Enhanced transdermal delivery of indomethacin using combination of PLGA nanoparticles and iontophoresis in vivo.

Abstract

Nanoparticles effectively deliver therapeutic agent by penetrating into the rat skin in vivo. Indomethacin (IM) and coumarin-6 were loaded in PLGA nanoparticles with an average diameter of 100 nm. Indomethacin (IM) and coumarin-6 were chosen as a model drug and as a fluorescent marker, respectively. The surfaces of the nanoparticles were negatively charged. Permeability of IM-loaded PLGA nanoparticles through rat skin was studied in vivo. Higher amount of IM was delivered through skin when IM was loaded in nanoparticles than IM was free molecules. Also, iontophoresis was applied to enhance the permeability of nanoparticles. When iontophoresis was applied at 0.05 mA/cm(2), permeability of IM was much higher than that obtained by simple diffusion of nanoparticles through skin. The combination of charged nanoparticle system with iontophoresis is useful for effective transdermal systemic delivery of therapeutic agents.

Full text

Page 1

This article appeared in a journal published by Elsevier. The attached
copy is furnished to the author for internal non-commercial research
and education use, including for instruction at the authors institution
and sharing with colleagues.
Other uses, including reproduction and distribution, or selling or
licensing copies, or posting to personal, institutional or third party
websites are prohibited.
In most cases authors are permitted to post their version of the
article (e.g. in Word or Tex form) to their personal website or
institutional repository. Authors requiring further information
regarding Elsevier’s archiving and manuscript policies are
encouraged to visit:
http://www.elsevier.com/copyright

Page 2

Author's personal copy
Colloids
and
Surfaces
B: Biointerfaces
92 (2012) 50–
54
Contents
lists
available
at SciVerse
ScienceDirect
Colloids
and
Surfaces
B:
Biointerfaces
j our
na l ho
me
p age:
www.elsevier.com/locate/colsurfb
Enhanced
nanoparticles
transdermal
delivery
of
indomethacin
using
combination
of
PLGA
and
iontophoresis
in
vivo
Keishiro
Kimiko
Tomodaa,b,c,
Hiroto
Terashimaa,
Kenichi
Suzukid,
Toshio
Inagie,
Hiroshi
Teradaa,b,c,
Makinoa,b,c,∗
aFaculty
bCenter
cCenter
dPharmaceutical
ePharmaceutical
of
Pharmaceutical
Sciences,
Tokyo
University
of
Science,
2641
Yamazaki,
Noda,
Chiba
278-8510,
Japan
for
Drug
Delivery
Research,
Faculty
of
Pharmaceutical
Sciences,
Tokyo
University
of
Science,
2641
Yamazaki,
Noda,
Chiba
278-8510,
Japan
for
Physical
Pharmaceutics,
Tokyo
University
of
Science,
2641
Yamazaki,
Noda,
Chiba
278-8510,
Japan
Division,
Kowa
Company,
Ltd.,
332-1
Ohnoshinden,
Fuji,
Shizuoka
417-8650,
Japan
Division,
Kowa
Company,
Ltd.,
4-14
Nihonbashi-honcho
3-chome,
Chuo-ku,
Tokyo
103-8433,
Japan
a
r
t
i
c
l
e

