Pentacyclic triterpenes from Terminalia arjuna show multiple benefits on aged and dry skin.

BACKGROUND: Pentacyclic triterpenoids improve epidermal barrier function and induce collagen production. Here, their effects on cutaneous aging by means of objective instrumental measurements were elucidated. METHODS: Reconstituted human epidermis, cultivated keratinocytes and fibroblasts were incubated with Terminalia arjuna triterpenes (T. arjuna bark extract), and mRNA and protein expression of various genes was determined using microarray analysis, qRT-PCR and ELISA techniques. Clinical efficacy of T. arjuna bark extract versus vehicle control cream was elucidated in 30 patients and transepidermal water loss (TEWL), skin hydration and elasticity were measured. Another 30 female patients in their postmenopausal phase were treated with a similar regime, and skin sebum content, cutaneous blood microcirculation and skin density/echogenicity were assessed. RESULTS: Incubation with T. arjuna triterpenes increased FGF-2, TSP-1, TGF-ß and CTGF expression, and VEGF secretion in vitro. Elevated lactate dehydrogenase release upon sodium dodecyl sulphate challenge was reversed by the application of T. arjuna bark extract. T. arjuna bark extract decreased TEWL, improved skin moisturization, reduced scaliness and led to significantly improved skin elasticity. Also, increases in blood microflow and skin sebum content as well as improved skin thickness/echogenicity were noted on postmenopausal skin, resulting in visible reduction of sagging skin on the jowls as demonstrated by digital photography. CONCLUSION: T. arjuna bark extract appears as an innovative active ingredient that exerts versatile antiaging properties in vitro and in vivo.

Methods for the assessment of the efficacy of products and slimming treatments for cellulite according to the Italian Interdisciplinary Group for the standardization of efficacy tests on cosmetic products.

Cellulite is a very common skin alteration with a complex pathogenesis; different degrees of severity of cellulite can be observed in most part of people after puberty, and numerous cosmetic or more invasive treatments have been proposed, with variable efficacy. Since reproducible methods of evaluation of the effectiveness of cellulite treatments are lacking, the purpose of our group was to define and set general testing principles for evaluating the efficacy of slimming products and treatments/remodeling methods for cellulite, to achieve a delineation of reliable and reproducible research steps following a well-designed and scientifically valid methodology. After a careful review of literature and textbooks and according to personal experience, we defined assessment protocols based on clinical and instrumental tools. In order to make studies reliable, reproducible and safe, a protocol standardization is needed. The sponsor is responsible for assuring quality and information concerning the product under investigation; moreover, investigators should be experienced on cellulite evaluation and treatment, and, finally, the duration and modalities of application of the product should be specified. A treated VS non treated area comparison can be performed, to evaluate the severity of cellulite and the clinical outcomes of the treatment. Besides clinical evaluation, instrumental methods should always be implemented to provide objective data for treatment outcome.

Lifting properties of the alkamide fraction from the fruit husks of Zanthoxylum bungeanum.

The fruits of various Zanthoxylum species are used as a spice in the Chinese and Japanese cuisine because of their delicate flavour and tingling properties. The lipophilic hydroxyalkamides hydroxy α- and ß-sanshools (1a,b) have been identified as the tingling principles of these plants, and previous studies have validated a sanshool-rich lipophilic extract from the fruit husks of Z. bungeanum Maxim. (Zanthalene ® ) as an anti-itching cosmetic ingredient. Because tingling is a sort of 'paralytic pungency', and Zanthalene ® potently inhibits synaptic transmission, we have investigated its capacity to relax subcutaneous muscles and act as a topical lifting agent for wrinkles. An anti-wrinkles extract rich in spilanthol (2), a lipophilic alkamide having sensory properties similar to those of Zanthalene ® , was used as a reference. Short-term (lifting effect) and long-term (anti-wrinkle) improvements of skin roughness parameters were evaluated by both objectives' and subjectives' measurements. An immediate 'lifting' effect was observed with the sanshool-rich lipophilic extract, at dosages at which the reference alkamide extract was inactive in the objective assays. Limited desensitization after repeated application and good overall tolerability were observed, although a modest long-term anti-wrinkle effect was shown by both products. Taken together, these observations validate the use of sanshool-rich lipophilic extracts as an efficacious, immediate-action lifting agent, and exemplify the relevance of sensory observations to foster the development of innovative cosmetic ingredients.

