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1856 Vol. 38, No. 12Biol. Pharm. Bull. 38, 1856–1863 (2015)
© 2015 The Pharmaceutical Society of Japan
Regular Article
Safety and Efficacy of Rice Bran Supercritical CO2 Extract for Hair
Growth in Androgenic Alopecia: A 16-Week Double-Blind Randomized
Controlled Trial
Jae-Suk Choi,a,# Jae Beom Park,b,# Woi-Sook Moon,c Jin-Nam Moon,c Sang Wook Son,*,b and
Mi-Ryung Kim*,a
a Major in Food Biotechnology, Division of Bioindustry, College of Medical and Life Sciences, Silla University; 140
Baegyang-daero, 700 Beon-gil, Sasang-gu, Busan 617–736, Republic of Korea: b Department of Dermatology, Korea
University Ansan Hospital; 123 Jeokgeum-ro, Danwon-gu, Ansan 425–701, Republic of Korea: and c Department
of R&D, ECOMINE Co., Ltd.; Busan Innobiz Center #402, 1 Mandeok 3-ro, 16 Beon-gil, Buk-gu, Busan 608–736,
Republic of Korea.
Received May 6, 2015; accepted August 25, 2015
We conducted a 16-week double-blind randomized controlled single-center trial to evaluate the safety
and efficacy of dermal rice bran supercritical CO2 extract (RB-SCE) in the treatment of androgenic alopecia.
Fifty alopecia patients were randomly assigned to the experimental and placebo groups. The experimental
group received a dermal application of 0.5% RB-SCE (8 mL/d) to the head skin for 16 weeks while the
control group received a dermal application of placebo. Changes in hair count, diameter, and density were
evaluated with a Folliscope®. Patient satisfaction was evaluated via questionnaire and clinical photographs
were rated by dermatologists. The results showed that RB-SCE significantly increased hair density and hair
diameter in male subjects. Patient satisfaction and the evaluation of photographs by dermatologists also con-
firmed the effectiveness of RB-SCE in the treatment of alopecia. No adverse reactions related to RB-SCE
were reported. Therefore, RB-SCE shows promise for use in functional cosmetics and pharmaceuticals.
Key words rice bran supercritical CO2 extract; hair growth-promoting activity; alopecia; clinical study
A daily loss of about 50–60 scalp hairs is considered nor-
mal, but a loss exceeding approximately 100 hairs will result
in alopecia. Alopecia has been estimated to affect between
0.2 and 2% of the world’s population.1–3) Although medically
viewed as a relatively minor dermatological condition, alope-
cia may have a significant negative impact on quality of life
based on the psychological and symbolic importance of hair.
The demand for drugs that alter hair growth and appear-
ance has spawned a multi-billion dollar industry. To date, only
two alopecia t reatments, mi noxidil and finaste ride, have been
approved by the U.S. Food and Drug Administration. Topical
minoxidil solution (Rogaine for Men and Women; Pharmacia
Corp., Peapack, NJ, U.S.A.) has been shown to stimulate new
hair growth and to help prevent further hair loss in affected
areas in both men and women with alopecia,4,5) although the
specific mechanism of action remains unclear. Finasteride
(Propecia; Merck Co., Rahway, NJ, U.S.A.) is a synthetic azo-
steroid that is a potent and highly selective non-competitive
antagonist of 5α-reductase type 2. It binds irreversibly to the
enzyme and inhibits the conversion of testosterone to dihy-
drotestosterone.6)
Generally, minoxidil is well tolerated for long-term daily
use. Side effects of minoxidil are uncommon, but contact der-
matitis,7) skin irritation,8 ,9) dizziness, and tachycardia9) have
been repor ted. Finasteride is also generally well tolerated,
with few adverse effects reported over 5 years. In finasteride-
treated patients, 1.9% reported loss of libido and 1.4% report-
ed erectile dysfunction in the first year. The placebo-treated
group reported the same events with frequencies of 1.3 and
0.6%, respectively. These events appeared to resolve on ces-
sation of the treatment and, in some cases, during continued
treatment.10) Both drugs were discovered as the result of seren-
dipity rather than rational hair drug design. Thus, to develop
more precise therapies for alopecia, the mechanisms of hair
growth-promoting effects as well as new drugs and natural
hair growth enhancers require exploration.
In previous research, rice bran supercritical CO2 extract
(RB-SCE) was found to be a potent inducer of hair growth
in mice11) via 5-alpha-reductase inhibition.12) In addition,
the toxicological safety of RB-SCE was investigated using
the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-
2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay in RAW264.7
cells. Further safety evaluations included single oral dose tox-
icity in rats,13) an acute dermal and ocular irritation test,14) a
single dose and 4-week repeated dose dermal toxicity study,15)
and a genotoxicity assessment.16) Thus, RB-SCE is thought to
be safe. Despite its therapeutic potential and safety, no reports
have described a clinical study of RB-SCE for alopecia.
