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EURO COSMETICS
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New data confirm skin revitalizing and
stress protection by Glycoin® natural
By Silke Karin Schagen, Sabrina Overhagen, Andreas Bilstein *
Abstract
Glycoin® natural (INCI: Glyceryl Glucoside) is an ECOCERT certied multifunctional active ingredient. In nature
this strong extremolyte is produced by the desert resurrection plant Myrothamnus abellifolia and blue-green algae
(cyanobacteria). For cosmetic use the natural glyceryl glucoside isomer (2-O-a-D-glucopyranosyl glycerol) is manu-
factured with high purity by an enzymatic process. Glyceryl glucosides are mentioned for increasing skin elasticity,
moisturizing, and reduction of symptoms like itching, burning, tightness, tingling, and feeling of dryness. In vitro
assays in cell cultures and skin models demonstrate strong activity of Glycoin® natural on aged and stressed skin
cells with rejuvenating, recovering and radical scavenging properties. In vivo treatments conrm positive effects on
increased skin elasticity, smoothness and thickness; the epidermis and dermis density regenerates.
Introduction
Glycoin® natural (INCI: Glyceryl Glucoside) is used as a multi-
functional anti-aging, cell-stimulating and skin moisturizing agent
in cosmetics. In skin, glyceryl glucoside treatment enhanced gly-
cerol derivatives and increased the aquaporin 3 expression in vi-
tro and ex vivo1). Glyceryl glucosides are mentioned for increas-
ing skin elasticity, moisturizing, the reduction of symptoms like
itching, burning, tightness, tingling, and feeling of dryness 2).
Glycoin® natural belongs to the group of glyceryl glucosides
and the chemical isomer is called “2-O-a-D-glucopyranosyl glycer-
ol”. This special glyceryl glucoside isomer is a strong extremo-
lyte 3). In nature 2-O-a-D-glucopyranosyl glycerol is produced by
the resurrection plant Myrothamnus abellifolia and Spirulina
subsalsa “marine blue-green algae”4, 5). Glyceryl glucoside is an
osmoregulatory and stress protecting molecule, which protects
plants or bacteria under extreme conditions. The presence of
glyceryl glucoside diminishes stresses and strains in the mem-
branes at low or zero hydration level 6).
The plant Myrothamnus abellifolia for example is known for
its survival strategies under extreme conditions. It could remain
in the dry state under simulated eld conditions for many years,
without losing viability7). The described glyceryl glucoside iso-
mer is synthesized during dry season (dehydrating process) in the
desert plant, to protect the plant’s structure from damage due to
total desiccation5). In this context, Ferjani et al. study results de-
scribe glyceryl glucoside action as an osmoprotectant 8).
As soon as water is available again the glyceryl glucoside acts
as energy booster and the Myrothamnus abellifolia revives and
re-greens 9).
Cyanobacteria like marine blue-green algae (Spirulina subsalsa)
also synthesize and accumulate 2-O-a-D-glucopyranosyl glyce-
rol 4). Like the Myrothamnus flabellifolia, blue-green algae use
glyceryl glucoside as stress-protection molecule10). In general,
these algae favor fresh water habitats, but some strains prosper in
strongly alkaline salt lakes or marine environments11).These
blue-green algae are characterized by their impressive ability to
synthesize and accumulate glyceryl glucoside to acclimate in
habitats with high or fluctuating salinity12, 10). Glyceryl glucosides
are also naturally present in red, rosé and white wine, as well as
in Japanese food brewed with koji, like sake, miso and mirin 13).
Glycoin® natural production
bitop AG produces Glycoin® natural, the only natural glyceryl
glucoside (2-O-a-D-glucopyranosyl glycerol) currently available
for the cosmetics industry, in Germany with a high purity. The
patented production process was developed in cooperation with
the University of Graz, Austria. The Glycoin® natural production
Picture 1:
Myrothamnus flabellifolia during
dry season (left), Myrothamnus
flabellifolia after rain (right)
*) Contact: Eva Galik, bitop AG, galik@bitop.de, www.bitop.de
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is realized by a biocatalytical process, an enzymatic reaction,
using the enzyme “succrose phosphorylase”. The natural
substrates succrose and glycerine are transferred into Glycoin®
natural and fructose. After the downstream process, a highly pure
and all-natural glyceryl glucoside remains. In this production
process, the yield of Glycoin® natural is 90%14).
Cell-protection and stabilization properties
by Glycoin® natural treatment
Toxicological study results show that the use of Glycoin® natu-
ral (concentration 0.5% up to 3% w/w) as active ingredient in
cosmetic/cosmeceutical formulations exhibits no aberrant risks
for local toxicity or mutagenic properties. An additional in vivo
sensitization study (50 volunteers) of Glycoin® natural cosmetic
formulation revealed neither sensitizing properties nor any other
aberrant adverse effects. Furthermore Glycoin® natural did not
show any irritating effect to the skin, eyes or mucosa in different
in vitro and in vivo studies.
