A Randomized Study of Biomimetic Peptides Efficacy and Impact on the Growth Factors Expression in the Hair Follicles of Patients with Telogen Effluvium

Abstract

Currently, biomimetic peptides have been developed to treat alopecia, which consists of amino acid residues and are able to selectively interact with cell receptors. This article studies the expression of growth factors (VEGF, KGF, EGF, TGF-β1) in the hair follicle of patients with telogen effluvium and healthy individuals and evaluating the efficacy of therapy with biomimetic peptides in patients with telogen effluvium based on clinical data and the dynamics. The method of immunofluorescence study of scalp specimens, which is taken by the punch biopsy from the frontotemporal areas before and after the therapy, was used. 30 female patients with telogen effluvium aged between 20 and 56 years old were recruited for the research and randomly divided into two groups. Patients of group I were treated with lotion based on biomimetic peptides and those of group II were treated with 0.9% saline. The therapeutic efficacy of biomimetic peptides in women with telogen effluvium was assessed by trichoscopy and phototrichography. A change in the expression of VEGF, KGF, TGF-β1 growth factors was found in women with telogen effluvium as opposed to the healthy ones. After the therapy, a significant increase in the VEGF, EGF expression and a decrease in KGF and TGF-β1 expression were noted. The study proved the effectiveness of the proposed method of therapy for women with telogenic effluvium. The research findings can be useful to dermatologists and cosmetologists for increasing the effectiveness of therapy for patients with telogenic effluvium. Also, the data obtained in the article open up the prospect for the further study of the pathogenetic mechanisms of telogenic effluvium and provide an opportunity for an innovative approach to the therapy of patients with telogenic effluvium.

Full text

Journal of Applied Pharmaceutical Science Vol. 8(04), pp 015-022, April, 2018
Available online at http://www.japsonline.com
DOI: 10.7324/JAPS.2018.8403
ISSN 2231-3354
© 2018 Alexey Alekseevich Kubanov et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommer-
cial-ShareAlikeUnported License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
*Corresponding Author
Yulia Albertovna Gallyamova; Department of Dermatovenereology and
Cosmetology, Russian Medical Academy of Continuous Professional
Education, 2/1 Barrikadnaya St., Moscow, 123995, Russia.
E-mail: yulya.gallyamova.69 @ mail.ru
A Randomized Study of Biomimetic Peptides Efficacy and Impact on
the Growth Factors Expression in the Hair Follicles of Patients with
Telogen Effluvium
Alexey Alekseevich Kubanov1,2, Yulia Albertovna Gallyamova1,*, Olga Andreevna Korableva2
1Department of Dermatovenereology and Cosmetology, Russian Medical Academy of Continuous Professional Education, Moscow, Russia.
2State Research Center of Dermatovenereology and Cosmetology Ministry of Healthcare of the Russian Federation, Moscow, Russia.
ARTICLE INFO ABSTRACT
Article history:
Received on: 07/07/2017
Accepted on: 20/11/2017
Available online: 29/04/2018
Currently, biomimetic peptides have been developed to treat alopecia, which consists of amino acid residues and are
able to selectively interact with cell receptors. This article studies the expression of growth factors (VEGF, KGF, EGF,
TGF-β1) in the hair follicle of patients with telogen euvium and healthy individuals and evaluating the ecacy of
therapy with biomimetic peptides in patients with telogen euvium based on clinical data and the dynamics. The
method of immunouorescence study of scalp specimens, which is taken by the punch biopsy from the frontotemporal
areas before and after the therapy, was used. 30 female patients with telogen euvium aged between 20 and 56
years old were recruited for the research and randomly divided into two groups. Patients of group I were treated with
lotion based on biomimetic peptides and those of group II were treated with 0.9% saline. The therapeutic ecacy of
biomimetic peptides in women with telogen euvium was assessed by trichoscopy and phototrichography. A change
in the expression of VEGF, KGF, TGF-β1 growth factors was found in women with telogen euvium as opposed to
the healthy ones. After the therapy, a signicant increase in the VEGF, EGF expression and a decrease in KGF and
TGF-β1 expression were noted. The study proved the eectiveness of the proposed method of therapy for women
with telogenic euvium. The research ndings can be useful to dermatologists and cosmetologists for increasing
the eectiveness of therapy for patients with telogenic euvium. Also, the data obtained in the article open up the
prospect for the further study of the pathogenetic mechanisms of telogenic euvium and provide an opportunity for
an innovative approach to the therapy of patients with telogenic euvium.
