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Baicapil™

Stimulates hair growth and prevents hair loss

Baicapil™ Stimulates hair growth and prevents hair loss www.provitalgroup.com HAIR GROWTH

www.provitalgroup.com

HAIR GROWTH
HAIR
GROWTH

THE IMPORTANCE OF HAIR

THE IMPORTANCE OF HAIR Hair is a symbol of vitality and health, as well as being

Hair is a symbol of vitality and health, as well as being a key feature of the individuality and identity of a person. A gleaming hair in good condition is one of the main attributes of beauty and contributes to having high self-confidence and self-esteem.

In the year 4000 BC, in ancient Egypt, hair was already documented as being important at a social and psychological level. Nowadays it still plays this important role, and that is why hair and scalp disorders cause ample frustration in a great

part of society. People suffering from hair loss are generally perceived as older people who are physically and socially less attractive (Goh & Zippin, 2009).

Hair loss has been for many years a common problem for dermatologists consultation. For both men and women, it is extremely important that hair is healthy, full of plenitude, movement and freshness, as it directly influences our mood and character and it can define our professional, personal and even sentimental success.

Baicapil™, obtained from nature, is a combination of three botanical active ingredients that reduces hair
Baicapil™, obtained from nature, is a combination of three botanical active ingredients that reduces
hair loss, stimulates growth and increases hair density.
With Baicapil™, hair regains a healthy and strong condition.

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HAIR BIOLOGY

HAIR BIOLOGY STRUCTURE OF HAIR Hair is a complex organ that includes the external part, known

STRUCTURE OF HAIR

Hair is a complex organ that includes the external part, known as the hair shaft, and the internal part, called the hair follicle. The following parts can be distinguished inside the follicle, depending on the current phase of the hair cycle (fig. 1):

ANAGEN

TELOGEN

Shaft Sebaceous gland Hair erector muscle Bulge stem cells Bulge stem cells Dermal papilla Secondary
Shaft
Sebaceous gland
Hair erector
muscle
Bulge stem cells
Bulge stem cells
Dermal papilla
Secondary germ stem
cells
Outer root
sheath
Secondary germ
stem cells
Dermal papilla

Fig. 1. Anagen and telogen phases of the hair cycle.

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Multipotent stem cells are present in the follicle. These cells are self-renewing and differentiate into

Multipotent stem cells are present in the follicle. These cells are self-renewing and differentiate into more mature but less potent specialized cells. The division and differentiation of these cells can restore hair. For this reason, they are directly related to hair growth and absence of hair loss.

Recent studies have shown that this group of stem cells is not homogenous, because it consists of two biochemically and functionally distinct populations:

The first one, situated in the bulge, consists of the stem cells which are responsible for the differentiation and growth of various cell types to produce pigmented hair (Paus & Cotsarelis,

1999).

The second one, which niche is called the secondary germ, consists of the progenitor cells. They are the direct descendants of the bulge stem cells and are in contact with the dermal papilla, which activates them to initiate and control hair growth (Myung et al., 2013).

The dermal papilla (DP) is a conglomerate of specialized fibroblasts that is involved in the morphogenesis and hair cycle through the regulation of various cell types in the follicle. It contains stimulating factors for the proliferation and differentiation of follicular keratinocytes, which induce the formation of a new hair follicle (Kwack et al., 2013).

HAIR CYCLE

Each hair follicle is constantly subjected to cycles consisting of three stages: a stage of rapid growth and hair shaft formation (anagen), followed by a regression stage based on apoptosis (catagen) and a resting period of the hair follicle (telogen).

1.

Anagen

During the initial stages of hair regeneration (end of the telogen phase/beginning of the anagen phase), the hair follicle stem cells are quiescent (Rompolas et al., 2012).

The activation of the anagen phase starts with a signal from the dermal papilla towards the secondary germ stem cells, which activate and proliferate in order to initiate hair growth. The bulge stem cells are the next ones to activate and proliferate. These cells are responsible for extending the outer root sheath (it separates the hair follicle from the dermis) and maintaining the matrix that supports hair growth (Myung et al., 2013). These stem cells differentiate into matrix cells.

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Then, matrix cells quickly proliferate to produce the hair shaft. Hair pigmentation is due to

Then, matrix cells quickly proliferate to produce the hair shaft. Hair pigmentation is due to melanocytes inserted between these cells (Paus & Cotsarelis, 1999).

