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Biomolecules & Therapeutics,
17(3)
, 241-248 (2009) ISSN: 1976-9148(print)/2005-4483(online)
www.biomolther.org DOI: 10.4062/biomolther.2009.17.3.241
*Corresponding author
Tel: +82-53-810-2822 Fax: +82-53-810-4654
E-mail: byoo@ynu.ac.kr
Fig. 1.
The chemical structures of magnesium ascorbyl pho-
sphate (MAP) and methylsulfonylmethane (MSM). (A) Magnes-
ium ascorbyl phosphate (MAP). (B) Methylsulfonylmethane (MSM).
The Effect of Methylsulfonylmethane on Hair Growth Promotion of
Magnesium Ascorbyl Phosphate for the Treatment of Alopecia
Srinivasan S
HANMUGAM
, Rengarajan B
ASKARAN
, Santhoshkumar N
AGAYYA-SRIRAMAN
,
Chul-Soon Y
ONG
, Han-Gon C
HOI
, Jong Soo W
OO
, and Bong-Kyu Y
OO
*
College of Pharmacy, Yeungnam University, Kyungsan 712-749, Republic of Korea
(Received April 10, 2009; Revised May 16, 2009; Accepted May 23, 2009)
Abstract
- The purpose of this study was to evaluate the effect of methylsulfonylmethane (MSM) on hair
growth promotion of magnesium ascorbyl phosphate (MAP) for the treatment of alopecia. Aqueous solutions
of MAP 7.5% with or without MSM 1%, 5% or 10% were prepared and applied onto the depilated back skin of
the male mice once a day for 20 days. The degree of hair growth was evaluated by visual scoring using hair
growth quantification scale (0-5, 0 being initial state and 5 being complete hair growth). In vitro transdermal
penetration and intradermal retention studies of MAP were performed with Franz diffusion cell using hairless
mice skin. Hair growth in the group treated with the aqueous solution containing MAP 7.5% and MSM 10%
was comparable to or better than the result in the group treated with minoxidil 5% solution. Hair growth
promotion of MAP was dose-dependently increased by the presence of MSM used in combination with MAP
7.5% solution. The in vitro transdermal penetration of the MAP was decreased in proportion to the
concentration of MSM. However, intradermal retention of MAP was profoundly and dose-proportionally
increased as a function of MSM concentration, reaching 802 μg/cm
2
in the presence of MSM 10% (200-fold
increase). The effect of MSM on hair growth promotion of MAP was dose-proportional to the concentration of
MSM due to the enhanced intradermal retention of MAP in the presence of MSM. Therefore, topical
application of MAP together with MSM appears to be useful for the treatment of alopecia.
Keywords:
Magnesium ascorbyl phosphate, Methylsulfonylmethane, Alopecia, Hair growth, Skin per-
meation/accumulation, Permeation enhancer
INTRODUCTION
Magnesium ascorbyl phosphate (MAP) (Fig. 1A), a sta-
ble phosphate derivative of L-ascorbic acid has been re-
ported to stimulate growth of dermal papilla cells in vitro
(Hata and Senoo, 1989; Takamizawa et al., 2004) and in-
duce early conversion of telogen phase to anagen phase
of hair cycle in experiment with mice (Sung et al., 2006).
Furthermore, MAP treatment resulted in significant elonga-
tion of hair shafts in isolated hair follicles. Mechanism re-
sponsible for the growth stimulation of dermal papilla cells
and elongation of hair shafts has been suggested that
MAP induces proliferative and anti-apoptotic effects on
dermal papilla cells (Sung et al., 2006). This finding pro-
vided a potential that MAP might be useful for hair growth
promotion in the treatment of alopecia. However, because
of its hydrophilicity MAP has lower ability to penetrate the
skin barrier stratum corneum to produce its therapeutic ef-
fect (Spiclin et al., 2002; Yoo et al., 2008).
242
Srinivasan Shanmugam et al.
Table I.