Investigation of new vehicles to patch test corticosteroids: our experience with ethoxydiglycol to detect contact allergy to hydrocortisone butyrate.

Topical corticosteroids (CS) are widely used in dermatology because of their anti-inflammatory, anti-proliferative and immuno-suppressive properties. On the other hand, the prolonged application of corticosteroids may induce adverse reactions such as allergic contact dermatitis (ACD). Patch testing CS often poses methodological issues correlated to their drug properties that may hide the manifestations of a positive reaction. Furthermore, the ideal concentration to patch test corticosteroid is still a matter of study and some vehicles have some well-known limitations. This article is divided into two parts: the first one investigated vehicles that may efficiently dissolve the corticosteroids, according to the polarity of the latter; the second part compared the results of the patch tests with hydrocortistone-17-butyrate using two different vehicles: ethanol, which is the standard one, and another vehicle selected as suitable from our CS solubility test.

Allergic and photoallergic contact dermatitis from ketoprofen: evaluation of cross-reactivities by a combination of photopatch testing and computerized conformational analysis.

Allergic contact dermatitis (ACD) and photo-ACD are cell-mediated delayed hypersensitivity reactions of the skin caused by a wide range of substances. Topical ketoprofen (KP), a nonsteroidal anti-inflammatory drug (NSAID), can induce ACD and photo-ACD. Patients with ACD and/or photo-ACD to KP frequently show concomitant sensitization to other substances. The aim of this study was to identify the substances most frequently associated with sensitization to KP, and to evaluate, by means of computerized conformational analysis, whether this association could be due to cross-allergy. 15 subjects with ACD and photo-ACD to KP were tested with the SIDAPA (Società Italiana di Dermatologia Allergologica Professionale ed Ambientale) patch test standard series, including fragrance mix and its components (eugenol, isoeugenol, oak moss, geraniol, hydroxycitronellal, amylcinnamaldehyde, cinnamyl alcohol and cinnamaldehyde) and with the SIDAPA photopatch test series. Allergic reactions to cinnamyl alcohol were noted in all patients, whereas some patients also showed positive reactions to fenticlor, octocrylene and benzophenone-10. Computerized conformational analysis demonstrated that the structure of cinnamyl alcohol is similar to that of KP, whereas the structures of benzophenone-10, octocrylene and fenticlor are completely different. These results suggest that in patients with contact allergy to KP, concomitant positive reactions to cinnamyl alcohol are due to cross-sensitization, whereas simultaneous allergic reactions to fenticlor, octocrylene and benzophenone-10 should be regarded as co-sensitizations.

Aminoethylethanolamine: a new allergen in cosmetics?

Amphoacetates are organic compounds used in many industrial applications and in cosmetic formulations for the skin, hair and mucosa, as surfactants, mild foaming and cleansing agents in concentrations ranging from 0.1% to 50%. Despite the fact that they have been in use for many years, cases of contact allergy to them are extremely rare. We describe 4 patients who developed an eczematous reaction after use of detergents containing amphoacetates. Patch testing showed positive reactions to sodium lauroamphoacetate (Miranol HM Special, Rhodia, England) as is or diluted at decreasing concentration (10%, 5% and 1%) in water and to aminoethylethanolamine (AEE) at the concentration of 1% in various vehicles (ethanol, acetone, and sodium laurylethersulfate 1% aqueous solution) and at decreasing concentrations ranging from 1% to 0.005% in water. AEE is one of the reagents used in the synthesis of amphoacetates. This molecule, that is structurally an ethylenediamine derivative, has sensitizing power and is reported as a cause of occupational contact allergy in cable jointers. Combined eczematous reactions to AEE and sodium lauroamphoacetate can be consequent to the presence of the former substance as an impurity in amphoacetates-containing products, as demonstrated by ion chromatography-mass spectrometry analysis.

Occupational sensitization to epoxy resin and reactive diluents in marble workers.