An in vitro model was established to demonstrate that glyceryl
glucoside protects cell membranes. Keratinocytes were pre-
treated with Glycoin® natural or Myrothamnus plant extract and
then stressed with 0.001% SDS. Only the pure Glycoin® natural
protected the cell membranes of keratinocytes from damage
(gure 1).
Glycoin® natural in vitro results
show anti-aging effects on skin cells
Different in vitro models were used to investigate anti-aging
effects of Glycoin® natural on human skin cells.
Cell vitality by glucose consumption
Cell vitality was determined by investigation of glucose con-
sumption in human epidermal keratinocytes. Glucose plays a role
in the energy metabolism of keratinocytes. It is associated with
cell regeneration and renewal.
These results show the cell viability of human epidermal ke-
ratinocytes cultivated with Glycoin® natural (1%) versus untreated
control. The Glycoin® natural treatment signicantly increases
cell vitality compared to control. The more active cells are, the
more glucose they need for their metabolic activity.
Glycoin® natural increases the vitality and metabolic activity of
aging skin cells by up to 170% (gure 3). These effects may be
useful in reversing dried, aged skin conditions to promote healthy
regenerated skin.
Figure 1: Keratinocytes were pre-treated with Glycoin® natural or Myrothamnus
plant extract (concentration: 0.03%, 0.04% and 0.05%) Then cells were stressed
with SDS and cell membrane stability was measured.
Figure 3:
Measured cell viability during
cell aging per treatment days
(measured by glucose consump-
tion).
Figure 2: ATP metabolism in aged human keratinocytes after pre-treatment with
Glycoin® natural and a Myrothamnus plant extract at different concentrations.
In human keratinocyte cell line (HaCaT) treatment only Glycoin®
natural stimulates the ATP production in aged skin cells compared
to plant extract (gure 2).
Adenosine triphosphate increase by
Glycoin® natural treatment –
leads to stimulation of cell vitality
In two different studies, keratinocyte cell models were treated
with Glycoin® natural (1%). Afterwards adenosine triphosphate
expression was measured. Adenosine triphosphate (ATP) trans-
ports energy within cells for metabolism. It is responsible for
cellular activity and vitality, DNA replication, regeneration and
healing as well as collagen and elastin synthesis. Skin cells syn-
thesize great amounts of ATP. During the skin aging process the
amount of ATP in skin cells decreases constantly 15).
The results of these in vitro models show that cell treatment
with Glycoin® natural stimulates ATP expression. An increase of
ATP leads to a stimulation of cell vitality and cell renewal. With
chronological aging (passage of time) and extrinsic skin aging
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(external stress), skin cells show a clear reduction of ATP produc-
tion, followed by a reduced metabolic activity and cell vitality.
In an in vitro human keratinocytes (HaCaT cells) model, HaCaT
cells were aged over 2 weeks and then treated with Glycoin®
natural, glucose, fructose and glycerin (Concentration: 0.75%,
0.5% and 0.1%). After 6h incubation ATP concentration was meas-
ured with luciferase assay. Beside the increase of the cell metabo-
lism, Glycoin® natural also signicantly increased the ATP con-
centration in aged skin cells (gure 4).
DNA array study confirms skin cell recovery
A DNA array study presented results which indicate that skin
treatment with Glycoin® natural may improve aged skin, deliver
anti-oxidant, anti-stress, healing and maybe whitening effects.
Glycoin® natural cell treatment induces the expression of growth
factors like broblast growth factor 7 (FGF7) and transforming
growth factor beta 1 (TGF-beta 1) which are involved in the
wound healing process and initiate the proliferation of new der-
mal cells. FGF7 is also mentioned for potent regenerative, cyto-
protective effects against oxidative stress, toxic compounds and
UV irradiation, and usually is up-regulated after injury and during
healing.
Additionally, the expression of ROS scavenger molecules (cata-
lase (CAT), superoxide dismutase 2 (SOD2), metallothionein-1
(MT1H)) are enhanced by Glycoin® natural 17).
Viability-enhancing effect by Glycoin® natural
With an epidermis model the positive effect of Glycoin® natural
treatment was investigated. The Glycoin® natural treatment on
non-irradiated epidermis models (SkinEthic®) was able to achieve
a viability-enhancing effect. The cells, which were in good condi-
tion and not stressed, experienced a “vitality boost” by adding
Glycoin® natural.
In addition, an increased viability versus untreated control cells
could also be demonstrated in an UV-irradiated epidermis model.
In both study models, a reduction of IL-1 alpha levels after Gly-
coin® natural treatment was achieved. This correlated with the
viability results. The increase in vitality, measured by the release
of lactat dehydrogenase (LDH), could be shown in both experi-
ments.
