Effects of cosmetics therapy using isoflavone and pine bark extract on the skin and QOL : A double-blind placebo-controlled trial

Anti-Aging Medical Research. Vol. 1, No. 1:48-58, September 2004
Effects of cosmetics therapy using isoflavone and pine bark
extract on the skin and QOL: A double-blind placebo-controlled
trial
Yoshikazu Yonei 1, Yoshio Mizuno 1, and Eri Katagiri 2
1Department of Internal Medicine, Nippon Kokan Hospital
2Eri Clinic, Japan
Abstract
Objective: The efficacy and safety of anti-aging cosmetics that use isoflavone and pine bark extract (Flavangenol(r)) as the
primary ingredients were studied in a double-blind, non-crossover manner on 21 female volunteers in the control group
(age: 46.6
6.7 years) and 20 female volunteers in the cosmetics-treated group (age: 45.8
4.9 years).
Methods: The subjects used a sample product that was indistinguishable between the groups, and conducted a self facial
skincare twice a day, in the morning and at night, for 4 weeks. Physical examinations were conducted and a questionnaire
was administered before and after this skincare regimen.
Results: The cosmetics-treated group showed significant improvements in parameters related to physical symptoms such
as "tendency to gain weight" and "excessive sensitivity to cold," and mental symptoms such as "irritability," "loss of
motivation," "reluctance to talk with others," "pessimism," and "lapse of memory." In the control group, diastolic blood
pressure dropped significantly (121.3/77.0 mmHg to 121.4/73.2 mmHg), while both diastolic and systolic blood pressures
lowered significantly (127.7/79.0 mmHg to 119.6/73.9 mmHg) in the cosmetics-treated group. In a study of oxidative
stress markers, the cosmetics-treated group showed significant improvements in serum lipid peroxide and 8-isoprostane
production rate. In a study on skin measurements, the amount of melanin on the forehead had markedly increased in the
control group, but this was suppressed in the cosmetics-treated group. No adverse events were seen throughout the study
period.
Conclusion: These findings confirm the efficacy and safety of this product.
Key words: Anti-aging medicine, cosmetics, stress, pine bark extract, estrogen, quality of life
Introduction
options including diet, exercise, and other lifestyle
therapies, drug therapies including nutritional supplement
The objective of anti-aging medicine is to help people live
therapies, and hormone replacement therapies. Lifestyles
a long, healthy life. This is a medical treatment aimed not
have been shown to have a major influence on health and
simply at prolonging life but at preventing physical and
longevity.1 However, among the different lifestyle
mental deterioration caused by aging, and also maintaining
activities, very few studies have thus far been conducted on
a high quality of life (QOL).1 Anti-aging drugs are
the use of cosmetic creams, especially on whether such use
classified as preventive medicine. After undergoing a
would favorably influence QOL, health, and longevity,
medical evaluation or screening to assess the degree of
besides direct effects on the skin. Estrogen, a female
aging, one may choose from a range of anti-aging treatment
hormone whose secretion declines with age, is closely
Received: 10 May 2004
Accepted: 13 July 2004
Reprint requests to: Yoshikazu Yonei, Department of Internal Medicine, Nippon Kokan Hospital, Japan
5-21-26 Ryokuen,Izumi-ku, Yokohama, Kanagawa 245-0002, Japan
Email: yyonei-gi@umin.ac.jp
48

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Effects of cosmetics therapy using isoflavone and pine bark extract on the skin and QOL: A double-blind placebo-controlled trial
related not only to health and longevity but also to the
methylpolysiloxane, polyglycerol stearate-10, xanthan gum,
aging of the skin.2-4 Recent reports that isoflavone contained
carbomer, octyl dodecanol, sodium polyacrylate, sodium
in soybean extract possesses an estrogen-like activity are
hydroxide, titanium oxide, silica, alumina, mica, ethanol,
drawing strong interest from the perspective of estrogen
phenoxyethanol, methylparaben, and propylparaben; active
secretion and the aging of the female skin.5
test cream-the same ingredients as the control cream, plus
In our previous study, via an open, non-blinded test, we
soybean extract (isoflavone), extract of the bark of a pine
investigated the safety and efficacy of cosmetics (skincare
tree found on the French sea coast (Flavangenol),
products) that contained soybean extract (eterrite,
tocopherol, ascorbyl phosphate Mg, Saxifraga stolonifera
manufactured by Charle Corporation, Kobe, Japan), as well
extract, peony extract, kudzu root extract, chlorella extract,
as their possible relation to QOL and longevity.6 We found
hydrolase extract, grapefruit extract, and eglantine extract
the cosmetic product improved several subjective
(these are all considered to be active ingredients).
