Provislim™
CareActives
Body Sculpture
ProvislimTM
PROVISLIMTM was awarded with the 3rd rank in the category of Innovative Raw Materials - naturals/actives, since it is an
innovation for the anti-cellulite treatment.
The BSB European Innovation Prize for cosmetics and raw materials was awarded in the March 30 th of 2011 during the In-
Cosmetics Exhibition in Milano.
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The Innovation Prize, a prize brought to life by BSB`s founder Dr. Jan-H. Riedel in 2003, is supported by business and aims
to promote the global dissemination of up-to-date knowledge.
PROVISLIMTM, the ultimate solution in anti-cellulite: multi-active ingredient that
works in any situation
Eight out of ten women suffer from cellulite. Genetic inheritance, lack of physical activity and an inadequate caloric balance
promote its formation and development. The end result is an unsightly fat accumulation that needs to be reduced in order
to have a smooth silhouette.
Adipocytes, cells that lodge in the subcutaneous tissue, are responsible for storing energy as triglycerides and facilitating
its availability when the body needs them. When this fat accumulation located under the epidermis (along with other fluids)
infiltrates the dermis locally, it causes an increase in the volume of fat tissue and irregularities on the skin surface, known
as cellulite or orange peel skin.
To globally reduce the adipose tissue and associated cellulite, it is necessary to act on the different processes that cause
this fat accumulation in the tissue. Provislim™ is a combination of two natural origin active ingredients (fisetin and
frambinone) that meets this need:
• It activates basal lipolysis (degradation of triglycerides) at rest
• It promotes adrenaline-induced lipolysis, during exercise or stress situations
• It inhibits lipogenesis (formation of triglycerides) after food intake
• It inhibits adipogenesis (differentiation into mature adipocytes) as a global preventive effect
In addition, the anti-cellulite cosmetic application of the main pathway of action of one of its active ingredients is exclusive
to Provital.
As an added advantage, Provislim™ does not contain preservatives, so the final product can be labeled as 100% of
vegetable origin (except water).
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As a result of its multiple actions, Provislim™ is an active ingredient that helps reduce localized fat and cellulite 24 hours
a day, acting under any circumstances. It reduces orange peel skin and cellulite manifestations, reducing the
accumulation of triglycerides.
MECHANISMS IN ADIPOSE TISSUE
Lipolytic balance
Adipose cells constantly absorb and store fat from the bloodstream
(lipogenesis) while at the same time they are degradating stored fat
to release it back into the bloodstream (basal lipolysis).
These two processes together form the so-called fat accumulation-
release cycle. Depending on the messages received by the adipocyte
from the outside, this cycle tends towards net fat accumulation or
towards fat release.
Basal lipolysis (at rest)
The degradation of triglycerides that occurs in the adipocytes in the absence of lipolytic agents takes place constantly.
Even if it is a moderate lipolysis (except in situations of fasting, in which it is increased, Morimoto, C., 1998), as it is a
constant process, it can impact body fat levels.
Lipolysis during exercise or stress
Catecholamines (adrenaline and noradrenaline) are the "fight or flight" hormones that the body produces when it is in
danger and in cases of exercise or stress. When due to these situations the body needs extra energy, the adipocytes are
stimulated by these hormones in order to increase its lipolysis and, consequently, the release of fatty acids. This lipolysis
is much more pronounced than basal lipolysis (up to 50 times more), but it only occurs during specific periods.
Lipogenesis after food intake
After food intake, adipocytes receive the message to store triglycerides (to increase lipogenesis) through the activation of
the lipoprotein lipase (LPL) and the insulin pathway.
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Insulin pathway activation causes the phosphorylation of phosphatidylinositol 3-kinase (PI3K). When activated, this
enzyme causes the translocation of the GLUT4 transporter from intracellular vesicles to the cell membrane surface, allowing
a greater influx of glucose into the cell. This glucose is converted into glycerol and it combines with the free fatty acids
produced by the activated lipoprotein lipase, forming triglycerides.
At the same time, insulin pathway activation inhibits basal lipolysis through the inhibition of hormone-sensitive lipase
(HSL), which also leads to an accumulation of triglycerides in the adipocyte.