Finally, the stem cells are silenced and return to a state of quiescence. In addition, during the transition from the anagen phase to the catagen phase, proliferating cells in the matrix are induced into a coordinated apoptosis (Plikus, 2012).

Approximately 85-90% of the follicles are in the anagen phase (Rittié et al., 2009), which usually lasts from 2 to 8 years (Paus & Cotsarelis, 1999). The duration of the anagen phase determines hair length, which varies between individuals and declines with age.

2.

Catagen

It is the end of the hair fiber production phase, when the follicle is subjected to a controlled process of regression. Most follicular keratinocytes undergo apoptosis. Cell growth and pigmentation stop, the bulb is separated from the dermal papilla and follicular shortening occurs (Paus & Cotsarelis, 1999).

It is the shortest phase of the cycle and it only lasts two to three weeks. Therefore, at any given time only between 1% and 2% of the follicles are in the catagen phase (Restrepo, 2010).

At the end of this phase, the follicle retracts into the surface of the scalp (it does not protrude from the dermis) and its size is significantly reduced.

3.

Telogen

The telogen phase typically lasts from 2 to 4 months, before the follicles reenter the anagen phase and the cycle starts again. Therefore, it determines when new hair is originated.

During this phase, the hair shaft matures into fully keratinized hair that detaches from the follicle (usually due to hair combing or washing). Most people lose 50 to 150 hairs a day.

The percentage of follicles in the scalp in the telogen phase is from 5 to 15%. An increase in this percentage leads to excessive hair loss (Paus & Cotsarelis, 1999).

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HAIR LOSS

HAIR LOSS Approximately 5 million hair follicles cover the human body at birth. New follicles are

Approximately 5 million hair follicles cover the human body at birth. New follicles are not formed after birth, but hair and follicle size may vary over time (Paus & Cotsarelis, 1999).

The daily loss of an average of 50 to 150 hairs is considered as normal. It is a cycle under constant renovation. The same way a person is born, grows and dies, hair follows a very similar pattern: it grows, rests and falls. The problem arises when this life cycle of hair is altered and the common problem of alopecia appears.

hair is altered and the common problem of alopecia appears. Hair loss is caused by an

Hair loss is caused by an alteration in the hair growth cycle, due to several factors (androgen metabolism, genetics or stress). It is characterized by:

Changes in the ratio of anagen and telogen hair: the number of hairs in the anagen phase is reduced and more hair remains in the telogen phase.

Shortening of the anagen phase: hair stops growing earlier than expected; it is shorter and thinner.

Extension of the telogen phase.

Some alopecias are considered as reversible, because the hair cycle is disturbed, but the hair follicles are still present and they follow the hair cycle, even in bald scalps (Paus & Cotsarelis, 1999).

Baicapil™

in bald scalps (Paus & Cotsarelis, 1999). Baicapil™ Taking into account the life cycle of hair,

Taking into account the life cycle of hair, in order to prevent hair loss it is desirable that the active ingredient used has the ability to maintain a long anagen period and a short telogen period, or the ability to change the hair cycle gently from the telogen phase to the anagen phase.

In addition, it is also very important that it improves the

condition of the follicle cells and their microenvironment, as this is a crucial factor in maintaining the balance between the anagen and telogen phases.

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Baicapil ™ is a combination of 3 plants ( Scutellaria baicalensis , Triticum vulgare sprout

Baicapilis a combination of 3 plants (Scutellaria baicalensis, Triticum vulgare sprout and Glycine max sprout) that increases cellular energy and activates follicle stem cells, while protecting them from senescence. This way, it visibly fights hair loss by:

Stimulating hair growth.

Increasing hair density.

Reducing hair loss.

Recovering a healthy and strong appearance.

COMPOSITION

1. CHINESE SKULLCAP

Botany

Scutellaria baicalensis Georgi, known as Chinese skullcap or baical skullcap, is a source of baicalin, one of the main actives of Baicapil™.

of baicalin, one of the main actives of Baicapi l™. It is a biennial perennial plant

It is a biennial perennial plant that can reach up to 1.2 m in height, with bluish purple flowers. Flowering is from May to August. The English name skullcap describes the shape of the flower calyx, which resembles the helmets that were used in the Middle Ages. The fruits appear in late August.

It grows mainly in China, Russia, Mongolia and the Korean Peninsula. The Lamiaceae family, to which S. baicalensis belongs, includes more than 350 species, many of them medically active. It is one of the 50 fundamental herbs for traditional Chinese medicine, and it is also used in Nepal, Japan and Korea (Stutte et al., 2008).