Key words:
hair loss, telogen euvium,
hair follicle, hair cycle, hair
growth, growth factors,
peptides.
INTRODUCTION
Diuse cyclic hair loss in women has been described for
the rst time by W. Guy and W. Edmundson in 1960 (Guy and
Edmundson, 1960). In 1961, A.M. Kligman introduced the term
of telogen euvium to dene a condition that is accompanied
by intensied telogen hair loss as a result of hair follicle cycle
abnormalities (Kligman, 1961).
A vast number of triggers to induce hair loss were
discovered. These include hypo- and hyperthyroidism (Sperling,
2001), chronic systemic disorders, such as amyloidosis, hepatic
failure, chronic renal failure, inflammatory bowel disorders,
or lymphoproliferative disease, which may cause telogen
effluvium (Kligman, 1961; Fiedler and Gray, 2003). Telogen
effluvium is also observed with autoimmune diseases, such
as systemic lupus erythematosus or dermatomyositis (Fiedler,
2003), infectious diseases, such as human immunodeficiency
virus type 1 (Sperling, 2001), and secondary syphilis (Rook
and Dawber, 1982). Chronic skin disorders, such as psoriasis
or seborrheic dermatitis, can also cause telogen effluvium
(Apache, 1986; Bergfeld, 2008).
A number of drugs are known to lead to hair loss and
these include combined oral contraceptives, androgens, retinoids,
beta-blockers, anticonvulsants, antidepressants, anticoagulants,
antithyroid agents, ACE inhibitors, and lithium medications

Kubanov et al. / Journal of Applied Pharmaceutical Science 8 (04); 2018: 015-022016
(Fiedler and Gray, 2003; Tosti and Pazzaglia, 2007; Patel et al.,
2013; Watras et al., 2016).
Many authors highlight the association between
telogen euvium and decits of micronutrients, such as zinc,
Ferrum, Vitamin D, Vitamin H, etc. (Trüeb, 2016; Cheung et
al., 2016). There is research available to prove insuciency of
essential micronutrients (zinc and selenium) and excess of toxic
microelements (cadmium, lead, beryllium, and stibium) in those
suering from telogen euvium (Fiedler and Gray, 2003; Rook
and Dawber, 1982; Rushton, 2002; Abdel-Aziz et al., 2015).
The eect of the psychoemotional state on hair loss
intensity was mentioned multiple times in last-century publications
(Habit, 1990; Pasricha, 1991). Ebling suggested yet back in 1987
that hypothalamic cells that are capable of transforming nerve
impulses into hormonal and inuence hair follicles indirectly
(Ebling, 1987). Modern authors, however, believe these arguments
are weak since everyday stress is not enough to cause intensive hair
loss (Harrison, 2002; Fiedler, 2003). Despite many publications
attaching great importance to psychoemotional stress in telogen
euvium development, this matter is still disputed (Fiedler and
Gray, 2003; Rook and Dawber; 1982).
One or another provoking factor aects keratinocyte
division and dierentiation, metabolic processes in the bulge
region of the hair follicle, which leads to premature anagen
termination. The hair follicle gradually passes into the phase of the
catagen, and then the telogen, the ratio of telogen and anagen hair
changes, respectively. This cycle takes a certain period of time,
usually about two to four months. Patients notice massive hair
fallout in two to four or even six months; therefore, the causality
between hair thinning and the etiologic factor is not obvious
(Harrison, 2002; Headington, 1993).
The authors believe telogen euvium to be acute (last
up to 6 months), chronic (6 months and more), or chronic with
relapses (Bergfeld, 2001; 2008). If the exposure to the triggering
factor is short, hair fallout regresses spontaneously or against
appropriate treatment. If the pathological eect on the hair follicle
lasts for a long time, or dierent triggering factors are in action by
turn, telogen euvium grows strong (Bergfeld, 2008).
Considering the pathologic mechanism of telogen
euvium, potential therapeutic possibilities include catagen
inhibition, which requires anagen prolongation; anagen induction
in telogen follicles and exogene suppression in order to stop hairs
from falling out. Neither drug that is currently used to treat alopecia
is eective in inhibiting catagen or inducing anagen (Paus, 2008).
