HyaCare® 50 HyaCare® 50

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					HyaCare® 50
Very low molecular weight Hyaluronic Acid
- A breakthrough in anti-ageing properties -

             •   50 KDa Hyaluronic Acid

             •   Increased permeation through the skin compared to
                 high molecular weight hyaluronic acid

             •   Rejuvenates the skin by improving its viscoelastic

             •   Significantly decreases deep wrinkles
                                                            Personal Care
INCI Name (CTFA name)                                     In parallel, intensive developments have been made
                                                          to check if molecular weight reduction would provide
Under evaluation by the CTFA
                                                          even more skin care benefits. This lead to the
                                                          introduction of HyaCare® 50, a new and unique very
                                                          low molecular weight hyaluronic acid (50Kda)
Chemical and physical properties                          produced by fermentation of Bacillus subtilisis using
(not part of specifications)                              an environmentally friendly aqueous recovery
Form                                 Powder
Solubility in water                10g/100g
                                                          In-vitro penetration study
Hyaluronic acid (HA) is a non-sulfated
glycosaminoglycan (GAG), naturally distributed in         The penetration of different molecular weight
human body. Hyaluronic acid is the major                  fractions has been evaluated using side by side
component of the extra cellular matrix. It is found in    diffusion cells and tritiated hyaluronic acid on
high quantity in the skin where it is produced mainly     dermatomed skin from the porcine ear. The receptor
by both fibroplasts and keratinocytes.                    was filled with PBS buffer and the donor with
                                                          radiolabeled HA solution. After incubation for 5 h
One of its main functions is to hold the water in the     and 22 h at room temperature, the receptor phase
intercellular matrix of the connective tissue. This       was removed and radioactivity was determined by
water-binding capacity contributes significantly to       liquid scintillation counting.
the elasticity of the skin. Hyaluronic acid plays the
role of water pump maintaining the elasticity by
acting as a water reservoir. Hyaluronic acid is also        150
involved in tissue repair. While it is abundant in
extracellular matrices, it also contributes to tissue
hydrodynamics, movement and proliferation of cells,         100                     First bar: flux at 5 hr
and participates in a number of cell surface receptor
                                                                                    Second bar: flux at 22 hr
interactions, notably those including its primary
receptor, CD44.                                              50

When skin is excessively exposed to UV-B rays, it
becomes inflamed and the cells in the dermis stop              0
producing hyaluronic acid and increase the rate of its                50K        300K           800K            1500K
degradation. With aging, the quantity of hyaluronic                         HA molecular weight (Da)
acid and its degree of polymerisation decrease
leading to a decrease in the water content hold by        Figure 1: Influence of HA MW on percutaneous transport
the connective tissue and a loss of elasticity that       As seen in figure 1, HyaCare® 50 shows a much
manifests itself in wrinkles.                             stronger permeation through the skin in comparison
                                                          to higher molecular weight products.
In normal human skin, hyaluronic acid exists as a
polymer of medium molecular weight (600-1,000
Kda). The physiological degradation of hyaluronic         In-      DNA-
                                                          In-vitro DNA-Chip analysis
acid in the skin is done via internalisation by the       The study was performed at the University of
keratinocytes and fragmentation by hyaluronidases.        Regensburg (D).
                                                          The effect of HyaCare® 50 on gene expression was
High molecular weight hyaluronic acid has been used       analyzed using DNA-chip technology and SkinEthics
for many years as natural moisturizer in the cosmetic     models. It was demonstrated that HyaCare® 50
industry. In 2003 the FDA has approved hyaluronic         changed the regulation of more than 40 genes,
acid injections for filling soft tissue defects such as   namely upregulation of detoxification and cell-cell
facial wrinkles. Hyaluronic acid injections temporarily   adhesion genes. From this result it can be
smooth wrinkles by adding volume under the skin,          anticipated that HyaCare® 50 has positive effects on
with effects typically lasting for six months.            detoxification mechanisms and tautness of the skin.
This FDA approval and the significant trend for botox
like cosmetics reinforced the demand for hyaluronic
acid. Hyaluronic acid is now not only considered a
moisturizer but as a relevant anti-aging ingredient.
Our in vivo data on our medium molecular weight
HyaCare® confirmed this statement.
In-vivo efficacy study
Long-term evaluation of skin elasticity and