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The root of S. baicalensis has been used for thousands of years to prepare Huang

The root of S. baicalensis has been used for thousands of years to prepare Huang Qin, a traditional medicine to cure nervousness, high blood pressure and respiratory problems, and to alleviate and detoxify fever episodes (Stutte et al., 2008). Nowadays, Huang Qin is extensively used in the treatment of inflammatory diseases, hepatitis, tumors and diarrhea in Southeast Asia. S. baicalensis is officially registered in the JPXIII Japanese Pharmacopoeia and in the Chinese Pharmacopoeia.

Chemistry

Pharmacopoeia and in the Chinese Pharmacopoeia. Chemistry The chemical composition of the roots of Asian Scutellaria

The chemical composition of the roots of Asian Scutellaria is characterized by its high content in flavonoids. Out of a total of 26 identified flavonoids, five of them are C- glycosides, twelve are O-glycosides and the remaining nine are aglycones (Han et al., 2007). The characteristic components of this root are baicalin (fig. 2), which is the glucuronic derivative of baicalein (6-9%), wogonin-7-O- glucuronideglucurinide (wogonoside) (2-8%), baicalein (0.1- 1.6%) and wogonin (0.01-0.3%).

Fig. 2. Chemical structure of baicalin.

The presence of other flavonoids in lower concentrations has also been described, such as oroxylin A, visidulin, skullcapflavone II, neobaicalein, acteoside, chrysin and their corresponding glucosides.

2. SOY AND WHEAT SPROUTS

Soy

Soy (Glycine max L.) is an annual herbaceous plant that belongs to the Leguminosae family (Fabaceae).

plant that belongs to the Leguminosae family ( Fabaceae ). Soy is characterized by being up

Soy is characterized by being up to 1.5 m tall, with trifoliate and hairy leaves that usually detach before the seeds are ripe. The flowers are white-yellowish or blue-violet in color and small in size. The fruit is a

hairy arched pod containing 2-6 smooth subglobose seeds, variable in color from white-yellowish to brown.

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Soy originated in the Far East, where it has always been a basic food constituent.

Soy originated in the Far East, where it has always been a basic food constituent. It is currently cultivated in many temperate-warm regions of the planet.

cultivated in many temperate-warm regions of the planet. germ together with the reserve substance. Wheat Triticum

germ together with the reserve substance.

Wheat

Triticum vulgare Vill., called wheat, belongs to the Poaceae family, also known as Gramineae. It is a vivacious herbaceous plant. The stems are simple and hollow, with pointed leaves. The inflorescence is a panicle or spikelets spike. The flowers produce a single fruit, called grain or seed, which carries an embryo or

Wheat originated in ancient Mesopotamia, but it is cultivated worldwide, from the limits of the Artic to areas close to the Equator. It is adaptable to different conditions, from xerophytic to coastal conditions.

Sprouts

Baicapilcontains soy and wheat sprouts, rich in sugars. Germination comprises a number of physiological processes that occur in the seeds of higher plants and that result in the transformation of the embryo into a mature plant.

The process starts when the optimal conditions to ensure the proper use of the reserve material and the obtaining of the expected biomolecules exist.

Reserve substances include:

Carbohydrates: starch, hemicelluloses, amyloids and galactomannans.

Proteins: peptides and free amino acids, in particular proline, glutamine, glutamic acid and asparagine.

Lipids: the main reserve in the seeds are triglycerides, which hydrolyze transforming into glycerol and unsaturated fatty acids, such as oleic acid, linoleic acid and linolenic acid.

Mineral elements: phosphate in the form of phytin and other cations (potassium, calcium and magnesium).

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The sprouts in Baicapil™ are obtained from seeds, using only water for germination, without pesticides

The sprouts in Baicapil™ are obtained from seeds, using only water for germination, without pesticides or other chemical additives. This process is stopped after 3 days, which is when active biomolecules reach peak levels (peptides, amino acids, oligosaccharides, glucose).

MECHANISM OF ACTION OF Baicapil™

The innovative mechanism of action of Baicapil™ offers effective results in both men and women.

1. METABOLISM OF GLUCOSE

Growing follicles use glucose almost two times faster than resting follicles in order to obtain cellular energy (ATP). Glucose is degraded through glycolysis and the oxidative metabolism to provide the energy material, but during this process oxygen is consumed and cellular respiration increases.

