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Microsoft word - new_studies_demonstrate_anti-cellulite_efficacy.doc
Slimming and Tissue Strengthening: Soy Isoflavone for a Perfect Body
Dr. Daniel Schmid, Esther Belser, Sandra Meister
An ingredient comprising the soy isoflavone genistein, caffeine, carnitine and an extract of the Spirulina
platensis algae was found in a clinical trial to be very effective against cellulite. In vitro studies in a 3D-full skin
model, showed the capacity of genistein to inhibit differentiation of preadipocytes into adipocytes and to
stimulate production of collagen IV. Thus, genistein counteracts cellulite on two levels; it reduces storage of
lipids and it strengthens the skin epithelium.
Inhibition guarantees a high pool of cAMP. This in turn
activates the enzyme lipase which is responsible for the
Dimpling of the skin of the buttocks and thighs is
hydrolysis of triglycerides. The long chain fatty acids
known as cellulite, a typical problem for women. They
are then oxidized to CO2 and H2O to produce energy.
have a characteristic depot of subcutaneous adipose
Caffeine also stimulates microcirculation because the
tissue in the gluteal-femoral region (pear shape). The
increased release of catecholamines will induce
adipose tissue is a type of connective tissue that is
vasodilation. The improved blood circulation in the
specialized for the storage of fat (triglycerides). There
cellulite area helps in the breakdown of fat.
is also a gender-related difference in the anatomy of
the adipose tissue. In women, the organization of the
Carnitine, a quaternary ammonium compound, is
subcutaneous fat cell chambers and of the connective
another group A active. Carnitine’s role is to carry long
tissue septa that divide them, allows vertical stretching
chain fatty acids into the mitochondria. These are the
of the fat chambers. After stretching, the chambers
cellular organelles where fatty acids are oxidized.
protrude into the reticular dermis, leading to the
Spirulina platensis is a filamentous blue-green algae
species that naturally lives in tropical and subtropical
Cosmetic ingredients to treat cellulite can be
lakes of high alkalinity. The extract of Spirulina cells is
a rich source of carotenoids, unsaturated fatty acids,
essential amino acids and minerals. The carotenoids
protect the skin against oxidative stress. The algae
extract is known to stimulate elasticity and firmness of
the skin and thus contributes to skin strengthening
Compounds that stimulate microcirculation
(group B active). A firmer skin resists better the vertical
stretching of the fat chambers into the reticular dermis
This article describes an anti-cellulite ingredient
comprising the soy isoflavone genistein, caffeine,
carnitine and an extract of the Spirulina platensis
The use of the soy isoflavone genistein as an anti-
cellulite active seems at first glance not to be logical
because soy isoflavones are known as phytoestrogens.
Cellulite is a typical female phenomenon and thus
Caffeine is the best known representative of group A.
primarily regulated by estrogen. The structures of soy
Like theobromine (chocolate) and theophylline (tea) it
isoflavones, heterocyclic phenols, are very similar to the
belongs to the group of xanthine derivatives. They
steroidal estrogen. Thus they can bind to some extent to
stimulate mobilisation of lipids by increasing the
estrogen receptors. Compared with the principal
release of catecholamines and by inhibiting the
circulating estrogens in humans, isoflavones are bound
activity of the enzyme phosphodiesterase. This
at a much lower rate (103-fold less). Genistein, the
enzyme normally degrades cyclic AMP (cAMP), an
predominant isoflavone in soy, is also a well-known
important signaling molecule in the lipolysis process.
inhibitor of protein tyrosine kinases (1).
Kinases are involved in the signaling process inside
Compared to the cells in the differentiation medium
cells. In the skin, they are essential components of the
without genistein, labeling of the lipid droplets showed
signal transduction pathway that is activated by
that genistein at 25 µM reduced adipogenesis by 31%
radicals and inflammatory cytokines and leads to the
after 3 days and by 42% after 5 days incubation in the
expression of collagen- and elastin-degrading enzymes
differentiation medium. This strong inhibitory effect is
(2;3). These enzymes, called matrix clearly visible in the fluorescence pictures (figure 2). metalloproteinases, are key players in skin aging.
Genistein can block this signaling pathway and as a
Figure 2: Fluorescence labelling of lipid droplets (right: differentiation medium
consequence, radicals and inflammatory cytokines can
with genistein; left: control without genistein)
no longer induce the breakdown of collagen and
elastin. In this way, genistein helps to strengthen the skin (group B active). Several scientific reports show a further physiological activity of genistein. It is an effect that is specifically directed at adipocytes. Genistein was shown to act as a negative regulator of adipogenesis (4;5). Thus, genistein seems to be an efficient anti-cellulite active, belonging to the categories A and B. In the next two
chapters, in vitro studies are described that
Also the results of the gene array analysis reflected the
demonstrate the efficacy of genistein to reduce the
efficacy of genistein to inhibit adipogenesis (figure 3).
formation of adipose tissue and to enhance the concentration of collagen in a 3D-full skin model.
Inhibtion of the Differentiation of Preadipocytes
The mass of the adipose tissue can be controlled either by the recruitment of new adipocytes (adipogenesis)
or by the volume of the existing adipocytes (Figure 1).
