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HYDRANOV A New Look at Hydration HYDRANOV A New Look at Hydration COSMOS EPIDERMIS SYNTHESIZES 4 TIMES MORE HYALURONIC ACID THAN THE DERMIS. Epidermal hyaluronic acid: a still overlooked potential. The rate of hyaluronic synthesis is higher in the epidermis than in the dermis. Since the dermis is much thicker than the epidermis, it contains 4 to 9 times more hyaluronic acid, but for equivalent tissue quantities, the epidermis synthesises 4 times more hyaluronic acid than the dermis (1). In the epidermis hyaluronic acid is localised in the intercellular space of the basal and spinous layers. In the same way as in the dermis the hygroscopic properties of hyaluronic acid are of great importance in hydrating the deep layers of the epidermis, but its function goes further than conventional hydration. It has been shown that the presence of high concentrations of hyaluronic acid in the intercellular space tends to renew desmosomes and tight junctions, and so reorganise the keratinocytes not only within the basal layer, but also throughout the process of differentiation as far as the stratum corneum (2). Hyaluronic acid acts therefore like an overall epidermal restructuring agent by initiating and maintaining constant cellular remodelling from the basal layer to the cornified layer. « HYALURONIC LIKE », HYDRANOV HAS HYGROSCOPIC PROPERTIES ALLOWING THE CAPTURE OF WATER MOLECULES AT THE SKIN SURFACE. « EPIDERMAL HYALURONIC ACTIVATOR », IT ALSO STIMULATES THE SYNTHESIS OF EPIDERMAL HYALURONIC ACID FOR AN OVERALL EPIDERMAL RESTRUCTING AND HYDRATING ACTION. HYDRANOV : High Tech Concentrate of Oligofurcellaran The oligofurcellaran is obtained by the depolymerisation of a sulfated furcellaran (galactose and anhydrous galactose) coming from the red alga Furcellaria lumbricalis, widespread in Scandinavian waters. Depolymerization is made using supercritical CO2 technology in anhydrous conditions. After depolymerization, the oligofurcellaran is rehydrated with an enriched sea salt solution. This final intake of marine nutrients, and mainly potassium, induces a double helicale reorganization of the polymer that gives it hygroscopic and gelling properties comparable to hyaluronic acid. The resulting oligofurcellaran has a molecular weight >200kDa. This depolymerisation method, water and acid free, complies with the policy of energetic resources managment developped by Codif R&N. HYDRANOV CHRONOLOGICAL ACTION 1 ‐ « hyaluronic like » action at the surface of the skin 2 ‐ « hyaluronic activator » action : stimulates the synthesis of epidermal hyaluronic acid 3 ‐ Reinforcement of tight junctions – New target : Claudin 1 4 ‐ Stimulation of ceramids synthesis 5 ‐ Reinforcement of NMF – New target : Filaggrin “HYALURONIC LIKE” ACTION OF HYDRANOV IN‐VITRO TEST The oligofurcellaran concentrated in Hydranov has high molecular weight. Therefore in the same way as hyaluronic acid it acts as a macromolecule with surface properties. PROTOCOL: Pure samples of Hydranov and hyaluronic acid are put in specific cells with a relative humidity that will vary from 0 to 95%. For each condition of relative humidity, the polymers capture water molecules to reach a maximum called « gain mass at sorption equilibrium ». The value of gain mass reflects the ability of each polymer to trap water molecules. 