During the transformation of resting follicles into growing follicles, glycolysis increases by 200% and ATP production through the respiratory chain augments by 270% (Adachi et al., 1970). It was also showed that during the transition from the telogen to the anagen phase, DNA content and follicle size increase (Adachi et al., 1999).

Baicapil™ provides additional sugars to the follicles, which enter the citric acid cycle and increase cellular respiration. In in vitro assays with Baicapil™, it can be seen that both isolated mitochondria and intact cells increase oxygen consumption. This probably increases ATP production, providing the cellular energy required for hair growth, development and maintenance. Baicapil™ also protects against oxidative stress, which may appear as a result of high cellular energy production (ATP).

Therefore, the nutrients provided by Baicapil™ lead to more active and proliferating follicles. This represents an induction to hair growth, increasing hair density and extending the anagen phase.

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2. TERT (HUMAN REVERSE TRANSCRIPTASE)

2. TERT (HUMAN REVERSE TRANSCRIPTASE) TERT (human reverse transcriptase) is one of the characteristic proteins of

TERT (human reverse transcriptase) is one of the characteristic proteins of stem and progenitor cells, which is essential for their activation and functioning.

Activation of stem cells

As we mentioned earlier, quiescent stem cells are activated to produce a new hair.

It has been observed that TERT induces a rapid transition from the telogen to the anagen phase, facilitating hair growth through the induction of quiescent stem cells to proliferation and mobilization. The result is the growth of strong and resilient hair (Flores et al., 2005; Sarin et al., 2005; Choi et al., 2008).

Baicapil™, through the activation of stem cells by inducing TERT expression, accelerates the initiation of the anagen phase that originates a new hair strand.

Mitochondria protection

Oxidative stress is one of the mechanisms contributing to hair loss. Lipid peroxides induce apoptosis of the hair follicle cells and the catagen phase is initiated prematurely (Trüeb, 2009).

It has been demonstrated that TERT protects neurons, fibroblasts and stem cells against oxidative stress. Thus, in cells were TERT is overexpressed, mitochondrial DNA (mtDNA) is protected against oxidative damage and fewer reactive oxygen species (ROS) are produced. All this is indicative of a better mitochondrial function (Ahmed et al., 2008).

When mitochondria are damaged, a communication occurs towards the nucleus that results in the expression of genes typical of cell senescence. By reducing mitochondrial damage, it can be stated that TERT action leads to the rejuvenation of fibroblasts (Ahmed et al., 2008). All this helps to increase metabolic energy and resistance against stress (Ahmed et al., 2008; Haendeler et al., 2009).

As a summary, it can be concluded that the induction of TERT expression originated by Baicapil™ protects the follicle against oxidative stress and increases metabolic energy, thus extending the anagen phase of hair, improving follicular activity and reducing hair loss.

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Protection against senescence

Protection against senescence It has been observed in vitro that dermal papilla (DP) cells in bald

It has been observed in vitro that dermal papilla (DP) cells in bald scalps suffer premature senescence, compared to DP cells in normal scalps (Upton et al., 2013).

Baicapil™ induces TERT overexpression, improves mitochondrial function and protects against ROS, so fibroblasts are protected against senescence and remain young and active.

KEY POINTS IN THE MECHANISM OF ACTION OF Baicapil™

Baicapil™ provides nutrients and induces TERT overexpression in cells, leading to:

1. An increase in glycolysis and ATP production more active and vigorous follicles.

2. The activation of stem cells initiation of a new anagen phase, leading to hair growth and greater hair density.

3. Mitochondrial DNA protection against oxidative stress and ROS reduction in stem cells and fibroblasts more active and healthier cells, leading to a longer anagen phase and thicker and denser hair.

4. The preservation of follicle fibroblasts from senescence for longer time, resulting in an extension of the anagen phase.

IN VITRO EFFICACY

1. SOY AND WHEAT SPROUTS: OXYGEN CONSUMPTION

Thanks to the respiration process as a component of cell metabolism, aerobic cells obtain energy from the oxidation of biomolecules (amino acids, glucose, fatty acids).

In order to evaluate the action of wheat and soy sprouts on cellular metabolism, especially on mitochondrial respiration, two different models have been used: isolated mitochondria and cell culture of human fibroblasts.

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Results

Results Wheat and soy sprouts activate mitochondrial respiration in both models.  Mitochondrial model The following

Wheat and soy sprouts activate mitochondrial respiration in both models.