Global adipogenesis
The expansion of adipose tissue in adults is not only caused by the enlargement of adipocytes as they store triglycerides,
but also by the increase in the number of adipocytes. Although mature adipocytes have lost the ability to replicate, adipose
tissue contains precursor cells (similar to fibroblasts), that can differentiate into new adipocytes. This process is promoted
in cases of imbalance in favor of energy intake versus energy consumption.
Insulin pathway activation produces an increase in adipogenesis through a complementary dual mechanism (Klemm, D.J.,
2001): activation of the phosphatidylinositol 3-kinase (PI3K) (inhibitors of this enzyme, such as Wortmannin, have anti-
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adipogenic effects) and activation of the Ras-MAP kinase-CREB pathway, where CREB is the last responsible for initiating
the cascade of differentiation into adipocytes.
CONSTANT COMBINED ACTION
Provislim™ is an effective combination of two compounds (fisetin and frambinone) that reduces the accumulation of fat
in adipose tissue, cellulite and orange peel skin all day long. It acts under any circumstances, 24 hours a day.
What is Fisetin?
This natural flavonoid isolated from Buxus Sinica (Rehd. et Wils.) Chen can be found in fruits (strawberries, apples, kiwis)
and vegetables (tomatoes, cucumbers) in the normal human diet (Arai, Y., 2000). Fisetin has interesting physiological effects
on adipose tissue.
Fisetin structure
Benefits of Fisetin
Fisetin pathway of action on adipocytes with an anti-cellulite action is exclusive to Provital. Fisetin, like other flavonoids,
inhibits the phosphorylation of phosphatidylinositol 3-kinase (Nomura, M., 2008). Preventing the activation of this
enzyme reduces the influx of glucose into the cell and as a result lipogenesis decreases. Moreover, there is increased
activity of hormone sensitive lipase, due to the non-activation of the kinase that inhibits its action, and therefore moderate
lipolysis activation is produced.
Recently it has been accepted that calorie restriction in several organisms causes a delay in aging processes and an increase
in life expectancy. In mammals, these effects are mediated by a molecule belonging to the Sirtuin family (Sirt1 sirtuin).
This family, so fashionable lately, also has interesting benefits to remodel and reduce body adipose tissue. Fisetin is an
active ingredient capable of activating Sirt1 sirtuin, which through the modulation of PPAR- leads to a reduction of
adipogenesis, an increase in lipolysis and a mobilization of fatty acids in adipocytes (Picard, F., 2004).This flavonoid acts
as an anti-inflammatory and antioxidant agent in itself (Wang, L., 2006), but it also increases other intracellular
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antioxidants (glutathione). In addition, this molecule maintains the mitochondrial function during oxidative stress, reduces
the production of lipid peroxides and their pro-inflammatory derivatives (Maher, P., 2009), significantly decreases some
inflammatory cytokines (TNF-, IL-6 or IL-8) and shows some anti-inflammatory activity (Lee, J.D., 2009). Due to the link
between cellulite and inflammatory processes, fisetin is interesting to reduce inflammatory processes and cellulite.
What is Frambinone?
This naturally occurring phenolic compound (derived from the anise plant, Pimpinella anisum) is responsible for the
characteristic aroma of fruits such as raspberries. It is widely known and used in perfumery.
Frambinone structure
Benefits of Frambinone
Frambinone helps prevent obesity and activates lipid metabolism.
Morimoto, C. (2005) compared animals fed a high-fat diet with other
animals which also took frambinone. Frambinone prevented the body
weight gain induced by the high fat diet while considerably increasing
noradrenaline-induced lipolysis, associated to the activity of hormone-
sensitive lipase (HSL). In a subsequent study (Park, K.S., 2010), it was
found that adipocytes treated with frambinone significantly increased
lipolysis as well as the expression and secretion of adiponectin, a
protein involved in lipid metabolism which is reduced in obese people.
In the same field of action, frambinone also produces an increase in the -oxidation of fatty acids, a step in the catabolic
process to produce energy, preventing their accumulation. On the other hand, topical application of frambinone causes a
significant increase in skin elasticity due to the activation of sensory neurons (Harada, N., 2008).
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IN VITRO STUDY
In vitro efficacy study on subcutaneous adipocytes obtained from liposuctions conducted on adult women with some degree of
overweight. The effects of Provislim™ and its active ingredients separately on basal lipolysis and catecholamine-induced lipolysis,
lipogenesis and adipogenesis in fatty tissue were analyzed to confirm the efficacy of the combination of the two active
ingredients.
