Cellulosomes ® Eryngium – Global skin restructuring

1. Cellulosomes® Eryngium – Global skin restructuring

2. HALOPHYTE PLANTS

3. DIFFERENCES BETWEEN HALOPHYTES & GLYCOPHYTES Salty soil Marine wind spray aggressions HALOPHYTE PLANTS SPECIFICITIES Adaptative water management (leaves, stem…) Ion management and control

4. UNIQUE PROPERTIES OF HALOPHYTE PLANTS: Specific molecules: Lipids Amino-acids such as proline Polyols Polyphenols Soluble sugars Specific properties: Powerful protection against degradation from free radicals Anti-inflammatory properties HALOPHYTE PLANTS PROPERTIES

5. Cellulosomes® Eryngium - Characteristics CTFA/INCI Name: Eryngium Maritimum Callus culture Filtrate CAS N° : 90028-44-7 EINECS/ELINCS N° : 289-835-1 RECOMMENDED USE: 0.05% ASPECT: powder COLOR: beige ODOR: none SOLUBILITY: water dispersible PRESERVATIVE: none CERTIFICATIONS: Ecocert

6. Cellulosomes® Eryngium- Sea plant specific answer Induction of stem cells from Eryngium maritimum. Sea plants are the first & original organisms to have come out of the water. Sea plants, with their roots in the marine original medium have, at the origin, to face the constraints of the terrestrial conditions (dehydration - light - UV- and additional stress from salt) They developed an unique answer in the adaptation phenomenon to the terrestrial environmental conditions. FAMILY: APIACAE PROTECTED SPECIES LOCATED ON DUNE AND SHINGLE SEA SHORE

7. Cellulosomes® Eryngium- halophyte stem cell / patent We take an explant from meristematic area Stem cell* culture Lyophilisation (Freeze dry) Filtration from the aqueous medium Bioreactor Stem cell production in suspension *Mix with growth factors, minerals, trace elements…/ sterilized batches

8. Cellulosomes® Eryngium - Sea plant stem cell Advantages of this new technology based on stem cells Each dedifferentiated cell contains all of the plant’s components – Pluripotent Product properties are optimized and enlarged Regeneration - Rejuvenation Integration by biomimetism with cutaneous cell “Energetic seed, a pure concentrate of life” Pollution free - Solvent free - Preservative free No cytotoxicity => Sustainable development & resources protection

9. Cellulosomes® Eryngium -Targets Physiological phenomena occurring at all skin levels EPIDERMIS • Epidermis gets thinner with atrophic, irregular & disorganized epidermis • Modifications of the epidermal basal layer (morphology and operation) DERMO-EPIDERMAL JUNCTION • Dermo-epidermal junction is flattened, affected by some deterioration • Permeability with less effective adhesion, permeability disorders DERMIS • Most important changes observed with dermis thickness reduction • Extra Cellular Matrix (ECM) disrupts itself (Collagen, GAG…) • Less fibroblasts & disorganized elastic fibers

10. Cellulosomes® Eryngium– Dermo Epidermal Junction, the skin structure

11. Cellulosomes® Eryngium– Dermo Epidermal Junction Integrins B1: Adherence & transduction of the matrix and cells Laminin 5: Consolidation of the construction of the adherent structure of hemidesmosomes. Elastin: Support the skin elasticity Fibulin 5: Reinforce the interconection between elastin / integrins and the organisation of the elastic fibers in the ECM Fibrillin: Orientation of the elastic fibers Collagens IV & VII: Reinforcement of the anchorage fibres and the organisation of the architecture of the basal layer and its communication

12. Cellulosomes® Eryngium - Summary ACTIONS ON EPIDERMIS Regulation of the keratogenesis Proliferation + differentiation ACTIONS ON DERMO-EPIDERMAL JUNCTION Protection & stimulation ACTIONS ON DERMIS Stimulation of the ECM components Protection of the ECM components PROTECTION Against free radicals degradation Protection against inflammation disorders

13. Evaluation of Cellulosomes® Eryngium on mediators and molecules involved on communication and junction between epidermis and dermis without irradiation under irradiation UVA (10 J/cm2) + UVB (150 mJ/cm2) (immunohistochimical study on reconstitued epidermis) Markers analysed: Integrin α3 B1 - Laminins 5 - Elastin Fibulin 5 – Fibrillin - Collagens Cellulosomes® Eryngium– Dermo Epidermal Junction

14. Without irradiation Cellulosomes® Eryngium– Dermo Epidermal Junction – Results 1 Adherence & transduction of the matrix and cells Consolidation of the construction of the adherent structure of hemidesmosomes. Support the skin elasticity Reinforce the interconection between elastin / integrins and the organisation of the elastic fibers in the ECM Orientation of the elastic fibers Reinforcement of the anchorage fibres et the organisation of the architecture of the basal layer and its communication • MARKERS STIMULATED • Integrins B1 • Laminin 5 • Elastin • Fibulin 5 • Fibrillin • Collagens IV et VII ACTIONS Cellulosomes® Eryngium stimulates the neosynthesis of the macromolecules located on Dermo Epidermal Junction, basal layer and upper dermis