symptoms related to QOL, reduced the level of cortisol, a
Clinical test parameters were examined twice, once before
stress hormone, and occasionally elevated the estradiol
the skincare creams were used and again after completing
level. However, we were unable to identify if these effects
the period of using the creams. Subjective symptoms were
were attributable to the application of the makeup, the
divided into physical and mental symptoms. They were
activity of common components of the makeup, or to the
investigated by using an Anti-Aging QOL Common
characteristic ingredients contained in eterrite, principally
Questionnaire (AAQOL).7 The numerical scale is a
isoflavone and pine bark extract (Flavangenol).
subjective one for each physical symptom queried; however,
In this study, via a controlled, double-blind, non-
the degree is related. For example, for the assignment of
crossover test, we investigated the efficacy and safety of an
value 3, the subject might begin to consider a visit to a
anti-aging skin cream (eterrite Brightening Force; hereafter
doctor, or self medicate with over the counter remedies.
referred to as "the Product"). Double-blind means that
Values of 1 and 2 might be perceived as within a normal
neither the subjects nor the clinicians conducting the study
range over the population. In addition to the general blood
are aware of the particular treatment each subject enrolled
and biochemical tests, the values of estradiol, progesterone,
in the study is receiving.
insulin-like growth factor-I (IGF-I), dehydroepiandrosterone
sulfate (DHEA-s), and cortisol were measured.
Lipid peroxide, 8-hydroxy-2'deoxyguanosine (8-OHdG),
Materials and Methods
and 8-isoprostane F-2 (isoprostane) were used as
indicators of the degree of oxidative stress.8-10 These
The subjects in our study were women aged 40 to 59 years,
indicators were measured at the Japan Institute for Control
who had used other companies' cosmetics products
of Aging. The level of lipid peroxide in the blood was
(skincare) 3 or more times a week, and who gave their
measured. In addition, early-morning urine (initial urine for
informed consent to participate in this study. Patients with
the day) was sampled and 8-OHdG, isoprostane, and
mental disorders, severe hepatic, renal, cardiac and/or
creatinine levels were measured. The rate of 8-OHdG
hematological diseases, and personnel related to the
production, the rate of isoprostane production, and the
cosmetics business were excluded from the study. This was
creatinine correction amount were calculated based on the
in accordance with ethical guidelines set forth by the Japan
urine amount and urine-collection period (the time from the
Anti-Aging foundation.
final urination the previous evening to the time of the first
Forty-one female subjects were randomly divided into 2
urination in the morning).
groups: a control group A (21 subjects; 46.6
6.7 years)
Facial skin was evaluated at 2 sites-the forehead and the
and an active test group B (20 subjects; 45.8
4.9 years).
left cheek-to investigate the blood flow and the volume of
The subjects used samples that were indistinguishable in
moisture, sebum, melanin, and erythema. We used changes
appearance between the 2 groups to carry out a skincare
in the electrical capacitance of the skin to estimate the
program that included a 15- to 30-minute self facial skincare
amount of water in the keratin layer, and adapted a
twice a day, in the morning and at night, for 4 weeks.
corneometer (CM825SPANC; Courage and Khazaka
The ingredients of each sample type are as follows:
Electric GmbH, Germany) for this purpose.11,12 To measure
control cream-water, glycerin, BG (butylene glycol), DPG
the volume of sebum on the skin's surface, we used the
(propylene glycol), sorbitol, licorice extract, PEG-40
changes in the penetration of exclusive film, and adapted a
hydrogenated castor oil (Polyethylene glycol 2000),
sebumeter (SM810SPANC; Courage and Khazaka Electric
49

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Y. Yonei et al.
GmbH, Germany) for this purpose.13,14 To measure the skin's
Results
melanin volume and the degree of erythema, we used a
mexameter (MX18; Courage and Khazaka Electric GmbH,
Tables 1 to 4 show the results of the investigation on
Germany). With this device, light of a specific wavelength was
subjective symptoms, based on the QOL Common
irradiated onto the skin, the reflected light was detected and
Questionnaire. The physical symptoms that improved
measured by using a light diode, and the melanin and erythema
significantly among the group A subjects were "blurry
indices were calculated.11,15,16 SKInfo (Integral Co., Ltd.),
eyes," for which the score changed significantly from 2.4
which integrates various measurement devices, was used to
0.9 to 2.0
0.8 (
= 0.001), "skin problems," for
record these parameters. The skin's blood flow was measured
which the score changed significantly from 2.6
1.0 to
5 times using PeriScan PIMII (PERIMED, Sweden), a laser
2.2
0.8 (
= 0.014), and "constipation," for which the
Doppler blood flow measurement device, and the mean
score changed significantly from 3.0
1.3 to 2.6
1.2
values were used as the measurement.17,18
(
= 0.008) (Table 1). Among scores for the group B
The test was to be promptly discontinued if severe
subjects, "tendency to gain weight" changed significantly
adverse reactions or events occurred, and appropriate
from 3.4
1.1 to 3.1
1.2 (
= 0.015), "skin problems"
treatment, based on the judgment of the physician in charge
changed significantly from 3.2
0.8 to 2.4
0.8 (
=
of the test, would be provided.
