HYDRANOVA New Look at Hydration

HYDRANOV A New Look at Hydration

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 theepidermis, it contains 4 to 9 times more hyaluronic acid, but for equivalent tissue quantities, the epidermis synthesises 4times 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 inthe 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 ofhyaluronic acid in the intercellular space tends to renew desmosomes and tight junctions, and so reorganise the keratinocytesnot only within the basal layer, but also throughout the process of differentiation as far as the stratum corneum (2). Hyaluronicacid acts therefore like an overall epidermal restructuring agent by initiating and maintaining constant cellular remodellingfrom the basal layer to the cornified layer.

« HYALURONIC LIKE », HYDRANOV HAS HYGROSCOPIC PROPERTIES ALLOWING THE CAPTURE OFWATER MOLECULES AT THE SKIN SURFACE. « EPIDERMAL HYALURONIC ACTIVATOR », IT ALSOSTIMULATES THE SYNTHESIS OF EPIDERMAL HYALURONIC ACID FOR AN OVERALL EPIDERMALRESTRUCTING AND HYDRATING ACTION.

HYDRANOV :High Tech Concentrate of OligofurcellaranThe oligofurcellaran is obtained by the depolymerisation of a sulfated furcellaran (galactose and anhydrous galactose) comingfrom the red alga Furcellaria lumbricalis, widespread in Scandinavian waters.

Depolymerization is made using supercritical CO2 technology in anhydrous conditions. After depolymerization, theoligofurcellaran 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 tohyaluronic acid. The resulting oligofurcellaran has a molecular weight >200kDa.

This depolymerisation method, water and acid free, complies with the policy of energetic resources managment developpedby Codif R&N.

CCOSMOS

“HYALURONIC LIKE” ACTION OF HYDRANOVIN‐VITRO TEST

The oligofurcellaran concentrated in Hydranov has high molecular weight. Therefore in the same way as hyaluronic acid itacts 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 atsorption equilibrium ». The value of gain mass reflects the ability of each polymer to trap water molecules.

RESULT:Hydranov is able to trap atmospheric water molecules following a kinetic totally comparable to hyaluronic acid. The moreatmosphere 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 ANDTHUS MAINTAINS A MOISTURE FILM ON THE SURFACE OF THE SKIN.

HYDRANOV CHRONOLOGICAL ACTION1 ‐ « hyaluronic like » action at the surface of the skin2 ‐ « hyaluronic activator » action : stimulates the synthesis of epidermal hyaluronic acid3 ‐ Reinforcement of tight junctions – New target : Claudin 14 ‐ Stimulation of ceramids synthesis5 ‐ Reinforcement of NMF – New target : Filaggrin

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“HYALURONIC ACTIVATOR” ACTION OF HYDRANOVGiven the size of the oligofurcellaran and the low probability that it can penetrate into the epidermis, the effect of Hydranovhas been evaluated by topical application on human skin explants or human reconstructed epidermis.

HYDRANOV INCREASES THE EXPRESSION OF HYALURONAN SYNTHASE 3EX‐VIVO TEST

Hyaluronan synthase 3 is an enzyme involved in hyaluronicacid synthesis.

PROTOCOL:Reconstructed human epidermis ‐ Topical application of1% Hydranov for 72 hours. Gene expression analysis usingRT‐qPCR.

RESULT:+64% HYALURONAN SYNTHASE 3

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 epidermalhyaluronic acid synthesis using immuno‐labelling (fuchsiacolour).

RESULT:+211%** HYALURONIC ACIDX3 : TOTAL SURFACE AREA OCCUPIED BY HYALURONIC ACID**p<0.01 Student test

INCREASE IN EPIDERMAL HYALURONIC ACID SYNTHESIS WILL PROVIDE A BETTER HYDRATION OF THEDEEPER LAYERS OF THE EPIDERMIS BUT ALSO STIMULATE THE RENEWAL OF TIGHT JUNCTIONS IN THEMIDDLE AND UPPER LAYERS.

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RESTRUCTURING ACTION OF HYDRANOV :CLAUDIN 1 ET CERAMIDSThe function of tight junctions is to block circulation of fluidsbetween cells and to ensure in this way that there is a tight sealbetween two tissue compartments. They were identified inhuman skin explants in the medium layers (granular and sub‐basal layers)(3). The main factor contributing to the formationof these tight junctions is a protein called Claudin 1.In 2002, Furuse and al. showed that mice which did notexpress Claudin 1 died of massive Trans‐Epidermal Water Lossdue to a deficient barrier function in the granular layer (4).

HYDRANOV STIMULATES THE SYNTHESISOF CLAUDIN 1EX‐VIVO TEST

PROTOCOL:Human skin explants – Topical application of 1.5% Hydranovfor 9 days. Quantification of Claudin 1 synthesis usingimmuno‐labelling.

RESULT:+45% CLAUDIN 1

HYDRANOV STIMULATES THE SYNTHESISOF CERAMIDSEX‐VIVO TEST

PROTOCOL:Human skin explants – Topical application of 1.5% Hydranovfor 9 days. Quantification of Ceramids synthesis usingimmuno‐labelling.

RESULT: :+56%** CERAMIDS**p<0.01 Student test

INCREASE IN THE SYNTHESIS OF CLAUDIN 1 AND CERAMIDS HELPS TO STRENGTHEN THE SEALINGOF THE EPIDERMIS. THE HYDRATION RATE PREVIOUSLY INCREASED VIA EPIDERMAL HYALURONICACID IN THE DEEPER LAYERS IS PRESERVED.

