Chitin–glucan,a natural cell scaffold for skin moisturization and rejuvenation

Two clinical studies were conducted to evaluate the tolerance and effects of a copolymer of chitin and beta‐glucan, forming the exoskeleton of fungal cell walls, now supplied for cosmetic applications. A 6‐week randomized, double‐blind, placebo‐controlled study was conducted on 13 volunteers with sensitive skin to compare with 0.5–2% formulations chitin–glucan applied twice daily. Biometrological evaluations showed that erythema did not develop, the water retention capacity of the stratum corneum increased and the transepidermal water loss moderately decreased. Another 16‐week randomized, double‐blind, placebo‐controlled study was conducted on 20 men showing signs of ageing skin. A 1.5% chitin–glucan formulation was applied twice daily. Objective biometrological assessments showed a progressive increase in skin firmness and stratum corneum hydration when desquamation and skin roughness decreased. In conclusion, the chitin–glucan formulations appear safe. They significantly mitigate some signs of skin ageing and improve both stratum corneum hydration and skin barrier function.     

Formulating for efficacy

Active ingredients have been around in cosmetics for a long time but have they really resulted in active cosmetic products? In order to achieve this, the right active needs to be delivered to the right location at the right concentration for the correct period of time. And the extent (and therefore the concentration) of this delivery depends on the formulation. From a rather theoretical approach based on the polarity of the active ingredient, the stratum corneum and the oil phase, the Relative Polarity Index was established that enables the selection of a suitable emollient for ensuring skin penetration of the active ingredient. Practical examples subsequently show the validity of this approach that demonstrates that one can regulate the delivery of an active molecule (and therefore the efficacy of a cosmetic formulation) by selection and control of the emollient system. Cosmetic formulations are generally quite complex mixtures and subsequent experiments using different emulsifier systems indicated that this component of a cosmetic formulation could also have an impact on steering the active ingredient to the right layer of the skin, although it is too early to be able to derive general rules from this.     

Future developments in cosmetic formulations

In recent years, there have been a great deal of interest in applications of microemulsions, liposomes (vesicles) and multiple emulsions in cosmetic formulations. These systems will provide the cosmetic industry with new types of formulations which are easier to apply, better functional benefit and potentially safer formulations. Microemulsions are thermodynamically stable systems and hence shelf life is no problem. Many cosmetic ingredients can be adequately solubilized in the swollen micelles of the microemulsions. Such solubilized systems may enhance transport and diffusion through various barriers, eg., the skin, thus enhancing the efficacy of the formulations. However, microemulsions may cause skin irritation by disrupting the liquid crystalline structure of the stratum corneum. This problem may be overcome by formulating microemulsions, which on evaporation produce lamellar liquid crystalline structures. The problem of skin irritation is certainly reduced or eliminated using liposomes or vesicles, which offer an alternative to microemulsions. The principles for formation and stabilization of vesicles are discussed in this paper and research work is needed to produce nanocapsules from liposomes, using polymerizable surfactants. Multiple emulsions of the water/oil/water (w/o/w) or oil/water/oil (o/w/o) types are also valuable systems for formulating cosmetics. In the first place, they offer a means of sustained release of the various ingredients. Secondly, they allow one to separate the various ingredients in the formulation, thus preventing their possible interaction. The basic principles required for preparation of stable multiple emulsions are summarised. Developments of polymeric surfactants led to the formulation of stable multiple emulsions. An example of a recently formulated stable w/o/w multiple emulsion is given in this paper The stability of the system was investigated using optical microscopy. Creaming occurred on storage, particularly at high temperature (40°C) and this was significantly reduced by addition of Kelzan (a polysaccharide with high molecular weight). The final formulation was studied rheological techniques.     