Assessment of lipolysis at rest
To assess basal lipolysis, adipocytes were incubated for 3 hours with the active ingredients to be tested (fisetin,
frambinone and Provislim™) at two concentrations, plus a control group. Later, nonesterified fatty acids (NEFA) that had
been released to the medium as a result of the degradation of triglycerides in the adipocyte were quantified.
The following chart (chart1) shows the results of the three products tested at two concentrations, plus control. Basal
lipolysis in the presence of Provislim™ significantly increases, well above control and even fisetin, which also has some
stimulating activity. The high activation of basal lipolysis produced by frambinone by itself is highly interesting.
Basal lipolysis
14.00
12.00
Nonesterified fatty acids (uM)
10.00
8.00
6.00
4.00
2.00
0.00
Fisetin 14 ppm Frambinone 12 ppm Provislim™ 22 ppm Control
Chart 1. Basal lipolysis results
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Stimulation vs control (%)
Fisetin 14 ppm 4.8
Fisetin 29 ppm 23.8
Frambinone 12 ppm 109.4
Frambinone 25 ppm 123.7
Provislim™ 22 ppm 33.3
Provislim™ 45 ppm 57.1
Considering the stimulation of basal lipolysis versus control, the treatment with Provislim™ is able to increase this basal
lipolysis by 57% at a concentration of 45 ppm of active matter (0.2% of Provislim TM) thanks to the combined effect of its
two active principles. Frambinone is the active ingredient that most contributes to achieve this relevant effect, as
individually it increases basal lipolysis by more than 123% (at 25 ppm).
Assay on adrenaline-induced lipolysis
Catecholamines (such as adrenaline) increase lipolysis in adipocytes in states
of stress or exercise. Therefore, the aim was to determine whether the
treatment with Provislim™ managed to further enhance adrenaline-induced
lipolysis.
The adipocytes were incubated (3 hours) with the compounds to be assessed
in the presence of adrenaline (0.05 ppm). Next, the released nonesterified
fatty acids were quantified, as in the previous study.The following chart (chart 2) shows the results obtained for the
products that had an effect on lipolysis in the presence of adrenaline. This excludes fisetin, which did not act in this process.
It can be clearly seen that Provislim™ and frambinone significantly enhance the activation of adrenaline-stimulated
lipolysis, surpassing the effect of adrenaline alone and control.
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Adrenaline-induced lipolysis
100.00
Nonesterified fatty acids (uM)
80.00
60.00
40.00
20.00
0.00
Frambinone Frambinone Frambinone Provislim™ Provislim™ Provislim™ Adrenaline Control
4 ppm 12 ppm 25 ppm 11 ppm 22 ppm 45 ppm
Chart 2. Adrenaline-induced lipolysis results
Considering the results of this stimulation versus control, Provislim™ increases adrenaline-induced lipolysis by 169% at a
concentration of 11 ppm of active matter. This action is due in part to the action of Provislim™ itself and in part to the
action of adrenaline added to the culture, which stimulates lipolysis by itself.
If the effect of adrenaline is considered as standard, Provislim™ enhances its lipolytic effects by 36% (at 11 ppm) and
frambinone by 56% (at 25 ppm). With these values it is evidenced that frambinone is the Provislim™ component that
causes the stimulation of induced lipolysis. These results show that during the practice of sport and in stressful situations
(adrenalin), Provislim™ further activates the process of lipolysis and lipid degradation, which is already stimulated by
adrenalin itself.
Lipogenesis inhibiting effects after glucose intake
After food intake, adipocytes receive the message to store triglycerides and allow a greater influx of glucose into the cells.
This assay measured the incorporation of a radioactively labeled glucose derivative (3H-2-deoxyglucose) in the presence
of insulin, in adipocytes treated with the active ingredients compared to control. Results were expressed as percentage of
inhibition of insulin induced glucose incorporation (10 nM) and as radioactivity due to incorporated glucose, in CPM (counts
per minute). The following chart (chart 3) shows the results of the active ingredients tested with effect on lipogenesis and
insulin (control). Frambinone had no detectable effect in this study.