0.004), and "excessive sensitivity to cold" changed
The t-test, paired t-test, or rank sum test, where appropriate,
significantly from 2.7
1.3 to 2.3
1.0 ( .029) (Table
were performed to statistically analyze the survey results. The
3). The parameters that worsened were scores for
results were shown in terms of mean
standard deviation,
"headache" in group A, which changed significantly from
and a probability value of less than .05 was considered to be
2.4
1.0 to 2.7
1.1 (
= 0.048), and "tinnitus" in
significant. The Wilcoxon Matched-Pairs Signed-Ranks Test
group B, which changed significantly from 1.7
1.1 to 1.9
was not applied in the statistical analysis.
1.1 (
= 0.048).
Table 1. Physical symptoms-group A
Variable
Before
4 weeks later
– level
Tired eyes
2.9
1.1
2.5
1.0
0.083
Blurry eyes
2.4
0.9
2.0
0.8 0.001
Eye pain
2.0
0.7
1.8
0.7 0.164
Stiff shoulders
3.3
1.3
3.4
1.1 0.288
Muscular pain/stiffness
2.7
1.0
2.9
1.0
0.226
Palpitations
1.9
0.9
2.1
0.8 0.102
Dyspnea
1.8
0.8
1.8
0.7 0.374
Tendency to gain weight
3.0
0.9
3.0
1.0
0.288
Weight loss; thin
1.3
0.7
1.4
0.6 0.214
Lethargy
2.6
1.0
2.5
1.1
0.417
No feeling of good health
2.5
1.0
2.3
1.0
0.206
Thirst
1.9
1.0
1.9
0.8
1.00
Skin problems
2.6
1.0
2.2
0.8
0.014
Anorexia 1.8
0.8
1.9
0.7
0.358
Heavy stomach
2.0
0.7
2.0
0.8
1.00
Gastric pain
2.0
0.7
1.9
0.8 0.165
Liable to catch a cold
2.1
0.8
2.1
0.8
0.288
Coughing and sputum
2.0
0.9
2.1
1.2
0.226
Diarrhea
2.0
0.9
1.9
0.8 0.189
Constipation
3.0
1.3
2.6
1.2 0.008
Headache
2.4
1.0
2.7
1.1
0.048
Dizziness
2.0
1.0
2.2
0.9
0.068
Tinnitus
1.8
1.0
2.0
0.9
0.129
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Effects of cosmetics therapy using isoflavone and pine bark extract on the skin and QOL: A double-blind placebo-controlled trial
Lower back pain
2.4
0.9
2.5
0.9
0.270
Joint pain
2.2
1.2
2.5
0.9 0.134
Swelling
2.0
0.8
2.0
0.7 0.374
Easy to perspire
2.3
1.0
2.4
0.7
0.189
Polakisuria
1.8
0.7
2.0
0.7
0.068
Hot flash
2.0
0.9
2.1
0.8 0.189
Excessive sensitivity to cold
2.7
1.2
2.8
1.2
0.314
Table values are mean
SD score from Anti-Aging QOL Common Questionnaire,7 range of values, 1 to 5; 1 =
none, 2 = almost none, 3 = a little, 4 = moderate, 5 = severe. n = 21, paired t-test.
Table 2. Mental symptoms-group A
Variable
Before
4 weeks later
– level
Irritability
2.8
0.9
2.7
0.8 0.247
Easily angered
2.6
0.9
2.5
0.8 0.093
Loss of motivation
2.4
0.9
2.4
0.4 0.374
No feeling of happiness
2.3
1.0
2.3
0.8
1.00
Nothing to look forward to in life
2.1
1.1
2.0
0.7
0.324
Daily life is not enjoyable
2.3
0.9
2.2
0.7
0.270
Loss of confidence
2.1
0.8
2.2
0.8
0.252
Reluctance to talk with others
2.0
0.9
2.0
0.7
0.417
Depressed
2.2
0.8
2.0
0.7
0.240
A sense of uselessness
1.9
0.7
1.9
0.6 1.00
Shallow sleep
2.5
0.9
2.8
1.0 0.081
Difficulty falling asleep
2.0
1.0
2.1
0.8
0.413
Pessimism
2.8
0.9
2.7
0.9
0.247
Lapse of memory
2.9
1.2
3.0
1.0
0.386
Inability to concentrate
2.1
1.0
2.2
0.8
0.165
Inability to solve problems
2.1
0.8
2.1
0.6 1.00
Inability to make judgments readily
2.0
0.7
2.1
0.6
0.041
Inability to sleep because of worries
2.2
1.0
2.3
0.8
0.374
A sense of tension
2.2
1.0
2.2
0.8
1.00
Feeling of anxiety for no special reason
2.2
1.2
2.1
0.9
0.362
Feeling of vague fear
2.0
1.1
1.7
0.7
0.142
Table values are mean
SD score from Anti-Aging QOL Common Questionnaire,7 range of values, 1 to 5; 1 =
none, 2 = almost none, 3 = a little, 4 = moderate, 5 = severe. n = 21, paired t-test.