Mitochondrial model

The following chart shows how mitochondrial activity rises in a preparation of mitochondria, with an

increase in wheat and soy sprouts concentration (ppm) (chart 1). In this experiment, mitochondria were

pre-incubated with the active ingredient for 15-18 hours.

Mitochondria

40 35 30 25 20 15 10 5 0 0.6 1.2 2.4 4 Mitochondrial activity
40
35
30
25
20
15
10
5
0
0.6
1.2
2.4
4
Mitochondrial activity (%)

Sprouts extract (ppm)

Chart 1. Mitochondrial activity in mitochondria.

Model of human skin fibroblasts

Chart 2 shows the rise of mitochondrial activity in cultured fibroblasts due to the application of wheat and

soy sprouts. In this experiment, fibroblasts were pre-incubated with the active ingredient for 15-18 hours.

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Mitochondrial activity (%)

200

150

100

50

0

Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.

Fibroblasts

Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.
Mitochondrial activity (%) 200 150 100 50 0 Fibroblasts 0.42 Sprouts extract (ppm) 1.38 Chart 2.

0.42

Sprouts extract (ppm)

1.38

Chart 2. Mitochondrial activity in fibroblasts.

Sprouts increase cellular activity

2. BAICALIN: SENESCENCE

The effect of baicalin on cell cultures of primary human fibroblasts was assessed to observe:

TERT activation.

The effect on cell senescence.

Results

TERT activation

Fibroblast cultures were treated with baicalin for 24 hours. The results are expressed as the percentage of luciferase activity relative to untreated cells (control). It is demonstrated that baicalin activates TERT in primary fibroblasts.

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300 250 200 150 100 50 0 Control 1 µM baicalin 3 µM baicalin 10
300 250 200 150 100 50 0 Control 1 µM baicalin 3 µM baicalin 10
300
250
200
150
100
50
0
Control
1 µM baicalin
3 µM baicalin
10 µM baicalin
TERT activation

Chart 3. TERT activation due to baicalin (luminescence).

Cell senescence

To demonstrate the effect on cell senescence, human fibroblasts cultures were treated with baicalin, beginning with cultures that were in an exponential growth phase until they stopped growing upon reaching senescence.

It was demonstrated that 5 additional cell divisions took place in treated cells, compared to control, before entering senescence. Therefore, baicalin has the ability to delay the entry into senescence of human fibroblasts, as it increases the number of duplications by 10%, considering that the total number of replications of a fibroblast is 50 (Schneider & Mitsui, 1976).

Baicalin delays cellular senescence

Due to the combination of the sprouts and baicalin, Baicapilis a product capable of:

Increasing hair growth.

Increasing hair density.

Reducing hair loss.

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IN VIVO EFFICACY

IN VIVO EFFICACY PROTOCOL The efficacy in regeneration and against hair loss of Baicapil™ has been

PROTOCOL

The efficacy in regeneration and against hair loss of Baicapil™ has been demonstrated in an in vivo efficacy study. To this end, a hair lotion at 3% of Baicapil™ was tested versus another lotion without active ingredient (placebo).

This double-blind study involved 61 volunteers with Mediterranean Caucasian hair, 17 men and 44 women aged between 18 and 60, randomly distributed between the placebo and Baicapil™ groups.

30 volunteers applied the placebo and 31 applied Baicapil™ on the scalp daily for 6 months of study. Volunteers washed their hair every other day using a neutral formula shampoo. No other product against hair loss was applied on the scalp during this study.

To evaluate the efficacy in regeneration and against hair loss of Baicapil™, the following assessments were made:

1. Assessment of the ratio between the anagen and telogen phases using a trichogram.

2. Assessment of hair density using scalp microphotography.

3. Assessment of hair loss using the washing and combing test.

4. Assessment of overall hair condition using photography.

The tests have been carried out by specialists in hair treatments and the assessments have been performed by a specialist in trichology. All assessments have been performed at day 0 and day 180 of the study. Additionally, the washing and combing test was performed on day 90 of the study.

1. RATIO BETWEEN THE ANAGEN AND TELOGEN PHASES

The assessment of the anagen and telogen phases of hair follicles consists in the direct observation of follicles under the microscope and in identifying the phase according to the characteristics of each follicle (sheath hydration and development, matrix condition and pigmentation, follicle appearance).

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To assess the percentage of follicles in anagen phase versus follicles in telogen phase, a

To assess the percentage of follicles in anagen phase versus follicles in telogen phase, a certain number of hairs (between 15 and 25 hairs) is randomly extracted from the scalp using traction (always from the same area) to be subjected to microscopic study (observation and photography).