Today’s fundamental cytogenetic and biochemical
research highlights the role of regulatory substances in hair
cycling. These include growth factor families and receptors,
transcription factors, cytokines, neurotrophins, and intracellular
signaling pathways. Special consideration is given to polypeptide
growth factors that are together referred to as trophic regulatory
substances. These factors exert a variety of eects on various
cells: stimulate or inhibit cell proliferation chemotaxis, and
dierentiation. Growth factors capable of regulating the hair
growth cycle have been established recently (Takakura, 1996;
Yano, 2001; Rishikaysh et al., 2014).
Growth factors are capable of inducing cell proliferation
of vascular endothelium and dermal broblasts, prolonging
anagen and delaying catagen onset in a hair follicle (Böhlen,
1985; Kawano, 2005; Kimura-Ueki et al., 2012). Hence, anagen
can be prolonged and hair growth can be stimulated with the help
of these growth factors (Ozeki, 2003; Tomita, 2006; Jain and
De-Eknamkul, 2014).
Vascular endothelial growth factor (VEGF) is the
most studied growth factor. VEGF is of great importance in the
development and life of hair. It promotes growth, determines the
dierentiation, structure, and duration of the hair follicle and hair
shaft growth in vivo (Yano, 2001). VEGF exerts a huge impact
on vascularization and angiogenesis, thus stimulating hair growth
(Ozeki and Tabata, 2003; Bartels et al., 2011; Kim et al., 2013).
The study of the mechanisms of apoptosis in the hair follicle
has determined the important role of the transforming growth
factor (TGF). Many studies have shown that TGF-β has unique
multidirectional eects on the physiology of the hair follicle – both
stimulating and inhibiting eects on hair growth (Stenn, 2001;
Inoue, 2009). Controversial data was obtained in the study of the
role of epidermal growth factor (EGF) (Cohen, 1963; Moore,
1981; Kashiwagi, 1997) and keratinocyte growth factor (KGF)
(Guo and Fuchs, 1993; Danilenko, 1995; Kawano et al., 2005) in
the hair cycle. Studies have shown a dose-dependent eect of KGF
and EGF on hair growth (Guo and Fuchs, 1993; Danilenko, 1995;
Kashiwagi, 1997).
Currently, numerous studies are of experimental nature,
and the role of growth factors in the pathogenic mechanism of
various types of alopecia and their eect on lesion severity is still
undisclosed.
Growth factors are studied not just for theoretical
purposes, but out of practical necessity as well. It was established
that physiological processes inuence just small fragments of growth
factors, i.e. oligopeptides, instead of whole molecules. With view to
the above, biomimetic peptides have been presently synthesized (Lat.
bios – life and mimesis – imitation). They are composed of amino acid
residues that are capable of selective interaction with cell receptors and
can be used topically (Lupo and Cole, 2007).
We have studied the eect biomimetic peptides have on
growth factor expression in a hair follicle and assessed therapeutic
ecacy in female patients with telogen euvium.
MATERIALS AND METHODS
This study involved 30 female subjects with telogen
euvium at average age of 37.1 ± 9.29 years. Eight healthy females
at average age of 38.5 ± 7.4 years constituted the control group.
Inclusion criteria included Informed Consent and age of 18 to 60
years. Exclusion criteria included thyroid disorders, active skin lesions,
infectious diseases, hyperandrogenism, severe somatic comorbidities,
and other types of alopecia. The study protocol was approved by the
Ethical Committee of the Russian Medical Academy of Postgraduate
Education. The selected patients were randomly divided into two
groups; group I included 15 patients, who were treated with lotion
based on biomimetic peptides, and group II included 15 control
patients who were treated with 0.9% saline.
Intervention technique
A lotion based on biomimetic peptides (2 ml) was
topically applied on the scalp of group I patients and followed
by microneedle therapy (needle size of 0.2 mm) (China, Beijing,

Kubanov et al. / Journal of Applied Pharmaceutical Science 8 (04); 2018: 015-022 017
Beijing Peak Beauty Technology Co., Ltd., Micro Needle Roller
System). The patients of group II were treated with normal
saline followed by microneedle therapy. The major components
of the topical solution used for treatment were: Decapeptide-18,
Oligopeptide-54, Decapeptide-10, Octapeptide-2, Decapeptide-19,
Oligopeptide-71, and Decapeptide-28. Each patient received 16
treatments at 3-4 days intervals.