Therefore, in the present study, we evaluated the safety of
RB-SCE and its effects on markers of hair growth in alopecia.
We expect that the results of this open-label trial can provide
basic clinical information regarding changes in biochemical
markers after RB-SCE treatment.
MATERIALS AND METHODS
Test Material RB-SCE (rice bran, Oryza sativa L. var.
japonica) was prepared in a semi-continuous flow-type ap-
paratus with a 3-L extractor.11) Test tonic products with or
without RB-SCE were prepared and kindly supplied by Kol-
mar Korea (Sinjeong-ri, Yeongi-gun, Chungcheongnam-do,
Korea). The studied formulations (Table 1) were prepared
* To whom correspondence should be addressed. e-mail: skin4u@korea.ac.kr; haha7kmr@silla.ac.kr
# These authors contributed equally to this work.
Vol. 38, No. 12 (2015) 1857
Biol. Pharm. Bull.
in a PRIMIX RM homomixer (PRIMIX Co., Ltd., Japan) at
3000 rpm within 10 min and supplemented with 0.5% (w/w)
RB-SCE. A placebo formulation was prepared without RB-
SCE.
Study Design The study design and a flow chart of study
subjects are shown in Fig. 1. We enrolled 50 patients who
were diagnosed with alopecia according to the criteria for the
diagnosis of alopecia in Korean patients. Informed consent
was obtained from all study participants. To assess the degree
of hair loss, we used the Hamilton–Norwood classification
for men and the Ludwig classi fication for women. This was a
double-blind randomized controlled clinical trial. The patient
group comprised a total of 50 subjects including 22 women
and 28 men. They were enrolled in either the RB-SCE group
or the placebo group by the table of random numbers. They
were randomized 1 : 1 to either RB-SCE or placebo treatment
(25 patients each; Table 2). The RB-SCE group received a
dermal application of a test tonic product containing 0.5%
RB-SCE to bald scalp skin and the placebo group received a
dermal application of a test tonic product that contained no
RB-SCE. At the baseline visit, subjects were given a plastic
bottle containing a 16-week supply of their assigned test tonic
(with or without RB-SCE). They were instructed to treat the
scalp with 4 mL of solution once or twice a day at approxi-
mately 12-h intervals (total daily dose of 8 mL).
Scalp photography and phototrichography (Folliscope 2.5;
LeadM Co., Seoul, Korea) were performed at baseline (0
week) and after 8 and 16 weeks of treatment. The study pro-
tocol was approved by the institutional review board (IRB)
of the Korea University Ansan Hospital (IRB No. AS13141).
All procedures were conducted in accordance with the ethics
standards of the Declaration of Helsinki and Good Clinical
Practice Guidelines.
To ensure compliance and monitor adverse effects, subjects
underwent diagnostic tests conducted by a dermatologist after
8 and 16 weeks. To assess treatment compliance, subjects
were asked to bring in their solution bottle at the end of the
study and the remaining test tonic volume was measured by
research staff. Restricted food and drugs during the study
period included any hair growth-promoting agents, alopecia
treatments, prostatic disease treatments, hormone products,
and skin and hair health supplements.
For all site visits, subjects were asked to visit at the same
time in the morning. Physical examinations including vital
sign and hair measurements were performed at baseline and
after 8 and 16 weeks. The patient questionnaire assessment
and skin tolerance and safety evaluations by clinical observa-
tion were also performed after 8 and 16 weeks of treatment.
Inclusion Criteria The study inclusion criteria were as
follows: age greater than 18 years; a diagnosis of alopecia
more than 2 week s prior to the study; t reatment requ irement;
and agreement to participate in this clinical trial after receipt
of an explanation of the objectives, methods, and efficacy of
the study drug.
Table 1. Formulation of the Test Tonic Product Containing RB-SCE
INCI Percentage of components (w/w)
Water (aqua), demineralized as 100%
Glycerin 1.0
Hydroxyethyl cellulose 0.5
Tetrasodium EDTA 0.03
C12–14 Pareth-12 2.0
Hyaluronic acid 0.5
Alcohol 15.0
RB-SCE 0.5
PEG-40 castor oil 2.0
Triethanolamine 0.3
INCI=International Nomenclature of Cosmetic Ingredients; RB-SCE=rice bran
supercritical CO2 extract.
Fig. 1. The Study Design a nd Flow Char t of Study Subjects
Of the 50 en rolled pe rsons, 43 completed t he study visits an d inter vention . Six
men and one woma n we re excluded fr om the effica cy analysi s d ue to poor compl i-
ance (<80%).
