Sensory neuron activation from topical treatments modulates the sensorial perception of human skin.

Neural signaling of skin sensory perception from topical treatments is often reported in subjective terms such as a sensation of skin "tightness" after using a cleanser or "softness" after applying a moisturizer. However, the mechanism whereby cutaneous mechanoreceptors and corresponding sensory neurons are activated giving rise to these perceptions has not been established. Here, we provide a quantitative approach that couples in vitro biomechanical testing and detailed computational neural stimulation modeling along with a comprehensive in vivo self-assessment survey to demonstrate how cutaneous biomechanical changes in response to treatments are involved in the sensorial perception of the human skin. Strong correlations are identified between reported perception up to 12 hours post treatment and changes in the computed neural stimulation from mechanoreceptors residing deep under the skin surface. The study reveals a quantitative framework for understanding the biomechanical neural activation mechanism and the subjective perception by individuals.

Quantitative nanohistology of aging dermal collagen.

The skin is the largest organ in the body and is essential for protecting us from environmental stressors such as UV radiation, pollution, and pathogens. As we age, our skin undergoes complex changes that can affect its function, appearance, and health. These changes result from intrinsic (chronological) and extrinsic (environmental) factors that can cause damage to the skin's cells and extracellular matrix. As higher-resolution microscopical techniques, such as Atomic Force Microscopy (AFM), are being deployed to support histology, it is possible to explore the biophysical properties of the dermal scaffold's constituents, such as the collagen network. In this study, we demonstrate the use of our AFM-based quantitative nanohistology, performed directly on unfixed cryosections of 30 donors (female, Caucasian), to differentiate between dermal collagen from different age groups and anatomical sites. The initial 420 (10 × 10 µm2) Atomic Force Microscopy images were segmented into 42,000 (1 × 1 µm2) images before being classified according to four pre-defined empirical collagen structural biomarkers to quantify the structural heterogeneity of the dermal collagen. These markers include interfibrillar gap formation, undefined collagen structure, and registered or unregistered dense collagen fibrillar network with evident D-banding. The structural analysis was also complemented by extensive nanoindentation (∼1,000 curves) performed on individual fibrils from each section, yielding 30,000 indentation curves for this study. Principal Component Analysis was used to reduce the complexity of high-dimensional datasets. The % prevalence of the empirical collagen structural biomarkers between the papillary and reticular dermis for each section proves determinant in differentiating between the donors as a function of their age or the anatomical site (cheek or breast). A case of abnormal biological aging validated our markers and nanohistology approach. This case also highlighted the difference between chronological and biological aging regarding dermal collagen phenotyping. However, quantifying the impact of chronic and pathological conditions on the structure and function of collagen at the sub-micron level remains challenging and lengthy. By employing tools such as the Atomic Force Microscope as presented here, it is possible to start evaluating the complexity of the dermal matrix at the nanoscale and start identifying relevant collagen morphology which could be used toward histopathology standards.

Reduction of wrinkles: From a computational hypothesis to a clinical, instrumental, and biological proof.

BACKGROUND: Facial wrinkles are clear markers of the aging process, being chronological, photo-induced, or reflecting repetitive facial expressions. The aim of this study is to provide new insights into the biophysical and biological mechanisms involved in the formation, prevention, or elimination of the expression wrinkles. MATERIALS AND METHODS: We use a computational model to get a better understanding of the wrinkle mechanical behavior and evolution after skin softening and suggesting a possible antiaging mechanism. Then, we provide a clinical demonstration of the anti-wrinkle effect of a long-term application of a 20% glycerol in a moisturizer formula (GBM) versus its vehicle on crow's feet. Skin hydration, elasticity, and wrinkles visibility were evaluated by a combination of clinical and instrumental in vivo data, inverse finite element analysis, and proteomic data. RESULTS: The computational model shows a predominantly compressive stress beneath the wrinkle and its significant decrease by the softening of stratum corneum. The associated clinical study confirmed a significant increase of skin hydration and elasticity as well as a decrease of wrinkle visibility after 2 and 4 months as application for both formulas; this effect being stronger for GBM. A softening effect on stratum corneum and dermis was also observed for the GBM. Furthermore, proteomic data revealed an effect of upregulation of four proteins associated with desquamation, cell-glycan extracellular interactions, and protein glycation/oxidation, functions related to the tissue mechanics and adhesion. CONCLUSIONS: We provide an in vivo demonstration of the anti-ageing benefit of glycerol at high dose (20%) reflected by a cumulative skin surface softening effect. The use of high moisturizing potent formulations should bring additional performance to other conventional moisturizing formulations.