15. Under irradiation UVA (10 J/cm2) + UVB (150 mJ/cm2) Cellulosomes® Eryngium– Dermo Epidermal Junction – Results 2 ACTIONS Adherence of the matrix and cells Support the skin elasticity Orientation of the elastic fibers Reinforcement of the anchorage fibres et the organisation of the architecture of the basal layer and its communication • MARKERS STIMULATED • Integrins B1 • Elastin • Fibrillin • Collagens IV et VII Cellulosomes® Eryngium has a positive effect on the regulation of the neosynthesis of macromolecules located on Dermo Epidermal Junction, basal layer and upper dermis

16. Regenerating activity on skin Proliferation ( KI67) Differentiation (K10 & Involucrin) Protection against Free Radicals (MDA measurement) Inflammatory activity (IL1 - IL6 & PGE2) Cellulosomes® Eryngium – Integral protection of the epidermis / In-vitro tests

17. Cellulosomes® Eryngium–Skin renewal Application of a cream at 0.05% on reconstitued epidermis (72H): Proliferation of the new keratinocytes from the basal layer immunohistochemical treatment to measure KI 67(mitotic index) - Increase by +19% Control Cellulosomes Eryngium stimulates cell proliferation & skin renewal

18. Cellulosomes® Eryngium–Skin renewal Application of a cream at 0.05% on reconstitued epidermis (72H): Epidermis differentiation by K10 early granular layer & involucrin late markers Result: Reinforcement K10 Control With Cellulosomes Eryngium Involucrine Cellulosomes Eryngium stimulates cell proliferation & differentiation for a better skin radiance & luminosity

19. Cellulosomes® Eryngium- Keratogenesis regulation Immunohistochemical marking of the HB-EGF: (Heparin binding epidermal growth factor) HB-EGF Untreated skin treated skin with Cell. Eryngium Acid vitamin A Cellulosomes Eryngyium stimulates HB–EGF formation

20. IN VITRO TEST ON DERMIS COMPONENTS Stimulation of the collagen Stimulation of the proteoglycans : membrane (peri & trans) matrice Cellulosomes® Eryngium – Dermis

21. Marker - Matricial proteoglycans* (biglycan, decorin) - Transmembranar proteoglycans (syndecan) - Perimembranar proteoglycans - Collagen Cellulosomes® Eryngium – stimulation of the ECM * receptor and actor Stimulation of proteoglycans and collagen on fibroblasts culture

22. Cellulosomes® Eryngium- Anti radical protection • Protocol & material: • Epidermis on culture • MDA measurement after 24 hours in contact • MDA was extracted then measured by HPLC (fluorescence) • Lipoperoxidation initiated by UVB (150mJ/cm²) MDA: Malondialdehyde, final by-product and tracer of lipoperoxidation Cellulosomes ® Eryngium shows a global protection against UVB induced free-radical

23. Cellulosomes® Eryngium- Decrease of UVB induced inflammation & irritation • Protocol & material: • Human Keratinocyteson culture • Cytokines measurement under physiological conditions and UVB exposure Cellulosomes ® Eryngiumstrongly limits keratinocytes activation & neutralizes biological consequences of stresses - no variationunder physiological conditions

24. Cellulosomes® Eryngium - In vivo results Duration of the test: 28 days 20 volunteers from 35 to 60 years old with skin sagging Application of a cream formulated with 0.05 % twice a day Evaluation by dermatologist Clinical evaluation • Do & D28, the dermatologist measured different clinical signs (erythema, dryness, roughness, desquamation, hyperkeratose, œdema, vesicles) on face (cheeks, forehead, nose, chin). • After the treatment, the dermatologist noticed a reduction of the : • Erythematic stage • Dryness stage

25. Cellulosomes® Eryngium - In vivo results / firmness Self evaluationby questionnaire (scale from 1 to 9) of the skin from day 0 to day 28 controlled by dermatologist. • The efficacy has been perceived by 90% of the volunteers with improvement on: • skin grain • skin tonicity & firmness • skin hydration (osmolites…sucre acide amine) 90% of the volunteers satisfied

26. Cellulosomes® Eryngium- Global properties Cellulosomes® Eryngium Global skin restructuring RESULTS Skin radiance Skin firmness Global protection Hemidesmosomes filaments COLLAGEN PROTEOGLYCANS FIBULIN ELASTIN FIBRILLIN MATRICIELS PERI MEMBRANAIRES TRANS MEMBRANAIRES

27. Cellulosomes® Eryngium - Formulation guide line Better for cream applications No incompatibility Need some viscosity Ph range: 3.5 - 8