■ Markers of adipogenesis process■ Markers of preadipocytes
Figure 3: Effect of genistein on gene expression of preadipocytes in the differentiation medium
Figure 1: Differentiation of preadipocytes leads to the formation of new adipocytes
Compared to the cells in the differentiation medium
without genistein, typical markers for mature adipocytes
Adipogenesis is the differentiation process of
were reduced whereas markers for preadipocytes were
preadipocytes (fibroblast-like) into adipocytes. The
conversion is normally regulated by various hormones
and is associated with the coordinate induction of key
Stimulation of Collagen IV in the Basement
enzymes of the lipid metabolism. In this study, the
effect of genistein on adipogenesis was investigated.
Human preadipocytes were cultured until the cells
Most important for the strength of the skin tissue are the
reached confluence. Then the medium was replaced
collagen proteins. There are at least 14 types of
by a specific differentiation medium with genistein.
collagens that differ in their structural organization.
The controls were preadipocytes kept in the normal
Collagen IV forms sheet-like networks and is specific
medium (not differentiated control) and preadipocytes
for basement membranes. The skin basement membrane
cultured in the differentiation medium without
is a thin matrix between the epidermis and the dermis
genistein. The conversion of preadipocytes into
and serves as an adherent connection between the two
mature adipocytes was analyzed using two methods:
skin layers. The basement membrane is very important
1. Demonstration of accumulation of lipids in large fat
for the attachment of the basal keratinocytes and for
droplets (adipocyte-specific phenotype). The lipid
their proliferation. A natural basement membrane is also
droplets were labeled with a fluorescent dye
formed in 3D-full skin models. This model was used to
(AdipoRed®). The number of total cells was
analyze the effect of genistein on the synthesis of
controlled by Hoechst staining of the cell nuclei. 2.
collagen IV. The in vitro skin model was incubated for
Analysis of adipocyte-specific metabolism by gene
several days with a medium containing 5 µM genistein.
array. A chip containing 96 genes related to
The concentration of collagen IV was analyzed with
immunofluorescence microscopy using anti-collagen IV
antibodies. The number of total keratinocyte and
fibroblast cells was controlled by Hoechst staining of
The immunofluorescence pictures (figure 4) show the
localization of collagen IV in the basement membrane.
Incubation of the skin model in a medium with
genistein clearly stimulated the synthesis of collagen
IV (+ 53% after 6 days). The analysis of cell nuclei
showed that this stimulation was specific and not only
Figure 5: Improvement of skin elasticity and smoothness after 6 weeks application of a cream with 4% Iso-SlimComplex
Figure 4: Immunofluorescence labelling of collagen IV (upper row) and cell nuclei (lower row). The pictures on the right are from 3D-full skin models
incubated with genistein. On the left side are pictures from skin models
Figure 6: Reduction of cellulite appearance after 6 weeks application of a cream
Clinical Study with an Anti-Cellulite Formulation
A cosmetic ingredient to treat cellulite (Iso-
SlimComplex), comprising the soy isoflavone
genistein, caffeine, carnitine and an extract of the
Spirulina platensis algae, was tested in a clinical study
(1) Akiyama T., Ishida J., Nakagawa S., Ogawara H., Watanabe S.,
over 6 weeks with 20 women aged from 39 to 58 with
Itoh N., Shibuya M. and Fukami Y. (1987) Genistein, a specific
light to heavy cellulite. A cream containing 4%
inhibitor of tyrosine-specific protein kinases. J. Biol. Chem. 262: 5592-5595
IsoSlimComplex was applied once daily on the right
thigh. The other thigh stayed untreated and served as a
(2) Kajanne R., Miettinen P., Mehlem A., Leivonen S.-K., Birrer M.,
control. The following skin parameters were
Foschi M., Kähäri V.-M. and Leppä S. (2007) EGF-R Regulates MMP
measured: smoothness (PRIMOS system), elasticity
Function in Fibroblasts Through MAPK and AP-1 Pathways. J. Cell. Physiol. 212: 489 – 497
(cutometer), thigh circumference and cellulite degree
(3) Xu Y., Shao Y., Voorhees J.J. and Fisher G.J. (2006) Oxidative
inhibition of receptor-type protein-tyrosine phosphatase kappa by
The results clearly demonstrated the efficacy of Iso-
ultraviolet irradiation activates epidermal growth factor receptor in human keratinocytes. J. Biol. Chem. 281: 27389 – 27397
SlimComplex. Compared to the untreated areas,
elasticity and smoothness improved after 6 weeks’
(4) Park H.J., Yang J.Y., Ambati S., Della-Fera M.A., Hausman D.B.,
application by 57% and 41% respectively (figure 5).
Rayalam S. and Baile C.A. (2008) Combined effects of genistein,
The circumference of the right thigh was reduced by
quercetin, and resveratrol in human and 3T3-L1 adipocytes. J. Med.
3.8 cm after 6 weeks whereas the circumference of the
untreated thigh was only reduced by 0.8 cm. There
(5) Liao Q.C., Li Y.L., Qin Y.F., Quarles L.D., Xu K.K., Li R., Zhou
was a visible improvement of the cellulite appearance.
H.H. and Xiao Z.S. (2008) Inhibition of adipocyte differentiation by
The cellulite degree dropped from 3.7 to 2.5 after 6
phytoestrogen genistein through a potential downregulation of
weeks treatment (Figure 6). There was no visible
extracellular signal-regulated kinases 1/2 activity. J. Cell. Biochem.
change in cellulite on the untreated thigh.
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