120 ‐ Hyaluronic Acid mass gain % at equilibrium 100 ‐ Hydranov 80 ‐ 60 ‐ 40 ‐ 40 60 % of relative humidity ‐ 20 ‐ 0 ‐ ‐ 0 ‐ 20 ‐ 80 100 RESULT: Hydranov is able to trap atmospheric water molecules following a kinetic totally comparable to hyaluronic acid. The more atmosphere is saturated with water, the more Hydranov traps water molecules. For 95% of relative humidity, 86% mass gain is measured. HYDRANOV IS CAPABLE TO TRAP A QUANTITY OF WATER ALMOST EQUAL TO ITS INITIAL MASS AND THUS MAINTAINS A MOISTURE FILM ON THE SURFACE OF THE SKIN. “HYALURONIC ACTIVATOR” ACTION OF HYDRANOV Given the size of the oligofurcellaran and the low probability that it can penetrate into the epidermis, the effect of Hydranov has been evaluated by topical application on human skin explants or human reconstructed epidermis. HYDRANOV INCREASES THE EXPRESSION OF HYALURONAN SYNTHASE 3 PROTOCOL: Reconstructed human epidermis ‐ Topical application of 1% Hydranov for 72 hours. Gene expression analysis using RT‐qPCR. RESULT: +64% HYALURONAN SYNTHASE 3 180 ‐ 160 ‐ 140 ‐ +64% 120 ‐ 100 ‐ 80 ‐ 60 ‐ 40 ‐ 20 ‐ 0 ‐ Hyaluronan synthase 3 is an enzyme involved in hyaluronic acid synthesis. expression % compared to untreated control EX‐VIVO TEST Control 1% Hydranov HYDRANOV INCREASES THE SYNTHESIS OF EPIDERMAL HYALURONIC ACID EX‐VIVO TEST PROTOCOL: Human skin explants ‐ Topical application of 1.5% Hydranov for 9 days. Quantification of epidermal hyaluronic acid synthesis using immuno‐labelling (fuchsia colour). RESULT: +211%** HYALURONIC ACID X3 : TOTAL SURFACE AREA OCCUPIED BY HYALURONIC ACID Untreated explant **p<0.01 Student test Explant treated with 1.5% Hydranov INCREASE IN EPIDERMAL HYALURONIC ACID SYNTHESIS WILL PROVIDE A BETTER HYDRATION OF THE DEEPER LAYERS OF THE EPIDERMIS BUT ALSO STIMULATE THE RENEWAL OF TIGHT JUNCTIONS IN THE MIDDLE AND UPPER LAYERS. RESTRUCTURING ACTION OF HYDRANOV : CLAUDIN 1 ET CERAMIDS The function of tight junctions is to block circulation of fluids between cells and to ensure in this way that there is a tight seal between two tissue compartments. They were identified in human skin explants in the medium layers (granular and sub‐ basal layers)(3). The main factor contributing to the formation of these tight junctions is a protein called Claudin 1. In 2002, Furuse and al. showed that mice which did not express Claudin 1 died of massive Trans‐Epidermal Water Loss due to a deficient barrier function in the granular layer (4). Tight junction: protein complex Lipid bilayer Claudine Tight junction HYDRANOV STIMULATES THE SYNTHESIS OF CLAUDIN 1 EX‐VIVO TEST PROTOCOL: Human skin explants – Topical application of 1.5% Hydranov for 9 days. Quantification of Claudin 1 synthesis using immuno‐labelling. RESULT: +45% CLAUDIN 1 expression % versus untreated control Claudin 1 160 ‐ 140 ‐ +45% 120 ‐ 100 ‐ 80 ‐ 60 ‐ 40 ‐ 20 ‐ ‐ 0 Control 1.5% Hydranov HYDRANOV STIMULATES THE SYNTHESIS OF CERAMIDS EX‐VIVO TEST PROTOCOL: Human skin explants – Topical application of 1.5% Hydranov for 9 days. Quantification of Ceramids synthesis using immuno‐labelling. Untreated explant RESULT: : +56%** CERAMIDS **p<0.01 Student test Explant treated with 1.5% Hydranov INCREASE IN THE SYNTHESIS OF CLAUDIN 1 AND CERAMIDS HELPS TO STRENGTHEN THE SEALING OF THE EPIDERMIS. THE HYDRATION RATE PREVIOUSLY INCREASED VIA EPIDERMAL HYALURONIC ACID IN THE DEEPER LAYERS IS PRESERVED. NMF BOOSTER EFFECT OF HYDRANOV Filaggrin Procaspase 14 maturation EX‐VIVO TEST PROTOCOL: Reconstructed human epidermis ‐ Topical application of 1% Hydranov for 72 hours. Gene expression analysis using RT‐ qPCR. RESULT: +43% FILAGGRIN Hygroscopic AA Caspase 14 activation KLK5 160 ‐ 140 ‐ 120 ‐ +43% 100 ‐ 80 ‐ 60 ‐ 40 ‐ 20 ‐ 0 ‐ HYDRANOV STIMULATES THE SYNTHESIS OF FILAGGRIN degradation KLK7 expression % versus untreated control Numerous studies report the essential role of Filaggrin which, when it is broken down in the stratum corneum, adds hygroscopic amino acids (AA) to the NMF (Natural Moisturizing Factors). An absence of filaggrin expression leads to an almost complete lack of stratum corneum (ichtyosis) and major skin dehydration (xerosis) (5). Filaggrin is broken down in the stratum corneum by Caspase 14 which is itself matured under the control of a serine protease called kallikrein‐related peptidase 7 (KLK7). Activation of KLK7 