Compositions of aqueous solution of MAP containing
various concentrations of MSM
MAP MSM Solvent
MAP 7.5% only 7.5% - Distilled water
MAP-MSM 1% 7.5% 1% Distilled water
MAP-MSM 5% 7.5% 5% Distilled water
MAP-MSM 10% 7.5% 10% Distilled water
MAP: magnesium ascorbyl phosphate, MSM: methylsulfonylme-
thane, MAP-MSM: MAP 7.5% in the presence of designated
concentration of MSM.
In transdermal drug delivery arena, several approaches
have been employed to lessen the barrier function of the
stratum corneum for enhanced penetration of drug of
choice, and one popular approach has been the use of
trandermal permeation enhancers (Godwin and Michniak,
1999; Asbill and Michniak, 2000). These are chemical
compounds that reversibly alter the barrier function of the
skin and enhance penetration. Among various classes of
permeation enhancers, those that enhance local skin-drug
concentrations (intradermal) and do not produce sig-
nificant transdermal enhancement (across the skin) are
considered ideal permeation enhancers for topical delivery
(Asbill and Michniak, 2000; Im et al., 2008). Myriad of re-
searchers reported the possible mechanisms and uses of
various penetration enhancers. However, despite major in-
terest in this research arena, very few penetration en-
hancers have been made commercially available because
of the limitations on selection based on their efficacy, lack
of toxicity, and compatibility with other components of the
transdermal system (Skelly et al., 1987). One of the ear-
liest and widely investigated penetration enhancers is
DMSO (Williams and Barry, 1992).
Methylsulfonylmethane (MSM), also known as dimethyl
sulfone or methyl sulfone (Fig. 1B), is an organic sul-
fur-containing compound that occurs naturally in a variety
of plants and animals including humans (Pearson et al.,
1981; Usha and Naidu, 2004; Ameye and Chee, 2006).
MSM is a normal oxidative metabolite product of dime-
thylsulfoxide (DMSO) and has been widely used as a diet-
ary supplement for the treatment of several indications
such as osteoarthritis, seasonal allergic rhinitis, interstitial
cystitis, and snoring (Childs, 1994; Barrager et al., 2002;
Blum and Blum, 2004; Kim et al., 2006). Like DMSO, MSM
is an aprotic solvent that can dissolve wide range of sol-
utes and is miscible with many solvents, and therefore
could be easily incorporated into pharmaceutical for-
mulations (Clark et al., 2008).
It is well known that much of the unique properties of
DMSO are related to its characteristic polar O-S bond,
which enables the compound to interact strongly with wa-
ter by forming hydrogen bonds. This interaction with water
is thought to be one of the key elements in the mechanism
of action of DMSO as a penetration enhancer (Franz et al.,
1995). Due to the structural similarity with DMSO, MSM is
expected to act as a skin permeation enhancer for various
medications by similar mechanisms (Engelen and Engelen,
2002). Structural similarity coupled with non-toxicity and
safety (GRAS Notice Number: GRN 229) as an oral nutri-
tional additive as well as unavailability of prior work, put
forth MSM a rightful candidate to explore its permeation
enhancing potential among many other permeation
enhancer.
In this study, we have hypothesized that hair growth pro-
motion effect of MAP can be improved by co-application
with MSM because of its skin permeation enhancing effect.
The purpose of this study was (1) to evaluate in vivo hair
growth promotion effect of MAP (in the absence and pres-
ence of MSM) using alopecia model of C57BL/6N mice
and (2) to substantiate the in vivo result with transdermal
penetration and intradermal retention of MAP in vitro.
MATERIALS AND METHODS
Materials
Magnesium ascorbyl phosphate, methylsulfonylmethane,
and methyl paraben were obtained from Wako Pure
Chemical Industries (Osaka, Japan). Thioglycolic acid
80% cream (Niclean Cream
Ⓡ
, Ildong Pharmaceutical Inc.,
Korea) and minoxidil 5% solution (Minoxyl Solution
Ⓡ
,
Hyundai Pharmaceutical Inc., Korea) were purchased from
a local pharmacy. Sodium phosphate monobasic mono-
hydrate and sodium phosphate dibasic heptahydrate were
purchased from Sigma-Aldrich, Inc., (St. Louis, USA).
