Procyanidin B-2 is a compound we have identified in apple which acts as a
growth-promoting factor on murine hair epithelial cells. This report
describes our investigation of the hair-growing effects of 1% procyanidin
B-2 tonic after sequential use for 4 months. A
double-blind clinical trial
was performed, involving a total of 29 subjects (procyanidin B-2, 19 men;
placebo, 10 men). No adverse side effects were observed in either group. In
the procyanidin B-2 group, 78.9% showed an increased mean value of hair
diameter, whereas only 30.0% in the placebo group showed any increase (p <
0.02, Fisher's exact probability test). The increased ratio of hairs
measuring more than 40 microm in diameter after 4 months of procyanidin B-2
treatment was significantly higher than that of the placebo controls (p <
0.05, two-sample-t-test). The increase in number of total hairs in the
designated scalp area (0.25 cm(2)) of procyanidin B-2 subjects after a 4
month trial was significantly greater than that of the placebo controls
(procyanidin-B-2, 3.67 +/- 4.09 (mean +/- SD)/0.25 cm(2); placebo, -2.54 +/-
4.00/0.25 cm(2); p < 0.001, two-sample t-test).
Procyanidin B-2 therapy
shows potential as a promising cure for male pattern baldness. Copyright
2001 John Wiley & Sons, Ltd.
topical application of procyanidin B-2 from apple to identify its potential
use as a hair growing agent.
Kamimura A, Takahashi T, Watanabe Y.
Tsukuba Research Laboratories, Kyowa Hakko Kogyo Co., Ibaraki, Japan.
Procyanidin B-2 is a polyphenol compound we have identified in apple which
acts as a hair-growing factor in the murine model both in vitro and in vivo.
This report describes our investigation of the effects of 1% procyanidin B-2
tonic on human hair growth after sequential use for 6 months. A
clinical test involving a total of 29 subjects was performed. Nineteen men
in the procyanidin B-2 group and 10 men in the placebo control group were
subjected to analyses. No adverse side effects were observed in either
group. The hair-growing effect was evaluated using a macrophotography
technique combined with measurements of the hair diameter of clipped hairs.
The increase in number of total hairs in the designated scalp area (0.5 cm
square = 0.25 cm2 area) of procyanidin B-2 group subjects after the 6-month
trial was significantly greater
than that of the placebo control group
subjects (procyanidin B-2, 6.68 +/- 5.53 (mean +/- SD)/0.25 cm2; placebo,
0.08 +/- 4.56 (mean +/- SD)/0.25 cm2; P < 0.005, two-sample t test). The
increase in number of terminal hairs, which are defined as hairs more than
60 microm in diameter, in the designated area (0.5 cm square = 0.25 cm2
area) of the procyanidin B-2 group subjects after the 6-month trial was
significantly greater than that of the placebo control group subjects (procyanidin
B-2, 1.99 +/- 2.58 (mean +/- SD)/0.25 cm2; placebo, -0.82 +/- 3.40 (mean +/-
SD)/0.25 cm2; P < 0.02, two-sample t test).
Procyanidin B-2 therapy shows
potential as a safe and promising cure for male pattern baldness.
Journal of Investigative Dermatology
112, 310-316 (1999)
Procyanidin Oligomers Selectively and
Intensively Promote Proliferation of Mouse Hair Epithelial Cells In Vitro
and Activate Hair Follicle Growth In Vivo1
Tomoya Takahashi, Toshikazu Kamiya, Atsuhiro
Hasegawa,* and Yoshiharu Yokoo
Tsukuba Research Laboratories, Kyowa Hakko
Kogyo, Tsukuba, Ibaraki, Japan; *Tokyo Research
Laboratories, Kyowa Hakko Kogyo, Machida, Tokyo, Japan
Reprint requests to: Dr. Tomoya Takahashi,
Tsukuba Research Laboratories, Kyowa Hakko Kogyo Co., 2, Miyukigaoka,
Tsukuba, Ibaraki 305-0841, Japan. 1The authors disclosed
conflict of interest. 2Wilson C, Walkden V, Powell S, Shaw S,
Wilkinson J, Dawber R: Contact dermatitis in reaction to 2% topical
minoxidil solution. J Am Acad Dermatol 24:661–662, 1991 (brief
We have previously reported that proanthocyanidins
extractedfrom grape seeds possess growth-promoting activity
toward murinehair epithelial cells in vitro and stimulate
anagen inductionin hair cycle progression in vivo. This
report constitutes acomparison of the growth-promoting activity
of procyanidin oligomersand the target cells of procyanidins in
the skin. Results showthat procyanidin dimer and trimer exhibit
higher growth-promotingactivity than the monomer.