Table 3. Physical symptoms-group B
Symptom
Before
4 weeks later
– level
Tired eyes
2.9
0.7
2.8
1.1
0.395
Blurry eyes
2.3
0.9
2.6
0.9
0.109
Eye pain
1.8
0.8
2.1
1.1 0.134
Stiff shoulders
3.3
1.2
3.3
1.1 1.00
Muscular pain/stiffness
2.5
0.9
2.8
1.1
0.165
Palpitations
1.8
0.6
1.9
0.7
0.270
Dyspnea
1.8
0.6
1.8
0.6
1.00
Tendency to gain weight
3.4
1.1
3.1
1.2
0.015
Weight loss; thin
1.3
0.6
1.6
0.7
0.011
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Y. Yonei et al.
Lethargy
2.4
0.9
2.4
0.9
0.417
No feeling of good health
2.2
0.8
2.1
0.8
0.303
Thirst
2.0
0.9
1.9
1.0
0.214
Skin problems
3.2
0.8
2.4
0.8
0.004
Anorexia 1.6
0.8
1.7
0.6
0.374
Heavy stomach
1.8
0.7
2.0
0.9
0.129
Gastric pain
1.8
0.7
2.0
0.9
0.068
Liable to catch a cold
2.0
0.6
1.9
0.8
0.134
Coughing and sputum
1.8
0.7
1.8
0.9
0.386
Diarrhea
1.9
0.7
2.0
0.8 0.214
Constipation
2.7
1.1
2.8
1.0
0.247
Headache
2.2
0.7
2.2
0.7
1.00
Dizziness
2.2
1.1
2.1
1.1
0.358
Tinnitus
1.7
1.1
1.9
1.1
0.048
Lower back pain
2.9
1.3
2.7
1.1
0.224
Joint pain
2.0
1.3
2.0
0.9 1.00
Swelling
2.3
1.2
2.2
1.0
0.363
Easy to perspire
2.9
1.0
2.7
1.2
0.107
Polakisuria
2.1
0.7
2.2
0.7
0.214
Hot flash
2.2
1.3
2.3
1.1
0.226
Excessive sensitivity to cold
2.7
1.3
2.3
1.0
0.029
Table values are mean
SD score from Anti-Aging QOL Common Questionnaire,7 range of values, 1 to 5; 1 =
none, 2 = almost none, 3 = a little, 4 = moderate, 5 = severe. n = 20, paired t-test.
Regarding the mental symptoms in group A, scores for
(
= 0.018), "pessimism" score changed significantly from
"inability to make judgments easily" changed significantly
2.4
0.7 to 2.0
0.8 (
= 0.008), and "lapse of memory"
for the worse, from 2.0
0.7 to 2.1
0.6 (
= 0.041)
score changed significantly from 3.0
0.7 to 2.7
0.7
(Table 2). In group B, "irritability" score changed
(
= 0.028), all in the direction of improvement (Table 4).
significantly from 2.4
0.8 to 2.0
0.8 (
= 0.004), "loss
In other words, although group A showed no improvement
of motivation" score changed significantly from 2.6
1.0
in any of the scores for mental symptoms, group B showed
to 2.0
0.7 (
= 0.012), "reluctance to talk with others"
significant improvements in 5 parameters.
score changed significantly from 2.3
1.1 to 1.7
0.7
Table 4. Mental symptoms-group B
Symptom
Before
4 weeks later
– level
Irritability
2.4
0.8
2.0
0.8 0.004
Easily angered
2.3
0.7
2.2
0.9 0.190
Loss of motivation
2.6
1.0
2.0
0.7
0.012
No feeling of happiness
2.3
1.1
2.0
0.9
0.074
Nothing to look forward to in life
2.3
1.2
1.9
0.9
0.084
Daily life is not enjoyable
2.3
1.1
1.9
0.9
0.080
Loss of confidence
2.3
1.0
2.1
0.8 0.117
Reluctance to talk with others
2.3
1.1
1.7
0.7 0.018
Depressed
2.1
0.9
2.0
0.8
0.134
A sense of uselessness
2.0
0.8
1.9
0.6 0.165
Shallow sleep
2.3
1.1
2.2
1.0 0.270
Difficulty falling asleep
2.2
1.1
2.0
1.0
0.093
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Effects of cosmetics therapy using isoflavone and pine bark extract on the skin and QOL: A double-blind placebo-controlled trial
Pessimism 2.4
0.7
2.0
0.8
0.008
Lapse of memory
3.0
0.7
2.7
0.7
0.028
Inability to concentrate
2.6
0.6
2.4
0.7
0.107
Inability to solve problems
2.2
0.8
2.3
0.8
0.303
Inability to make judgments readily
2.3
0.7
2.2
0.7
0.165
Inability to sleep because of worries 2.0
0.6
2.0
0.7
1.00
A sense of tension
2.5
0.5
2.3
0.8
0.165
Feeling of anxiety for no reason
2.0
0.6
1.9
0.6
0.214
Feeling of vague fear
1.6
0.7
1.7
0.5
0.374
Table values are mean
SD score from Anti-Aging QOL Common Questionnaire,7 range of values, 1 to 5; 1 =
none, 2 = almost none, 3 = a little, 4 = moderate, 5 = severe. n = 20, paired t-test.