depends on a second kallikrein coded by the gene KLK5 Control 1% Hydranov HYDRANOV FAVOURS FILAGGRIN DEGRADATION EX‐VIVO TEST PROTOCOL: Reconstructed human epidermis – Topical application of 1% Hydranov for 24 hours (KLK5 and KLK7 analysis) and 72 hours (caspase 14 analysis). Analysis of genes expression using RT‐ qPCR. Visualization of Caspase 14 using immuno‐labelling (fuchsia colour) in human skin explant treated with 1.5% Hydranov. Visualization of Caspase 14 in untreated explant RESULT: +60% KLK5 / +39% KLK7 +65% CASPASE 14 Visualization of Caspase 14 in explant treated with 1.5% Hydranov BY FAVOURING FILAGGRIN BREAKDOWN, HYDRANOV BOOSTS NMF AND REINFORCES THE HYDRATION OF CORNIFIED LAYER. THIS LATEST PROPERTY COMPLETES HYDRANOV ACTION: A GLOBAL HYDRATION FROM THE BASAL LAYER TO THE STRATUM CORNEUM. NMF COMPARATIVE HYDRATING PROPERTIES OF HYDRANOV AND HYALURONIC ACID IN‐VIVO TEST PROTOCOL: 15 volunteers apply a solution containing 1.5% HYDRANOV (equivalent to 0.009% pure oligofurcellaran) 15 volunteers apply a solution containing 0,025% pure HYALURONIC ACID (% recommended by the supplier) of high molecular weight: >200kDa. Twice daily applications for 2 weeks on forearms Measure of skin hydration level using corneometer Hydranov hydrates the skin with an immediate and continuous efficacy during the 24 hours following its application. Its hydrating action is faster and higher than thus of Hyaluronic acid. HYDRATION INCREASE INDUCED BY HYDRANOV IS 3 TIMES HIGHER THAN THUS TRIGGERED BY HYALURONIC ACID. *p<0.05 Student test Hydration variation versus T0 (%) COMPARATIVE EFFICACY AFTER A SINGLE APPLICATION 18 ‐ Hydranov 16 ‐ Hyaluronic acid 14 ‐ 12 ‐ * 10 ‐ 8‐ 6‐ * * 4‐ ‐ 0 ‐ 2‐ 4H 6H 24H *p<0.05 Student test Hydranov 16 ‐ Hyaluronic acid * 14 ‐ 12 ‐ 10 ‐ * * * 8‐ 6‐ 4‐ 2‐ 0 D1 ‐ AFTER A 2 WEEKS TREATMENT, THE HYDRATION PROVIDED BY HYALURONIC ACID IS STILL LOWER THAN THUS PROVIDED BY HYDRANOV AFTER ONLY 24H. 18 ‐ ‐ Hydranov has a cumulative hydrating effect. Day after day, skin hydration is increased. Hydration variation versus T0 (%) COMPARATIVE EFFICACY AFTER 2 WEEKS D7 D14 VISUALIZATION OF THE RESTRUCTURING ACTION OF HYDRANOV EX‐VIVO TEST PROTOCOL: Human skin explants. Topical application of 1.5% Hydranov for 1 week. Staining, rinsing, drying and observation of the surface using Scanning Electron Microscopy. Untreated explant Explant treated with Hydranov THE EXPLANT TREATED WITH HYDRANOV HAS AN HOMOGENEOUS AND SMOOTHED SURFACE WITH ONLY FEW DESQUAMING CELLS. THIS OBSERVATION ILLUSTRATES THE RESTRUCTURING ACTION OF HYDRANOV ON THE WHOLE EPIDERMIS, FROM THE BASAL LAYER TO THE CORNIFIED LAYER. CONCLUSION : HYDRANOV targets a molecule with outstanding potential and little known: the epidermal hyaluronic acid. Hydranov reinvents hydration by targeting new molecules with unexploited potential. By increasing the synthesis of epidermal hyaluronic acid, Hydranov promotes not only hydration of the deepest layers of the epidermis, but also renewal of tight junctions to reorganise the keratinocytes in all the layers of the epidermis. In the corneal layer, Hydranov activates metabolism of filaggrin to enrich NMF with hygroscopic amino acids while on the surface, in the same way as hyaluronic acid, it captures water molecules to maintain a hydrating film. Hydranov is a Hi‐Tech concentrate of oligofurcellaran, faster and 3 times more efficient than hyaluronic acid to hydrate and restructure the skin. Marine plant origin Hydrosoluble active ingredient In‐vitro test ex‐vivo test In‐vivo test COSMOS Cosmos Validation FORMULATION GUIDELINE HYDRA‐RESTRUCTURING FLUID This fluid is formulated with HYDRANOV: hydra‐restructuring ingredient, HYDROFILTRAT MENYANTHES G: dermo‐ restructuring active and a moisturizing ingredient: PHEOHYDRANE. Phase Raw material / commercial name INCI name % NIKKOMULESE LC (1) Behenyl Alcohol & Stearyl Alcohol & PEG‐20 Phytosterol & Cetyl Alcohol & Phytosterols & Glyceryl Stearate & Hydrogenated Lecithin & Caprylic/Capric Triglyceride 5 CRODAMOL GTCC / MIGLYOL 812 / WAGLINOL (2) Caprylic/Capric Triglyceride 3 SILICON (DIMETHICONE (100CS) (3) Dimethicone 0.5 PHENOXYETHANOL (4) Phenoxyethanol 0.8 ARLAMOL HD (5) Isohexadecane 4 LANOL 99 (6) Isononyl Isononanoate 5 DEMINERALIZED WATER Aqua AVICEL PC 611 (7) Microcrystalline Cellulose & Cellulose Gum 1 DIPROPYLENE GLYCOL (8) Dipropylene Glycol 3 KELTROL CGSFT (9) Xanthan Gum 0.5 ELESTAB CPN (10) Chlorphenesin 0.27 HYDRANOV P (11) Aqua & Phenoxyethanol & Sodium Carrageenan & Maris Sal 1.5 PHEOHYDRANE P (11) Aqua & Hydrolyzed Algin & Maris Aqua & Phenoxyethanol & Chlorella Vulgaris Extract 1 HYDROFILTRAT MENYANTHES G (11) Glycerin & Aqua & Menyanthes Trifoliata Extract 2 FRAGRANCE FLOWER AND FRUIT 0217350 (12) Fragrance A 72.33 B B’ B’’ C (1) IMCD, (2) Croda, (3) Quimdis, (4) Laserson, (5) Quimasso, (6) Seppic, (7) FMC BioPolymer, (8) Brenntag, (9) CP Kelco, (10) Cognis, (11) CODIF Recherche et Nature, (12) Expressions parfumées Product Aspect: White Glitter Fluid cream Product Properties: pH = 6.54 ± 0.5 Operating Method: • Heat A at 75°C • Heat the phase B at 75°C under emulsifier 2000 rpm during 10 minutes. • Add the mixture B´ under emulsifier 1500 rpm during 5 minutes. • Add the mixture B’’ under emulsifier 1500 rpm during 5 minutes. • Add A into B+B’+B’’ under emulsifier 2500 rpm, then let shake in these conditions during 10 minutes. • Around 25‐30°C, add the phase C. 0.1 BIBLIOGRAPHIC REFERENCES (1) Tammi M, Saamanen AM : Degradation of Newly Synthesized High Molecular Mass Hyaluronan in the Epidermal and Dermal Compartments of Human Skin in Organ Culture. J Invest Dermatol 97: 126‐130, 1991 (2) Tammi R, Tammi M: Hyluronan in the Epidermis ‐ http://glycoforum.gr.jp/science/hyaluronan/HA04/HA04E.html (3) Adherens Junctions, Desmosomes and Tight Junctions in Epidermal Barrier Function Johanna M. Brandner, Marek Haftek and Carien M. Niessen. The Open Dermatology Journal, 2010, 4, 14‐20 (4) Claudin‐based tight junctions are crucial for the mammalian epidermal barrier: a lesson from claudin‐1‐deficient mice. Furuse M, Hata M, Furuse K, et al. J Cell Biol 2002; 156: 1099‐111. (5) Filaggrin in the frontline: role in skin barrier function and disease. Sandilands A, Sutherland C, et al. J Cell Science, 2009, 122, 1285‐1294 HYDRANOV - A NEW LOOK AT HYDRATION Cosmetic activities • « hyaluronic like » action: traps water molecules at the surface of the skin • « hyaluronic activator » action : stimulates the synthesis of epidermal hyaluronic acid • Reinforces tight junctions via Claudin 1 • Increases ceramids synthesis • Reinforces the NMF via Filaggrin • Hydrating efficacy faster and higher than hyaluronic acid • Restructuring action INCI name Hydranov P: Water (and) Phenoxyethanol (and) Sodium Carrageenan (and) Sea salt Hydranov PA : Water (and) Phenethyl alcohol (and) Sodium Carrageenan (and) Sea salt COSMOS Recommended % of use: Hydranov P : 1.5% ‐ Hydranov PA : 1.5% Technical department : infotech@codif‐recherche‐et‐nature.com Commercial department : commercial@codif‐recherche‐et‐nature.com 70, rue du Commandant l’Herminier ‐ CS11781 ‐ 35417 Saint‐Malo cedex ‐ FRANCE Tel : +33‐2‐23‐18‐31‐07 Fax : +33‐2‐23‐18‐31‐01 www.codif‐recherche‐et‐nature.com CODIF Recherche & Nature is committed to sustainable development. Iris, water mint, flowering rush, willows etc… now form part of the image of the brand which is recognized for its commitment to the preservation of natural resources. RPDOC133GB All our wastewater is processed and purified using filtrating gardens. A succession of airtight horizontal and vertical filters is used to depollute wastewater by the roots of plant species chosen for their draining properties.