Metaphosporic acid and trichloroacetic acid were pur-
chased from Junsei Chemical Company Ltd., (Tokyo,
Japan). Methanol used was HPLC grade and all other
chemicals were of analytical grade and used without fur-
ther purification.
Preparation of MAP solutions
MAP (7.5 g) was accurately weighed and dissolved into
100 ml distilled water. Aliquots of this solution (10 ml) were
taken and added with 100, 500, or 1,000 mg of MSM to
make 1%, 5%, or 10% MSM in the MAP 7.5% solution. The
resultant aqueous solutions of MAP 7.5% alone or with
MSM 1%, 5% or 10% were thoroughly mixed with mag-
netic bar and used for in vitro or in vivo studies thereafter.
Compositions of aqueous solution of MAP containing vari-
ous concentrations of MSM are shown in Table I.
Hair Growth Promotion of Magnesium Ascorbyl Phosphate
243
Fig. 2.
Hair growth quantification scale. 0: initial state, 1: grey to
black coloration, 2: short visible hair, 3: sparse long hair, 4:
dense long hair, 5: complete hair growth.
HPLC analysis
The concentration of MAP in transdermal penetration
and intradermal retention studies, was assayed by HPLC
system (Shimadzu, Japan) equipped with Class VP com-
puter software, LC 10 AD VP pump, and SPD 10A UV-VIS
detector at 240 nm using methyl paraben as internal
standard. Column used was Inertsil CN-3 (4.6
×250 mm,
GL Science Inc, Japan) and mobile phase consisted of a
mixture of 0.02 M phosphate buffer and methanol (60:40,
v/v) adjusted to pH 2.3 with phosphoric acid. Flow rate was
0.8 ml/min and the injection volume of the sample was 20 μl.
The validation of the HPLC assay was performed by re-
peating five times a day for five consecutive days for MAP
concentration range of 50 ng/ml-100 μg/ml.
Animals
Animals care and procedures were conducted accord-
ing to the guidelines for animal use in toxicology (Society of
Toxicology USP 1989) and the study protocol was ap-
proved by the Animal Care and Use Committee, College of
Pharmacy, Yeungnam University. Six-week-old male mice
(C57BL/6N) and six-week-old male hairless mice were ob-
tained from Orient Bio (Seoul, Korea) and housed in
groups not exceeding six per cage and maintained under
standard conditions. The acclimation period was two
weeks before the experimental procedure with a dark/light
cycle of 12 h/12 h at the temperature of 23 ± 2
o
C. Food and
tap water were available ad libitum during the acclimation
period.
Evaluation of hair growth promotion using alopecia
model in mice
The hair on the back of male C57BL/6N mice was re-
moved by electric clipper and depilated using thioglycolic
acid 80% cream (Niclean cream
Ⓡ
, Ildong Pharmaceutical
Inc., Korea) one day before applying the samples. About
0.5 g of the cream was applied on the hair-clipped back of
the mice and allowed for 2 min. The cream and hair were
wiped off using cotton for depilation, and the back skin was
rinsed with plenty of distilled water to remove remaining
thioglycolic acid. The mice without significant wound on the
back were selected, randomly grouped, and used for hair
growth study. Freshly prepared aqueous solution (0.2 ml)
of MAP 7.5% alone or with MSM 1%, 5% or 10% was ap-
plied onto the depilated back of the mice (n=5) and allowed
to dry. The sample application was done once a day for
twenty days. Saline solution (0.9%) and Minoxyl
Ⓡ
(minoxidil
5% solution) were also used as negative and positive con-
trol, respectively. The degree of hair growth was evaluated
by visual scoring of hair growth quantification scale as re-
ported by Chen and his colleagues (0: initial state, 1: grey
to black coloration, 2: short visible hair, 3: sparse long hair,
4: dense long hair, 5: complete hair growth) (Fig. 2) (Chen
et al., 2005). Photographs of all mice in this study were tak-
en every five days for 20 days.