Results of the physical and blood examinations are
seen in terms of changes in the erythrocyte count, hemoglobin
shown in Tables 5 and 6. Diastolic blood pressure changed
amount, platelet count, gamma-glutamyltranspeptidase ( -
significantly from 121.3/77.0 mmHg to 121.4/73.2 mmHg
GTP), uric acid, and triglycerides in group A, as well as in
in group A (systolic/diastolic;
= 0.012), while both
the platelet count and urea nitrogen in group B, they were
diastolic and systolic blood pressures changed significantly
all within the range of physiological variations, and thus,
from 127.7/79.0 mmHg to 119.6/73.9 mmHg in group B
were assessed as having no clinical significance.
( = 0.002, = 0.004). While significant differences were
Table 5. Clinical test results-group A
Variable
Unit
Before
4 weeks later
– level
Weight
kg
59.4
11.3
59.6
11.0
0.282
Body mass index
23.8
3.9
23.9
3.7
0.292
Percentage body fat
%
29.5
5.7
29.9
6.4
0.303
Systolic BP
mmHg
121.3
14.9
121.4
17.9
0.474
Diastolic BP
mmHg
77.0
13.4
73.2
11.3
0.012
Leukocyte count
/mm3
5805
1912
5565
1575
0.178
Erythrocyte count
104x/mm3
431.5
25.5
421.0
25.5
0.004
Hemoglobin
g/dl
12.4
1.6
12.1
1.5
0.009
Platelet count
104x/mm3
25.1
4.4
26.6
6.3
0.027
GOT
U/l
20.1
3.5
19.8
4.0
0.278
GPT
U/l
18.1
5.6
16.8
7.7
0.082
ALP
U/l
200.5
51.2
197.8
50.7
0.330
-GTP
U/l
28.4
26.2
23.4
15.6
0.032
Creatinine
mg/dl
0.602
0.085
0.602
0.106
1.00
Urea nitrogen
mg/dl
13.7
3.2
13.2
2.5
0.099
Uric acid
mg/dl
4.63
1.05
4.32
0.93
0.018
Total cholesterol
mg/dl
205.5
34.7
201.2
34.7
0.106
Triglyceride
mg/dl
85.0
44.3
73.4
37.1
0.020
Sodium
mEq/l
141.1
1.7
141.0
1.9
0.362
Potassium
mEq/l
4.1
0.4
4.2
0.3
0.126
Chlorine
mEq/l
103.3
1.5
103.3
1.5
0.118
Lipid peroxide
nmol/ml
0.37
0.09
0.35
0.08
0.165
Fasting blood sugar
mg/dl
94.7
10.8
93.5
11.6
0.186
IGF-I
ng/ml
224.0
63.0
185.1
63.7
0.001
Cortisol
g/dl
9.3
6.2
9.5
4.9
0.432
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Y. Yonei et al.
DHEA-s
ng/ml
1326.9
777.1
1273.4
629.3
0.234
Estradiol
pg/ml
80.6
59.5
63.0
67.4
0.196
Progesterone
ng/ml
3.3
6.7
2.8
4.7
0.221
ALP, alkaline phosphatase; BP, blood pressure; IGF-I, insulin-like growth factor-I; GOT, glutamic-oxaloacetic
transaminase; GPT, glutamic-pyruvic transaminase; -GTP, gamma-glutamyltranspeptidase; DHEA-s,
dehydroepiandrosterone sulfate. n = 21, mean
SD, paired t-test.