10 out of 22 marble workers handling a bicomponent resin, based on epoxy resin and ortho-cresyl glycidyl ether (CGE), developed contact dermatitis and airborne contact dermatitis within 20 days to 2 months of exposure. All 22 subjects were patch tested with epoxy resin, bisphenol A, epichlorohydrin and a series of reactive diluents. The 10 symptomatic subjects were all positive to the reactive diluent CGE, and 4 of them also to epoxy resin. The other reactive diluents that gave positive reactions were phenyl glycidyl ether (7 cases), cyclohexanedimethanol glycidyl ether (2 cases), 1,6-hexanediol diglycidyl ether (2 cases) and allyl glycidyl ether (1 case). The findings of our study suggest that allergic potential is directly proportional to the electronic charge available, for all electron-rich molecules (solvents, high and low polymers, reagents) that interact with the glycidyl ether group. Lesser, but still noticeable, effects are detected when activation of the glycidyl group is related to the possible formation of intramolecular hydrogen bonds. In practice, the occupational problem was partly solved by changing the type of glycidyl ether.

Contact allergy to impurities in surfactants: amount, chemical structure and carrier effect in reactions to 3-dimethylaminopropylamine.

Since finding that all subjects with contact allergy to cocamidopropylbetaine give positive reactions to 3-dimethylaminopropylamine (DMPA), we wished to verify whether sensitization to other industrially-used tensioactives might also be due to content of DMPA as an impurity. We also investigated the possible "carrier action" that tensioactives might exert on minimal quantities of DMPA. Finally, we analyzed the relationship between the structure of DMPA and other chemically-correlated molecules and their sensitizing potential, with particular reference to the structure of alkylamidopropylbetaines. For this purpose, in 34 patients with contact allergy to DMPA, we tested: (i) DMPA in concentrations below the threshold limit in water and in different tensioactives; (ii) substances that employ DMPA as a reagent in their synthesis; (iii) substances similar to DMPA as regards chemically reactive groups. The study showed that: (i) DMPA remains as a quantitatively detectable impurity in all tensioactives employing it in their synthesis; (ii) some common anionic (SLES) and non-ionic (polysorbate 20) tensioactives enhance the risk of sensitization from very low doses of DMPA, presumably due to a "carrier effect;" (iii) the sensitizing chemical structures in DMPA and related molecules are the primary amine and the tertiary (dimethyl-substituted) amine groups, when separated by either 2 or 3 carbon atoms; (iv) no sensitizing action can be attributed to the functional groups present in alkylamidopropylbetaine molecules.

3-Dimethylaminopropylamine: a key substance in contact allergy to cocamidopropylbetaine?

In the past year, 1200 consecutive eczematous patients were tested with cocamidopropylbetaine 1% aq. Contact allergy was evinced in 46 subjects (3.8%), while irritant reactions (slight erythema only) were observed in 15 cases (1.25%). 30 out of 46 patients with allergic reactions were subsequently tested with the substances used in the synthesis of cocamidopropylbetaine, together with a sample of cocamidopropylbetaine declared by the supplier to possess a greater purity. In all 30 subjects, positive reactions were obtained to 3-dimethylaminopropylamine (DMPA) 1% aq., while the cocamidopropylbetaine defined of purer grade, at 0.5% and 1% aq., gave positive reactions in 10% and 53% of cases, respectively. These results suggest that the DMPA present at various levels as an impurity in the commercial product is responsible for cocamidopropylbetaine allergy. Owing to the inconsistency of positive reactions to cocamidopropylbetaine of variable purity, and to the consistency of positive reactions to DMPA, it seems likely that these reactions may also be connected with the presence in the product, defined of purer grade, of unknown amounts of DMPA as impurity. Structural similarities between the 2 molecules cannot be considered in this case, because the DMPA structure is radically changed in its transformation to the betaine structure. Further experiments with other molecules related to the above structures are in hand.

Protective effect evaluation of free radical scavengers on UVB induced human cutaneous erythema by skin reflectance spectrophotometry.