Assessment of the response
Assessment of the response was done two months after
treatment commencement and two and six months after the last
treatment. Photography Digital photos were taken for the scalp
before therapy and during subsequent visits.
Physical examination of the scalp covered the
following: assessment of visible hair thinning, thinning of a hair
shaft, intensity of sebaceous excretions, and scalp rash, if any.
Assessment of hair density, diameter, proportion of vellus and
terminal hairs, ratio and distribution of hairs in follicular units,
proportion of anagen and telogen hairs in the androgen dependent
and androgen independent scalp regions was done for all patients,
using a specialized Aramo SG microcamera (Rep. Korea, Seoul,
Bundang Seoul National University Hospital Health Care
Innovation Park 401-ho, Aram HUVIS Co., Ltd.) in combination
with Trichoscience 1.7 application (Russia, Moscow, Trilogic) in
the xed zone, and a permanent mark was made.
Immunouorescence
Punch biopsy (4 mm) was done under local anesthesia in
patients with alopecia. For the local anesthesia, the anesthetic lidocaine
(Hungary, Budapest, Egis Pharmaceuticals PLC) was administered
subcutaneously around the site of biopsy. Parietal bioptates were
taken before the treatment. In 2 months, bioptates were taken from the
regions in close proximity to the rst punch biopsy.
Eight control scalp samples were taken from the patients
that did not complain of hair loss and did not present any signs of
alopecia during a surgery or other routine intervention for benign
lesions, always by voluntary informed consent of the patient.
VEGF, KGF, EGF, and TGF-β1 expression was studied
by immunouorescence, using standard technique using antibodies
directed against VEGF, KGF, EGF (all UK, Cambridge, Abcam),
TGF-β1 (UK, Abingdon, Novus Biologicals).
For the assessment of the results, the morphometric
research was conducted using the system of computer analysis
of microscopic images, which comprised a confocal Olympus
FlueView1000 microscope and a personal computer on the basis
of Intel Pentium 4, and “Videotest Morphology 5.0” (Russia). The
expression of growth factors was assessed in 5 visual elds at
magnication of x 200.
Statistical analysis
A statistical analysis was performed using R version v3.2.0.
The Shapiro–Wilk test was used to determine patient distribution by
parameter values. Descriptive statistic values were calculated, using
standard techniques. If distribution of quantitative parameters was
recognized normal, mean deviation and standard deviation were
calculated. If the distribution deviated from normal, median and
quartiles were calculated. Mode and 25-75% quartiles were calculated
for qualitative parameters. The Wilcoxon test was used for time
comparison before and after the treatment with p < 0.05. The null
hypothesis for the groups was rejected.
RESULTS
Under our supervision, there were 30 patients with the
diagnosis of telogen euvium and the average age of the patients
was 37.1 ± 9.29 years old, average age of disease onset was 26
(22-33.25) years old, and the average disease duration was 5
(1.25-16) years.
According to patient histories, the patients associated
their disease onset with childbirth (n = 6; 20%), or stress (n = 12;
40%), and one patient mentioned diet deviation (n = 1; 3.33%).
Eleven women (36.67%) were not able to associate their disease
onset with any certain provoking factor. It was determined from
family histories that eleven (30%) patients with telogen euvium
had hereditary diseases, passed by their father: ten (33.33%),
passed by their mother: four (13.33%), including both mother and
father (in three patients), all by rst relation degree.
The majority of patients used self-treatment with topical
hair products and oral medicines: 13 (43.33%) and 10 (33.33%),
respectively. A variety of lotions and shampoos, folk remedies
were used topically, and combined vitamins and minerals were
taken orally. Among them, seven patients (23.30%) underwent
mesotherapy. Despite their treatment, temporary eect was
observed in 4 (21%) patients, and no positive changes were found
in 11 (57.89%) of patients with telogen euvium. Eleven (36.67%)
patients among the studied women did not receive any treatment.
Trichoscopy and phototrichography assessment
According to trichoscopy and phototrichography,
normal ratio of anagen and telogen hairs was observed in the
patients before treatments towards the increased proportion of
telogen hairs. At the same time, the number of hairs per square
centimeter, ratio of terminal and vellus hairs in the parietal and
occipital regions was within normal.