Table 6. Clinical test results-group B
Variable
Unit
Before
4 weeks later
– level
Weight
kg
56.6
6.8
56.3
6.4
0.124
Body mass index
22.9
2.9
22.7
2.7
0.118
Percentage body fat
%
28.0
5.2
27.8
4.0
0.389
Systolic BP
mmHg
127.7
18.1
119.6
18.9
0.002
Diastolic BP
mmHg
79.0
11.4
73.9
10.5
0.004
Leukocyte count
/mm3
5064
1186
5300
1452
0.236
Erythrocyte count
104x/mm3
430.0
30.3
428.6
29.2
0.379
Hemoglobin
g/dl
12.6
1.5
12.5
1.6
0.283
Platelet count
104x/mm3
24.8
6.4
26.0
7.4
0.049
GOT
U/l
20.0
4.0
19.7
4.8
0.332
GPT
U/l
16.2
6.1
15.8
5.6
0.315
ALP
U/l
182.5
61.7
179.5
56.0
0.253
-GTP
U/l
17.6
12.2
17.1
8.1
0.307
Creatinine
mg/dl
0.589
0.065
0.594
0.058
0.324
Urea nitrogen
mg/dl
11.8
2.2
12.7
2.5
0.018
Uric acid
mg/dl
4.60
0.78
4.43
0.89
0.092
Total cholesterol
mg/dl
197.7
27.4
200.2
29.8
0.138
Triglyceride
mg/dl
76.7
46.8
66.4
30.3
0.093
Sodium
mEq/l
141.3
2.0
141.5
1.9
0.180
Potassium
mEq/l
4.0
0.3
4.1
0.3
0.080
Chlorine
mEq/l
103.4
2.1
103.4
2.0
0.449
Lipid peroxide
nmol/ml
0.44
0.11
0.39
0.08
0.006
Fasting blood sugar
mg/dl
92.8
6.0
91.6
5.9
0.182
IGF-I
ng/ml
201.9
49.5
176.0
43.8
0.001
Cortisol
g/dl
10.1
6.2
9.5
5.3
0.327
DHEA-s
ng/ml
1179.1
560.5
1140.7
572.7
0.245
Estradiol
pg/ml
71.7
57.8
70.0
50.6
0.438
Progesterone
ng/ml
3.9
6.7
4.4
6.2
0.267
ALP, alkaline phosphatase; BP, blood pressure; IGF-I, insulin-like growth factor-I; GOT, glutamic-oxaloacetic
transaminase; GPT, glutamic-pyruvic transaminase; - GTP, gamma-glutamyltranspeptidase; DHEA-s,
dehydroepiandrosterone sulfate. n = 20, mean
SD, paired t-test.
In endocrine examinations associated with anti-aging
estradiol, or progesterone.
medicine, IGF-I fell significantly in both group A and
Tables 7 and 8 show the results of the measurements of
group B, from 224.0
63.0 ng/ml to 185.1
63.7 ng/ml
the skin. No significant changes were seen in the sebum
(
= 0.001), and from 201.9
49.5 ng/ml to
levels in either group A or group B. Moisture had increased
176.0
43.8 ng/ml (
= 0.001), respectively. No
significantly on the forehead and the left cheek for both
significant changes were seen in cortisol, DHEA-s,
groups A and B. Blood flow had decreased significantly,
54

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Effects of cosmetics therapy using isoflavone and pine bark extract on the skin and QOL: A double-blind placebo-controlled trial
from 0.630
0.256 U to 0.481
0.179 U in group A
the forehead. The volume increased significantly in group
( = 0.001), and from 0.621
0.203 U to 0.471
0.168 U
A, from 160.3
33.1 U to 171.7
36.4 U (
= 0.003) and
in group B ( = 0.001). A significant difference was seen
did not show any significant differences in group B, between
between the groups in terms of the volume of melanin in
156.6
35.3 U and 159.2
31.1 U ( = 0.308).
Table 7. Measurements of the skin-group A
Variable
Unit
Before
4 weeks later
– level
Forehead: sebum
U
0.865
1.592
1.170
1.830
0.230
Forehead: moisture
U
49.86
10.70
58.41
10.80
0.008
Forehead: melanin
U
160.3
33.1
171.7
36.4
0.003
Forehead: erythema
U
310.1
60.7
281.5
49.0
0.002
Left cheek: sebum
U
0.325
0.862
0.430
0.542
0.303
Left cheek: moisture
U
52.8
11.0
62.5
8.0
0.003
Left cheek: melanin
U
149.5
35.0
151.0
46.4
0.392
Left cheek: erythema
U
301.6
57.7
283.9
45.2
0.026
Blood flow
U
0.630
0.256
0.481
0.179
0.001
n = 21, mean
SD, paired t-test. U; units are proper and depend on the measurement apparatus
Table 8. Measurements of the skin-group B
Variable
Unit
Before
4 weeks later
– level
Forehead: sebum
U
0.535
0.707
0.965
2.55
0.211
Forehead: moisture
U
51.34
11.12
56.77
11.92
0.049
Forehead: melanin
U
156.6
35.3
159.2
31.1
0.308
Forehead: erythema
U
302.8
65.1
273.8
61.7
0.002
Left cheek: sebum
U
0.321
1.116
0.347
0.313
0.461
Left cheek: moisture
U
55.2
10.6
61.6
8.8
0.010
Left cheek: melanin
U
151.8
33.1
155.8
35.1
0.278
Left cheek: erythema
U
292.6
47.4
277.8
53.5
0.114
Blood flow
U
0.621
0.203
0.471
0.168
0.001
n = 20, mean
SD, paired t-test. U; units are proper and depend on the measurement apparatus.