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Glucose incorporation inhibition (%)
110.00
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
Fisetin 7 Fisetin 14 Fisetin 29 Provislim™ Provislim™ Provislim™ Insulin
ppm ppm ppm 11 ppm 22 ppm 45 ppm
Chart 3. Glucose incorporation inhibition results
According to these results, Provislim™ and fisetin individually have a powerful dose dependent anti-lipogenic effect.
Provislim™ completely inhibits glucose incorporation in human adypocites at doses over 22 ppm of active matter and
fisetin also completely inhibits its incorporation from a similarly low dose.
Radioactivity due to Glucose uptake
glucose (average CPM) inhibition (%)
Insulin 5846 0.00
Provislim™ 11 ppm 843 85.57
Provislim™ 22 ppm 216 96.30
Provislim™ 45 ppm - 150 102.57
Fisetin 7 ppm 584 90.01
Fisetin 14 ppm 296 94.94
Fisetin 29 ppm - 189 103.24
These results demonstrate that the treatment with Provislim™ clearly reduces the incorporation of glucose in adipocytes,
similarly to the action of fisetin alone. It is confirmed that fisetin is the component that causes the important effect of
Provislim™ reducing lipogenesis, as it clearly inhibits glucose incorporation.
Evaluation of adipogenesis inhibition
This assay assesses the ability to inhibit the differentiation of human subcutaneous pre-adipocytes into mature adipocytes,
analyzing total triglycerides and an Oil red O tincture. Pre-adipocytes were incubated in a differentiation medium in the
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presence of Provislim™ at different concentrations. Should it be active, this would result in a decrease in total accumulated
lipids, compared to control.
For the Total triglycerides assay, cells were lysed at the end of the incubation period and were treated with enzymes that
released glycerol that was then quantified. Control culture represents the complete adipocyte differentiation and its
glycerol content is considered as 100%.
Results clearly show that Provislim™ drastically reduces the differentiation of pre-adipocytes into mature adipocytes,
with the subsequent decrease in the accumulation of triglycerides. This reduction is dose-dependent, as shown in the
following chart (chart 4).
Differentiation into adipocytes (%)
100
71.5%
80
60
40 32%
20
5%
0
Control Provislim™ Provislim™ Provislim™
22 ppm 45 ppm 67 ppm
Chart 4. Results of differentiation into adipocytes
Before taking the images of the Oil Red O tincture (triglycerides marker) and once the incubation was finished, the medium
was withdrawn and the cells were fixed and stained. The following images clearly show the important reduction in the
differentiation of pre-adipocytes caused by the treatment with Provislim™, since the presence of triglycerides decreases
as the active ingredient concentration increases.
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IN VIVO STUDY
The efficacy of Provislim™ was assessed in vivo on 15 female volunteers with an average age of 47 with grade 2-3 cellulite
according to the Pinch-test scale (scale 0-4, 4 being the most severe). An anti-cellulite cream was applied (with 5%
Provislim™) twice a day on one leg and the same cream with no active ingredient (placebo) on the other leg, during 56
days. Parameters assessed were thigh circumference, skin thickness, skin roughness and skin elasticity.
The applied formulas are listed below:
INCI Active formula Placebo
% (w/w) % (w/w)
Aqua (Water) c.s.p. c.s.p.
Behenyl Alcohol, Glyceryl Stearate, Lecithin, Glycine Soja 4.50 4.50
(Soybean) Sterols
Tetrasodium Glutamate Diacetate, Sodium Hydroxide, Aqua 0.20 0.20
(Water)
Tocopheryl Acetate 0.30 0.30
Cetyl Alcohol 1.00 1.00
Helianthus Annuus (Sunflower) Seed Oil, Rosmarinus Officinalis 20.00 20.00
(Rosemary) Leaf Extract
Dehydroxanthan Gum 0.90 0.90
Dimethicone 2.00 2.00
Phenoxyethanol, Methylparaben, Butylparaben, Isobutylparaben, 0.80 0.80
Ethylparaben, Propylparaben
Parfum (Fragrance) 0.20 0.20
PROVISLIM™ 5.00 ---
CI 42090 0.02 0.02
Centimeter reduction
Centimeter measurements were taken immediately below the buttocks and it was observed that the treatment with
Provislim™ caused a reduction of thigh volume compared to the initial measurements, unlike the placebo (not significant).
The difference between the effect of Provislim™ and placebo in the average centimeter reduction on the thigh was 0.3
cm, with Provislim™ causing an interesting volume reduction. The maximum difference between active ingredient and
placebo was a reduction of up to 1 cm approximately. These results are statistically significant.