i
n
f
o
Article
Received
Received
Accepted
Available online 18 November 2011
history:
31 August
2011
in revised
form
7 November
2011
8 November
2011
Keywords:
Iontophoresis
Transdermal
PLGA
Indomethacin
nanoparticle
a
b
s
t
r
a
c
t
Nanoparticles
(IM)
Indomethacin
tively.
nanoparticles
IM
the
was
The
systemic
effectively
deliver
therapeutic
agent
by
penetrating
into
the
rat
skin
in
vivo.
Indomethacin
and
coumarin-6
were
loaded
in
PLGA
nanoparticles
with
an
average
diameter
of
100
nm.
(IM)
and
coumarin-6
were
chosen
as
a
model
drug
and
as
a
fluorescent
marker,
respec-
The
surfaces
of the
nanoparticles
were
negatively
charged.
Permeability
of IM-loaded
PLGA
through
rat
skin
was
studied
in
vivo.
Higher
amount
of
IM
was
delivered
through
skin
when
was
loaded
in
nanoparticles
than
IM
was
free
molecules.
Also,
iontophoresis
was
applied
to
enhance
permeability
of
nanoparticles.
When
iontophoresis
was
applied
at
0.05
mA/cm2,
permeability
of
IM
much
higher
than
that
obtained
by
simple
diffusion
of
nanoparticles
through
skin.
combination
of charged
nanoparticle
system
with
iontophoresis
is
useful
for
effective
transdermal
delivery
of
therapeutic
agents.
© 2011 Elsevier B.V. All rights reserved.
1.
Introduction
Skin
has
been
studied
as
administration
site
of
drug
for
its
sys-
temic
for
effect
dosage
stratum
dermal
permeability
[4–22].
In
ticles
rat
an
faces.
was
tophoresis
of
effects,
since
systemic
therapeutic
agents
can
be
delivered
long
time
with
a
controlled
ratio,
escaping
from
the
first
pass
by
liver
by
the
transdermal
delivery,
which
can
decrease
the
form
[1–3]. The
low
permeability
of
drug
molecules
through
corneum
has
been
the
limiting
factor
for
developing
trans-
delivery
system
of
therapeutic
agents.
To enhance
the
of
drug
molecules,
many
studies
have
been
reported
a previous
paper
[23], we
have
reported
that
PLGA
nanopar-
effectively
deliver
therapeutic
agent
by
penetrating
into
the
skin.
Indomethacin
(IM)
was
loaded
in PLGA
nanoparticles
with
average
diameter
of
100
nm and
with
negatively
charged
sur-
Higher
amount
of
IM was
delivered
through
skin
when
IM
loaded
in nanoparticles
than
IM was
free
molecules.
When
ion-
with
3 V/cm
was
applied
to enhance
the
permeability
nanoparticles
in vitro,
permeability
of
IM was
much
higher
than
∗Corresponding
of
fax:
E-mail
author
at:
Faculty
of
Pharmaceutical
Sciences,
Tokyo
University
Science,
2641
Yamazaki,
Noda,
Chiba
278-8510,
Japan.
Tel.:
+81
4 7121
3662;
+81
4 7121
3662.
address:
makino@rs.noda.tus.ac.jp
(K.
Makino).
that
combination
was
in vitro
In
cent
through
iontophoresis
were
plasma
obtained
by
simple
diffusion
of
nanoparticles
through
skin.
The
of
charged
nanoparticle
system
with
iontophoresis
useful
for
effective
transdermal
delivery
of
therapeutic
agents
[23].
this
study,
the
permeability
of
IM and
coumarin-6
(fluores-
marker)-loaded
PLGA
nanoparticles
with
diameter
of 100
nm
rat
abdominal
skin
was
studied
in vivo.
The
effects
of
on
permeability
of the
nanoparticles
through
skin
evaluated
by
analyzing
IM concentration
in skin,
muscle,
and
in vivo.
2.
Materials
and
methods
2.1.
Materials
Poly(lactide-co-glycolide),
the
polyvinyl
500,
purchased
6
was
Other
available.
PLGA,
having
Mw
of
10,000
with
monomer
composition
of lactic
acid/glycolic
acid
=
75/25,
alcohol
(PVA),
with
the
degree
of polymerization
of
indomethacin,
trehalose
dehydrate
and
flufenamic
acid
were
from
Wako
Pure
Chemical
Industry,
Japan.
Coumarin-
was
purchased
from
Sigma–Aldrich,
Japan.
Physiological
saline
purchased
from
Otsuka
Pharmaceutical
Factory,
Inc.,
Japan.
chemicals
were
of
the
highest
reagent
grade
commercially
0927-7765/$
doi:10.1016/j.colsurfb.2011.11.016
– see
front
matter ©