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NMF BOOSTER EFFECT OF HYDRANOVNumerous studies report the essential role of Filaggrin which,when it is broken down in the stratum corneum, addshygroscopic amino acids (AA) to the NMF (NaturalMoisturizing Factors). An absence of filaggrin expression leadsto an almost complete lack of stratum corneum (ichtyosis) andmajor skin dehydration (xerosis) (5). Filaggrin is broken down in the stratum corneum by Caspase 14 which is itself matured under the control of aserine protease called kallikrein‐related peptidase 7 (KLK7).Activation of KLK7 depends on a second kallikrein coded bythe gene KLK5

HYDRANOV STIMULATES THE SYNTHESISOF FILAGGRINEX‐VIVO TEST

PROTOCOL:Reconstructed human epidermis ‐ Topical application of 1%Hydranov for 72 hours. Gene expression analysis using RT‐qPCR.

RESULT:+43% FILAGGRIN

HYDRANOV FAVOURS FILAGGRIN DEGRADATIONEX‐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.

RESULT:+60% KLK5 / +39% KLK7+65% CASPASE 14

BY FAVOURING FILAGGRIN BREAKDOWN, HYDRANOV BOOSTS NMF AND REINFORCES THEHYDRATION OF CORNIFIED LAYER. THIS LATEST PROPERTY COMPLETES HYDRANOV ACTION:A GLOBAL HYDRATION FROM THE BASAL LAYER TO THE STRATUM CORNEUM.

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Visualization of Caspase 14 in untreatedexplant

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COMPARATIVE HYDRATING PROPERTIESOF HYDRANOV AND HYALURONIC ACIDIN‐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 molecularweight: >200kDa.Twice daily applications for 2 weeks on forearmsMeasure of skin hydration level using corneometer

COMPARATIVE EFFICACY AFTER A SINGLE APPLICATION

Hydranov hydrates the skin with an immediate andcontinuous efficacy during the 24 hours following itsapplication. Its hydrating action is faster and higher than thus ofHyaluronic acid.

HYDRATION INCREASE INDUCED BY HYDRANOV IS 3 TIMESHIGHER THAN THUS TRIGGERED BY HYALURONIC ACID. *p<0.05 Student test

COMPARATIVE EFFICACY AFTER 2 WEEKS

Hydranov has a cumulative hydrating effect. Day after day,skin hydration is increased.

AFTER A 2 WEEKS TREATMENT, THE HYDRATION PROVIDEDBY HYALURONIC ACID IS STILL LOWER THAN THUS PROVIDEDBY HYDRANOV AFTER ONLY 24H.*p<0.05 Student test

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VISUALIZATION OF THE RESTRUCTURING ACTION OF HYDRANOVEX‐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.

THE EXPLANT TREATED WITH HYDRANOV HAS AN HOMOGENEOUS AND SMOOTHED SURFACE WITH ONLY FEWDESQUAMING CELLS. THIS OBSERVATION ILLUSTRATES THE RESTRUCTURING ACTION OF HYDRANOV ON THE WHOLEEPIDERMIS, FROM THE BASAL LAYER TO THE CORNIFIED LAYER.

CONCLUSION :HYDRANOV targets a molecule with outstanding potential and little known: the epidermal hyaluronicacid.

Hydranov reinvents hydration by targeting new molecules with unexploited potential. By increasingthe synthesis of epidermal hyaluronic acid, Hydranov promotes not only hydration of the deepestlayers of the epidermis, but also renewal of tight junctions to reorganise the keratinocytes in all thelayers of the epidermis. In the corneal layer, Hydranov activates metabolism of filaggrin to enrich NMFwith hygroscopic amino acids while on the surface, in the same way as hyaluronic acid, it captureswater molecules to maintain a hydrating film.

Hydranov is a Hi‐Tech concentrate of oligofurcellaran, faster and 3 times more efficient than hyaluronicacid to hydrate and restructure the skin.

Untreated explant Explant treated with Hydranov

Marine plant origin In‐vivo test Cosmos ValidationHydrosoluble activeingredient

ex‐vivo test CCOSMOSIn‐vitro test

FORMULATION GUIDELINEHYDRA‐RESTRUCTURING FLUIDThis fluid is formulated with HYDRANOV: hydra‐restructuring ingredient, HYDROFILTRAT MENYANTHES G: dermo‐restructuring active and a moisturizing ingredient: PHEOHYDRANE.

(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.

Phase Raw material / commercial name INCI name %

A

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

BDEMINERALIZED WATER Aqua 72.33

AVICEL PC 611 (7) Microcrystalline Cellulose & Cellulose Gum 1

B’DIPROPYLENE GLYCOL (8) Dipropylene Glycol 3

KELTROL CGSFT (9) Xanthan Gum 0.5

B’’ ELESTAB CPN (10) Chlorphenesin 0.27

C

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 0.1

BIBLIOGRAPHIC REFERENCES

(1) Tammi M, Saamanen AM : Degradation of Newly Synthesized High Molecular Mass Hyaluronan in the Epidermal andDermal 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, MarekHaftek 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 saltHydranov PA : Water (and) Phenethyl alcohol (and) Sodium Carrageenan (and) Sea salt

Recommended % of use: Hydranov P : 1.5% ‐ Hydranov PA : 1.5%

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