Following the combined treatment, there was observed
a statistically signicant increased proportion of anagen hairs in
group I (from 62.98 ± 11.26% to 82.09 ± 10.18% and from 75.51 ±
6.52% to 85.77 ± 6.08%, respectively) and a statistically signicant
decreased proportion of telogen hairs (from 37.02 ± 11.26% to 17.64
± 10.09% and from 24.49 ± 6.52% to 14.23 ± 6.08%, respectively),
juvenile hairs also emerged in the parietal region. Positive changes
were observed 2 and 6 months after treatment (Table 1).
Therefore, a statistically signicant increase in total
number of hairs per square centimeter in the parietal region,
decrease in the proportion of telogen hairs and increase in the
proportion of anagen hairs, and a statistically signicant increase
in the hair diameter in the parietal and occipital regions were
observed, when comparing trichoscopy and phototrichography
data for group I patients with telogen euvium before and after
treatment (р < 0.05). A statistically signicant increase in the
total number of hairs and follicular units per square centimeter
of the parietal region was observed, when comparing trichoscopy
and phototrichography data before treatment and 2 months after
treatment.

Kubanov et al. / Journal of Applied Pharmaceutical Science 8 (04); 2018: 015-022018
Table 1: Trichoscopy and phototrichogram analysis before and after treatment.
Localization Parameter
group I (n = 15) group II (n = 15)
before treat-
ment
after
treatment
2 months after
treatment
6 months after
treatment
before
treatment
after treat-
ment
2 months after
treatment
6 months after
treatment
Androgen-dependent
(parietal) region
Hair density
[number of
hairs per sq.cm]
242.4 (231.9-
289.6)
289.65 ±
82.551 303.22 ± 87.72289.2 (243.7-
334.4)3,4
310.85 ±
86.75 310.87 ± 88.58 321.31 ± 85.942320.09 ± 88.684
[follicular units
per sq.cm]
165.7 (124.4-
179.5)
168.07 ±
54.44 181.86 ± 57.152188.37 ±
66.144
197.75 ±
53.98 191.64 ± 63.3 188.91 ± 55.79 196.71 ± 57.94
average hair
diameter [μm] 66 ± 8.57 80 (73-80)177 (66;77.5) 72.73 ± 12.67 63.13 ± 7.47 65.87 ± 8.98165 (62-69) 64 (59-67.5)3
anagen hairs
[%] 62.98 ± 11.26 82.09 ±
10.18185.95 ± 4.08 89.1 (85.1-
90.1)3,4
64.09 ±
12.01 72.84 ± 5.77181.87 ± 4.2282.05 ± 4.984
Telogen hairs
[%] 37.02 ± 11.26 17.64 ±
10.09112.67 ± 4.03 10.9 (9.9-14.9)435.91 ±
12.01 27.16 ± 5.63118.13 ± 4.2217.95 ± 4.984
juvenile [%] 0 0 0 (0-1.1) 0 0 0 0 0
vellus hairs [%]
(<30 μm) 12 (6-16) 11.93 ±
5.71 7.93 ± 5.32239.6 ± 1.78 13 ± 6.15 12.4 ± 5.78 6 (4-15.5)29 (3-10)
Androgen-independent
(occipital) region
Hair density
[number of
hairs per sq.cm]
242.75 ±
34.81
249.75 ±
41.89
220.9 (210.95-
267.6) 252.62 ± 52.35
275.5
(217.5-
276.9)
269.6 (228.55-
280.25) 241.89 ± 47.29 248.07 ± 41.983
[follicular units
per sq.cm]
115.3 (105.9-
172.2)
124.6
(108.15-
167)
135.27 ± 44.74 139.8 (59.08) 137.59 ±
36.5 133.05 ± 50.55 142.05 ± 51.11 147.4 ± 44.97
Average hair
diameter [μm] 61 ± 9.33 71 (68.5-
75.5)173 (72-75) 72 (67-75.5)461.53 ± 8.02 63.87 ± 6.63 63.27 ± 7.89 65.93 ± 6.864
anagen hairs
[%] 75.51 ± 6.52 85.77 ±
6.08189.44 ± 5.97 89.11 ± 3.82475.9 (69.55-
78.6)
80.4 (77.25-
81.7)184.07 ± 4.35284.39 ± 2.264
Telogen hairs
[%] 24.49 ± 6.52 14.23 ±
6.08110.56 ± 5.97 10.89 ± 3.82424.1 (21.4-
30.45)
19.6 (18.3-
22.75)115.93 ± 4.35215.61 ± 2.264
Vellus hairs
[%] (<30 μm) 12.6 ± 7.31 13 ± 8.4 6 (2.5-12)28 (4.5-13) 14 ± 5.45 13.33 ± 8.25 10.4 ± 5.95 10.73 ± 7.27
Values are mean (SD) or median and range.