Dispersions of lamellar phases of non-ionic lipids in cosmetic products

Although aqueous dispersions of lipids in the form of particles having a lamellar structure (liposomes) are already known as excellent vehicles for pharmaceutical substances, their usefulness in cosmetic formulations has not been demonstrated. The present work shows the advantages obtained by application of such systems to the skin, and in particular the use of non-ionic lipids in aqueous dispersions. Thus, in comparison with classical formulations such as emulsions, these systems exhibit lower toxicity and permit closer control of the availability of active substances at the stratum corneum. As examples, compositions suitable for skin moisturising and for tanning products are presented. Dispersions de phases lamellaires de lipides non-ioniques en cosmétique     

J. Cosmet. Sci., 59, 217–224 (May/June 2008) In vivo spectrophotometric evaluation of skin barrier recovery after topical application of soybean phytosterols

The skin's uppermost thin layer, stratum corneum, plays a crucial role in protecting the body against unwanted influences from the environment. Disruption of the stratum corneum, by tape stripping or chemical injury, results in epidermal recovery of the skin barrier. Soy phytosterols are widely used in the cosmetic field as active ingredients in creams and lipsticks. Furthermore, they deserve an important place among nutracosmeceuticals; in fact, after their absorption from the diet they are transferred from the plasma to the skin, playing an important role in the constitution of skin surface lipids. The aim of the present work was to study the effect of the topical application of soybean phytosterols on skin barrier recovery in human volunteers using the extent of methyl nicotinate (MN)-induced erythema in damaged skin as a parameter to evaluate the rate of stratum corneum recovery. MN was chosen as an erythematogenous substance for its capability to cause an erythema whose intensity and duration are proportional to the quantity of the substance that has entered the living epidermis over time. MN-induced erythema was monitored using reflectance spectrophotometry as a non-invasive instrumental technique. The results show clearly that soy phytosterols exert positive results on skin repair; in fact, 3 days after tape stripping, the sites treated with a formulation containing phytosterols showed an appreciable recovery of barrier function compared to those treated with a vehicle control without soy phytosterols.     

A review on the extensive skin benefits of mineral oil

This review was initially prepared in 2011 before Professor Johann Wiechers tragically passed away. It has been updated and is being published in his memory. It discusses the importance of mineral oil and its benefits to skin. Its source, structure, properties and efficacy are discussed. Mineral oil has been shown to improve skin softness and barrier function better than some other emollients using the gas‐bearing dynamometer and standard water vapour transmission testing as well as in vivo studies showing its effects on suppressing transepidermal water loss (TEWL). It has also been subjected to the rigour of the newer in vivo confocal microscopic measurements now used for testing the performance of moisturizers by following the swelling characteristics of the stratum corneum and been found favourable compared with many vegetable oils. Its introduction as a cosmetic oil was in the late 1800s, and still today, it is used as one of the main components of moisturizers, a true testament to its cost to efficacy window. Naturally, it has physical effects on the stratum corneum, but it is expected that these will translate into biological effects simply through its mechanism of hydrating and occluding the stratum corneum from which many benefits are derived.     

Skin barrier structure, function and formation - learning from cryo-electron microscopy of vitreous, fully hydrated native human epidermis

The hydration level of the stratum corneum largely depends on the physical state and molecular organization of the stratum corneum intercellular lipid matrix. A better understanding of stratum corneum lipid organization may thus aid the development of more rational cosmetic formulations. Several experimental and theoretical problems of a fundamental character remain, however, unresolved. These are, e.g. that precise quantitative skin barrier compositional data are difficult to obtain and that in vitro experimental skin barrier models usually are of limited value because of the prevailing non-equilibrium conditions in vivo. However, new experimental methods have recently been developed, which may help to overcome some of these limitations. These are, e.g. direct electrospray ionization-mass spectrometry (ESI-MS) of intact long-chain skin ceramides and direct high-resolution cryo-transmission electron microscopy (cryo-TEM) of vitreous sections of native, fully hydrated epidermis. Here, we show that cryo-transmission electron micrographs of vitreous normal human skin often dramatically differ from those obtained by conventional electron microscopy of resin-embedded skin. Our epidermal cryo-TEM data are subsequently discussed in relation to central problems of present conceptions of skin barrier structure, function and formation.     