2011 Elsevier B.V. All rights reserved.

Page 3

Author's personal copy
K. Tomoda
et al.
/ Colloids
and
Surfaces
B:
Biointerfaces
92 (2012) 50–
54
51
2.2.
Preparation
of
IM-loaded
PLGA
nanoparticles
The
nanoparticles
evaporation
940
of
was
emulsified
SON
The
allow
pension
80WX,
ticles
residual
at
the
added
freeze
[23].
were
prepared
by using
emulsion
solvent
method.
Briefly,
the
amounts
of
60
mg
of
IM and
mg
of
PLGA7510
were
dissolved
in 20
mL
of
the
mixed
solution
10 mL
of
acetone
and
10
mL
of
dichloromethane.
The
solution
added
to 100
mL
of
2.0%
(w/v)
PVA
aqueous
solution
and
was
using
a probe
sonicator
(Digital
Sonifier
S-250D,
BRAN-
Co.
Ltd.)
at 200
W
of
energy
output
for
5 min
on
ice
bath.
emulsion
was
stirred
overnight
on
a magnetic
stir
plate
to
organic
solvents
to be evaporated.
The
nanoparticle
sus-
was
ultracentrifugated
at 40,000
rpm
for
30
min
(Himac
Hitachi
Koki
Co.
Ltd.).
And
then,
the
precipitated
nanopar-
were
washed
three
times
with
distilled
water
to remove
PVA.
Finally,
the
obtained
nanoparticles
were
centrifuged
10,000
rpm
for
10
min
to remove
aggregated
nanoparticles
and
supernatants
were
collected.
To the
supernatant,
trehalose
was
to be 0.05
(w/v%)
as
a cryoprotectant
and
the
suspension
was
dried
at −50◦C using
freeze
dryer
(FD-1000,
EYELA,
Japan)
2.3.
iontophoresis
Transdermal
delivery
of IM-loaded
PLGA
nanoparticles
using
Sprague-Dawley
used.
solution
64.8
square
shown
persed
added
side
were
permeation
whereas
600
resistance
MEMORY
After
600
heparinized
10
the
using
to remove
samples
Pure
cles)
study
rats
aging
8 weeks
(Japan
SLC
Co.,
Ltd.)
were
Pentobarbital
sodium
salt
was
dissolved
in saline
and
the
was
subcutaneously
injected
to the
rat
thigh
to be
mg/kg
rat
weight.
The
abdominal
skin
was
shaved
and
two
gauze
(2 cm × 2 cm)
sheets
were
placed
on
the
skin,
as
in Fig.
1.
IM-loaded
PLGA
nanoparticles
(6.4
mg)
were
redis-
in 5 mM
NaCl
solution
(500
?L)
and
the
suspension
was
on
the
cathode
(donor)
side
gauze.
To the
anode
(receptor)
gauze,
500
?L of
saline
solution
was
added.
Carbon
electrodes
used
and
iontophresis
was
carried
out
at 0.05
mA/cm2. The
study
with
iontophoresis
was
carried
out
until
360
min,
the
study
without
iontophoresis
was
carried
out
until
min.
In the
permeation
study
with
iontophoresis,
electrical
of
skin
was
measured
by
using
a data
logger
(8807/8808
HiCORDER,
Hioki
Co.,
Nagano,
Japan).
certain
time
period
(30,
60,
90,
120,
240,
360,
480,
and
min),
systemic
blood
(1 mL)
was
collected
from
heart
using
syringe
and
was
centrifuged
at 3000
rpm
at 4◦C for
min.
The
supernatant
was
collected
in heparinized
tube.
Also
skin
and
muscle
under
the
set
area
of
gauze
were
excised
by
scissors.
The
skin
was
wiped
three
times
with
fresh
gauze
nanoparticles
accumulated
on
the
surface
of
skin.
All
were
kept
at −30◦C.
IM (0.25
mg:
same
amount
of
IM loaded
in PLGA
nanoparti-
was
dissolved
in PBS
(pH
7.4,
I = 154
mM)
and
the
permeation
was
also
done
as
a control.
2.4.
Analysis
of IM concentration
in plasma
The
plasma
concentration
of
IM was
measured
using
HPLC
(SIL-20A
CTO-10ASvp,
with
Ltd.).
acid
2
internal
supernatant
solution/acetonitrile.
(pH
for
the
prominence,
SPD-20A
prominence,
LC-20AD
prominence,
DGU-20A3prominence,
SHIMADZU
co.)
at 254
nm
an
ODS
column
(Develosil
ODS-HG-5,
Nomura
Chemicals
Co.,
The
mobile
phase
was
the
mixed
solution
of
0.1
M of
acetic
aqueous
solution
and
acetonitrile
with
a volume
ratio
of
9:11.
× 10−6g of
flufenamic
acid
was
dissolved
in 1 mL
of acetonitrile
as