From decoding the perception of tightness to a clinical proof of soothing effects derived from natural ingredients in a moisturizer.

OBJECTIVE: To decode the feeling of skin tightness after application of a cosmetic product and how to soothe this discomfort. To pursue this aim, we considered the ingredient's effect on stratum corneum (SC) biomechanics to differentiate between consumers prone to tightness from those that are not and correlate these effects with mechanoreceptor activation. METHODS: In vivo clinical trials were used to assess the tightness perception dichotomy between groups of Caucasian women; in vitro experiments were used to measure the mechanical stresses induced in the SC after cleanser and moisturizer application; and in silico simulations were used to illustrate how the measured mechanical stresses in the SC result in the development of strains at the depth of cutaneous mechanoreceptors, triggering tightness perceptual responses. RESULTS: Before any cream application, women prone to tightness tend to have a more rigid SC than their less sensitive counterparts, however cleanser application increases SC stiffness in all women. Surprisingly, no correlation was found between tightness perception and hydration measurements by the Corneometer or barrier function, as evaluated by transepidermal water loss. Self-declared tightness and dryness scores were strongly associated with a self-described sensitive skin. After application of the optimized moisturizing formula, Osmoskin® containing natural waxes with good filming properties, consumers report a strong decrease in tightness and dryness perception. These results match with laboratory experiments where the cleanser was shown to increase SC drying stresses by 34%, while subsequent application of Osmoskin® decreased stresses by 48%. Finite element modelling, using experimental results as input, elucidates the differences in perception between the two groups of women. It makes clear that Osmoskin® changes the mechanical status of the SC, producing strains in underlying epidermis that activates multiple cutaneous mechano-receptors at a level correlated with the self-perceived comfort. CONCLUSION: Integration of the in vivo, in vitro and in silico approaches provides a novel framework for fully understanding how skin tightness sensations form and propagate, and how these sensations can be alleviated through the design of an optimized moisturizer.