Investigation of interactions between silicones and stratum corneum lipids

Ingredients of topically applied skin care formulations have not only positive effects on the appearance of human skin but can also disturb the Stratum corneum (SC) lipid barrier. In the present study, the influence of silicones (PDMS), as often used cosmetic ingredients, on the microstructure of SC lipids was investigated. For this purpose the interactions of four different PDMS with excised human SC were examined first using differential scanning calorimetry (DSC) and wide angle X-ray diffraction for physical characterization. Because the physical properties of human stratum corneum strongly depend on the lipid composition, showing inter-and intra-individual differences, the interactions with an in vitro model lipid system containing SC fatty acids were also studied, using polarized light microscopy, transmission electron microscopy, small angle X-ray diffraction and DSC. The results revealed that the investigated PDMS do not change either the microstructure of excised human SC or the biphasic lamellar/inverse hexagonal structure of the in vitro model. We concluded that PDMS will not cause any side-effects when topically applied and that our simplified in vitro model could be helpful for estimating interactions between cosmetic ingredients and other topically applied substances and the skin barrier at an early moment of formulation development.     

Role of proteins in cosmetics

The review critically evaluates the safety and effectiveness of proteins or peptides in cosmetic formulations designed for skin and hair care. Special attention is paid to soluble collagen and the use of this fibrillar protein in combination with detergents.
The authors indicate various misconceptions on the mechanism of the effect of collagen and possible risks if the formulations containing collagen are not used on intact skin. Besides the moisturizing effect of collagen on the stratum corneum, the formation of a protective layer on the surface of the skin or hair, and the formation of complexes with detergents and the mechanisms of the effectiveness of this protein are considered.
Le role des proteines dans les cosmetiques     

Human skin equivalents are an excellent tool to study the effect of moisturizers on the water distribution in the stratum corneum

In this study, the mode of action of moisturizers on the level of water in the stratum corneum was studied using cryo‐scanning electron microscopy. As model for dry skin, we used human skin equivalents (HSEs) generated at 93% or 60% relative humidity (RH). During the generation of the HSEs, the moisturizers were applied during a period of maximal 2 weeks. In HSEs generated under normal culture conditions (93% RH), application of 10% glycerol or 5% urea formulations resulted in increased water levels. Whereas the 5% urea formulations resulted mainly in the formation of intercellular water domains, after 10% glycerol both swelling of corneocytes and formation of intercellular water domains were noticed. A reduction in RH to 60% during treatment reduced the stratum corneum water levels drastically. Treatment with the non‐occlusive lipophilic moisturizer isopropyl isostearate resulted in increased water level in the central part of the stratum corneum compared with the untreated control. Our results show that HSEs can be used as a model to study the water distribution.     

Original semiologic standardized evaluation of stratum corneum hydration by Diagnoskin stripping sample

In a normal and healthy skin, the regular elimination of the superficial corneocytes, called desquamation, is a fundamental physiologic process intended to protect the barrier function of the skin. This invisible loss of corneocytes, individually or in small groups, is incessantly compensated by the divisions of the proliferative layer and the upward cellular maturation in order to maintain the harmonious renewal of the epidermis and the integrity of the stratum corneum. The harmony of this desquamation process is intimately conditioned by a sufficient hydration of the stratum corneum: (i) an abnormal desquamation leads to a disruption of the water barrier function and consequently to a dehydration tendency of the stratum corneum, and (ii) a cutaneous dryness (whatever the cause) is able to disturb the desquamation process. Protecting the water content of the stratum corneum has always been a major preoccupation of the cosmetic industry scientists. Consequently, the moisturizing properties of a cosmetic product are objectively measured by various explorations directly targeted on the hydration (corneometry) and on the level of the water barrier function (transepidermal water loss (TEWL) measurements), which depends directly on the skin hydration state. This intimate linkage of the desquamation process and the water content of the stratum corneum enable us to suggest an indirect assessment of the hydration from a direct study of the desquamation by examining a skin-stripping sample (D-Squames) by an optical microscope (linked to a computer). We will describe this already known technique and mainly its new and unpublished semiologic exploitation, named Diagnoskin, whose advantages are its simplicity and its reproducibility particularly interesting in the case of sequential appraisal of dermatologic or cosmetic treatments.     