The maximum growth-promoting
activityfor hair epithelial cells with procyanidin B-2, an
epicatechindimer, reached about 300% (30 µM) relative to
controls(=100%) in a 5 d culture. Optimum concentration of
procyanidinC-1, an epicatechin trimer, was lower than that of
procyanidinB-2; the maximum growth-promoting activity of
procyanidin C-1was about 220% (3 µM).
No other flavonoid compounds
examinedexhibit higher proliferative activities than the
procyanidins.In skin constituent cells, only epithelial
cells such as hairkeratinocytes or epidermal keratinocytes
respond to procyanidinoligomers. Topical application of 1%
procyanidin oligomers onshaven C3H mice in the telogen phase led
to significant hairregeneration [procyanidin B-2, 69.6% ± 21.8%
(mean ±SD); procyanidin B-3, 80.9% ± 13.0%; procyanidin C-1,78.3% ± 7.6%] on the basis of the shaven area; application
of vehicle only led to regeneration of 41.7% (SD = 16.3%).
Inthis paper, we
demonstrate the hair-growing activity of procyanidinoligomers
both in vitro and in vivo, and their potential for
use as agents to induce hair growth.
Dermatology, Toyama Medical and Pharmaceutical University
The Hair-Growing Activity of Procyanidin Oligomers
Department of Dermatology, Toyama Medical and Pharmaceutical University,
Sugitani, Toyama, JP.
Procyanidins are a family of condensed tannins we have
identified in apples, which act as a hair-growing factor in the
murine model both in vitro and in vivo. We have previously reported that the
growth-promoting effect on murine hair epithelial cells attributable to
procyanidin B-2, one species of procyanidin oligomer, reaches about
300% relative to controls;
and have also shown that procyanidin B-2 possesses intensive anagen-inducing
activity in the C3H in vivo mouse model. This presentation describes our
investigations during a 12-month clinical trial of highly purified
procyanidin oligomers isolated
from unripe apples, chiefly comprising procyanidin B-2,
procyanidin B-1, and procyanidin C-1. The clinical trial was performed in a
total of 21 subjects showing male pattern baldness on the head. The test
agent (about 1.8 ml per dose) was applied to the subjects’ affected scalp
area twice a day, giving a daily dose of 16 mg of procyanidin oligomers.
During the 12 months of twice-daily application of the agent, the
hair-growing effects were evaluated according to the following parameters:
the macrophotographically recorded change in the number of hairs in the
designated scalp area, the changes in the diameter of hairs clipped from the
designated scalp area, and the changes in the photographically recorded
global view of the subjects’ heads. No side effects were observed in any
subjects. After 12 months of use,
71% of the subjects showed an increased number of hairs in the
designated scalp area relative to pre-trial measurements. The numbers of
total hairs in the designated scalp area after the 12-month trial were
significantly greater than
the measured values at the start of the trial (paired t-test, p < 0.005). We
also observed a clear trend towards increased number of non-vellus hairs (>
40 µm) in the designated scalp area after the 12-month trial compared to the
values measured at the start of the test. A number of the subjects showed
cosmetically satisfactory changes.
Procyanidin therapy shows promise as a potential cure for male pattern
Br J Dermatol.
Procyanidin B-2, extracted from apples, promotes hair
growth: a laboratory study.
BACKGROUND: We have previously reported that several selective protein
kinase C (PKC) inhibitors, including
promote hair epithelial cell growth and stimulate anagen induction.