Regarding the investigation into the degree of oxidative
hour) in group A, between 1.80
1.16 and 1.39
0.72
stress(Tables 9 and 10), no significant differences were
(
= 0.072); however, a significant decrease was seen in
seen in blood lipid peroxide in group A, from 0.37
0.09
group B, from 2.83
2.46 to 2.30
1.95 (
= 0.044).
nmol/ml to 0.35
0.08 nmol/ml (
= 0.165); however, a
No significant differences were seen in either group A or
significant decrease was seen in group B, from 0.44
0.11
group B in terms of the rate of urinary 8-OHdG production,
nmol/ml to 0.39
0.08 nmol/ml (
= 0.006). Moreover,
which is an indicator for oxidative DNA damage.
no significant differences were seen in terms of the rate of
No adverse reactions or events were seen in either group
isoprostane production (in nanograms per kilogram per
A or B throughout the course of treatment.
Table 9. Degree of oxidative stress-group A
Variable
Unit
Before
4 weeks later
– level
Urinary 8-OHdG
ng/ml
5.08
2.28
5.07
3.05
0.494
Urinary creatinine
mg/dl
68.7
37.5
66.5
28.6
0.378
55

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Y. Yonei et al.
Urinary isoprostane
ng/ml
2.06
1.62
1.53
0.88
0.075
Urine volumeml
341.0
114.7
359.5
128.0
0.289
Urine-collection period
hour
5.99
1.48
6.29
1.19
0.266
8-OHdG production rate
ng/kg/h
4.80
2.03
4.65
2.33
0.363
Isoprostane production rate
ng/kg/h
1.80
1.16
1.39
0.72
0.072
8-OHdG/CRE
ng/mg CRE
8.69
3.82
7.80
2.79
0.078
Isoprostane/CRE
ng/mg CRE
2.97
1.16
2.37
1.08
0.023
Blood lipid peroxide
nmol/ml
0.37
0.09
0.35
0.08
0.165
8-OHdG, 8-hydroxy-2'deoxyguanosine; CRE, creatinine. n = 21, mean
SD, paired t-test.
Table 10. Degree of oxidative stress-group B
Variable
Unit
Before
4 weeks later
– level
Urinary 8-OHdG
ng/ml
7.52
5.57
6.43
4.34
0.150
Urinary creatinine
mg/dl
87.9
56.1
93.9
66.0
0.344
Urinary isoprostane
ng/ml
3.08
2.20
2.33
1.89
0.072
Urine volume
ml
288.5
114.6
311.3
105.7
0.148
Urine-collection period
hour
5.53
1.92
5.23
1.22
0.238
8-OHdG production rate
ng/kg/h
6.50
4.23
6.24
4.20
0.280
Isoprostane production rate
ng/kg/h
2.83
2.46
2.30
1.95
0.044
8-OHdG/CRE
ng/mg CRE
8.46
2.14
7.48
3.47
0.078
Isoprostane/CRE
ng/mg CRE
3.70
1.97
2.51
0.81
0.002
Blood lipid peroxide
nmol/ml
0.44
0.11
0.39
0.08
0.006
8-OHdG, 8-hydroxy-2'deoxyguanosine; CRE, creatinine. n = 20, mean
SD, paired t-test.
Discussion
collagenase, hyaluronidase, and elastase. Some of its
characteristic ingredients are other antioxidative
No appropriate assessments have thus far been made on the
substances such as tocopherol (vitamin E) and ascorbyl
efficacy of cosmetic products containing ingredients that
phosphate Mg (stabilized vitamin C derivative), which are
have estrogen-like activities such as isoflavone, or those
believed to help the activity of pine bark extract.
containing antioxidative substances such as pine bark
Isoflavone is believed to have estrogen-like activity.
extract. In our present study, to clear up these and other
Estrogen maintains femininity,2-4 as well as youthful and
doubts, we conducted 2 double-blind tests in a clinical
healthy skin. In addition, it is said to prevent bone density
setting. The results showed that, compared with a control
from declining, prevent arteriosclerosis from advancing,
group, women given the active Product saw improvements
and, in the long-term, it reportedly works to prevent the
in both mental and physical symptoms. Favorable effects
advancement of senility.3,4 There were 5 mental symptom
were also seen in the latter group in the oxidative stress
parameters in which group B saw favorable effects:
tests and in their skin data. Whether these test results can be
"irritability," "loss of motivation," "reluctance to talk with
explained by the presence of isoflavone and pine bark
others," "pessimism," and "lapse of memory." Estrogen is
extract will be examined hereunder.