OBJECTIF: Décoder l'impression de tiraillement de la peau après l'application d'un produit cosmétique et la manière d'apaiser cette sensation désagréable. Pour poursuivre cet objectif, nous avons pris en compte l'effet de l'ingrédient sur la biomécanique de la couche cornée afin de différencier les consommatrices sujettes à un tiraillement de celles qui ne le sont pas et de corréler ces effets avec l'activation des mécanorécepteurs. MÉTHODES: Des essais cliniques in vivo ont été utilisés pour évaluer la dichotomie de perception de tiraillement entre des groupes de femmes de race caucasienne; des expériences in vitro ont été utilisées pour mesurer les contraintes mécaniques induites dans la couche cornée après application d'un produit nettoyant et d'un produit hydratant; et des simulations in silico ont servi à illustrer comment les contraintes mécaniques mesurées dans la couche cornée entraînent le développement de souches à la profondeur des mécanorécepteurs cutanés, qui déclenchent les réponses perceptives de tiraillement. RÉSULTATS: Avant toute application de crème, les femmes sujettes au tiraillement tendent à avoir une couche cornée plus rigide que leurs homologues moins sensibles, mais l'application d'un produit nettoyant augmente la raideur de la couche cornée chez toutes les femmes. Étonnamment, aucune corrélation n'a été observée entre la perception de tiraillement et les mesures d'hydratation réalisées par le cornéomètre ou la fonction barrière, évaluée par la perte d'eau transépidermique. Les scores de tiraillement et de sécheresse auto-déclarés étaient fortement corrélés à une peau décrite par les sujets elles-mêmes comme sensible. Après application de la formule hydratante optimisée, Osmoskin®, qui contient des cires naturelles ayant de bonnes propriétés de dépôt de film, les consommateurs rapportent une forte diminution de la sensation de tiraillement et de sécheresse. Ces résultats concordent avec les expériences en laboratoire où le produit nettoyant s'est avéré augmenter les contraintes de séchage de la couche cornée de 34 %, tandis que l'application ultérieure d'Osmoskin® a réduit les contraintes de 48 %. La modélisation à éléments finis, en utilisant les résultats expérimentaux comme données, élucide les différences de perception entre les deux groupes de femmes. Il est clair qu'Osmoskin® modifie l'état mécanique de la couche cornée, et produit des souches dans l'épiderme sous-jacent qui activent plusieurs mécano-récepteurs cutanés à un niveau corrélé au confort perçu par la patiente. CONCLUSION: La combinaison des approches in vivo, in vitro et in silico fournit un nouveau cadre pour comprendre pleinement comment les sensations de tiraillement de la peau se forment et se propagent, et comment elles peuvent être soulagées en mettant au point une crème hydratante optimisée.

Turning autophobic wetting on biomimetic surfaces into complete wetting by wetting additives.

Autophobicity or pseudo partial wetting, a phenomenon of a liquid not spreading on its own monolayer, is characterized by an energy barrier that prevents the growth of a wetting film beyond the monolayer thickness. Applying a molecularly detailed self-consistent field theory we illustrate how autophobic wetting can be overcome by wetting additives. More specifically we use an emulsifier which keeps the interfacial tension between the wetting component and the majority solvent low, and a co-solvent additive which partitions inside the film and then destroys the molecular order in it so that the barrier for film growth is cleared. An application wherein it is believed that autophobic wetting is counteracted by such a set of wetting additives is found in an antidandruff shampoo formulation. We have experimental results that show thick deposits onto hydrophobic hair surfaces by administration of the antidandruff shampoo. The complementary modeling of such a system suggests that the active ingredient plays the role of the co-solvent additive. As significant amounts of the co-solvent additives are needed to approach the completely wet state, the formulation naturally brings large amounts of active ingredient to the root of the hair where its presence is required.

In vivo quantitative molecular absorption of glycerol in human skin using coherent anti-Stokes Raman scattering (CARS) and two-photon auto-fluorescence.

The penetration of small molecules through the human skin is a major issue for both safety and efficacy issues in cosmetics and pharmaceutic domains. To date, the quantification of active molecular compounds in human skin following a topical application uses ex vivo skin samples mounted on Franz cell diffusion set-up together with appropriate analytical methods. Coherent anti-Stokes Raman scattering (CARS) has also been used to perform active molecule quantification on ex vivo skin samples, but no quantification has been described in human skin in vivo. Here we introduce and validate a framework for imaging and quantifying the active molecule penetration into human skin in vivo. Our approach combines nonlinear imaging microscopy modalities, such as two-photon excited auto-fluorescence (TPEF) and coherent anti-Stokes Raman scattering (CARS), together with the use of deuterated active molecules. The imaging framework was exemplified on topically applied glycerol diluted in various vehicles such as water and xanthan gel. In vivo glycerol quantitative percutaneous penetration over time was demonstrated, showing that, contrary to water, the xanthan gel vehicle acts as a film reservoir that releases glycerol continuously over time. More generally, the proposed imaging framework provides an enabling platform for establishing functional activity of topically applied products in vivo.

Combining nano-physical and computational investigations to understand the nature of "aging" in dermal collagen.