                                                                                                    % decrease in viscoelastic ratio
roughness                                                                                                                              40                          **
The study was performed at ISPE srl., Milan (I).                                                                                       35
Skin elasticity and skin roughness of an O/W cream

                                                                                                            relative to t0
containing 0.1 % HyaCare® 50 in comparison to a                                                                                        25

placebo formulation were evaluated. The                                                                                                20                                   *                Placebo
measurements were carried out in a climatic room
(24°C; 50 % rh) in order to keep the temperature and
the humidity during the measurements constant.                                                                                          0
Each of the 12 volunteers was required not to wash                                                                                              4                       8
their face for at least 3 hours before performing the                                                                                               Time (weeks)                * significant
measurements. The assessment was performed on                                                                                                                                   ** highly significant

the face, where half of the face was treated with the                                              3:
                                                                                            Figure 3: Decrease in the skin viscoelastic ratio after 4
cream containing HyaCare® 50 and the other half                                                                             formulations
                                                                                            and 8 weeks application of test formulations
with the placebo cream. The side of application on
the face (left or right) of the two products was                                            Skin image analysis showed a significant decrease of
randomised.                                                                                 maximum skin roughness of 10% of the area treated
                                                                                            with HyaCare® 50 formulation (figure 4).
At the beginning of the study instrumental
evaluation of skin elasticity and roughness were
carried out onto the left and right peri-ocular areas,

                                                                                             roughness value relative to t0
marked out in a reproducible way. The subjects

                                                                                               % decrease in maximum
applied the products on the face at home once a day                                                                                    11   *                       *
for 8 weeks. After 4 weeks and at the end of the                                                                                        9
treatment the subject returned to the laboratory for                                                                                    7                                                   HA50
                                                                                                                                        5                                                   Placebo
the instrumental measurement.
The area treated with the formulation containing                                                                                       -1
HyaCare® 50 showed a significant increase of the                                                                                                4                       8
overall elasticity value, equal to 14% after 4 and 8                                                                                                Time (weeks)
                                                                                                                                                                                         * significant
weeks of application (figure 2).
                                                                                            Figure 4: Maximum Roughness ratio after 4 and 8 weeks
                                                                                            application of test formulations

                                                                                            These data confirmed the anti-wrinkle activity of
   % increase in overall elasticity

                                       20                                                   HyaCare® 50 mainly in terms of decrease of deep
                                       15   *                      *                        wrinkles as illustrated in figures 5 and 6.
           relative to t0

                                       10                                         HA50
                                        0                                         Placebo
                                                4                      8
                                                                           * significant
                                                    Time (weeks)

Figure 2: Overall skin elasticity after 4 and 8 weeks
application of test formulations

The area treated with HyaCare® 50 showed as well a
                                                                                            Figure 5: Skin image analysis before and after 4 weeks of
highly significant decrease in the viscoelastic ratio
                                                                                            treatment with HyaCare® 50
(R6) equal to 36% after 8 weeks. This reflects the
rejuvenation of the skin through the increase of its
elastic component (figure 3).