The results are expressed as the number of follicles in the anagen (A) or telogen (T) phase, their percentage in relation to all the hairs extracted from the volunteer, and the ratio of hair in anagen and telogen phase (A/T).

Nowadays, experts observe that the ratio of hair in anagen and telogen phase (A/T) on healthy scalp is rarely above 6. Therefore, many efficacy studies consider a result of A/T ≥ 4 as indicative of normal condition.

The observation of follicles under the microscope shows that their condition substantially improves in the group treated with Baicapil™. An example of this improvement can be seen in fig. 3. This figure shows the follicles collected from the same volunteer at the beginning (T0) and at the end (T180) of the study. Both follicles are in the anagen phase, but much more intense activity in the matrix area and good sheath development (more robust and thicker) can be observed in the follicle extracted at the end of the study, compared to the image taken at the beginning of the study. Moreover, the bulb removed at the end of the study is visibly larger and hair is clearly thicker compared to the initial picture.

T0 T180 Fig. 3. Microscopic images of two follicles in anagen phase collected from the
T0
T180
Fig. 3. Microscopic images of two follicles in anagen phase collected from the same volunteer at the beginning and at the end of
the application of Baicapil™. A visible improvement in the condition and activity of the follicle can be seen, together with an
increase in hair thickness.

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At the end of the treatment, the number of follicles in the anagen and telogen

At the end of the treatment, the number of follicles in the anagen and telogen phase was determined:

Volunteers who applied Baicapil™ have presented an average of 12.7% more anagen hair compared to the beginning of the study. Placebo treatment has also resulted in a slight increase, although in this case the difference has only been 2.0% compared to the beginning of the study. Therefore, at the end of the study the difference in improvement between Baicapil™ and placebo has been 10.7 percentage points, a statistically significant improvement (chart 4).

14

12

10

8

6

4

2

0

% anagen T180 vs. T0

12.7%

10.7 2.0%
10.7
2.0%

PLACEBO

BAICAPIL™

Chart 4. Improvement of the percentage of the anagen hairs as a result of the application of placebo and Baicapil™.

In the case of telogen hair count, volunteers who applied Baicapil™ have presented an average of 27.2% less telogen hair at the end of the study, compared to baseline. In the placebo group, the number of the telogen hair has increased by 6.9%. Therefore, at the end of the study the difference in improvement between Baicapil™ and placebo has been 34.1 percentage points (chart 5).

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10

5

0

-5

-10

-15

-20

-25

-30

10 5 0 -5 -10 -15 -20 -25 -30 % telogen T180 vs. T0 PLACEBO BAICAPIL™

% telogen T180 vs. T0

PLACEBO

BAICAPIL™

6.9% 34.1
6.9%
34.1

-27.2%

Chart 5. Improvement in the percentage of the telogen hair as a result of the application of placebo and Baicapil™.

Finally, the anagen/telogen hair ratio (A/T ratio) has increased after the application of Baicapil™ from 3.5

at the beginning of the study to 5.9 at the end. This change has been virtually undetectable in the placebo

group (A/T was 2.8 at the beginning and 2.9 at the end of the study) (chart 6). This means a 68.6% increase

in the A/T ratio compared to the beginning of the study with Baicapil™ and only a 3.6% increase in the

placebo group (chart 7).

Changes in the A/T ratio

7 5.9 6 5 3.5 4 2.9 2.8 3 2 1 0 Day 0 Day
7
5.9
6
5
3.5
4
2.9
2.8
3
2
1
0
Day 0
Day 180
PLACEBO
BAICAPIL™
Chart 6. Changes detected in the A/T ratio during the six months
of application of Baicapil™ and placebo.

% A/T T180 vs. T0

80 68.6% 70 60 50 40 30 20 10 3.6% 0 PLACEBO BAICAPIL™ Chart 7.
80
68.6%
70
60
50
40
30
20
10
3.6%
0
PLACEBO
BAICAPIL™
Chart 7. A/T ratio percentage improvement as a result of
placebo and Baicapil™ application after six months of study.

As shown in chart 6, Baicapil™ has increased the A/T ratio to 5.9, indicating the activation of telogen hair

follicles, the extension of the anagen phase, and leading to a healthier scalp.