standard
substance
in the
measurement
using
HPLC.
The
of
blood
(0.2
mL)
was
added
to 1 mL
of
flufenamic
acid
One
millilitre
of
0.5
M
citrate
buffer
solution
5.0)
and
5 mL
of
toluene
were
added
to the
sample,
and
shaken
15
min,
the
sample
was
centrifuged
at 3000
rpm
for
10 min,
and
supernatants
(toluene
phase)
were
collected.
The
supernatants
then
added
using
1.0
were
dried
in nitrogen
atmosphere.
Mobile
phase
(0.5
mL)
was
to the
residues
and
IM in the
samples
were
quantified
by
HPLC.
HPLC
measurement
was
carried
out
at
a flow-rate
of
mL/min
and
80
?L of
samples
was
applied.
2.5.
Analysis
of
IM concentration
in skin
and
muscle
The
frozen
samples
were
broken
into
fractions
by using
a ham-
mer.
the
ples
for
atmosphere
(pH
were
for
and
to 2 mL
by
rate
(Develosil
conditions
tration
almost
was
muscle.
The
samples
were
then
added
to the
mixture
of
1 mL
of
internal
standard
solution
and
10
mL
of
methanol.
The
sam-
were
shaken
for
60 min
and
were
centrifuged
at 3000
rpm
10
min.
The
supernatants
were
collected
and
dried
in nitrogen
at 60◦C.
One
millilitre
of
0.5
M citrate
buffer
solution
5.0)
and
5 mL
of
toluene
were
added
to the
sample,
and
they
shaken
for
15
min.
The
sample
was
centrifuged
at 3000
rpm
10 min,
and
the
supernatants
(toluene
phase)
were
collected
dried
in nitrogen
atmosphere
at
60◦C.
The
residues
were
added
of
mobile
phase
and
IM in the
samples
were
quantified
using
HPLC.
HPLC
measurement
was
carried
out
at a
flow-
of
1.0
mL/min
and
80 ?L of samples
was
applied.
ODS
column
ODS-7,
Nomura
Chemicals
Co.,
Ltd.)
was
used
and
other
are
the
same
as
those
for
measuring
plasma
concen-
of
IM.
Extraction
ratio
was
also
studied
and
the
ratio
was
100%.
That
means
above
mentioned
extraction
procedure
appropriate
and
indomethacin
was
extracted
from
skin
and
2.6.
PLGA
Transdermal
delivery
route
of
IM and
coumarin-6-loaded
nanoparticles
After
2 h permeability
study,
the
cross
section
of the
skin
and
paraformaldehyde
a
Germany)
were
Germany)
and
oview
of
also
microscope.
muscle
was
observed.
The
sample
was
fixed
using
4% (w/v)
solution
and
the
sample
was
embedded
in
medium
(Jung
tissue
freezing
medium,
Leica
Instruments,
and
sections
of the
sample
with
the
width
of
100
?m
prepared
using
a
cryostat
(CM3050S,
Leica
Instruments,
at −20◦C.
The
samples
were
mounted
on
a slide
glass
were
observed
using
confocal
laser
scanning
microscope
(Flu-
FV1000,
Olympus
Co.,
Ltd.,
Japan)
at
excitation
wavelength
488
nm and
emission
wavelength
of 519
nm.
The
samples
were
stained
with
hematoxylin–eosin
(HE)
and
was
observed
using
2.7.
Statistical
analysis
Two-tailed
Student’s
t tests
were
performed
on
original
data
to
levels
assess
statistical
differences
between
the
groups.
Significance
were
set
at P < 0.05.
3.
Results
and
discussion
3.1.
nanoparticles
Characterization
of
IM and
coumarin-6-loaded
PLGA
Nanoparticles
prepared.
nanoparticles,
dure,
in
marized
are
ionized
ological
molecules
with
the
average
size
of
101.76
± 2.59
nm were
They
were
almost
spherical.
The
characteristics
of
mean
diameter,
yield
during
the
preparing
proce-
IM concentration
in nanoparticles,
coumarin-6
concentration
nanoparticles
and
zeta
potential
of
the
nanoparticles
are
sum-
in Table
1 in Ref.
[23]. The
surfaces
of the
nanoparticles
negatively
charged,
since
terminal
carboxyl
groups
of
PLGA
are
in neutral
pH [23]. The
nanoparticles
were
stable
in physi-
saline
for
8 h at 32◦C [23]. In this
preparation
method,
PVA
absorb
on
the
surfaces
of
PLGA
nanoparticles.
Hydrated

