REVITALIZING POWER OF NATURE CONCENTRATED IN A PURE ACTIVE INGREDIENT

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1 revitalizing of aged cells stimulation of cell viability boost of anti-oxidative power (SOD1) moisturizing stimulation of procollagen I synthesis increase of skin elasticity soothing and calming wound healing reduction of sunburn (anti-erythema) THE CELL ENERGIZER REVITALIZING POWER OF NATURE CONCENTRATED IN A PURE ACTIVE INGREDIENT is the protection and re-activation substance of the resurrection plant Myrothamnus flabellifolia. These fascinating plants live in the deserts of Africa under extreme conditions like long-term dryness, high temperatures and extreme UV-radiations. They have the unique ability to survive severe dehydration - up to the complete loss of water, also known as desiccation. during dry period plant after rain is responsible for the recurring re-activation and re-vitalization of these plants after total desiccation for months. With the beginning of the rains, the plant starts to green again. During dry season, the stress protection molecule keeps the structures of the plant alive and protects it from dying despite total desiccation. 1 - Factsheet. bitop AG. 04/2015

2 Resurrection plants have to face the loss of all of their cellular water during the dry season. For every other plant this normally means total damage of the cell structure and death. The stress protection molecule enables the fascinating existence of the Myrothamnus plant by protecting the cell membrane during total desiccation and by "storing" water for as long as possible. With the start of the rains, as soon as water is available again, boosts the cell functions of the plant for a fast re-greening. This mechanism is transferable to our skin with convincing effects. Water retention capacity - hyaluronic acid like has a comparable capacity of holding/storing water like hyaluronic acid, but it is around times smaller and has no tacky skin feeling. % Water retention capacity water HA (0,1%) HEC G (0,1%) HEC M (0,1%) Glycoin natural (1%) fully soaked(%) 30min(%) 60min(%) 90min(%) 120min(%) 150min(%) 180min(%) Test method: Filter paper was weighed and soaked in the test substances and again weighed. Afterwards the papers were dried with interruptions to measure the weight of the drying filter papers. Figure 1: Water retention capacities of HA (high molecular weight), HEC (Hydroxyethyl Cellulose G and M type), and H20. Membrane stabilization protection of cell structure Test protocol: Ÿ in vitro, human epidermal keratinocytes Ÿ pre-treatment with and an extract from resurrection plant (0,03%, 0,04% and 0,05%) Ÿ cells stressed with 0,001% SDS Ÿ measured parameter: cell membrane stability Membrane stabilisation in comparsion to PBS (%) Figure 2: Membrane stabilization properties of compared to the membrane stabilization properties of an extract from a resurrection plant. 0,03% 0,04% 0,05% plant extract 0,03% plant extract 0,04% plant extract 0,05% 2 - Factsheet. bitop AG. 04/2015

3 Stimulation of cell vitality - increase of ATP in aged cells ATP (adenosine triphosphate) is the energy currency of the cell. An increase of ATP leads to a boost of cell viability and cell renewal. Test protocol: Ÿ in vitro, human keratinocytes from adult skin (HaCaT-cells); aging of cells for 2 weeks Ÿ treatment with, glucose fructose, glycerin (0,5%, 0,75%, 0,1%); 6h incubation time Ÿ measured parameter: ATP concentration in cells with luciferase assay Increase of ATP in aged cells 6 4 vitality ,75% 0,5% 0,1% glucose 0,75% glucose 0,5% Figure 3: Increase of ATP concentration in aged human keratinocytes after pre-treatment with and fructose at different concentrations. concentration The boosting effect of on the vitality and activity of aged cells was measured in various studies. Next to the increase of the cell metabolism ( gure 4), was able to also signi cantly increase the ATP concentration in aged cells ( gure 3). Even at an advanced cell age stage, when other substances like fructose, glucose or glycerin did not have any positive effect on the cell vitality any more. Stimulation of the viability of aging skin cells The cell viability of NHEKs cultivated in the presence and absence of (1%) was measured. Cells which had been cultivated in the presence of showed higher metabolic activity than the untreated control. 1,2E-05 1,0E-05 without with viability/cell 8,0E-06 6,0E-06 4,0E-06 2,0E-06 0,0E cuture [h] increases the cell viability and metabolic activity in aging skin cells by 170%. Figure 4: Viability of cells during aging. 3 - Factsheet. bitop AG. 04/2015