Baicapil™ stimulates hair growth

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2. HAIR DENSITY

2. HAIR DENSITY Hair density (number of hairs per surface unit) has been assessed using the

Hair density (number of hairs per surface unit) has been assessed using the scalp images taken with the

microcamera. These images are automatically transferred to specialized software that allows visible hair

counts in the scalp surface defined unit.

Baicapil™ has increased hair density by 18% compared to the beginning of the study. This result has been

6.7 percentage points average better than that obtained for placebo (chart 8).

20

18

16

14

12

10

8

6

4

2

0

% density T180 vs. T0

18% 6.7 11.3%
18%
6.7
11.3%

PLACEBO

BAICAPIL™

Chart 8. Improvement in hair density obtained during the 6 months of the study.

Below are microphotographs of the scalp of volunteers who applied Baicapil™ during 6 months. The first

row shows photographs taken at the beginning (T0) of the study. Photographs taken at 6 months of study

(T180) are shown on the second row.

We can see the visible improvement in hair density and thickness after the application of Baicapil™

observed in several volunteers (fig. 4):

Volunteer A 22% increase in hair density, from 182.98 hairs cm 2 to 222.75 hairs cm 2 .

Volunteer B 12% increase in the number of hairs, from 99.44 hairs cm 2 to 111.28 hairs cm 2 .

Volunteer C 28% increase in the number of hairs, from 155.13 hairs cm 2 to 198.89 hairs cm 2 .

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+22% VOLUNTEER A +12% VOLUNTEER B +28% VOLUNTEER C Fig. 4. Microphotographs of the scalp
+22% VOLUNTEER A +12% VOLUNTEER B +28% VOLUNTEER C Fig. 4. Microphotographs of the scalp
+22% VOLUNTEER A
+12% VOLUNTEER B
+28% VOLUNTEER C
Fig. 4. Microphotographs of the scalp of volunteers who applied Baicapil™.
Baicapil™ increases hair density and thickness with volumizing effect
increases hair density and thickness with volumizing effect 3. HAIR LOSS The efficacy of Baicapil™ against

3. HAIR LOSS

The efficacy of Baicapil™ against hair loss has been finally confirmed using the combing and washing test. In this assay, the number of hairs that are shed from the scalp during combing and its subsequent washing under standardized conditions is determined.

After 3 months of Baicapil™ application, a 60.6% reduction in hair loss compared to the beginning of the study has been observed (chart 9). This result has been slightly more moderate after 6 months of treatment (50.9% reduction in hair loss, chart 10) as a result of the adaptation of the hair follicle metabolism to the daily contribution of active ingredient. Placebo also improved the condition of the follicles, although the reduction in hair loss was much less pronounced than with Baicapil™ (chart 9 and 10).

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Baicapil™ substantially improves the condition of follicles, resulting in better follicle anchoring to the scalp.

Baicapil™ substantially improves the condition of follicles, resulting in better follicle anchoring to the

scalp.

% hair loss T90 vs. T0

% hair loss T180 vs. T0

PLACEBO

BAICAPIL™

PLACEBO

BAICAPIL™

0

-10

-20

-30

-40

-50

-60

-70

-37.8% 22.8 -60.6%
-37.8%
22.8
-60.6%

0

-10

-20

-30

-40

-50

-60

-37.4% 13.5 -50.9%
-37.4%
13.5 -50.9%

Chart 9. Improvement in hair loss observed after three months of placebo and Baicapilapplication.

Chart 10. Improvement in hair loss observed after six months of placebo and Baicapil™ application.

Baicapil™ reduces hair loss

4. OVERALL HAIR CONDITION

The objective of this assessment was to check whether the application of Baicapil™ leads to a visible

improvement of hair density in volunteers.

Several pictures of the top of the scalp of each volunteer were taken at day 0 and 180 of the study. The

images show the hairline and crown.

On volunteers who applied Baicapil™ considerable improvements in hair quantity and quality have been

observed (fig. 5).

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T0 T180 Fig. 5. Example of improvement in overall hair condition in a volunteer after
T0
T0
T180
T180
Fig. 5. Example of improvement in overall hair condition in a volunteer after the application
Fig. 5. Example of improvement in overall hair condition in a volunteer after the application of Baicapil™ for six months.

Baicapil™ visibly improves hair quantity and quality

* The final formula of Baicapil™ contains arginine, used as a stabilizing agent. However, arginine is also known for its efficacy against hair loss. In this case, it was decided that, in order not to distort the results of our in vivo study, arginine would be removed from the hair lotion with Baicapil™ used for these tests.

