Soliance - Hydreïs
3 in 1 active ingredient to rehydrate,
restructure & repair
INCI China approved
Ecocert and Cosmos approved
Hydreïs Product File – May 2015          1
Summary
Vertisol ............................................................................................................................................... 3
Biological context ............................................................................................................................... 4
Production Process ............................................................................................................................. 5
Mechanism of action .......................................................................................................................... 6
Tests .................................................................................................................................................... 7
  Stimulation of the synthesis of filaggrin ........................................................................................ 7
  Stimulation of the synthesis of aquaporin .....................................................................................8
  Stimulation of the synthesis of involucrin ..................................................................................... 9
  Restoration of the skin’s structure and homeostasis ................................................................... 10
  Restoration of the skin’s structure with filaggrin .........................................................................11
  Restoration of hydric homeostasis with AQP-3 ........................................................................... 12
  Reparative effect on the barrier function ..................................................................................... 13
Ecological profile .............................................................................................................................. 14
Characteristics .................................................................................................................................. 14
Cosmetic uses ................................................................................................................................... 14
Conclusion ........................................................................................................................................ 15
Hydreïs Product File – May 2015                                                                                                                        2
Vertisol
The island of Martinique is a veritable treasure trove. If some of
its treasures are known by all, like the wealth of its creole
culture, its lush nature and its idyllic beaches, other treasures
remain hidden…
In the depths of the tropical jungle and
hidden away from the eyes of man, if you
look carefully, you can find a unique biotope
: vertisol.
Vertisol is a type of soil with a high content of
iron- and manganese-rich clay. It is the result of
thousands of years of volcanic irruptions in this
land of fire.
The particularity of this soil is that it is subject to
geological variations of extreme hydric stress.
During the dry season, the earth cracks and
opens, revealing the stigmata of drought.
Then, when the rains return, the water violently          Soliance’s biotechnology experts have managed
fills the different layers of the vertisol.               to extract its quintessence in order to enable you
                                                          to imagine the cosmetics of tomorrow, today.
Maintaining the structure of this type soil seems
impossible, yet recent research shows that it is          Just like nature, the skin needs water and
essentially ensured by the presence of exo-               essential components to repair, strengthen, and
polysaccharides.                                          rehydrate itself.
These polymers are produced naturally by a                It was to meet this need that Soliance developed
strain of bacteria that, once isolated and                Hydreïs, an oligosaccharide derived from
identified, was named Burkholderia caribensis in          Burkholderia caribensis.
homage to the Caribbean people.
                                                          Mimicking its original function, Hydreïs
This microorganism has a real benefit in terms of         restructures the skin’s architecture, acting on the
structuring the different layers of the vertisol.         formation of the stratum corneum.
Indeed, the metabolites that it produces are              Hydreïs thereby helps to maintain the skin’s role
essential to maintaining its organic stock and            as a barrier and to minimise trans-epidermal
water level, thereby contributing to fighting             water loss (TEWL).
erosion.
Hydreïs Product File – May 2015                                                                           3
Biological context
The stratum corneum                                    Involucrin
Epidermal differentiation is a continuous process      Involucrin is a major component in the cornified
that renews the upper layer of the skin surface        envelope formation.
on the stratum corneum.
                                                       It is an early differentiation marker as it is
This layer consists of a mass of flattened             synthesised by keratinocytes on the stratum
anuclear cells, the corneocytes.                       spinosum.
The corneocytes are the result of the last phase
of keratinocyte differentiation in the base layer of   On the stratum corneum, it binds to the
the epidermis.                                         membrane proteins and is crosslinked by the
                                                       action of transglutaminases.
The primary function of the stratum corneum is
to limit the loss of water from the dermis to the      This process leads to the stiffening of the
outside.                                               corneocyte membrane and the formation of the
It is also a physical barrier against external         cornified envelope.
attacks.
                                                       Involucrin and filaggrin are complementary in the
Keratinocyte differentiation is, therefore, a          establishment and the protection of the skin
fundamental process because, over time, any            barrier allowing to resist external aggression.
disturbance can affect the "barrier" role.
The stratum corneum and the maintenance of its         Aquaporin-3
structure are essential for the skin.                  Aquaporins (AQP) are a class of membrane
                                                       proteins that form pores that are permeable by
In this knowledge, Soliance carried out                water molecules and glycerol in biological
research on proteins that it needs to carry out        membranes.
its role and repair itself: filaggrin, involucrin      Aquaporin-3 (AQP3) has the highest expression
and aquaporin-3.                                       in normal human keratinocytes.
                                                       Furthermore, recent research (Li, 2010) has
Filaggrin
                                                       proven that its expression reduces with age.