Table 2. Age and Sex Distribution of Alopecia Areata Patients
Enrollment Completion
NAge (years) NAge (years)
RB-SCE treated group Male 12 39.0±8.6 9 41.0±9.2
Female 13 42.6±12.8 12 42.2±13.3
Sub-total 25 — 21 —
Mean — 40.1±11.0 — 41.7±11.5
Placebo group Male 16 41.9±8.2 13 41.3±6.6
Female 9 50.0±15.7 9 50.0±15.7
Sub-total 25 — 22 —
Mean — 44.8±11.9 — 44.9±11.8
Sum (male/female) Total 50 (28/22) —43 (22/21) —
Range 25–68 28–68
Mean — 42.0±11.37 — 43.3±11.58
RB-SCE=Test tonic containing RB-SCE; Placebo=Test tonic with no RB-SCE. The patient group comprised a total of 50 subjects including 22 women and 28 men. Of
these, 43 subjects (12 men and 11 women) completed the study visits and intervention. Six men and one woman were excluded from the efficacy analysis due to poor compli-
ance (<80%).
1858 Vol. 38, No. 12 (2015)
Biol. Pharm. Bull.
Exclusion Criteria The study exclusion criteria were as
follows: treatment with other products or medications within
2 weeks before study initiation; allergy to any component of
RB-SCE; medication for another disease; pregnancy or breast-
feeding; enrollment in another clinical study within 3 months
of this study; inability to understand the objectives and meth-
ods of this clinical trial; and inappropriateness for study par-
ticipation as deemed by the clinician.
Phototr ichography Hair measurement was performed via
phototrichography according to the methods of Oh and Son17)
using a computerized hand-held USB camera (Folliscope 2.5)
at baseline and after 8 and 16 weeks. Briefly, the primary site
of scalp baldness was tattooed and examined for hair density
(number of hairs per square centimeter) and hair diameter (mi-
crometer). Hair density per square centimeter was determined
manually under 50-fold magnification. Hair diameter was
measured under 100-fold magnification.
Expert Panel Assessment of Global Photographs Stan-
dardized global photographs of the primary site of scalp bald-
ness were obtained at baseline, 8 weeks, and 16 weeks. The
patient’s head was placed in a stereotactic device to maintain
consistency of patient positioning and photographic distance.18)
Subjects were instructed to maintain their hairstyle throughout
the study and to avoid dyeing their hair or using any hair en-
hancement procedures.
Global photographs were reviewed in a blinded manner by
an expert panel of three dermatologists (Jae Beom Park, Sung
Kyu Jung, and Sang Wook Son) at the end of the trial using a
7-point scale.19) Assessments included the percentage of scalp
affected by alopecia and a scoring of hair regrowth from −3
to +3 using a 7-point scale as follows: greatly, moderately, or
slightly decreased; no change; slightly, moderately, or greatly
increased.
Patient Questionnaire Assessment After 8 and 16
weeks of treatment, each subject was asked to perform a self-
assessment of personal hair growth and scalp appearance. The
self-assessment questionna ire parameters included size of the
vertex spot, hair loss on top of the scalp, bitemporal reces-
sion, hair shedding, hair quality, and overall satisfaction.2 0)
Patient satisfaction was assessed using the 3-point rating scale
(improved, no change, or worse) most commonly used in
alopecia-related research.17)
Skin Tolerance and Safety Evaluations Product safety
was assessed by a dermatologist’s clinical observation as well
as subject feedback. Adverse effects were recorded using
WHO Adverse Reaction Terminology and evaluated for a
causal relationship to the treatment. The objective signs were
erythema, edema, scaling, and papule. The subjective signs
included itching, prickling, burning, stinging, stiffness, tight-
ness, burning of the eyes, weeping, etc. The frequency, dura-
tion, and intensity of each symptom and a possible or probable
relationship with the test samples were investigated.
Statistical Analysis All statistical analyses were per-
formed using SPSS software (version 12.0; SPSS Inc., Chi-
cago, IL, U.S.A.). A paired t-test was used to evaluate statisti-
cally significant differences in hair density, hair diameter, and
expert panel assessment. Student’s t-test was used to evaluate
increases in hair density and hair diameter. The chi-square
test was used to evaluate statistically significant between-
group differences in the questionnaire analysis. Statistical
significance was accepted at p-values less than 0.05.
RESULTS
Characterization of Patients This study took place be-
tween February 2014 and July 2014. Of the 61 people who
were screened for study inclusion, 50 subjects (28 men and
22 women) participated. Seven subjects (6 men and 1 woman)
withdrew after the baseline visit for personal reasons, and 43
subjects (22 men and 21 women) completed the study (Table
2). Patient ages ranged from 28 to 68 years (mean, 43.3±11.6
years). Mean ages were 41.7 years in the RB-SCE group and
44.9 years in the placebo group.