Synopsis This paper assesses the suitability of UVB induced skin erythema measured by reflectance spectrophotometry in humans as a model for differentiating topical efficacy of free radical scavengers. Two different formulations (aqueous gels and O/W emulsions) of each active compound (tocopherol, tocopherol acetate, superoxide dismutase (SOD), glutathione, ascorbyl palmitate) were tested on healthy human volunteers before and after skin exposure to UVB radiation. Skin erythema was monitored by calculating erythema index values from the skin spectral data obtained using a reflectance spectrophotometer. The free radical scavengers tested were not able to inhibit UVB induced skin erythema from both formulations when they were topically applied before UVB irradiation. Applying the free radical scavenger formulations after skin exposure to UVB radiation, glutathione and SOD showed the best ability in inhibiting the induced erythema (percentage inhibition 53.3 and 41.6%, respectively from gels). Tocopherol and tocopherol acetate inhibited UVB skin erythema by 27% while ascorbyl palmitate showed a poor efficacy. For all the active compounds tested, no significant difference was observed comparing the results obtained from gels to those from emulsions. Liposomal gel formulations containing the free radical scavengers which showed the best activity (SOD and glutathione) were prepared and topically applied after skin exposure to UVB radiation. SOD and glutathione liposomal formulations were more effective than the corresponding conventional gels. The proposed model, if validated by further studies, could be useful for differentiating the effectiveness of free radical scavengers in inhibiting photoaging due to long-term sunlight skin exposure.

Evaluation of predictable irritative power of surfactant mixtures by human fibroblast culture and its correlation to physico-chemical parameters.

Summary The predictable toxic effects of some surfactants, their blends and some preserving agents on human fibroblast cultures were investigated with in vitro tests, with the aim of finding a possible correlation between the biologic evaluations and some physical characteristics of detergent solutions. Lactate dehydrogenase release into the medium was used as a marker of the plasma membrane integrity, while the amount of (3)H-radiolabelled proteins in the fibroblasts was measured in order to assess the cell biosynthetic machinery function. Disodium-alkyl-semi-sulphosuccinate induced membrane damage in the lactate dehydrogenase test and decreased the protein synthesis, with an EC 50 around 1mm, while sodium lauryl ether sulphate had an EC 50 at about 100 mum, indicating that this compound is ten-fold more toxic, when measured by this method. An ethoxylated glyceride, on the contrary, was completely harmless on the plasma membrane and, surprisingly, activated fibroblast protein synthesis in a dose-response way up to two-fold. Mixtures of the three surfactants evidenced the protective effect of the non-ionic against the cellular functionality damage. Parabens do not influence this type of evaluation, while some influence was shown by the formaldehyde releaser 2-bromo-2-nitro-propandiol at the highest concentration. The comparison between critical micellar concentration measures of the different surfactants and their in vitro detected irritative power shows, for the two anionics, that in vitro toxicity is proportionally bound to the amount of micelles even if the structural differences between the two types of molecules are reflected into different damage values, while the non-ionic compound shows a not defined CMC and a very low toxicity profile. Blends of anionics with the non-ionic show an increased CMC and a reduced toxicity profile. Toxicity evaluations of complex finished foaming formulations, carried out with human fibroblast cultures evaluation show that a relationship between micelles amount and cell toxicity seems to exist, mainly when multiple surfactants blends are tested.

Use and properties of perfluoropolymethyl-isopropylethers in skin and hair cleaning; system stabilization and interference with sebum redistribution on skin and hair.

Synopsis Perfluoro-polymethyl-isopropyl ethers (PPIE) are known for their low surface tension and spreading characteristics. Their use in detergent systems was difficult because of their insolubility in most fluids (except fluorinated ones), but microdispersions have been obtained with surfactant blends, the compositions of which minimize the interfacial tension. Thickeners have been tried in order to improve suspension stability during shelf life. Spreading of PPIE has been studied by using in vivo and in vitro evaluations, both on hair and on skin. Results are discussed, as these systems could improve greasy skin conditions and decrease the rebound effect. Résumé Utilisation et proprietés des polyethers perfluorometyl-isopropyl (PPIE) dans le nettoyage de la peau et le lavage des cheveux Les PPIE sont connus pour leur basse tension superficielle et leur facilité d'étalement. Leur utilisation dans des produits détergents a été difficile du fait de leur insolubilité dans la plupart des fluides (excepté les fluides fluorés), mais on a pu obtenir des microdispersions en mélangeant des tensio-actifs dont la composition minimise la tension interfaciale. Des épaississants ont été essayés pour améliorer la stabilité de la suspension. La facilité d'étalement du PPIE aétéétudiée en effectuant des évaluations in vivo et in vitroà la fois sur des cheveaux et sur la peau. Les résultats sont discutés, puisque ces systèmes permettraient d'améliorer la condition des cheveux gras et de diminuer les effets secondaires.