In the skin, filaggrin, or filament aggregating
                                                       This result further highlights the role of
protein, is a late differentiation marker involved
                                                       aquaporin-3 in maintaining skin hydration.
in hydration and the barrier function of the
epidermis.
                                                       As well as hydration, aquaporin-3 could also
                                                       have a role in the mechanisms of the
Filaggrin is synthesized in the form of a giant
                                                       proliferation and differentiation of keratinocytes.
precursor - profilaggrin (>400 kDa) – in the
stratum granulosum.
It is the principal component of keratohyalin
granules.
In the last phases of keratinocyte differentiation,
profilaggrin is proteolysed into many filaggrin
monomers.Filaggrin has two essential biological
functions:
 It bonds with intermediary keratin filaments,
 causing them to aggregate into macrofibrils
 aligned in compact parallel rows.
 Furthermore, the filaggrin is hydrolysed into
 free amino acids that make up part of the
 natural moisturising factors (NMF). They bind
 the water molecules, thereby keeping the
 middle and upper layers of the stratum corneum
 moisturised.
Hydreïs Product File – May 2015                                                                         4
Production Process
The nature of Hydreïs                                 Production secrets
Hydreïs is an active ingredient whose proven          Hydreïs is produced industrially using an
activity is to repair, strengthen and moisturise      ecoresponsible biotechnological process.
the cutaneous barrier.
                                                      The raw materials used are renewable plant by
It is a oligosaccharide with a controlled molecular   products from crops grown regionally.
weight of between 1 and 15 kDa.
                                                      The functionalisation of these materials via
This molecular weight facilitates its penetration     Hydreïs’s production process is part of a policy of
and its activity in the skin.                         generating value from the entire plant.
Hydreïs’s principal chain is made up of 1 to 15       This concept is exploited by the Biorefinery of
monomer repeat units.                                 Pomacle Bazancourt in Champagne Ardenne.
This unit is composed of glucoses linked in β1-4      Selective purification makes it possible to obtain
and an original sugar: talose, a C2 epimer of         active oligomers weighing between 1 and 15
galactose.                                            kDa. It is specifically these oligomers that show
                                                      biological activity
   Figure 4: Hydreïs production process
Hydreïs Product File – May 2015                                                                         5
Mechanism of action
Hydreïs acts, first of all, by stimulating the repair         This biological activity makes it possible to:
of the involucrin and the filaggrin, responsible for           Structure the filaggrin-keratin matrix,
the structure of the stratum corneum and the                   Reinforce the cornified envelope,
natural moisturising factors (NMF).                            Generate sufficient quantities of natural
                                                               moisturising factors (NMF), to fix the water
Hydreïs also stimulates the synthesis of the                   molecules and restore intercellular flux.
components of intercellular hydration canals
called the aquaporin-3.                                       TEWL is reduced and hydration is maintained.
                     Figure 5: Mechanism of action of Hydreïs and metabolisms
Hydreïs Product File – May 2015                                                                                6
Tests
Stimulation of the synthesis of filaggrin
In vitro test
Protocol                                           Results
Normal human keratinocytes were inoculated and     Quantification of filaggrin
placed in a humid incubator at 37°C with 5% of
CO2 for five days (duration of the treatment).                                               **
Hydreïs was tested at 10 mg/ml (0.01% DM) in
the culture medium in the presence of cells.
Cells were left untreated in order to obtain a
control condition.
                                                     Average intensity
Filaggrin expression was revealed by
immunolabelling in green.
The nuclei were coloured in blue.
The cells were observed by optical fluorescence
microscopy with an enlargement of X10
(Olympus CK40 microscope).
                                                                            Control       Hydreïs
The quantification was carried out by analysing                                          10 mg/ml
the images with HistoLab software (Microvision).
                                                              ** = p<0,01
                                                   Figure 6: Intensity of filaggrin
                                                   Hydreïs causes a significant overexpression of
Immunolabelling after treatment:                   filaggrin compared to the control.
Figure 7 a. Untreated cells in control (x10)       b. Cells treated with Hydreïs (x10)
Hydreïs Product File – May 2015                                                                     7
Stimulation of the synthesis of aquaporin
In vitro test
Protocol                                           Results
Normal human keratinocytes were inoculated and     Quantification of aquaporin-3
placed in a humid incubator at 37°C with 5% of
CO2 for five days (duration of the treatment).                                             *
Hydreïs was tested at 10 mg/ml (0.01% DM) in
the culture medium in the presence of cells.
Cells were left untreated in order to obtain a
control condition.
                                                      Average intensity
The aquaporin-3 expression was revealed by
immunolabelling in red.