Hair Density At baseline, the average number of hairs
in male subjects in the RB-SCE group was 93.50 strands/
cm2. After RB-SCE treatment, hair density increased to 96.56
and 103.22 strands/cm2 at 8 and 16 weeks, respectively. The
hair density of the RB-SCE group was not significantly in-
creased after 8 weeks but was significantly increased after
16 weeks. In the placebo group, hair density increased from
88.19 s t rand s/cm2 at baseline to 88.00 and 90.89 strands/
cm2 at 8 and 16 weeks, respectively. The hair density of the
placebo group was not significantly increased after 8 a nd 16
weeks (Table 3). The average increase in the number of hairs
was 3.06 and 9.72 strands/cm2 in the RB-SCE group and only
−0.19 and 2.69 strands/cm2 in the placebo group at 8 and
16 weeks, respectively. The increase in hair density did not
significantly differ bet ween the RB-SCE and placebo groups
at 8 weeks but did significantly differ at 16 weeks (Table 3,
p=0.410 and p=0.034).
In female subjects, the average number of hairs in the RB-
SCE group was 81.96 strands/cm2 at baseline. After RB-SCE
treatment, hair density significantly increased to 89.38 and
95.59 strands/cm2 at 8 and 16 weeks, respectively. In the pla-
Table 3. Hair Density at 0, 8 and 16 Weeks after Treatment in Male and Female Subjects
Hair density (strand/cm2)Pairwise comparison p value Changes of hair density (strand/cm2)
0 wk 8 wk 16 wk 0 wk vs. 8 wk 8 wk vs. 16 wk 0 wk vs. 16 wk ΔX1 ΔX2 ΔX3
Male RB-SCE 93.50±11.8 96.56±11.2 103.22±10.1 0.106 0.000 0.000 3.06±5.04 6.67±3.35 9.72±4.62
Placebo 88.19±23.4 88.00±14.8 90.89±17.5 0.949 0.055 0.269 −0.19±10.7 2.89±4.90 2.69±8.37
p0.410 0.059 0.034
Female RB-SCE 81.96±13.6 89.38±8.4 95.59±8.4 0.024 0.005 0.006 7.42±9.82 6.21±6.10 13.63±13.87
Placebo 70.56±11.1 76.67±7.3 80.94±10.0 0.089 0.126 0.039 6.11±9.47 4.28±7.51 10.39±12.66
p0.280 0.148 0.198
RB-SCE=Test tonic with RB-SCE; Placebo=Test tonic without RB-SCE. ΔX1=hair density after 8 wk−hair density at 0 wk; ΔX2=hair density after 16 wk−hair density
after 8 wk; ΔX3=hair diameter after 16 wk−hair diameter at 0 wk.
Vol. 38, No. 12 (2015) 1859
Biol. Pharm. Bull.
cebo group, hair density increased from 70.56 strands/cm2 at
baseline to 76.67 and 80.94 strands/cm2 at 8 and 16 weeks,
respectively. The hair density of the placebo group was not
significantly increased after 8 weeks but was significantly
increased after 16 weeks (Table 3). The average increase in
the number of hairs was 7.42 and 13.63 strands/cm2 in the
RB-SCE group and only 6.11 and 10.39 strands/cm2 in the
placebo group at 8 and 16 weeks, respectively. The increase in
hair den sity did not sign ificantly differ betwee n the RB-SCE
and placebo groups at 8 and 16 weeks (Table 3, p=0.280 and
p=0.198).
Hair Diameter The average hair diameter in male sub-
jects in the R B-SCE group was 57 µm at baseline, increasing
to 66 and 75 µm after 8 and 16 weeks of treatment, respec-
tively. In the RB-SCE group, hair diameter was sign ificantly
increased after 8 and 16 weeks. In the placebo group, hair di-
ameter also increased from 59 µm at baseline to 66 and 69 µm
after 8 and 16 weeks, respectively. In the placebo group, hair
diameter was not significantly increased after 8 weeks but was
significantly increased at 16 weeks (Table 4). The change in
diameter did not differ significantly between the RB-SCE and
placebo groups at 8 weeks (8.6 vs. 6.8, p=0.711). However, the
change in hair diameter from 8 to 16 weeks in the RB-SCE
group was 3.1-fold higher than that in the placebo group (8.8
vs. 2.8, p=0.001), and the change from baseline to 16 weeks in
the RB-SCE group was 1.8-fold higher (17.3 vs. 9.7, p=0.127)
(Ta b le 4).