known to have neuropsychiatric effects such as boosting of
Pine bark extract is a polyphenol, which is extracted
sexual feelings, enhancement of cognitive ability, and
from the bark of a pine tree (Pinus pinaster Ait.) originating
improvement of depressive states.3-4 It is true that
in France. Its primary ingredient, oligomeric
replenishing estrogen to treat menopausal disorders can
proanthocyanidin, or OPC, is believed to have
improve the subjects' mental status after a relatively short
antioxidative and vascular endothelial protecting
period. Therefore, improvement of mental symptoms may
activities.19,20 It is also said to suppress proteases such as
be explained as the effects of estrogen. However, some
56

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Effects of cosmetics therapy using isoflavone and pine bark extract on the skin and QOL: A double-blind placebo-controlled trial
facts should also be taken into account, that is, the effects
or estradiol levels in both groups A and B. No significant
of isoflavone are only about several tenths those of
differences were seen in the value of estradiol itself
estrogen, and that it is unclear if isoflavone is absorbed by
between the 2 groups.
the skin. It is believed that pine bark extract is not involved
In the previous study, improvements were seen with
in this process.
respect to the following 4 mental symptom parameters: "no
Regarding oxidative stress, neither group saw
feeling of happiness," "reluctance to talk with others,"
improvements in 8-OHdG, although lipid peroxide in the
"pessimism," and "inability to sleep because of worries." In
blood and the rate of isoprostane production improved in
the present study, improvements were seen with respect to
group B. The substances contained in the Product, which
the following 5 mental symptom parameters: "Irritability,"
group B had used, had little or no effect on DNA oxidative
"loss of motivation," "reluctance to talk with others,"
damage, but were shown to significantly suppress the
"pessimism," and "lapse of memory." As seen in both tests,
hyperoxidation of lipids. This, however, is largely due to
"reluctance to talk with others" and "pessimism" were
the involvement of pine bark extract; it is believed that
improved. We all know that it is not easy to adequately
isoflavone is not involved in this process.
identify the changes in mental symptoms and as a result of
As for the skin measurement test, favorable effects were
comparing the 2 tests, we concluded that their
seen on the volume of melanin on the forehead. The
reproducibility was adequate.
question here is whether the interpretation, "Group A had
Regarding the cortisol value, our previous study found
increased volumes of melanin, while such increases were
that the mean for subjects aged 40 years and older was
suppressed in group B" is reasonable. If it is true that
relatively high, at 13.7
6.0 g/dl, and that, when the
estrogen works to maintain a youthful and healthy skin, it
numerical values were examined for each patient, those
could also have a favorable effect on the melanin deposits
with high cortisol levels saw a reduction, while those
in the skin.3 Because isoflavone is administered directly
whose levels were not high initially had no changes. In
on the skin along with the cosmetic base, it is definitely
other words, the activity of putting on makeup could be
possible that effective doses may have worked on the
regarded as working to maintain the body's homeostasis in
skin.
an auxiliary manner, rather than to reduce the cortisol
Then, what about indirect actions? These are derived
level. In the present study, the value of cortisol was not
from the fact that the Product used by group B improved
too high in the beginning, at 10.1
6.2 g/dl, and
the mental symptoms (which indicates feeling pleasant)
ultimately became 9.5
5.3 g/dl, which did not
when actually used. This was determined by survery
constitute a significant change. However, the figures are
question. If people actually use the Product and see their
consistent in terms of maintaining homeostasis or
skin conditions improve in addition to experiencing a
constancy.
pleasant feeling while using it, they may end up happier.
Although the same view could be also applied to
This may be the comprehensive effect of isoflavone and
estradiol, we determined the trial period to be 4 weeks, so
pine bark extract contained in the Product. However, it
as to eliminate as much as possible the influence of the
does not matter too much if there are aspects that cannot be
subjects' menstrual cycles.
fully explained by the pharmacological activity of
IGF-I and the amount of blood flow in the facial skin
isoflavone and pine bark extract. It is common to see
decreased in both group A and group B. Although there is a
favorable effects on a person's mental state, which in turn
possibility that IGF-I may have decreased because of the
induce favorable effects on their physical state.
activity of applying makeup, compared with ordinary
Let us examine the consistency between the previous test
biochemical tests, there still are many measurement errors,
and the present one. Results of the previous clinical test,
so the possibility of the effects of variations/discrepancies
conducted as an open, non-blinded study, showed that the
cannot be ruled out. Similar measurement errors must be
action of applying a skin cream 1 improved a number of
taken into account for the amount of blood flow in the
subjective symptoms related to QOL,2 reduced the level of
facial skin as well.
cortisol (a stress hormone) in subjects over the age of 40
In this paper, we have described the results of our present
years, and 3 elevated the level of estradiol.6 Meanwhile,
test and the interpretation thereof. In conclusion, the
results of the present study showed improvements in QOL
efficacy of anti-aging cosmetic products that contain
items that were different from those of the previous study.
characteristic ingredients (used by group B) was shown by
Further, no significant differences were seen in the cortisol
a double-blind test method.
57

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