The nuclei were coloured in blue. The cells were
observed by optical fluorescence microscopy with
an enlargement of X10 (Olympus CK40
microscope).                                                               Control        Hydreïs
                                                                                         10 mg/ml
                                                             * = p<0,05
The quantification was carried out by analysing
the images with HistoLab software (Microvision).
                                                    Figure 8: Intensity of aquaporin-3
                                                   Hydreïs causes a significant overexpression of
                                                   aquaporin-3 compared to the control.
Immunolabelling after treatment:
Figure 9: Untreated control cells (x10)            b. Cells treated with Hydreïs (x10)
Hydreïs Product File – May 2015                                                                     8
Stimulation of the synthesis of involucrin
In vitro test
Protocol                                             Results
Normal human keratinocytes were inoculated and       Quantification of involucrin
treated with 10mg/ml of Hydreïs for 5 days. Cell
pellets are rinsed with PBS and stored at -20 °C
before sonication.
Assays are performed with ELISA method at 450                                                                        *
nm.
                                                        [involucrine] (ng/ml/μg prot)
After 5 days of treatment with Hydreïs, we
observe the following effects: no involucrin
detected in the untreated control but significant
increase of involucrin for the pellet treated with
Hydreïs to 10mg/ml.
The involucrin expression was revealed by
immunolabelling in red.
The nuclei were coloured in blue.
                                                                                                     Control    Hydreïs
The cells were observed by optical fluorescence                                                                10 mg/ml
microscopy X10 (Olympus CK40 microscope).                                               * = p<0,05
                                                     Figure 8: Quantification of involucrin
                                                     Hydreïs causes a significant overexpression of
                                                     involucrin compared to the control.
Immunolabelling after treatment
Figure 9. Untreated control cells (x10)              b. Cells treated with Hydreïs (x10)
Hydreïs Product File – May 2015                                                                                           9
Restoration of the skin’s structure and
homeostasis
Ex vivo test
Protocol
Skin explants from the abdominal surgery of a 54                                                                  **
year-old Caucasian woman were left to stabilise
for four hours at 37°C in a DMEM medium with
10% of serum, L-glutamin and antibiotics.
The explants were delipidated in order to alter
the cutaneous barrier to then observe the effect
                                                        Average intensity
of Hydreïs on the restoration of the stratum
corneum.
The delipidation was done with a mixture of ether
and acetone for 2 minutes. The control explants
were not delipidated.
The other explants were treated with 20µl:
                                                                                       Control       Placebo    Hydreïs 1%
 of a 0% Hydreïs emulsion (placebo)
 of a 1% Hydreïs emulsion.                                                  ** = p<0.005               Delipidated skin
                                                      Figure 10: Intensity of filaggrin
The skin explants were then incubated at 37°C
for five days, then the expression of the filaggrin
and the aquaporin-3 was revealed by
immunolabelling.
The quantification was carried out by analysing                                                                   ***
images with HistoLab software (Microvision).
Results
                                                        Average intensity
 On non-delipidated skin, the intensity of
 filaggrin and AQP-3 marking was 2000 and
 1000 respectively.
  On delipidated skin treated with a placebo, the
 cutaneous barrier was altered and the marking                                                          **
 intensity of these proteins was significantly
 reduced.
                                                                                           Control    Placebo    Hydreïs 1%
On delipidated skin treated with Hydreïs, the
quantity of filaggrin and AQP-3 is higher than                                                       Delipidated skin
that of the non-delipidated control skin.                                   ** = p<0.005 *** = p<0.001
                                                      Figure 11: Intensity of aquaporin-3
Hydreïs Product File – May 2015                                                                                           10
Restoration of the skin’s structure with filaggrin
Ex vivo test
Immunolabelling after treatment                   NON-DELIPIDATED UNTREATED - CONTROL
The expression of filaggrin was revealed by
immunolabelling in green.
The nuclei were coloured in blue.
The skin samples were observed by fluorescent
optical microscopy with an enlargement of X10
(Olympus CK40 microscope).
Homogenous marking of filaggrin in the stratum
corneum.
Many marked cell layers at the base of the
stratum corneum.                                      DELIPIDATED – PLACEBO TREATMENT
Significant alteration of the stratum corneum.
Very heterogeneous epidermal marking.
Some cell layers of the upper layer are not
marked.
Homogenous marking in the form of a thin layer        DELIPIDATED – HYDREÏS TREATMENT
in the upper layer of the epidermis and the
stratum corneum.
Strong expression in several cell layers at the
base of the stratum corneum.
Better profile than that of the control.