1statistically signicant dierences between parameters before and after treatment in each group; P < 0.05
2statistically signicant dierences between parameters after treatment and 2 months after treatment in each group; P < 0.05
3statistically signicant dierences between parameters 2 months after treatment and 6 months after treatment in each group; P < 0.05
4statistically signicant dierences between parameters before treatment and 6 months after treatment in each group; P < 0.05.
A statistically signicant increase in the total number of
hairs by 19.3% and follicular units per square centimeter by 13.7%
in the parietal region, a statistically signicant increase in the
proportion of anagen hairs in the parietal region (by 26.12%) and
occipital region (by 13.6%), and a statistically signicant increase
in the diameter of hairs in the occipital region were established,
when comparing trichoscopy and phototrichography data before
treatment and 6 months after treatment (р < 0.05) (Table 1).
Fig. 1: Changes in anagen hairs in the parietal region (A) and occipital region (B) in group I patients against treatment.
Note: rectangles correspond to margins of 25% and 75% quartiles, black horizontal lines correspond to median. Time intervals are represented on the X-axis: before
treatment (1), after treatment (2), 2 months after treatment (3), 6 months after treatment (4); proportion of anagen hairs (%) is represented on Y axis.

Kubanov et al. / Journal of Applied Pharmaceutical Science 8 (04); 2018: 015-022 019
Fig. 2: Phototrichogram and panoramic photo before treatment (A), after treatment (B), two months after treatment (C), and six months after treatment (D).
According to our comparative analysis, the proportion
of anagen hairs increased statistically signicant by 26.12% in the
parietal region and by 13.6% in the occipital region in group I
patients, as a result of treatment. Furthermore, the eect of this
therapy was long lasting and retained 6 months after the treatment.
The proportion of growing hairs statistical increased by 8.5% more
in the parietal region and by 3.9% more in the occipital region for
this period of time (Table 1, Fig. 1, 2).
A statistical increase of anagen hairs in the parietal and
occipital regions (from 64.09±12.01 to 72.84±5.77 and from 75.9
(69.55-78.6) to 80.4 (77.25-81.7), respectively) was observed in
group II patients after treatment. Positive changes were observed
2 and 6 months after treatment.
A statistically signicant decrease in the proportion of
telogen hairs and statistical increase in the proportion of anagen
hairs in the parietal and occipital regions, and statistical increase
in the hair diameter in the parietal region were observed, when
comparing trichoscopy and phototrichography data in group II
patients before and after treatment. Comparison of total number of
hairs per square centimeter, follicular units per square centimeter
in the parietal and occipital regions, and hair diameter before
and after treatment in the parietal region did not produce any
statistically signicant dierences (Table 1).
Despite positive changes against treatment in group II
patients (Fig. 3), phototrichography data did not reach normal
parameter values in the majority of group II patients and were
statistical signicantly lower vs. the experimental group (Table
1), while the proportion of anagen hairs statistical increased by
17.96% in the parietal region and by 8.49% in the occipital region.
VEGF, KGF, EGF, and TGF-β1 expression assessment
Relative VEGF expression area was 68.53 ± 1.08% in
healthy patients. VEGF expression was signicantly lower on the
scalp of group I and group II patients vs. healthy patients (26.91
± 0.66 and 26.1 (25.57-26.88)%, respectively). A signicant
increase in VEGF expression on the scalp of group I patients was
observed after treatment up to 32.26 (32.22-32.84)% (p < 0.05);
therefore, VEGF expression increased by 19.8%. An increase in
VEGF expression merely by 2.8% (Table 2) was observed in the
control group.
Fig. 3: Changes in anagen hairs in the parietal region (A) and occipital region (B) in group II patients against treatment.
Note: rectangles correspond to margins of 25% and 75% quartiles, black horizontal lines correspond to median. Time intervals are represented on the X-axis: before
treatment (1), after treatment (2), 2 months after treatment (3), 6 months after treatment (4); proportion of anagen hairs (%) is represented on Y axis.