The extracellular matrix of the dermis is a complex, dynamic system with the various dermal components undergoing individual physiologic changes as we age. Age-related changes in the physical properties of collagen were investigated in particular by measuring the effect of aging, most likely due to the accumulation of advanced glycation end product (AGE) cross-links, on the nanomechanical properties of the collagen fibril using atomic force microscope nano-indentation. An age-related decrease in the Young's modulus of the transverse fibril was observed (from 8.11 to 4.19 GPa in young to old volunteers, respectively, P<0.001). It is proposed that this is due to a change in the fibril density caused by age-related differences in water retention within the fibrils. The new collagen-water interaction mechanism was verified by electronic structure calculations, showing it to be energetically feasible.

Cell surface glycans in the human stratum corneum: distribution and depth-related changes.

During the formation of the stratum corneum (SC) barrier, the extracellular spaces of viable epidermis, rich in glycans, are filled with a highly organized lipid matrix and the plasma membranes of keratinocytes are replaced by cornified lipid envelopes. These structures comprise cross-linked proteins, including transmembrane glycoproteins and proteoglycans, covalently bound to a monolayer of cell surface ceramides. Little is known about the presence and distribution of glycans on the SC corneocytes despite their possible involvement in SC hydration, cohesion and desquamation. In this work, we visualized ultrastructurally and quantified the distribution of glycans on the surface of native and delipidated corneocytes. The cells were harvested at different depths of the SC, allowing us to define the relationship between the distribution of various glycans, proteoglycans and glycoproteins, and other changes occurring in SC. At the cell periphery, we found a correlation between the depth-related alterations of corneodesmosome glycoproteins and α-d-mannosyl and N-acetyl-d-glucosamine-labelling patterns. Elimination of the terminal sugars, α-linked fucose and α-(2,3) linked sialic acid, was less abrupt, but also the initial extent of their peripheral distribution was overall lower than that of concanavalin A and wheat germ agglutinin lectin-detected glycans. Diffuse labelling of heparan sulphate glycosaminoglycans disappeared completely from the outermost corneocytes, whereas that of several simple carbohydrates could be detected at all SC levels. Our results suggest that specific glycan distribution may participate in the progressive changes of SC, as it evolves from the SC compactum to the SC disjunctum, towards desquamation.

Micron-scale assessment of molecular lipid organization in human stratum corneum using microprobe X-ray diffraction.

Lipid and protein components of the stratum corneum (SC) are organized in complex supramolecular arrangements. Exploring spatial relations between various possible substructures is important for understanding the barrier function of this uppermost layer of epidermis. Here, we report the first study where micro-focus X-ray scattering was used for assessing fine structural variations of the human skin barrier with micrometer resolution. We found that the scattering profiles were unchanged when scanning in the direction parallel to the SC surface. Furthermore, small-angle scattering profiles did not change as a function of depth in the SC, confirming that the lipid lamellar spacings remained the same throughout the SC. However, the wide-angle scattering data showed that the orthorhombic phase was more abundant in the middle layers of the SC, whereas the hexagonal phase dominated in the surface layers both at the external and the lowest part of the SC; i.e., the lipids were most tightly packed in the middle region of the SC. Taken together, our results demonstrate that microprobe X-ray diffraction provides abundant information about spatial variations of the SC lipid structure and thus may be a promising tool for assessing the effects of topical formulations on the barrier function of skin.

Impact of trochanteric heterotopic ossification on measurement of femoral bone density following cemented total hip replacement.