CONCLUSION

lotion with Baicapil™ used for these tests. CONCLUSION improving hair quantity and quality. Baicapil™ increases

improving hair quantity and quality.

Baicapil™ increases cellular energy and protects follicle fibroblasts against oxidative stress and senescence, thus improving follicular activity and extending the anagen phase of the hair cycle.

In addition, it has been demonstrated in vivo that it improves the ratio of anagen hair versus telogen hair and increases hair growth and density. Moreover, it reduces hair loss, visibly

Baicapil™ is an active ingredient that reduces hair loss, stimulating hair growth and improving its overall condition.

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COSMETIC APPLICATIONS

COSMETIC APPLICATIONS  Products against hair loss.  Hair care lines: tonics, serums, conditioners, masks,

Products against hair loss.

Hair care lines: tonics, serums, conditioners, masks, shampoos.

Stimulating products for hair growth.

Anti-aging hair product lines.

RECOMMENDED DOSAGE

The recommended dosage is between 2 and 4 %.

BIBLIOGRAPHY

Adachi K. et al. Activity of Glucose-6-phosphate 1-Dehydrogenase in Hair Follicles with Male-pattern Alopecia. Biosci. Biotechnol. Biochem. 1999, 63(12):2219-2221.

Adachi K. et al. Human Hair Follicles: Metabolism and Control Mechanisms. J. Soc. Cosmeti. Chem. 1970,

21:901-924.

Ahmed S. et al. Telomerase does not counteract telomere shortening but protects mitochondrial function under oxidative stress. J Cell Sci. 2008, 121(Pt 7):1046-53.

Choi J. et al. TERT Promotes Epithelial Proliferation through Transcriptional Control of a Myc- and Wnt- Related Developmental Program. PLoS Genet. 2008, 4(1):e10.

Flores I. et al. Effects of telomerase and telomere length on epidermal stem cell behavior. Science. 2005,

309(5738):1253-1256.

Goh C., Zippin J.H. Androgenetic alopecia: diagnosis and treatment with a focus on recent genetic implications. J Drugs Dermatol. 2009, 8(2):185-192.

Haendeler J. et al. Mitochondrial telomerase reverse transcriptase binds to and protects mitochondrial DNA and function from damage. Arterioscler Thromb Vasc Biol. 2009, 29:929-935.

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Kwack M.H. et al . Wnt5a attenuates Wnt/β -catenin signalling in human dermal papilla cells.

Kwack M.H. et al. Wnt5a attenuates Wnt/β-catenin signalling in human dermal papilla cells. Exp Dermatol. 2013, 22(3):229-31.

Myung P.S. et al. Epithelial Wnt ligand secretion is required for adult hair follicle growth and regeneration. J Invest Dermatol. 2013, 133(1):31-41.

Paus R., Cotsarelis G. The biology of hair follicles. N Engl J Med. 1999, 341(7):491-497.

Plikus M.V. New activators and inhibitors in the hair cycle clock: targeting stem cells' state of competence. J Invest Dermatol. 2012, 132(5):1321-1324.

Restrepo, R. Anatomía microscópica del folículo piloso. Rev Asoc Col Dermatol. 2010, 18(3):123-138.

Rittié L. et al. Hedgehog signaling maintains hair follicle stem cell phenotype in young and aged human skin. Aging Cell. 2009, 8(6):738-51.

Rompolas P. et al. Live imaging of stem cell and progeny behaviour in physiological hair-follicle regeneration. Nature. 2012, 487(7408):496-499.

Sarin K. et al. Conditional telomerase induction causes proliferation of hair follicle stem cells. Nature. 2005,

436:1048-1052.

Schneider EL, Mitsui Y. The relationship between in vitro cellular aging and in vivo human age. Proc. Natl. Acad. Sci. 1976, 73(10):3584-3588.

Stutte G. et al. Carbon Dioxide Enrichment Enhances Growth and Flavonoid Content of Two Scutellaria species. J. Amer. Soc. Hort. Sci. 2008, 133(5):631638.

Trüeb. R. Oxidative Stress in Ageing of Hair. Int J Trichology. 2009, 1(1):6-14.

Upton J.H. et al. Oxidative stress and cell senescence in androgenetic alopecia (AGA). Journal of Investigative Dermatology. 2013, 133:1398.

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PROVITAL. S.A. Pol. Ind. Can Salvatella Gorgs Lladó, 200 08210 Barberà del Vallès Barcelona (España) Tel. (+34) 93 719 23 50

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