Relative KGF expression area on the scalp of healthy
patients was 47.68 ± 0.93. KGF expression was signicantly
lower on the scalp of group I and group II patients vs. healthy
patients (35.65 ± 0.63% and 36 ± 0.75%). KGF expression on the
scalp of group I patients with telogen euvium decreased by 28%
after treatment (p < 0.05). An increase in KGF expression by 2.8%
was observed in the control group (Table 2).
Relative EGF expression area was 19.96 ± 3% in healthy
patients. Level of EGF expression in women of the I and II group
(18.89 ± 0.46% and 19.8 (19.19-20.24)%, respectively) did not
dier from the healthy values (p = 0.830). After treatment, EGF
expression increased in group I patients by 27.66 ± 0.6% (p <
0.05), and an increase in EGF expression was also observed in
group II patients EGF (21.37 (20.32-22.67)%) (p < 0.05) (Table 2).

Kubanov et al. / Journal of Applied Pharmaceutical Science 8 (04); 2018: 015-022020
Table 2: Growth factor expression in patients with telogen euvium (group I and II) before and after treatment.
Parameter
VEGF KGF EGF TGF-β1
Before treatment After treatment Before treatment After treatment Before treatment After treatment Before treatment After treatment
Relative expression area [%]
Group I 26.91 ± 0.66 32.26 (32.22-
32.84)* 35.65 ± 0.63 25.59 ± 0.83* 18.89 ± 0.46 27.66 ± 0.6* 64.67 ± 0.51 52.97 ± 0.65*
Group II 26.1 (25.57-
26.88) 26.83 ± 2.15* 36 ± 0.75* 37 ± 0.83* 19.8 (19.19-
20.24)
21.37 (20.32-
22.67)* 64 ± 0.86 55.27 ± 2.14*
Values are mean (SD) or median and range. *- statistically signicant dierences: p < 0.05.
Relative TGF-β1 expression area on the scalp of healthy
patients was 51.72 ± 2.21%. Study of TGF-β1 changes on the
scalp of group I and II patients, showed a signicant decrease in
TGF-β1 expression by 18.1% and 13.6%, respectively (p < 0.05)
(Table 2).
DISCUSSION
With this study, we determined that biomimetic peptides
are safe, as topical application followed by microneedle therapy.
Subjectively the patients tolerated the treatment well, no side
eects were identied, and none of the patients were excluded
from the observation group. After the end of treatment, the
withdrawal syndrome was not observed.
The analysis of the trichoscopy and phototrichogram
indices in the parietal and occipital regions (hair density, the
number of follicular units per square centimeter, the percentage
of hair in the anagen stage, the hair diameter) before treatment,
after treatment and in two and six months after treatment showed
a statistically signicant greater increase in the percentage of the
hair in the anagen stage in female patients of the main group,
compared to the control group. In patients treated with biomimetic
peptides, the percentage of the anagen and the telogen hair reached
of the normal values.
In addition, the patients of the main group had a
statistically signicant increase in follicular units in the parietal
region. These data indicate that the application of biomimetic
peptides in patients with telogen euvium leads to a decrease in
hair loss due to stimulation and prolongation of the anagen phase
and, consequently, reduction of the telogen phase. Initiation of the
anagen stage is conrmed by the objective fact of the appearance
of young hair, increase in the percentage of hair in the anagen
stage against the background of the increase in the total amount of
hair; and the increase in the number of follicular units during the
treatment period indicates the extension of the anagen period. In
both groups, throughout the entire observation period, the increase
in the hair diameter in the parietal and occipital areas was noted,
indicating that the increase in hair thickness was aected to a
greater extent by mechanical exposure of the dermoroller.
The analysis of growth factor expression showed a
change in VEGF, KGF, and TGF-β1 growth factors in female
patients with telogen euvium vs. healthy female population.
This study established that VEGF, KGF, and TGF-β1 expression
in a hair follicle was impaired in patients with telogen euvium.
EGF expression was equivalent in these patients and healthy
women and did not dier statistically.
Biomimetic peptide treatment ecacy in telogen
euvium patients has been assessed for the rst time in this study.
It was established earlier that VEGF regulates hair growth by
interacting with the VEGFR-2 receptor (Man, 2009; Wu, 2014).