In vivo methods for the analysis of the penetration of topically applied substances in and through the skin barrier

The efficacy of a drug is characterized by its action mechanism and its ability to pass the skin barrier. In this article, different methods are discussed, which permit this penetration process to be analysed non‐invasively. Providing qualitative and quantitative information, tape stripping is one of the oldest procedures for penetration studies. Although single cell layers of corneocytes are removed from the skin surface, this procedure is considered as non‐invasive and is applicable exclusively to the stratum corneum. Recently, optical and spectroscopic methods have been used to investigate the penetration process. Fluorescence‐labelled drugs can be easily detected in the skin by laser scanning microscopy. This method has the disadvantage that the dye labelling changes the molecular structures of the drug and consequently might influence the penetration properties. The penetration process of non‐fluorescent substances can be analysed by Raman spectroscopy, electron paramagnetic resonance, CARS and multiphoton microscopic measurements. Using these methods, the concentration of the topically applied formulations in different depths of the stratum corneum can be detected by moving the laser focus from the skin surface deeper into the stratum corneum. The advantages and disadvantages of these methods will be discussed in this article.     

Rational formulation design

To be efficacious and to satisfy the requirements for claim substantiation, a cosmetic formulation must achieve effective targeting of an active in the skin. Although the basic principles governing the skin permeation and disposition of molecules have been known for many years, attention has been far less focused on the role of the vehicle, particularly at cosmetically relevant doses. In this article, we discuss the necessity to understand the fate of the formulation components as well as the active once applied onto skin. Recent data confirm that the residence time of the formulation constituents can have a profound impact on the fate of the active. Approaches to identify the ideal vehicle for skin delivery are considered critically, specifically the recent work on ‘formulating for efficacy’ (FFE) by the late Johann Wiechers. Essentially, FFE aims to match the active with the optimal vehicle for skin delivery based on matching polarity/solubility values of the trinity of skin, active and vehicle. The emerging importance of techniques that provide insight to how the vehicle distributes in and on skin is highlighted.     

Lipid nanoparticles (SLN & NLC) for delivery of vitamin E: a comprehensive review

The antioxidative and photoprotective properties of vitamin E have caused it to be included as an active agent in various pharmaceutical and cosmetic products. However, its lipophilicity, chemical instability and poor skin penetration have limited the effectiveness of these formulations. For that reason, many attempts to include it in different drug delivery systems have been made. In recent decades, lipid nanoparticles have received special attention due to their advantages of compatibility with the skin, ability to enhance penetration of drugs in the stratum corneum, protection of the encapsulated substance against degradation induced by the external medium and control of drug release. This work reviews the current status of the encapsulation of vitamin E in lipid nanoparticles. We describe the most important methods for obtaining and characterizing lipid nanoparticles containing vitamin E (LNP‐VE), various techniques for the evaluation of vitamin E's properties after encapsulation, the main in vitro and in vivo studies of the potential effectiveness or toxicity of LNP‐VE, the formulations and stability studies of this delivery system, the commercial products based on LNP‐VE and the regulatory aspects related to lipid nanoparticles. Finally, we discuss the most relevant advantages of encapsulating vitamin E in such particles and critical aspects that still demand attention to enhance the potential of solid lipid nanoparticles to deliver vitamin E.     

In vitro and in vivo assessment of the effect of Laurus novocanariensis oil and essential oil in human skin

Laurus novocanariensis is an endemic plant from the Madeira Island forest that derives a fatty oil, with a strong spicy odour, from its berries that has been used for centuries in traditional medicine to treat skin ailments. This work aimed to investigate the effect of the application of both the oil and its essential oil on normal skin, to assess their safety and potential benefits. Diffusion studies with Franz cells using human epidermal membranes were conducted. The steady‐state fluxes of two model molecules through untreated skin were compared with those obtained after a 2‐h pre‐treatment with either the oil or the essential oil. Additionally, eleven volunteers participated in the in vivo study that was conducted on the forearm and involved daily application of the oil for 5 days. Measurements were performed every day in the treated site with bioengineering methods that measure erythema, irritation and loss of barrier function. Slightly higher steady‐state fluxes were observed for both the lipophilic and the hydrophilic molecule when the epidermal membranes were pre‐treated. Nevertheless, such differences had no statistical significance, which seems to confirm that neither the oil nor the essential oil impaired the epidermal barrier. Results collected with the Chromameter, the Laser Doppler Flowmeter and the visual scoring are in agreement with those established in the in vitro study. They indicate that the repeated application of the oil did not cause erythema, because the results observed in the first day of the study were maintained throughout the week. Application of the oil did not affect the skin barrier function, because the transepidermal water loss remained constant throughout the study. The stratum corneum hydration was slightly reduced on days 4 and 5. This work shows that both the oil and the essential oil were well tolerated by the skin and did not cause significant barrier impairment or irritation.     