Transdermal penetration and intradermal retention
studies
Hairless mice were sacrificed and full-thickness skin
was taken, followed by removal of subcutaneous fat.
Approximately 3 cm
2
of the trimmed skin was placed into
Franz diffusion cell with effective diffusion area of 2.27
cm
2
. For transdermal penetration study, freshly prepared
aqueous solution (0.4 ml) of MAP 7.5% alone or with MSM
1%, 5% or 10% was applied on the skin and allowed to
spread over the skin. Donor compartment (epidermal layer
of the skin) remained unoccluded, but covered with a lid,
and the receptor compartment (dermal layer of the skin)
was perfused with distilled water maintaining temperature
at 37
o
C. Serial sampling of the receptor compartment was
performed at predetermined intervals (1.5, 3, 6, 9, and 12
h). Cumulative amount of MAP penetrated through the skin
was plotted as a function of time. The flux of MAP was cal-
culated by the slope of the linear portion of the curve,
which was achieved in 6 h.
For intradermal retention study, the effective diffusion
area of the mounted skin was cut off after transdermal pen-
etration study and washed with distilled water, and stored
in the deep-freezer until HPLC determination. For analysis,
the skin was homogenized with isotonic phosphate buffer
(pH 7.4) for 10 min in an ice bath. To 0.45 ml of the skin ho-
mogenate, 0.05 ml of internal standard (100 μg/ml) sol-
ution and 0.5 ml of extraction solvent (10% metaphos-
phoric acid:20% trichloroacetic acid:distilled water=2:4:4)
were added, and was subjected to sonication for 10 min
and centrifugation at 16,000 g for 3 min. The clear super-
244
Srinivasan Shanmugam et al.
Fig. 3.
Hair growth quantification score after application of MAP
in alopecia model of C57BL/6N mice (n=5). MAP: magnesium
ascorbyl phosphate, * and ** represent p<0.05 and <0.01 com-
pared to negative control, respectively.
Fig. 4.
Hair growth quantification score after application of MAP
7.5% with or without MSM in alopecia model of C57BL/6N mice
(n=5). MAP: magnesium ascorbyl phosphate, MSM: methylsul-
fonylmethane, MAP-MSM: MAP 7.5% in the presence of desig-
nated concentration of MSM, * and ** represent p<0.05 and
<0.01 compared to MAP 7.5% only, respectively.
natant thus obtained was then analyzed for MAP by HPLC
utilizing the method described in HPLC assay section.
Data analysis
All the data obtained were analyzed using SPSS 12.0
for Windows and a significance level of p<0.05 was used
to indicate the statistically significant difference between
data sets.
RESULTS
Aqueous solution (0.2 ml) of MAP was investigated for
the ability to promote hair growth in the alopecia model of
C57BL/6N mice. There was no significant difference be-
tween the MAP-treated groups and control group up to 10
th
day (Fig. 3). However, in 15
th
day, hair growth quantifica-
tion score in the mice treated with MAP 7.5% or above was
significantly enhanced compared to control group (p
<0.05), and the hair growth status of the mice was “dense
long hair” in 20
th
day. MAP 5% also showed hair growth
promotion compared to control group in 20
th
day (p<0.05),
but hair growth status of the mice was in the level of
“sparse long hair”. Hair growth status in the group treated
with MAP 2.5% was not different from control group even
in 20
th
day.
