangement.

Lipid-coated membranes as skin surrogates for permeability assessment

There is a growing interest in developing in vitro skin models, particularly in the fields of skin care and drug delivery regarding optimization of topical drug formulations. The ban of animal experiments for cosmetic products testing has led to the development of alternative in vitro methods. The aim of this study was to develop and characterize simplified, cell-free, lipid-coated membrane models, exploring the usefulness and the potential application of these membranes in comparison to commercial synthetic membrane and to porcine skin. In these model membranes, lipids are deposited onto a polycarbonate membrane to create lipid-rich coated membranes. Two types of lipid-coated membranes were explored depending on the type of lipids used. In one type (phospholipidic), membranes were prepared depositing soybean phosphatidylcholine liposomes and in the other type (non-phospholipidic), major components of the stratum corneum lipid matrix were used to coat the polycarbonate membrane. Coating process was performed using drying and centrifugation steps. Under infinite dose conditions, and caffeine as permeant, high permeation correlation between developed membranes and pig skin was obtained, and similar permeation pro-files were obtained through phospholipidic and non-phospholipidic membranes. This study demonstrated the potential of the developed membranes to conduct permeation studies using Franz-type diffusion cells, and the simplication process to produce lipid-coated membranes.

Cerosomes as skin repairing agent: Mode of action studies with a model stratum corneum layer at liquid/air and liquid/solid interfaces

The stratum corneum (SC) is the largest physical barrier of the human body. It protects against physical, chemical and biological damages, and avoids evaporation of water from the deepest skin layers. For its correct functioning, the homeostasis of the SC lipid matrix is fundamental. An alteration of the lipid matrix composition and in particular of its ceramide (CER) fraction can lead to the development of pathologies such as atopic dermatitis and psoriasis. Different studies showed that the direct replenishment of SC lipids on damaged skin had positive effects on the recovery of its barrier properties.In this work, cerosomes, i.e. liposomes composed of SC lipids, have been successfully prepared in order to investigate the mechanism of interaction with a model SC lipid matrix. The cerosomes contain CER[NP], D-CER[AP], stearic acid and cholesterol. In addition, hydrogenated soybean phospholipids have been added to one of the formulations leading to an increased stability at neutral pH. For the mode of action studies, monolayer models at the air-water interface and on solid support have been deployed. The results indicated that a strong interaction occurred between SC monolayers and the cerosomes. Since both systems were negatively charged, the driving force for the interaction must be based on the ability of CERs head groups to establish intermolecular hydrogen bonding networks that energetically prevailed against the electrostatic repulsion. This work proved for the first time the mode of action by which cerosomes exploit their function as skin barrier repairing agents on the SC.

Effect of microplasma treatment on stratum corneum lipid molecule

The stratum corneum is a lipid matrix with embedded corneocytes. This layer protects the body against foreign substances and the outer environment. However, the lipid matrix is permeable to selective molecules and this permeability can be improved by several methods. One of these methods includes microplasma irradiation of the skin. Stearic acid is a model fatty acid from the lipid matrix of stratum corneum. In this study, the interaction between microplasma and stearic acid was investigated. Plasma was generated at a low voltage of about 800 V (0-Vpeak) using a thin dielectric (∼30 μm) as a barrier. The plasma electrode treated the lipids under an argon atmosphere at a frequency of 25 kHz. The results of the microplasma treatment were investigated by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) and X-ray photoelectron spectroscopy. The concentrations of C, O and N atoms were determined before and after the microplasma treatment. Also, functional groups of C–C/C–H, C–O, C=O, OH–C=O, N–C=O were identified in the carbon spectra. The structure of stearic acid was evaluated by analysis of CH2, CH3 and C=O bands in the ATR-FTIR spectrum.

Stratum corneum lipid matrix with unusual packing: A molecular dynamics study

The skin is an effective barrier against the external elements being the stratum corneum, with its lipid matrix surrounding the corneocytes, considered the major player responsible for its low permeability. The use of computational models to study the transdermal delivery of compounds have a huge potential to improve this research area, but requires reliable models of the skin components. In this work, we developed molecular dynamics models with a coarse-grained resolution, of the stratum corneum lipid matrix and the sebum. We developed the lipid matrix model with unusual lipid packing configuration as some recent works support. The simulation results show that this configuration is stable and may help to explain the low permeability of stratum corneum. The sebum simulations showed that this oily skin product can also play a significant role in the transdermal delivery of drugs.

The effect of hydrophilic penetration/diffusion enhancer on stratum corneum lipid models: Part II*: DMSO

To optimize dermal and transdermal administration of drugs, the barrier function of the skin, particularly the stratum corneum (SC), needs to be reduced reversibly. For this purpose, penetration/diffusion enhancers such as DMSO can be applied. However, there is the question whether DMSO is an aggressive penetration/diffusion enhancer in pharmaceutical and cosmetical relevant concentrations? Until now, it is unclear if this penetration/diffusion enhancement is caused by an interaction with the SC lipid matrix or related to effects within the corneocytes. Therefore, the effects of the hydrophilic enhancer DMSO on SC models with different dimensionality ranging from bilayers (liposomes) via oligo-layers to multilayers have been investigated in this study. The effects of DMSO should be compared to that of other relevant hydrophilic enhancers such as urea and taurine. An innovative spectrum of methods was applied to ascertain the mode of action of DMSO in relevant concentrations on a molecular scale.The experiments reveal that there is no specific interaction of 10% and 30% DMSO solutions with the SC model systems. Hence, if DMSO is applied in pharmaceutically and cosmetically relevant concentrations, it has no influence on the SC model systems used. Neither an additional water uptake in the head group region nor a decrease of the lipid chain packing density have been observed. The leakage studies on liposomes show that 10% DMSO is causing just a very slight leakage of 8%, lower than the leakage of 19.4% caused by 10% urea (Müller et al., 2016). Consequently, the interactions of DMSO with the SC model lipids used are very low in concentrations of 10% and 30%, respectively. Since the lipid composition in native SC lipid matrix is far more complex than this model mixture, the results can not be directly transferred to the native SC lipid matrix. However, the outcome of this study, together with various findings in the literature give rise to the assumption that the enhancing effect of DMSO concerning the diffusion of relevant hydrophilic drugs and actives appears to be realized via the corneocytes.