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Before applying Provislim™ (D0) After applying Provislim™ (D56)
Effects on skin thickness
Skin affected by cellulite has a thicker dermis due to the presence of edema, together with adipose infiltrations that can
almost reach the dermal-epidermal junction. Using ultrasounds structural changes that occur in the skin can be measured,
in particular the thickness of the epidermis, dermis and hypodermis.
Provislim™ reduced skin thickness by 0.03 mm, whereas placebo had no effect (very statistically significant results). The
maximum difference between the two treatments was a 0.07 mm reduction caused by the active ingredient.
Provislim™ caused a thickness reduction in 93% of volunteers. Ultrasound images below show the effect before (left
image) and after the treatment with Provislim™ (right image) on a volunteer.
Image before applying Provislim™ (D0) Image after applying Provislim™ (D56)
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Lighter areas indicate higher density and darker areas show adipose infiltrations, typical of cellulite. With Provislim™ there
is a redensification of the dermis (increase of light areas) and a decrease in dermal infiltrations (reduction in darker, hypo-
echogenic areas).
All this leads to a reduction of edema and adipose tissue infiltrated in the dermis, which causes the unsightly orange peel
skin effect.
Skin roughness reduction
The study on the effects of Provislim™ on skin roughness was conducted on silicone replicas. Using analysis techniques,
these replicas allow to determine average roughness (Ra) and maximum roughness (Rz).The thigh treated with Provislim™
presented a statistically significant reduction in average roughness of 10.9% versus the start of the treatment, compared
with a nonsignificant decrease in the placebo. Considering the average effect of the active ingredient versus the placebo,
there was a 4.6% global improvement, although the maximum reduction of Ra was 39.5%.
The thigh treated with Provislim™ presented a statistically significant decrease in maximum roughness of 7.0% versus the
start of the treatment, compared with a nonsignificant reduction in the placebo. Comparing the active ingredient versus
the placebo, a 3.7% global improvement was obtained, although the maximum reduction of Rz was 28.7%.A reduction in
average and maximum roughness reflects a smoothing of the skin, equivalent to a reduction and improvement of the
orange peel skin effect.
Skin elasticity improvement
Biomechanical properties of the dermis and epidermis have an influence on the appearance of cellulite, caused by the
pressure of subcutaneous adipose tissue on the dermis. Accumulated fat deposits generate tension on the skin, reduce its
flexibility and elasticity (Dobke, M.K. et al, 2002) and its elastic fibers can break, leading to the appearance of unwanted
marks and streaks.
Therefore, in this assay a Cutometer was used to measure the biological elasticity (R7 or Ur/Uf), indicator parameter of
skin elasticity whose value expresses greater elasticity the closer it is to 1. In addition, this parameter does not depend on
skin thickness of each individual and can be directly compared between different points of application and even individuals.
Provislim™ showed an average elasticity increase of 18.5% (statistically significant), compared with a nonsignificant
increase in the placebo. Comparing the active ingredient and the placebo, the elasticity improves by 13%, this being a
statistically significant improvement. The maximum improvement in elasticity was 48% (active ingredient vs placebo).
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CONCLUSION
Provislim™ is an effective combination of two natural origin active ingredients (Fisetin and Frambinone) that clearly
enhances the reduction of body fat tissue in any situation: at rest, during exercise or stress, after eating and at a general
level.
It reduces cellulite, reshapes your silhouette and improves skin elasticity, while decreasing skin roughness, edema and
skin thickness and volume.
APPLICATIONS
- Prevention and treatment of cellulite
- Slimming and body reshaping products
- Sport & cosmetic modeling products
- Complementary ingredient in moisturizing, firming and tanning body products
RECOMMENDED DOSE
The recommended dosage of Provislim™ is 2-5%.
BIBLIOGRAPHY
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Arai, Y., Watanabe, S., Kimira, M., Shimoi, K., Mochizuki, R., Kinae, N. Dietary intakes of flavonols, flavones and isoflavones
by Japanese women and the inverse correlation between quercetin intake and plasma LDL cholesterol concentration. J Nutr.,
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Dobke, M.K., Dibernardo, B., Thompson, R.C., Usal, H. Assessment of biomechanical skin properties: is cellulitic skin
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