Seasonal influences on stratum corneum ceramide 1 fatty acids and the influence of topical essential fatty acids

Ceramide 1 is the main repository of stratum corneum linoleic acid, and changes in the levels of ceramide 1 linoleate are associated with cutaneous abnormalities: essential fatty acid deficiency, atopic dermatitis and acne. We have previously reported seasonal variation in stratum corneum lipids where lipids, particularly ceramides, are very much reduced in winter which probably influences the appearance of skin xerosis in this season. However, investigations into the seasonal variation in ceramide 1 fatty acid levels and the effects of topical essential fatty acids on the composition of these lipid species have not been conducted in healthy subjects. We determined the composition of stratum corneum ceramide 1 esterified fatty acids in leg skin from Caucasian women in winter and summer and also investigated the effects of topical triglycerides rich in linoleic acid on stratum corneum ceramide 1 fatty acid levels in winter. A dramatic seasonal variation was observed. Whereas decreased levels of linoleic acid and some saturated fatty acids (C-15 and C-24) were observed in the winter months, increased levels of oleic acid were found. Nevertheless, it was possible to normalize the levels of stratum corneum ceramide 1 linoleate following topical application of formulations containing linoleic acid-rich triglycerides for 4 weeks. The reduction in ceramide 1 linoleate may lead to impaired stratum corneum function and increased susceptibility to skin xerosis in winter. However, by using formulations containing linoleic acid esters it is possible to correct these changes and possibly improve barrier resilience.     

Renewable sources: applications in personal care formulations

A global tendency for products considered environmentally sustainable, and ecologically obtained led the industry related to personal care formulations to fund the research and the development of personal care/cosmetics containing ingredients from natural resources. Furthermore, consumers are aware of environmental and sustainability issueans, thus not harming the environment represents a key consideration when developing a new cosmetic ingredient. In this study we review some examples of active ingredients or raw materials used in cosmetics/personal care/biomedical products that are coming from either through biotechnological systems, or as byproducts of several industries. A skin formulation containing biosynthetic actives, prepared by us and the study regarding its dermocosmetic properties are also described. The need for the standardization processes, the safety assessment tools, the improvement of the exploitation methods of these renewable sources in order the production to be ecologically and economically better are also discussed.     

Human synthetic sebum formulation and stability under conditions of use and storage

The human skin surface and hair are generally coated with a thin film of liquid phase sebaceous lipids. This surface lipid film contributes to the cosmetic properties of the skin. Synthetic sebum has been used for studies on properties of skin and hair. However, there has been no standardized formulation of synthetic sebum and many of the synthetic sebum formulations that have been used do not closely resemble actual sebum. In this study, a formulation for a standardized and inexpensive synthetic sebum is proposed, and the chemical stability of this lipid mixture is demonstrated under conditions of use and storage. The proposed synthetic sebum consists of 17% fatty acid, 44.7% triglyceride, 25% wax monoester (jojoba oil) and 12.4% squalene. This lipid mixture takes up approximately 6% of its weight in water when equilibrated in an atmosphere saturated with water vapour. It is stable on exposure to the atmosphere at 32°C for at least 48 h, and it is also stable on storage at 4 or −20°C, either dry or in chloroform : methanol solution for at least 6 months. This synthetic sebum could be useful in studies on cosmetic properties of the skin surface or hair, on penetration of chemicals into the skin or in development of standardized tests of laundry detergent performance.     