                                                                                            Figure 6: Skin image analysis before, after 4 and 8 weeks
                                                                                            of treatment with HyaCare® 50
Preparation                                            Guide Line Formulations
HyaCare® 50 is completely soluble in water and can
be processed cold. For the preparation of both O/W         Anti-
                                                       O/W Anti-Wrinkle Cream - Filler effect
or W/O emulsions it should be added to the water           534/3/3
                                                       Mac 534/3/3
phase. In O/W emulsions HyaCare® 50 might reduce       Phase A
the viscosity of the emulsion. The desired viscosity
                                                       TEGOSOFT® CI       (Cetearyl Isononanoate)        5.0%
of O/W emulsions can be adjusted by increasing the
amount of polyacrylates (e.g. TEGO® Carbomer) or by    TEGOSOFT® Liquid                                  5.0%
increasing the concentration of consistency             Cetearyl Ethylhexanoate)
enhancers, e.g. TEGO® Alkanol 18 (Stearyl alcohol),    TEGOSOFT® DC                (Decyl Cocoate)       4.0%
TEGO® Alkanol 1618 (Cetearyl alcohol), TEGIN® M or     TEGO®   Alkanol 1618       (Cetearyl Alcohol)     3.0%
TEGIN® 4100 (Glyceryl Stearate). HyaCare® 50 shows
                                                       TEGIN® 4100 Pellets        (Glyceryl Stearate)    1.0%
good compatibility with other ingredients.
                                                       Stearic Acid                                      1.0%
                                                       Tocopheryl Acetate                                2.0%
Recommended usage concentration                        Phase B
0.01 – 0.2% of HyaCare® 50                             TEGO® Care CG 90         (Cetearyl Glucoside)     1.0%
                                                       SK-Influx                                         5.0%
                                                        (Ceramide 3; Ceramide 6II; Ceramide 1;
                                                        Phytosphingosine; Cholesterol; Sodium
                                                        Lauroyl Lactylate; Carbomer;
HyaCare® 50 is suitable for O/W and W/O                 Xanthan Gum)
formulations of:                                       HyaCare® 50                                       0.2%
•   Moisturizing products (facial and body care)       Glycerin                                          3.0%
•   Anti-aging creams and lotions                      Allantoin                                         0.1%
•   Sun care applications                              Water                                            69.2%
                                                       Phase C
Storage                                                TEGO® Carbomer 134                (Carbomer)      0.1%
For short term (few weeks), the product can be         Mineral Oil (30 mPa s)                            0.4%
stored at room temperature. For long term (over        Phase D
several months), the product should be stored in a
                                                       Sodium Hydroxide (10% in water)                   q.s.
cool place (4 °C). Avoid freezing. The unopened
product is expected to be stable up to 36 months.      Phase Z
                                                       Preservative, perfume                             q.s.
Packaging                                              Preparation:
1kg package                                            1.  Heat phase A and B separately to approx. 80°C.
                                                       2.  Add phase A to phase B with stirring 1).
                                                       3.  Homogenise.
                                                       4.  Cool with gentle stirring to approx. 60°C and
Hazardous goods classification                             add phase C.
Information concerning                                 5. Homogenize for a short time.
                                                       6. Cool with gentle stirring and add phase D
•   classification and labelling according to              below 40°C.
    regulations for transport and for dangerous        1) Important : If phase A has to be charged into the
    substances                                         vessel first, phase B must be added without stirring.
•   protective measures for storage and handling
•   measures in accidents and fires
•   toxicity and ecological effects

is given in our material safety data sheets.
W/O Night Cream – Intensive Repair
Mac 534/1/2
Phase A
ABIL® EM 90                                        2.0%
(Cetyl PEG/PPG-10/1 Dimethicone)
ISOLAN® GI 34 (Polyglyceryl-4 Isostearate)         1.0%
TEGOSOFT®   OS          (Ethylhexyl Stearate)      5.0%
Paraffinum Perliquidum                            12.0%
Paracera W 80                                      1.2%
(Microcrystalline Wax, Paramelt, B.V)
Hydrogenated Castor Oil                            0.8%
Phase B
HyaCare® 50                                        0.2%

TEGO® Cosmo C 100                 (Creatine)       0.5%
Sodium Chloride                                    0.5%
Water                                             76.8%
Preservative, perfume                              q.s.
 1. Heat phase A to approx. 80°C.
 2. Add phase B (80°C or room temperature) slowly
    while stirring.
 3. Homogenise for a short time.
 4. Cool with gentle stirring below 30°C and
    homogenise again.

                                                C 05/07

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