4 In-vivo study: increase of skin moisture, elasticity and smoothness An in-vivo study conducted on 20 female test persons, aged 45 and older has shown extremely positive results of the revitalizing effects of. The table below shows results of a twice-daily application after 28 days of treatment with a cream containing 1%. Skin properties application, after 28 days treated area compared to placebo treated area Skin moisture (Corneometer) % % Skin elasticity (Cutometer) % % Skin smoothness (PRIMOS) % % Table 1: In-vivo study results Increase of procollagen I production in aged cells The stimulation of procollagen I synthesis was shown in an in-vitro study, using the replicative aging model (Hayflick) on aged dermal fibroblasts. Aged fibroblasts were treated with in different concentrations (0,17%, 0,5% and 1,5%). showed a significant stimulation of procollagen I synthesis at all concentrations. stimulates the procollagen I synthesis in aged cells by 36%. In-vivo study: reduction of skin redness (anti-erythema) The back of 20 male and female panelists with skin phototype II and III was irradiated with UVradiation. The irradiated areas were treated 3 times (after 7h, 24h and 32h after irradiation) with placebo cream, 2% cream and cream containing 0,5% Asiaticoside (as positive control). After 48h the skin redness (with chromameter) and skin microcirculation (with TiVi ) on the irradiated areas were measured. 76% of the test person treated with had a soothing effect on sunburn. Only 41% of the test persons treated with Asiaticoside and 35% treated with placebo cream had a positive effect on the sunburn. soothes sunburn after only 3 times of application. 4 - Factsheet. bitop AG. 04/2015

5 Boost of anti-oxidative power in aging skin cells (SOD1) An in-vitro study conducted on human epidermal keratinocytes (NHEKs) showed, that the presence of 1% Glycoin natural resulted in an increased production of the anti-oxidative skin protection enzyme Superoxide Dismutase 1 (SOD1). With an advanced aging process of the cells this effect even increased. Glycoin natural treated cells produced 280% more SOD1 than untreated cells. 3,00E-03 2,50E-03 without with SOD 1 [ng/cell] 2,00E-03 1,50E-03 1,00E-03 5,00E-04 0,00E cuture [h] Figure 5: Amount of SOD1 per cell during aging. Glycoin natural increases the amount of SOD1 in aging skin cells by 280%. Wound healing Test protocol: Ÿ ex vivo skin model; surface was mechanically reduced by 3mm (loss of epidermis and dermis) Ÿ immediately after wound making, wound spot was covered with 5µl PBS with 2,4% and without (placebo) Ÿ incubation for 48h Ÿ hematoxylin/eosin staining of skin model for evaluation treatment with Figure 6: Wound healing demonstration on ex vivo skin model after 48h. treatment with placebo (control) demonstrated excellent wound healing ef cacy after only 48h! 5 - Factsheet. bitop AG. 04/2015

6 Applications skin care (face, body) baby care sun care oral care color cosmetics suitable for extremely sensitive and allergic skin Formulation guidelines level of use 0,5-2% no incompatibility with other cosmetic active substances can be added directly to aqueous phase prior to emulsification 100% safe GMO free China compliant non allergenic Ecocert preservative free Bitop s is of 100% natural origin and the only enantiopure Glyceryl Glucoside in the market. It exclusively consists of the stereoisomer, which is also produced by the resurrection plant Myrothamnus flabellifolia and blue-green algae. INCI Glyceryl Glucoside CAS number Description Chemical name Molecular formula Molecular weight Concentration Appearance 100% natural origin, sugar derivate, small molecule, enantiopure 2-O-α-D-Glucopyranosylglycerine C H O g/mol ph (2%; water) Stability Solubility Shelf life Storage conditions 2 C 25 C Dosage 0.5-2% Pack sizes Production process Origin 50% Glyceryl Glucoside in aqueous solution slightly yellowish liquid, slightly characteristic odor ph 3-11, light and oxygen stable, <60 C excellent solubility in water 2 years, (repeated freezing and thawing is possible; frozen condition: shelf life >5 years), no preservatives ( auto-sterile ) 1kg, 5kg, 12,5 kg enzymatic reaction / biocatalytical process Ÿ component of the resurrection plant Myrothamnus flabellifolia for protection against dehydration Ÿ produced by salt tolerant cyano-bacteria (blue algae) Glycoin natural is a patented active ingredient. bitop AG Stockumer Str Witten Germany Contact: Phone: info@bitop.de The ingredients described herein are offered for the use in personal care products. bitop AG makes every possible effort to ensure that the information published is up to date and accurate but accepts no legal liability for errors or omissions and reserves the right to make changes without notice. Specifications, measurements and product data are for generalized informational purposes only. How each active ingredient would contribute to a particular product would be formulation specific. All material published here is copyright of bitop AG except for any material, which has been marked as being the copyright of a third party. Intellectual property rights in some of this material may be held by individual authors. Furthermore please note that this documentation is available for various countries all over the world and hence may contain statements not applicable for your country.

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