J. Cosmet. Sci., 68, 126–132 ( January/February 2017) 126 Unique natural exopolysaccharides for biomimetic protective effect against urban pollution MAGALI BOREL, ELISABETH LAMARQUE, and ESTELLE LOING, Lucas Meyer Cosmetics, IFF, ZA les Belles Fontaines, 91160 Champlan (M.B.), Lucas Meyer Cosmetics, IFF, Bioparc, 31036 Toulouse Cedex 1 (E.L.), France, and Lucas Meyer Cosmetics, IFF, Tour de la Cité, 2600, Québec, Canada (E.L.). Summary Through natural selection, living organisms have evolved well-adapted survival strategies over time. The shallow salt waters of Moorea lagoon are the site of accumulation of microbial mats called “Kopara,” in the native Polynesian language. This unique ecosystem is rich in fi lm-forming exopolysaccharides (EPSs) secreted by microorganisms within the biofi lm, as a mean to protect themselves from environmental stress (strong ultraviolet [UV], pH, salinity … ). Using blue biotechnology, a manufacturing process was developed to obtain an EPS with skin benefi ts. The active ingredient (EPS-229) protects against urban pollution, including free radicals, heavy metals, hydrocarbons, and PM2.5 (particulate matter with a size lower than 2.5 μm). Methods: The anti-lipid peroxidation action of EPS-229 was studied in an in vitro UVB-irradiated keratinocyte culture model, using lipophilic fl uorescent probe. The chelating properties of EPS-229 were evaluated in tubo in the presence of cadmium and lead. The protective effect of EPS-229 on pollution-exposed skin explants was investigated through quantifi cation of released malondialdehyde (MDA) and histological observation of skin morphology using optical microscopy. Clinical evaluation of the protective and cleansing effi cacy of a water solution containing EPS-229 (0.02% and 0.01% w/v, respectively) was performed, against placebo, on a panel of 18 volunteers. For these studies, the forearms of volunteers were treated with EPS-229 before (anti- adhesion affect) or after (cleansing effect) application of PM2.5 (iron particles of 1 μm). The presence of skin- adherent particles was observed and quantifi ed by image analysis, using specifi c digital masks. Results: In vitro, EPS-229 signifi cantly protected keratinocyte cell membranes from lipid peroxidation. A decrease of 28% was achieved when a concentration of 0.001% w/v EPS-229 was applied to the cell culture. In tubo, EPS-229 also presented strong chelating properties. Maximal adsorption was estimated at 154 mg/g (1.37 mmol/g) of EPS-299 for cadmium and at 250 mg/g (1.21 mmol/g) of EPS-229 for lead. In the skin explant model of pollution exposure, EPS-229 (0.03% w/v) reduced MDA production by 44%, preserved cell integrity, improved dermal–epidermal cohesion, and normalized the collagen network. In vivo, treatment of skin with EPS-229 before exposure to PM2.5 created a protective fi lm limiting particle adhesion. When used in a cleansing solution after exposure to PM2.5, EPS-229 formed a mesh that entrapped particles and removed them from the skin surface. Conclusion: Inspired by the French Polynesia Kopara unique ecosystem, a bioactive exopolysaccharide (EPS-229) has been developed that offers protection from environmental aggression. As a biomimetic shield at the surface of the skin, EPS-229 provides an immediate multiprotective action that effi ciently fi ghts the harmful effects of urban pollution and smog. Address all correspondence to Estelle Loing at estelle.loing@lucasmeyercosmetics.com.

EXOPOLYSACCHARIDES FOR BIOMIMETIC PROTECTIVE EFFECT 127 INTRODUCTION In French Polynesia, the shallow salt waters of lagoons are the site of accumulation of organic matter of microbial origin that forms mats called “Kopara” in the native Polynesian language (1). Under stress conditions, such as fl uctuations in temperature, water supply, pH, and salinity, microorganisms within the Kopara release exopolysaccharides (EPSs) as means of protection (2). EPS consist of high-molecular-weight carbohydrates that vary greatly in their sugar composition which impacts their chemical and physical properties (3). EPSs improve bacterial survival in many ways. They serve as a transport barrier to reactive chemicals. They can trap trace metals, thus reducing their toxicity (4). They strongly hold and redistribute water to prevent extreme cell dryness. They buffer against sudden osmotic changes. They contribute to oxidative defense (4). They also protect bac- terial cells form ultraviolet (UV) induced damage by their capacity to absorb UV rays (5). Several EPS found in the Kopara have been studied for skin care applications. Inspired by their natural shield function, research was oriented, early on, toward applications related to the protection of skin from environmental stress. One of these EPS (EPS-229) proved to be most promising in vitro, and was further characterized ex vivo and in vivo. EPS-229 is a highly ramifi ed polysaccharide produced by Alteromonas macleodii living in the Kopara. However, the coral reef and lagoon ecosystems being extremely fragile, it was primordial to develop ways of exploiting the properties of this EPS without compromising its natu- ral habitat. For industrial scale production, fermentation protocols were developed in collaboration with a local Polynesian biotech. Fermentation conditions, such as pH, tem- perature, oxygen concentration, agitation as well as the composition of the culture media, had to be carefully optimized to reproduce natural bacterial growth conditions. The cur- rent paper presents a brief summary of the research supporting the use of EPS-229 as an antipollution skin care ingredient, including new clinical data providing evidence of protection against PM2.5 particles (particulate matter with a size lower than 2.5 μm). EXPERIMENTAL ALTEROMONAS FERMENT EXTRACT (EPS-229) This extract (EPS-229) is a highly ramifi ed EPS produced by A. macleodii, as part of a protective shield against environmental aggressions. It is composed of neutral sugars (57%), uronic acids (25%), and sulfates (8%), presents a slight white color, and has a molecular weight of 1000 kDa. The bioactive EPS is obtained through biotechnology, using a fer- mentation process reproducing natural synthesis conditions. PROTECTION OF KERATINOCYTES FROM FREE RADICAL-INDUCED DAMAGES Confl uent human keratinocytes (NHEK) were incubated for 24 h with the test product EPS-229 (0.001% w/v) or butylated hydroxy anisole (BHA) (50 μM), a synthetic antioxi- dant used as a positive control. A C11-fl uor probe was then introduced into the culture media and unbound probe was removed by washing, 45 min later. Following reintroduc- tion of EPS-229 or BHA into the media, cells were challenged by exposure to UVB (201 mJ/cm2). Cells were further cultured for 1 h, then rinsed and trypsinized. Fluorescence of