During a study of bone mineral density changes around cemented femoral implants, we recognized heterotopic ossification occurring regularly in a position anterior to the greater trochanter and proximal femur. The aim of this study was to describe the incidence, distribution, and effect of this ossification on periprosthetic DXA scans following primary cemented total hip replacement. One hundred eleven patients underwent postoperative DXA examinations measuring changes in bone mineral density with heterotopic ossification identified and localized on standard radiographs with confirmation using DXA subtraction imaging. Male gender and age within the male group were significantly associated with occurrence of heterotopic ossification (p = 0.003 and 0.046, respectively). Femoral stem type, weight, and body mass index had no significant effect (p = 0.525, 0.372, and 0.243, respectively). Examining the Gruen zones in all patients suggested a median (plus interquartile range) zone 1 density drop of 4% (-12% to +7%). When separated and analyzed for the effect of heterotopic ossification, the 45 patients with heterotopic ossification showed a 2-year density gain of +6% (-5% to +15%), whereas those without heterotopic ossification showed a loss of 8% (-14% to 0%), a significant difference (p < 0.001). Zone 2 also showed a significant difference (p = 0.048). We therefore recommend that affected zones should be identified and excluded from analysis at all time points. Without this precaution, researchers risk underestimating periprosthetic bone loss in their studies and reporting misleading conclusions.

Recruitment to research studies in maternity hospitals: an example from the Early Births Study.

OBJECTIVES: to present issues associated with recruitment of women in maternity hospitals to a population-based case-control study of very preterm birth. DESIGN: a descriptive study of the recruitment process. SETTING: all maternity hospitals, including three providing neonatal intensive care services, in Victoria, Australia from April 2002 to April 2004. PARTICIPANTS: cases were women who had a singleton birth between 20 and 31+6 weeks of gestation. Controls were a random selection of women having a singleton birth of at least 37 completed weeks of gestation in the same time period as the cases. MEASUREMENTS AND FINDINGS: ethical approval was obtained from 73 of 77 maternity hospitals. Hospitals considered that privacy laws required that women should be approached initially by hospital staff for recruitment into the research study. Extensive effort was put into liaising with hospital personnel, determining hospital-specific protocols for approaching women and developing relationships with doctors, midwives and ward clerks. Recurrent reminders were provided to all hospitals. Of the 2785 women (cases and controls) ascertained as eligible, 13% of cases with surviving babies, 11% of controls and 74% of cases whose babies did not survive were not approached to participate in the study. Within these groups, there was variation by gestation and hospital. Once women were approached, 72% were interviewed. The interview response proportion was 50%. KEY CONCLUSIONS: recruitment to studies in the maternity setting in the postpartum period is a challenge. Barriers to recruitment that may have introduced selection bias in this study include: recruitment at many hospitals; short postnatal hospital stay; reliance on hospital staff to make the first approach to women; and low response from women whose babies did not survive. A dialogue between researchers and clinical midwives is proposed to explore ways of increasing researchers' understanding of the complex and demanding hospital environment, and to improve research awareness among clinical midwives.

Research interviewers' experience in the Early Births study of very preterm birth: qualitative assessment of data collection processes in a case-control study.

This paper describes an evaluation study that aimed to assess data collection processes in a population-based case-control study of very preterm birth. Semi-structured interviews were conducted with the 10 research interviewers to determine their perceptions of the hypotheses, the differences between interviewing cases and controls and between modes of interview, their reactions to questions which they had to ask in interviews and their training. Time and cost of the collection of data were also considered. None of the research interviewers identified which of the questions in the interview constituted the primary hypothesis. All interviewers interviewed cases and controls (including mothers of twins and singletons), and collected data face-to-face and by telephone. Whilst half of the interviewers had no issue with asking sensitive questions, hearing of intimate partner violence, risk-taking behaviour and inappropriate medical care were confronting for others. Training was judged as adequate, as was the continuing support of the project co-ordinator. On average, interviewing took 40% of research interviewers' time, and 25 interviews were completed per effective full-time month of interviewer time. Each interview cost approximately $170AU to complete. In relation to this case-control study, interviewer bias may be lessened since research interviewers had been unable to infer the hypothesis of the study. All interviewers had interviewed both cases and controls and any systematic differences can be adjusted for in the analysis.