The effect of MSM on the hair growth promotion of MAP
7.5% solution was investigated in the presence of MSM
1%, 5% or 10% (Fig. 4). MSM 1% did not result in sig-
nificant additive effect even after 20 days. However, MSM
10% demonstrated statistically significant additive effect
as early as in 10
th
day (p<0.05), showing 2.50 ± 0.33 of
hair growth quantification score compared to 1.92 ± 0.33 in
the group treated with MAP 7.5% solution without MSM.
After 20 days of treatment, hair growth status in the group
treated with MAP 7.5% solution with MSM 5% or 10% was
“complete hair growth”, while it was still in the level of
“dense long hair” in the group treated with MAP 7.5% sol-
ution without MSM (p<0.01). Furthermore, hair growth
rate in the mice treated with MAP 7.5% solution with MSM
10% was better than or similar to the mice treated with
minoxidil 5% solution. Photographs for hair growth status
in all groups tested are shown in Fig. 5.
Cumulative amount of MAP penetrated through the hair-
less mice skin during 12 h was 3.89 ± 0.41, 2.23 ± 0.39,
and 1.80 ± 0.24 μg/cm
2
in the presence of MSM 1%, 5%,
and 10%, respectively, while it was 4.17 ± 0.46 μg/cm
2
in
the absence of MSM (Fig. 6). Lag time for transdermal
penetration of MAP was about 1.5 h, and MAP was de-
tected in the receptor compartment of the Franz diffusion
cell after 3 h. When MAP was applied as 7.5% solution with
5% or 10% MSM, transdermal penetration of MAP was only
53.5% and 43.2% of the amount penetrated in the absence
of MSM, respectively (p<0.01). Average flux of MAP calcu-
lated by the slope of the linear portion of each curve was
0.08080 ± .04, 0.0359 ± 0.01, and 0.0313 ± 0.02 μg
×cm
−2
/h
in the presence of MSM 1%, 5%, and 10%, respectively,
while it was 0.2205 ± 0.02 μg
×cm
−2
/h in the absence of
MSM (Table II). The flux of MAP in the presence of MSM
5% or above was less than one fifth of the flux in the ab-
sence of MSM.
The amount of MAP remaining in the skin (intradermal
Hair Growth Promotion of Magnesium Ascorbyl Phosphate
245
Fig. 5.
Photographs of hair growth status during treatment with MAP in the absence or presence of MSM. MAP: magnesium ascorbyl
phosphate, MSM: methylsulfonylmethane, MAP-MSM: MAP 7.5% in the presence of designated concentration of MSM, NC: negative
control (0.9% saline solution), PC: positive control (minoxidil 5% solution).
retention) after 12 h of transdermal penetration study was
10.5 ± 12.1, 62.0 ± 34.6, and 801.7 ± 156.0 μg/cm
2
in the
presence of MSM 1%, 5%, and 10%, respectively. It was
only 4.2 ± 5.9 μg/cm
2
when MAP was applied without MSM
(Fig. 7). Intradermal retention of MAP was exponentially in-
creased in the presence of MSM, reaching about 200-fold
when MAP was applied with MSM 10%.
DISCUSSION
Hair growth in animals periodically undergoes a con-
tinuous hair cycle, during which the hair follicle switches
between phases of growth and regression (Paus, 2006).
During the hair cycle, the growth phase (anagen) lasts for
two to seven years followed by transition and involution
phase (catagen), which lasts for one to two weeks, and fi-
nally the resting phase (telogen) that lasts for five to 12
weeks (Ellis et al., 2002). MAP was reported to enhance
the hair growth through anagen induction, which is consid-
ered one of the most effective strategies for treating hair
loss (Paus, 2006).
As a preliminary study, we investigated hair growth pro-
motion effect of MAP in alopecia model using black-haired
mice (C57BL/6N). Concentrations of MAP tested in this
study were 2.5%, 5%, 7.5%, and 10%, because previous
researchers reported that MAP induced early conversion
from telogen phase to anagen phase of hair growth cycle
at 250 mM concentration, which is equivalent to about
7.25% of MAP (Sung et al., 2006). Hair growth quantifica-
tion score in the groups treated with MAP was in proportion
to the concentration of MAP up to 7.5% and plateaued
above this concentration. This result is consistent with the
246
Srinivasan Shanmugam et al.