The average hair diameter in female subjects in the RB-
SCE group was 72 µm at baseline, increasing to 80 and
89 µm after 8 and 16 weeks of treatment, respectively. In the
RB-SCE group, hair diameter was significantly increased
after 8 and 16 weeks. In the placebo group, hair diameter
also increased from 62 µm at baseline to 73 and 74 µm after
8 and 16 weeks, respectively. In the placebo group, hair di-
ameter was not sig nificantly increa sed after 8 weeks but was
significantly increased at 16 weeks (Table 4). The change in
diameter did not differ significantly between the RB-SCE and
placebo groups at 8 weeks (8.6 vs. 11.0 , p=0.586). However,
the change in hair diameter from 8 to 16 weeks in the RB-
SCE group was 21.5-fold higher than that in the placebo group
(8.6 vs. 0.5, p=0.004), and the change from baseline to 16
weeks in the RB-SCE group was 1.4-fold higher (16.6 vs. 11.5,
p=0.389) (Table 4).
Expert Panel Assessment of Global Photographs Global
photographs were reviewed in a blinded manner by expert
panels composed of three dermatologists using a 7-point scale
(Fig. 2). After 16 weeks of treatment, the experts observed im-
proved hair growth in the R B-SCE group (score=0.89±0.601,
p=0.002), while no improvement in hair growth was observed
in male subjects in the placebo group (0.08±0.862, p=0.753).
At 8 weeks of treatment, the average score was 0.67 in the
RB-SCE group and 0.00 in the placebo group, and the dif-
ference was not statistically significant ( p=0.061). However,
at 16 weeks of treatment, the average score in the R B-SCE
group (0.89±0.601) was clearly higher than that in the placebo
group (0.08±0.862) ( p=0.024) (Table 5).
After 16 weeks of treatment, the experts observed im-
proved hair growth in female subjects in both the RB-SCE
group (score=1. 25±0.452, p=0.000) and the placebo group
(1.11±0.601, p=0.013). At 8 weeks of treatment, the average
score was 0.67 in the RB-SCE group and 0.56 in the pla-
cebo group, and the difference was not statistically significant
(p=0.717). At 16 weeks of treatment, the average score in
the RB-SCE group (1.25±0.452) and in the placebo group
Table 4. Hair Diameter at Baseline and 8 and 16 Weeks after Treatment in Male and Female Subjects
Hair diameter (µm) pIncrease of hair diameter (µm)
0 wk 8 wk 16 wk 0 wk vs. 8 wk 8 wk vs. 16 wk 0 wk vs. 16 wk ΔX1 ΔX2 ΔX3
Male RB-SCE 57±18 66±14 75±16 0.013 0.000 0.001 8.6±8.1 8.8±3.6 17.3±10.2
Placebo 59±17 66±16 69±16 0.059 0.012 0.011 6.8±11.8 2.8±3.5 9.7±11.6
p0.711 0.001 0.127
Female RB-SCE 72±15 80±11 89±14 0.027 0.001 0.001 8.0±10.9 8.6±6.8 16.6±13.3
Placebo 62±14 73±10 74±11 0.046 0.672 0.027 11.0±13.9 0.5±3.4 11.5±12.8
p0.586 0.004 0.389
RB-SCE=Test tonic with RB-SCE; Placebo=Test tonic without RB-SCE. ΔX1=hair diameter after 8 wk−hair diameter at 0 wk; ΔX2=hair diameter after 16 wk−hair diam-
eter after 8 wk; ΔX3=hair diameter after 16 wk−hair diameter at 0 wk.
Fig. 2. Representative Photographs Used for Expert Panel Assessment of Global Photographs in a RB-SCE-Treated Male Subject at 0 (a) Week and
2 (b) and (c) 4 Weeks
1860 Vol. 38, No. 12 (2015)
Biol. Pharm. Bull.
(1.11±0.601) and the difference were not statistically signifi-
cant ( p=0.552) (Table 5).
Analysis of Subject Self-questionnaires The bitemporal
recession and hair shedding of male subjects in the RB-SCE
group were significantly higher than those in the placebo
group at 16 weeks based on the chi-square test for uniformity
of groups ( p=0.003, p=0.011, respectively; Table 6). No statis-
tically significant between-group differences in all titles in the
questionnaire analysis were observed in female subjects (Table
7). The overall satisfaction of all added male and female sub-
jects in the R B-SCE treatment group was higher at 16 weeks
than at 8 weeks ( p=0.033, data not shown). In addition, the
overall satisfaction of the RB-SCE group was significantly
higher than that of the placebo group at 16 weeks (p=0.005).