OBJECTIVES: We discuss the hypothesis that the hair-growing activity of
procyanidin B-2 is related to its downregulation or inhibition of
translocation of PKC isozymes in hair epithelial cells. METHODS: We examined
the effect of procyanidin B-2 on the expression of PKC isozymes in cultured
murine hair epithelial cells as well as PKC isozyme localization in murine
dorsal skin at different stages in the hair cycle. RESULTS: We observed that
procyanidin B-2 reduces the expression of PKC-alpha, -betaI, -betaII and -eta
in cultured murine hair epithelial cells and also inhibits the translocation
of these isozymes to the particulate fraction of hair epithelial cells. Our
immunohistochemical analyses demonstrated that PKC-alpha, -betaI, -betaII
and -eta are specifically expressed in the outer root sheaths of both anagen
and telogen hair follicles. The hair matrix at the anagen stage showed no
positive staining for these PKC isozymes. Moderate to intense staining for
PKC-betaI and -betaII in the epidermis and hair follicles was observed in a
telogen-specific manner; however, expression of PKC-alpha and -eta during
the telogen stage was not conspicuous. Go 6976, an inhibitor of
calcium-dependent (conventional) PKC, proved to promote hair epithelial cell
growth. CONCLUSIONS: These results suggest that PKC isozymes, especially
PKC-betaI and -betaII, play an important role in hair cycle progression and
that the hair-growing mechanisms of procyanidin B-2 are at least partially
related to its downregulation of PKC isozymes or its inhibition of
translocation of PKC isozymes to the particulate fraction of hair epithelial
PMID: 11841365 [PubMed - indexed for MEDLINE]
Skin Pharmacol Appl Skin Physiol 2000
Several selective protein kinase C inhibitors including
procyanidins promote hair growth.
Skin Pharmacol Appl Skin Physiol 2000 May-Aug;13(3-4):133-42
Takahashi T, Kamimura A, Shirai A, Yokoo Y. (Tsukuba Research Laboratories,
Kyowa Hakko Kogyo Co., Tsukuba, Ibaraki, Japan. firstname.lastname@example.org)
We have previously reported that procyanidin oligomers selectively promote
growth of murine hair epithelial cells in vitro and stimulate anagen
induction in vivo. We report here the possible relationship between the
protein kinase C-inhibiting activity of procyanidins and their hair-growing
activity. Of the procyanidins, procyanidin B-2 and procyanidin C-1, which
selectively inhibit protein kinase C, intensively promote hair epithelial
cell proliferation in vitro and stimulate anagen induction in vivo. On the
other hand, procyanidins, which inhibit both protein kinase C and A, showed
relatively low activity in in vitro and in vivo evaluations. We also found
that calphostin C, which is a selective inhibitor of protein kinase C,
possesses hair epithelial cell growth-promoting activity in vitro and anagen
phase-inducing hair-growing activity in vivo. Other selective protein kinase
C inhibitors, such as hexadecylphosphocholine, palmitoyl-DL-carnitine
chloride, and polymyxin B sulfate, also show marked anagen phase-inducing
hair-growing activity in vivo. Nonselective protein kinase inhibitors, such
as staurosporine and K252a, inhibit the growth of hair epithelial cells.
1,2-Dioctanoyl-sn-glycerol, a protein kinase C activator, dose-dependently
decreases the growth of hair epithelial cells.
Forskolin, an adenylate cyclase
activator, promotes hair epithelial cell growth and boosts the
growth-promoting effect of procyanidin B-2. It is speculated that the
hair-growing activity of procyanidins is related to their protein kinase
Copyright 2000 S. Karger AG, Basel
PMID: 10859531 [PubMed - indexed for MEDLINE]
Food Chem Toxicol.
Toxicological studies on procyanidin B-2 for external
application as a hair growing agent.
Takahashi T, Yokoo Y, Inoue T, Ishii A.
Tsukuba Research Laboratories, Kyowa Hakko Kogyo Co., Ibaraki, Japan.
Procyanidin B-2 [epicatechin-(4beta --> 8)-epicatechin] is one of condensed
tannin that exists widely in plants. We have reported previously that
procyanidin B-2 possesses
hair epithelial cell growth-promoting activity and stimulates anagen
induction in hair cycle progression. To evaluate the safety of topical
procyanidin B-2 as a hair growing agent, we examined the mutagenicity, acute
subcutaneous injection, primary irritation, skin sensitization, and eye
irritation of this compound. Mutagenicity tests using bacteria showed
procyanidin B-2 to be non-mutagenic. Chromosomal aberration tests using CHL
cells indicated that procyanidin B-2 caused polyploidy but no structural
aberrations. In micronucleus tests for mutagenicity using mice, procyanidin
B-2 was negative. Acute subcutaneous injection study using rats revealed no
symptoms of significant injury. The lethal dose of procyanidin B-2 is
greater than 2000 mg/kg (subcutaneous injection). Primary irritation tests
using rabbits indicated that procyanidin B-2 containing preparation shows no
primary irritation. In the guinea pig maximization test, there was no
evidence of sensitization to procyanidin B-2. In primary ocular irritation
tests using rabbits, procyanidin B-2 containing preparation and vehicle
showed slight irritation of conjunctivae which is assumed to be caused by
ethanol. It is suggested that
topical procyanidin B-2 is safe and acceptable from the series
of toxicological tests.