Fig. 7.
Amount of MAP accumulated in hairless mice skin (n=5).
MAP: magnesium ascorbyl phosphate, MSM: methylsulfonyl-
methane, MAP-MSM: MAP 7.5% in the presence of designated
concentration of MSM, ** and *** represent p<0.01 and <0.001
compared to MAP 7.5% only, respectively.
Table II.
Average flux of MAP in the skin penetration study (n=6)
MAP 7.5% only MAP-MSM 1% MAP-MSM 5% MAP-MSM 10%
Average flux (μg
×cm
−2
/h) 0.2205 ± 0.02 0.0808 ± 0.04
a
0.0359 ± 0.01
a
0.0313 ± 0.02
a
MAP: magnesium ascorbyl phosphate, MSM: methylsulfonylemethane, MAP-MSM: MAP 7.5% in the presence of designated concentration of
MSM,
a
p<0.01 compared to MAP 7.5% only.
Fig. 6.
Amount of MAP penetrated through hairless mice skin
(n=5). MAP: magnesium ascorbyl phosphate, MSM: methylsul-
fonylmethane, MAP-MSM: MAP 7.5% in the presence of de-
signated concentration of MSM, * and ** represent p<0.05 and
<0.01 compared to MAP 7.5% only, respectively.
previous researchers’ finding that MAP promotes hair
growth (Sung et al., 2006).
Hair growth status in the group treated with combination
of MAP 7.5% solution and MSM 5% or 10% was sig-
nificantly better than the result found in the group treated
with MAP 7.5% solution only (p<0.01). Notably, hair growth
in the group treated with MAP 7.5% solution and MSM
10% showed better than or similar to the group treated with
minoxidil 5% solution (positive control), which is consid-
ered a standard topical therapy for the treatment of
alopecia. Also remarkably, the hair growth status was in
proportion to the concentration of MSM, suggesting that
MSM played a role as a hair growth promotion agent. Due
to its high sulfur content, MSM has been believed to nour-
ish skin, hair, and finger nails (Richmond, 1986). So, we
performed a preliminary experiment to see if MSM itself
has hair growth promotion effect in the concentration
range of 1-10% using mice alopecia model. The experi-
ment, however, did not show any hair growth promotion ef-
fect of MSM in the concentration range tested. These find-
ings ruled out the probable hair growth ability of MSM and
demonstrated that the observed hair growth promotion in
MAP and MSM treated mice was only due to MAP.
However, the dose-dependent enhancement in hair
growth with increasing MSM concentration implied sig-
nificant role of MSM as a penetration enhancer, which
could have exerted its additive effect on the hair growth of
MAP 7.5% solution by enhancing transdermal delivery of
MAP to hair follicles in the dermis layer of the skin.
In the present study, based on the speculation above
(enhanced transdermal delivery of MAP with the aid of
MSM), we performed transdermal penetration study of
MAP 7.5% solution in the presence of MSM (1, 5, and
10%) using Franz diffusion cell. We expected that trans-
dermal penetration of MAP would be significantly in-
creased in the presence of MSM, because MSM is struc-
turally very similar to dimethyl sulfoxide, a very strong skin
permeation enhancer (van Engelen and van Engelen,
2002). Unexpectedly, in this experiment, the transdermal
penetration of MAP was significantly decreased when
MAP was applied with MSM concentrations of 5% or
above (Fig. 5) compared to control. This unexpected result
led us take into consideration that transdermal permeation
enhancers might be subdivided into several classes. In
fact, Asbill and his colleagues have categorized the en-
hancers into those that enhance transdermal penetration
across the skin, those enhance intradermal retention into
the skin, and those enhance both (Asbill and Michniak,
2000). In fact, accumulation of drug within skin without sig-
Hair Growth Promotion of Magnesium Ascorbyl Phosphate
247
nificant systemic appearance is the objective of any means
of topical delivery (Trommer and Neubert, 2006).