Skin Tolerance and Safety Evaluations On the basis of
both subject reports and clinical observation by the investiga-
tors, no adverse reactions such as itching, prickling, burning,
stinging, stiffness, tightness, burning of the eyes, weeping, er-
ythema, edema, scaling, papule, or any other RB-SCE-related
reaction were noted in all 43 subjects both at 8 and 16 weeks
of treatment utilization.
DISCUSSION
Alopecia is a common form of hair loss in both men and
women, affecting approximately 0.2 to 2% of the world’s pop-
ulation.1–3,21) Although topical minoxidil and oral finasteride
are available for the treatment of alopecia, these medicines
have sometimes been reported to have adverse effects.7–10) In
our previous st udy, RB-SCE showed hair growth-promoting
potential similar to that of 3% minoxidil, as evidenced by
findings of hair follicles induced in the anagen stage, and a
significant increase in the number of hair follicles in C57BL/6
mice.11) However, there have been no clinical studies of RB-
SCE. We therefore performed this clinical study to confirm
the ability of RB-SCE to improve hair loss in 43 patients
treated with or without RB-SCE.
Clinical research on topical alopecia treatments included
clinical studies of procyanidin B-222) and minoxidil.4,2 3) After
48 weeks of male pattern hair loss treatment, the parameter
known as non-vellus hair count (mean change from base-
line) in the 5% minoxidil, 2% minoxidil, and placebo groups
measured 18.6, 12.7, and 3.9 strands/cm2, respectively.4) The
change from baseline in the non-vellus hair count was sig-
nificantly superior with 5% topical minoxidil compared to
Table 5. Expert Panel Assessment of Global Photographs (7-Point Scale) in Male and Female Subjects
Expert panel assessment (score value)
p*
0 wk 8 wk 16 wk
Male RB-SCE 0 0.67±0.707 0.89±0.601 0.002
Placebo 0 0.00±0.817 0.08±0.862 0.753
p0.061 0.024
Female RB-SCE 0 0.67±0.779 1.25±0.452 0.000
Placebo 0 0.56±0.527 1.11±0.601 0.013
p0.717 0.552
RB-SCE=Test tonic with RB-SCE; Placebo=Test tonic without RB-SCE. * p=expert panel assessment value after 16 wk−expert panel assessment value at 0 wk.
Table 6. Questionnaire Analysis Results in Male Subjects
RB-SCE (%) Placebo (%) p Value
8 wk 16 wk 8 wk 16 wk 8 wk/16 wk RB-SCE/Placebo
Size of vertex spot Improved 44.4 44.4 15.4 23.1
No change 55.6 55.6 84.6 76.9 1 0.290
Worse 0.0 0.0 0.0 0.0
Hair loss on top of scalp Improved 44.4 66.7 23.1 46.2
No change 55.6 33.3 76.9 53.8 0.343 0.342
Worse 0.0 0.0 0.0 0.0
Bitemporal recession Improved 44.4 77.8 15.4 15.4
No change 55.6 22.2 84.6 84.6 0.147 1
Worse 0.0 0.0 0.0 0.0
Hair shedding Improved 66.7 77.8 23.1 23.1
No change 33.3 22.2 69.2 76.9 0.391 0.011
Worse 0.0 0.0 7.7 0.0
Hair quality Improved 33.3 55.6 15.4 30.8
No change 66.7 44.4 76.9 69.2 0.271 0.245
Worse 0.0 0.0 7.7 0.0
Overall satisfaction Improved 55.6 77.8 38.5 38.5
No change 44.4 22.2 61.5 61.5 0.371 0.069
Worse 0.0 0.0 0.0 0.0
The chi-square test was used to evaluate statistically significant between-group differences in the questionnaire analysis. RB-SCE=Test tonic with RB-SCE; Placebo=Test
tonic without RB-SCE.
Vol. 38, No. 12 (2015) 1861
Biol. Pharm. Bull.