Hydreïs restores the hydration barrier of
sensitive skin by creating a higher level of
filaggrin than that of normal skin
Hydreïs Product File – May 2015                                                   11
Restoration of hydric homeostasis with AQP-3
Ex vivo test
Immunolabelling after treatment                     NON-DELIPIDATED UNTREATED - CONTROL
                                                   NON- DELIPIDATED UNTREATED - CONTROL
The expression of the aquaporin-3 was observed
by Immunolabelling in red.
The nuclei were coloured blue. The skin samples
were observed by fluorescent optical microscopy
with an enlargement of X10 (Olympus CK40
microscope).
Homogenous marking of aquaporin-3 strongly
expressed in the stratum corneum.
                                                         DELIPIDATED – PLACEBO TREATMENT
                                                        DELIPIDATED – PLACEBO TREATMENT
Significant alteration of the stratum corneum.
Very heterogeneous epidermal marking.
Low level of marking: little aquaporin-3
expression.
                                                         DELIPIDATED – HYDREÏS TREATMENT
                                                        DELIPIDATED – HYDREÏS TREATMENT
Restoration of the stratum corneum.
Homogenous marking, in the upper layer of the
epidermis and the stratum corneum (with a
higher intensity in the stratum corneum).
Better profile than that of the control.
Hydreïs re-establishes cell homeostasis in
sensitive skin by reaching a level of aquaporin-
3 higher than that of normal skin.
Hydreïs Product File – May 2015                                                      12
                Reparative effect on the barrier function
                In vivo test
                Protocol
                The efficacy of Hydreïs was evaluated on a panel                  These values were measured the morning at D1
                of 19 volunteers with caucasian skin and with an                  and D3:
                average age of 50.                                                 T0 before stripping (TEWL0),
                                                                                   T immediately after stripping (TEWLi),
                They applied a placebo cream and a cream                           T1h after application of the cream (TEWL1)
                containing 1% of Hydreïs on their forearms twice
                a day (morning and evening) for three days.                       Using a Vapometer® (Delfin) on the zones
                                                                                  treated with placebo or with Hydreïs.
                A prior treatment of the skin using the stripping                 ∆TEWL = TEWL1-(TEWLi-TEWL0)
                technique with adhesive discs was carried out at
                D1 and D3 in order to reduce the effect of the                    At D1, the variation in TEWL caused is not
                natural barrier of the upper layers of the                        significantly different between the placebo cream
                epidermis.                                                        and Hydreïs.
                                                                                  At D3, we observe:
                The expected reparative effect of the products                      An increase in TEWL compared to D1 with the
                tested on the upper layers of the epidermis was                    placebo cream, showing its inefficacy.
                evaluated by comparing TEWL average values in                       A reduction in TEWL compared to D1 with the
                strictly controlled conditions of temperature and                  Hydreïs cream, which can be correlated to a
                humidity.                                                          reparative effect of the cutaneous barrier.
                                                                                  Hydreïs significantly restores the skin’s barrier
                                                                                  function compared to placebo, reducing the
                                                                                  variation of TEWL by 103%.
                                                                                       *                       Placebo
                                                                                                               Hydreïs 1%
                                                                                                               * = p<0.05
TEWL (g/m .h)
2
                                                                                              -103% TEWL
                                   Day 1                                          Day 3
                Figure 12: Variation of the TEWL after application of the cream
                Hydreïs Product File – May 2015                                                                                 13
Ecological profile
OCDE 301F Biodegradability
Figure 13: Biodegradability of Hydreïs after 28 days
Hydreïs is 100% biodegradable in relation to
its chemical oxygen demand after 10 days and
is therefore classed as easily biodegradable,
according to OCDE directive 301F.
OECD 202 Ecotoxicity
Hydreïs has no impact on aquatic life.
Indeed, the study of the inhibition of 50%
Daphnia magna mobilisation shows Hydreïs
ECi50/48h is superior to 1g/L.
Characteristics                                        Cosmetic uses
Hydreïs is approved by Ecocert and Cosmos.             Hydreïs is suited to mature skin or skin that is
                                                       sensitive because of cold or dry conditions,
Hydreïs INCI Name is Water (and) Hydrolyzed            pollution, or other environmental factors.
beta-glucan.
Preservative: Sodium benzoate and potassium
sorbate
Recommended dosage: 1%
Hydreïs Product File – May 2015                                                                           14
Conclusion
Hydreïs is a patented active produced through white biotechnology that
is 100% biodegradable after 10 days.
Its efficacy is proven by in vitro, ex vivo and in vivo tests.
It stimulates the synthesis and the restoration of essential components of
the hydration and repair of the cutaneous barrier; involucrin, filaggrin
and aquaporin-3.
Hydreïs Product File – May 2015                                         15