The VEGFR-2 receptor is located in human hair follicles (including
bulge in a dermal papilla), sebaceous glands, eccrines, and other
cells. It has been known that high VEGF mRNA expression in the
dermal papillary cells is registered in anagen.
Animal experiments demonstrated perifolicular vessels
enlarging by more than four times in anagen, which coincides
with cyclic follicle size change. At the same time, the perifolicular
angiogenesis correlates with activated VEGF mRNA expression
in folicular keratinocytes of the outer root sheath. All the above
allowed the authors to suggest improved vascularization and
increased hair follicle size in the experimental animals to be
associated directly with VEGF mRNA expression (Yano, 2001).
It was also established that VEGF mRNA count in a hair follicle
normally varies along the hair cycle: it increases in anagen and
decreases in catagen and telogen (Man, 2009). A signicant
increase of 19.8% in VEGF expression on the scalp was observed
after treatment in patients treated with biomimetic peptides, while
an increase of 2.8% in VEGF expression was observed in the
control group.
Increased EGF expression on the scalp of telogen
euvium patents was observed in the experimental and control
group as a result of the treatment. Investigation of EGF in the hair
cycle generated conicting data, since P.J. Miettinen et al., H.
Zhang et al. assumed EGF importance in hair follicle development
and hair growth (Miettinen, 1995; Zhang et al., 2016a; Zhang et
al., 2016b). Kashiwagi et al., Richardson et al. showed dose and
time dependent EGF eect on hair growth, using an animal model
(Kashiwagi et al., 1997; Richardson, 2009).
A signicant decrease in TGF-β1 expression on the scalp
was observed after treatment in patients treated with biomimetic
peptides by 18.1% vs. 13.6% in the control group. The study of
apoptosis mechanisms in a hair follicle determined an importance
of the transforming growth factor (TGF-β). K. Foitzik and X. Liu
with their colleagues determined on an animal model that TGF-β1
inhibits hair growth by shortening anagen and initiating follicle
catagen. The dependence of TGF-β1 expression on the hair follicle
phase was also shown (Foitzik et al., 2000; Liu et al., 2001; Lu et
al., 2016).
The KGF’s expression on the scalp of the patients
that were treated with biomimetic peptides lowered by 28%,
while an increase in the expression by 2.8% was observed in the
control group. D.M. Danilenko, et al., Richardson G.D., et al.,
and L.Guo, et al., showed the eect of the keratinocyte growth
factor (KGF) to depend on the dose and either to induce, or inhibit
hair growth. KGF is necessary for normal growth, development,

Kubanov et al. / Journal of Applied Pharmaceutical Science 8 (04); 2018: 015-022 021
and dierentiation of a hair follicle, while it binds to the KGFR
receptor (Guo et al., 1993; Danilenko, 1995; Richardson et al.,
2009).
CONCLUSION
Therefore, the following may be concluded that, the
use of biomimetic peptide treatment improves perifolicular
vascularization, which leads to improved blood ow and intensies
hair growth.
While TGF-β is a powerful growth inhibitor of dierent
cell types, including majority of epithelial cells, a decrease
in TGF-β expression following biomimetic peptide treatment
indicates lower apoptosis in patients after treatment, as compared
with the sample before treatment.
According to the research ndings, the role KGF and
EGF growth factors in alopecia development is poorly studied,
and their ambiguous expression can be associated with some
unknown molecular signaling mechanisms.
The authors have analyzed phototrichographys
parameters and concluded that telogen euvium patients
experience reduced hair thinning due to anagen stimulation
and prolongation and, respectively, telogen shortening. Anagen
initiation is objectively evidenced by the juvenile hairs present
and the increased number of units over the treatment period
indicates anagen prolongation.
CONFLICT OF INTERESTS
There are no conicts of interest.
FINANCIAL SUPPORT AND SPONSORSHIP
Nil.
ABBREVIATIONS
VEGF Vascular endothelial growth factor
KGF Keratinocyte growth factor
EGF Epidermal growth factor
TGF-β1 Transforming growth factor beta 1
VEGFR Vascular endothelial growth factor receptor
KGFR Keratinocyte growth factor receptor.
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How to cite this article:
Kubanov AA, Gallyamova YA, Korableva OA. A Random-
ized Study of Biomimetic Peptides Ecacy and Impact on the
Growth Factors Expression in the Hair Follicles of Patients with
Telogen Euvium. J App Pharm Sci, 2018; 8(04): 015-022.