2% topical minoxidil and placebo. After 48 weeks of female
pattern hair loss treatment, the non-vellus hair count (mean
change from baseline) in the 5% minoxidil, 2% minoxidil,
and placebo groups measured 24.5, 20.7, and 9.4 strands/cm2,
respectively.23) As in the male patients, 5% topical minoxidil
was significantly superior to 2% topical minoxidil and placebo
in the female patients. To investigate the effects of topical pro-
cyanidin B-2 (1%), which is purified from apples, on the scalp
and hair, a placebo controlled clinical trial was performed
in male pattern baldness.22) After 16 weeks, a significantly
greater increase in the total number of hairs in the designated
scalp area (1.0 cm2) was observed in subjects receiving procy-
anidin B-2 than in those receiving placebo (procyanidin B-2,
14.68 s t r a nds/cm2; placebo, −10.16 s t r a nds/cm2). In our study,
the non-vellus hair counts (mean change from week 0) were
9.72 and 2.69 strands/cm2 in male subjects in the RB-SCE and
placebo groups, respectively, showing a statistically significant
difference between the groups ( p=0.034; Table 3). In female
subjects, the hair counts were 13.63 and 10.39 strands/cm2 in
the RB-SCE and placebo groups, respectively. However, there
was no statistically signi ficant difference bet ween the groups
(p=0.198; Table 3). Although study duration, race, gender,
drug administration protocol, and other factors differ between
our study and the prior studies, the increase in hair count in
the 0.5% RB-SCE group after 16 weeks in our study (9.72
strands/cm 2 in male subjects) is similar to or slightly lower
than that reported for 2% minoxidil after 48 weeks of treat-
ment in men (12.7 strands/cm2)4) or for 1% procyanidin B-2
after 16 weeks of treatment in men (14.68 strands/cm2).22)
In terms of hair diameter, it was reported that 78.9% of
subjects treated with 1% procyanidin B-2 showed an increased
mean hair diameter, whereas only 30.0% in the placebo group
showed any increase. The change in hair diameter after 16
weeks was 2.68 and −1.08 µm, respectively, in the 1% procy-
anidin B-2 and placebo groups. The increased ratio (8.04%)
of hairs measuring more than 40 µm in diameter after 16
weeks of procyanidin B-2 treatment was significantly higher
than that in controls (−4.32%). In our study, the hair diameter
(mean change from week 8) after 16 weeks was 8.8 µm (from
66 to 75 µm) and 2.8 µm (from 66 to 69 µm) in male subjects
with RB-SCE and placebo, respectively, showing a significant
increase (p=0.001; Table 4). The hair diameter (mean change
from week 8) after 16 weeks was 8.6 µm (from 80 to 89 µm)
and 0.5 µm (from 73 to 74 µm) in female subjects with RB-
SCE and placebo, resp ectively, showing a signi ficant increase
(p=0.004; Table 4). The increase in hair diameter in the RB-
SCE group after 16 weeks (17.3 and 16.6 µm in male and fe-
male subjects, respectively) was higher than that reported for
1% procyanidin B-2 after 16 weeks (8.04 µm).
Regarding the expert panel assessment of global photo-
graphs in male subjects, at 16 weeks of treatment, the average
score in the RB-SCE group (0.89±0.601) was clearly higher
than that in the placebo group (0.08±0.862), and the dif-
ference was statistically significant ( p=0.024) (Table 5). In
female subjects at 16 weeks of treatment, the average score
in the RB-SCE group (1.25±0.452) and in the placebo group
(1.11±0.601) and the difference were not statistically signifi-
cant ( p=0.552) (Table 5).
Self-questionnaires have been reported useful for the evalu-
ation of new potential hair growth-promoting candidates or
medicines.4 ,17,2 3) In the study of Olsen et al. in male patients,4)
patient questionnaire hair growth composite scores at week 48
(efficacy-evaluable population) for 5% minoxidil , 2% m inoxi-
dil, and placebo were 60.4, 56.8, and 50.7, respectively. In the
study of Lucky et al. in female patients,23) patient question-
naire hai r growth compo site scores at week 48 (efficacy-eval-
uable population) for 5% minoxidil, 2% minoxidil, and pla-
cebo were 64.5, 60.5, and 56.4, respectively. In our study, the
population with improvement in bitemporal recession and hair
shedding i n the RB-SCE group of male subjects was signifi-
Table 7. Questionnaire Analysis Results in Female Subjects
RB-SCE (%) Placebo (%) p Value
8 wk 16 wk 8 wk 16 wk 8 wk/16 wk RB-SCE/Placebo
Size of vertex spot Improved 33.3 33.3 55.6 44.4
No change 66.7 66.7 44.4 55.6 1 0.604
Worse 0.0 0.0 0.0 0.0
Hair loss on top of scalp Improved 41.7 33.3 44.4 66.7
No change 58.3 66.7 55.6 33.3 0.673 0.130
Worse 0.0 0.0 0.0 0.0
Bitemporal recession Improved 25.0 16.7 55.6 44.4
No change 75.0 83.3 44.4 55.6 0.615 0.163
Worse 0.0 0.0 0.0 0.0
Hair shedding Improved 41.7 58.3 66.7 66.7
No change 50.0 33.3 33.3 22.2 0.693 —
Worse 8.3 8.3 0.0 11.1
Hair quality Improved 25.0 58.3 66.7 55.6
No change 75.0 41.7 33.3 44.4 0.098 0.899
Worse 0.0 0.0 0.0 0.0
Overall satisfaction Improved 50.0 83.3 55.6 66.7
No change 50.0 16.7 44.4 33.3 0.083 0.375
Worse 0.0 0.0 0.0 0.0
The chi-square test was used to evaluate statistically significant between-group differences in the questionnaire analysis. RB-SCE=Test tonic with RB-SCE; Placebo=Test
tonic without RB-SCE. —: not determined.