J. Cosmet. Sci., 59, 385–398 (September/October 2008)Evaluation of liposomal and conventional formulations of octyl methoxycinnamate on human percutaneous absorption using the stripping method

The objective of this study was to determine the influence of vehicles on the penetration of octyl methoxycinnamate (OMC), as a UV absorber, to the stratum corneum by the stripping method. The experimental formulations consisted of a conventional o/w emulsion and multilamellar and small unilamellar liposomes (MLVs and SUVs) containing OMC. MLVs containing OMC were prepared by the fusion method and then converted to SUVs by probe sonication. Various formulations were then applied onto the midvolar forearms of six volunteers at a dose of 2 mg cm^-2. After determined timepoints, the stripping method was conducted whereby 22 tape strips were applied and subsequently divided into different stripping groups. The sunscreen agent was assessed by HPLC while the SPF (sun protection factor) of the formulations was determined in human volunteers in accordance with the Australian standard. Overall the results indicate that skin accumulation of OMC in MLVs was significantly greater than in the o/w emulsion and SUVs. Furthermore, SUV's penetration into the deeper skin layers was significantly greater than MLV's and that of a conventional o/w emulsion. Also, higher amounts of OMC were recovered from the upper layers of the stratum corneum than from the deeper layers in all the formulations tested. Finally, the SPF of the liposomes containing OMC was slightly greater than that of the control lotions at a similar concentration of OMC. In conclusion, the result of this study indicates that an MLV prepared by the fusion method could be a better vehicle for OMC as a sunscreen since it has a slightly better SPF compared to a conventional formulation and more remains in the stratum corneum, reducing its penetration to the deeper layers.
Address all correspondence to M. R. Jaafari.     

Exfoliation for sensitive skin with neutralized salicylic acid?

Citation: IFSCC Magazine, 11 (2008) (2) 115–119 Abstract: Hydroxy acids enjoy tremendous interest in cosmetology thanks to their skin anti‐ageing and water barrier enhancing activities. One of their actions is to promote the natural stratum corneum (stratum corneum) desquamatory process. However, their use is limited due to an inherent pH‐related irritancy potential which is even more exacerbated on sensitive skin. Clearly there is an opportunity for improvement. In this research we evaluated in humans the efficacy of salicylic acid, and its salts, as a corneodesmolytic agent using the dihydroxyacetone method and measuring the reduction in skin staining with treatment over time using a chromameter. Salicylic acid at 2% in a preparation of pH 3.12 significantly increased exfoliation by 10.9% compared with placebo (P < 0.05), confirming its desquamatory enhancing properties. Then the effect of vehicle cream pH on salicylic acid activity was studied. Salicylic acid at close to neutral pH (mostly in its neutralized form as salicylate, pH 6.50) exerted a corneodesmolytic activity as good as that of salicylic acid in an acidic vehicle (pH 3.12) after only two days of application. Furthermore, the performance of glycolic acid and salicylic acid salts as exfoliants were compared at pH 6.50. When these two hydroxyl acids were formulated at the same molar level in a cosmetic base (14.47mmol L^‐1), the salicylic acid preparation gave an 8.2% increase in stratum corneum desquamation compared with the glycolic acid preparation (P < 0.05). The corneodesmolytic effects were confirmed using a tape‐stripping assay combined with a quantitative protein assay. Neutralized salicylic acid was found to enhance the removal of stratum corneum proteins significantly more than the vehicle after 25 sequential tape strippings (14%; P < 0.05). Finally, salicylic acid had no significant influence on skin water barrier properties after 22 days of treatment. In the second phase of this research we assessed the suitability of neutralized salicylic acid as an ingredient for sensitive skin. A stinging test was performed according to the Frosch & Kligman method to evaluate the influence of the formulation base‐pH on stinging potential. Salicylic acid formulated at pH 6.50 induced no stinging sensation (score 0) in contrast to salicylic acid at pH 3.12 (score 19; P < 0.05). In addition, a clinical study was conducted to assess the erythema induced on volunteers’ cheeks after a single application of a neutralized salicylic acid (1%) formulation compared with placebo. Visual redness was assessed by a dermatologist and then measured with a Mexameter. No significant differences were observed. Moreover, half of the panel had sensitive skin and no correlation could be established between redness and/or abnormal sensation and sensitive skin. In conclusion, neutralized salicylic acid at a 1% concentration is a suitable exfoliant agent for subjects with sensitive skin. Keywords: exfoliation, hydroxy acids, Salicylic acid, stratum corneum Paper presented at the IFSCC Conference 2007, Amsterdam, The Netherlands.