As a result of the UV irradiation, damage to the cell membranes
and the increase in the LDH values could be determined. In com-
parison to the untreated control, the addition of Glycoin® natural
resulted in a reduction in LDH release and thus an improvement
in membrane integrity and cell vitality (data not shown).
In vivo efficacy studies: Improved
skin elasticity and epidermis and dermis density
In a placebo controlled, blinded in vivo study with 20 Cauca-
sian volunteers a cream containing 1% Glycoin® natural was test-
ed. Cream was applied twice daily for 4 weeks. Parameters for
skin moisturizing, elasticity (cheek) and smoothness (lower leg)
were measured. The study documented a moisturizing rise of 23%
after application, an increase of skin elasticity by 93%, and of skin
smoothness by 62%. The cream containing 1% Glycoin® natural
was tolerated very well according to clinical dermatological crite-
ria and led to a reduction in the roughness of the skin.
Figure 4: ATP concentration in aged human keratinocytes after pre-treatment
with Glycoin® natural or glucose at different concentrations. Glucose was used
as control for a potential ATP increase induced by sugar treatment.
Glycoin® natural stimulates
anti-oxidant activity in aged skin cells
An in vitro study on human epidermal keratinocytes showed
that the cell treatment with 1% Glycoin® natural increased pro-
duction of the enzyme superoxide dismutase 1 (SOD1). SOD1 is
an important anti-oxidant which occurs naturally in the human
body. It plays a part in the skin’s self-defense system, reduces free
radical damage, and therefore prevents signs of aging, helps to
heal wounds, softens scar tissue and protects against UV induced
damage rays. Natural superoxide dismutase levels and activity
slow down during the aging process 16).
Glycoin® natural treated aged keratinocytes produce up to
280% more SOD1 compared to untreated cells. Glycoin® natural
increases the amount of SOD1, anti-oxidant activity and seems to
strengthen the skin’s own defense and immune system.
Figure 5: Amount of SOD1 per
cell during the cell aging pro-
cess. At day 5 Glycoin® natural
treated aged keratinocytes
produce up to 280% more SOD1
compared to untreated cells.
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Figure 6: Subepidermal low-echogenic band measured the increase of epidermal
and dermal thickness and density at the beginning and the end of the treatment
period. Glycoin® natural increased significantly epidermal and dermal thickness
and density compared to placebo.
An additional 4-week blinded, placebo-controlled cosmetic ef-
cacy study evaluated the anti-aging properties of a Glycoin®
natural containing cosmetic cream. Five Caucasian female volun-
teers were treated and assessed by ultrasonography on the inner
forearms. Epidermis and dermis density increased up to 9.9% af-
ter applying Glycoin® natural containing formulation (gure 6).
This increase was statistically signicant and better than the pla-
cebo. All volunteers reported easy and comfortable application
conditions of Glycoin® natural containing formulation. The topi-
cal Glycoin® natural containing formulation regenerates the epi-
dermis and dermis density.
Summary and discussion
Glycoin® natural is a 100% natural active anti-aging ingredient.
Glyceryl glucosides are already established in cosmetic formula-
tions, but have only been available as a synthetically produced
racemate. Glycoin® natural is the only natural alternative consist-
ing of the molecule preferentially formed in nature. Several in
vitro and in vivo studies showed the cell protecting and recover-
ing effect of Glycoin® natural treatments. In vitro models and skin
models demonstrate strong activity on aged and stressed skin
cells with rejuvenating, recovering and radical scavenging proper-
ties. In vivo treatments conrm positive effects on increased skin
elasticity, smoothness and thickness, the epidermis and dermis
density regenerate with no irritating effects.
A number of studies conrm Glycoin® natural results. Vehicle
controlled in vivo studies of two glyceryl glucoside formulations
demonstrated signicantly improved hydration versus controls.
Furthermore signicant reduction of transepidermal water loss
versus untreated site and basic vehicle was shown. Both formula-
tions (rich and light) signicantly decreased visible dryness and
tactile roughness. The rich formulation maintained signicant
moisturization and the light formulation reduced symptoms of
itching, burning, tightness, tingling, and feeling of dryness 2). In a
topical study by Harada et al. a cosmetic cream with alpha-D-
glucosyl glycerol was applied on 13 female volunteers. The re-
searchers observed an increase of elasticity in cheek-skin and
suggested that alpha D-glucosyl glycerol increases the production
of IGF-I in the skin through sensory neuron stimulation, thereby
increasing skin elasticity 17). This elasticity results may conrm
Glycoin® natural treatment observations.
In conclusion Glycoin® natural is useful for anti-aging skin care
and stress protecting formulations for mature, tired and stressed
skin.
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