1862 Vol. 38, No. 12 (2015)
Biol. Pharm. Bull.
cantly larger than that in the placebo group at 16 weeks based
on the chi-square test for uniformity of groups ( p=0.003,
p=0.011, respectively; Table 6), but it otherwise was not in
female subjects (Table 7). Although comparison with previous
results is difficult because of the use of different questionnaire
items,4,2 3) we confirmed that 2 items in questionnaire analy-
sis results in the RB -SCE group were significantly i ncreased
compared to those in the placebo group after 16 weeks, espe-
cially in male subjects.
Overall, the significant differences in hair density, hair
diameter, and expert panel assessment of global photographs
were shown in male subjects after 16 weeks of treatment with
RB-SCE. In addition, the significant differences bet ween the
RB-SCE and placebo groups after 16 weeks showed in hair
density, expert panel assessment of global photographs, and
questionnaire analysis in male subjects. In female subjects
after 16 weeks of treatment with RB-SCE, the significant
differences in hair density, hair diameter, and expert panel
assessment of global photog raphs were shown but not signi fi-
cantly different from those of the placebo group.
In a previous study,11) to examine the hair growth-promot-
ing activity of RB-SCE, we selected linoleic acid (LA), ory-
zanol (OZ), policosanol (PS), and tocotrienol (TT) as the main
components of RB-SCE. In particular, the unsaturated fatty
acids such as γ-LA, LA, and oleic acid as well as RB-SCE
have been shown to have anti-hair loss activity by inhibiting
the 5-α-reductase enzyme in androgen-responsive organs.12,24)
In this st udy, RB-SCE h ad a significant hair g rowth-promot-
ing effect in male subjects but not in female subjects, meaning
that RB-SCE may have an effect in androgen-dependent hair
loss. Recently, female patients with alopecia were reported to
be divided by androgen-dependent female pattern hair loss
(FPHL) and androgen-independent FPHL according to the
serum androgen levels of the female patients.25, 26) The analy-
ses of blood chemistr y tests including hormonal levels for
female patients are needed in further study.
Previous researchers have confirmed the toxicological
safety of RB-SCE in RAW264.7 cells. Additional safety evalu-
ations included single oral dose toxicity in rats,13) an acute
dermal and ocular irritation test,14) a single dose and 4-week
repeated dose dermal toxicity study,15) and a genotoxicity
assessment.16) As urinalysis, hematological tests, clinical bio-
chemistr y tests, necropsies, and histopathological examina-
tions were performed in male and female rats after 4-week
repeated-dose dermal administration of RB-SCE, no liver or
kidney damage was observed.15) In addition, in this study, no
specific side effects such as irritant contact dermatitis, allergic
contact der matitis, itching, pricking, burning, erythema, ooz-
ing, vesicles, skin rashes, and so on were observed in patients
receiving RB-SCE. Therefore, RB-SCE is thought to be safe
for topical application in humans.
However, the limitations of this study are the single-center
study design, small number of subjects, and lack of blood
chemistr y research including hormonal levels, extensive in-
terviews with patients about hormonal effects detected by
sexual dysfunction (e.g., loss of libido), and confirmation of
the absence of liver or kidney damage. Further studies, includ-
ing multicenter studies with longer treatment periods, larger
sample-size patient populations, blood chemistry research, and
extensive patient interviews, are needed to confirm the present
results.
RB-SCE is a mixture of fatty acids and oily materials and
not a purified compound. Nevertheless, the increase in hair
density and diameter as well as patient satisfaction with 0.5%
RB-SCE after 16 weeks were similar to or slightly lower than
those reported for 2% minoxidil after 48 weeks or 1% pro-
cyanidin B-2 after 16 weeks. Therefore, RB-SCE appears to
have considerable hair growth-promoting potential.
CONCLUSION
In conclusion, these results suggest that treatment with RB-
SCE can improve hair regrowth in human androgen-dependent
alopecia without side effects at a moderate dose. Therefore,
RB-SCE is a potentially promising source of functional cos-
metics and pharmaceuticals developed to treat male pattern
and androgen-dependent female pattern hair loss.
Acknowledgment This work was supported by a Grant
(No. 311014-03) from the Ministr y for Food, Agriculture, For-
estry and Fisheries, Republic of Korea.
Conflict of Interest The authors declare no conflict of
interest.
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