Accordingly, as an attempt to substantiate the possibility
of accumulation of MAP within the skin by MSM, we meas-
ured the amount of MAP remaining in the skin (intradermal
retention) after 12 h of transdermal penetration study.
Surprisingly, we found that the amount of MAP in the skin
was profoundly and dose-proportionally increased as a
function of MSM concentration. The increase was almost
about 200-fold in the presence of MSM 10%, compared to
the result in the absence of MSM (Fig. 7). Though, the ex-
act mechanism by which MSM enhances intradermal per-
meation of MAP is unclear now, MSM might have followed
the similar mechanism of DMSO due to the structural sim-
ilarity, principally with the possession of characteristic po-
lar O-S bond, which enables the compound to interact
strongly with water by forming hydrogen bonds. The hydro-
gen-bonding ability of a substance is a major determinant
of penetration. Therefore, the hydrogen-bonding groups of
the enhancers could indeed modify the penetration
(Abraham et al., 1995; Potts and Guy, 1995). Although
DMSO is considered a strong permeation enhancer, its
permeation enhancing effect is dose-dependent and in
general, more than 60% DMSO is needed for optimal per-
meation enhancing effect. Nevertheless, at such high con-
centrations, DMSO is reported to cause erythema and
wheals of the stratum corneum and may denature skin pro-
tein (Williams and Barry, 2003).
It is worth noting that MSM possesses two polar O-S
groups (Fig. 1B) that could have efficiently bound to water
of stratum corneum forming hydrogen bonds and aided
penetration of the solutes. Probably, the same reason
could be applicable for its penetration effectiveness even
at low concentration of about 10% or less compared
DMSO. It is possible that the high hydrogen bonding ability
of MSM might have perturbed the stratum corneum suffi-
ciently for the fast entry of MAP into the skin, while the low
concentration (and subsequent reduction in concentration
of MSM, due to its high penetration rate through the skin in
to the systemic/receptor compartment) may not be suffi-
cient to enhance the penetration of huge amount of MAP
accumulated in the skin to penetrate through the skin. It is
also possible that the strong interaction of MSM with water
molecules might have resulted in less hydrophilic environ-
ment for MAP which makes its partitioning difficult leading
to accumulation rather than further penetration (Kim et al.,
1999). Also, it appears that skin accumulation effect of
MSM was exponential to the concentration of MSM in the
formula applied to the skin. However, without further
mechanistic studies, we could not understand the exact
mechanism by which MSM exerts its skin accumulation
enhancing effect.
Despite the fact that the data reported do not directly re-
veal mechanistic information on percutaneous penetration
and enhanced intradermal accumulation, they do provide a
clear correlation of effect of MSM on the hair growth pro-
motion effect of MAP in vivo and substantial intradermal
accumulation of MAP in vitro, which was the primary ob-
jective of this study. Furthermore, this finding could be uti-
lized for optimization and exploitation of intradermal drug
delivery of MAP for the treatment of alopecia.
In conclusion, the results of the present study indicate
that topical application of magnesium ascorbyl phosphate
(MAP) and methylsulfonylmethane (MSM) is effective in
promoting hair growth in C57BL/6N mice in vivo. Authors,
first time to our knowledge report that, MSM could be used
as a penetration enhancer that augments hair growth pro-
motion of MAP in vivo through higher dermal accumulation
of MAP demonstrated by in vitro skin study. The pene-
tration enhancing effect of MSM was found to be dose-de-
pendent, both in vivo and in vitro. Overall, topical applica-
tion of MAP together with MSM appears to be useful for the
treatment of alopecia.
ACKNOWLEDGMENTS
This work was supported by Korea Research Foundation
Grant funded by the Korean Government (MOEHRD, Basic
Research Promotion Fund) (KRF-2007-314-E00246).
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