Page 4

Author's personal copy
52
K.
Tomoda
et al.
/ Colloids
and
Surfaces
B:
Biointerfaces
92 (2012) 50–
54
Fig.
1. Schematic
representation
of
animal
study.
Fig.
tration
PLGA
IM
2.
Effects
of
iontophoresis
and
nanoparticle
system
on
changes
in IM concen-
in skin
(mean
± S.D.,
n = 3) (?:
application
of
iontophoresis
on
IM-loaded
nanoparticle
suspension,
?: IM-loaded
PLGA
nanoparticle
suspension,
and
?:
solution).
PVA
[23]
layers
on
the
surfaces
work
as
a stabilizer
with
steric
hindrance
(Table
1).
3.2.
nanoparticles
In vivo
permeation
study
of
IM and
coumarin-6-loaded
PLGA
by
iontophoresis
Relationship
shown
increased
tion
in muscle
IM
until
loaded
the
not
after
that
nanoparticles
mis,
between
IM concentration
in skin
and
time
is
in Fig.
2.
In the
initial
60
min,
IM concentration
highly
by
applying
iontophoresis.
After
that,
IM concentra-
gradually
decreased
in skin,
whereas
the
IM concentration
gradually
increased
for
6 h as
shown
in Fig.
3.
Also
concentration
in plasma
apparently
increased
after
60 min
6 h,
as
shown
in Fig.
4.
From
these,
IM and
coumarin-6-
PLGA
nanoparticles
efficiently
delivered
IM to blood
by
combination
with
iontophoresis.
When
iontophoresis
was
applied,
IM was
detected
in skin,
muscle
and
plasma
6 h
from
the
start
of
the
experience,
which
were
slower
than
iontophoresis
was
applied.
IM and
coumarin-6-loaded
PLGA
are
considered
to be
delivered
to epidermis
and
der-
although
IM molecules
were
trapped
in stratum
corneum.
Also
Table
Properties
1
of
PLGA
nanoparticles
containing
both
IM and
coumarin-6.
Mean
Yield
IM
Coumarin-6
Zeta-potential
diameter
(nm)

101.76
55.44
3.93
0.0108
−2.12
± 2.59
(%)
± 1.18
concentration
in nanoparticles
(w/w%)
± 0.06
concentration
in nanoparticles
(w/w%)
± 0.000726
(mV)
± 0.31
IM and
tively
was
the
almost
and
permeability
result
was
Therefore
improved
coumarin-6-loaded
PLGA
nanoparticles
were
more
effec-
delivered
to epidermis
and
dermis
by
iontophoresis.
The
skin
kept
in a stable
condition
and
no damage
was
observed
during
experimental
time
period,
since
the
values
of
resistance
were
stable
for
6 h (15.83
±
1.04
k? at 2 h,
14.67
±
1.15
k? at
4 h
14.33
± 1.04
k? at 6 h).
Previously
we
examined
that
in vitro
of
IM solution
by applying
iontophoresis
and
the
showed
that
the
permeability
of
IM solution
through
rat
skin
not
affected
by
applying
iontophoresis
(data
are
not
shown).
the
combination
of iontophoresis
and
nanoparticles
the
delivery
of
IM beyond
skin,
as
has
been
reported
the
Fig.
tration
PLGA
IM
3. Effects
of
iontophoresis
and
nanoparticle
system
on
changes
in IM
concen-
in muscle
(mean
± S.D.,
n = 3) (?:
application
of
iontophoresis
on
IM-loaded
nanoparticle
suspension,
?:
IM-loaded
PLGA
nanoparticle
suspension,
and
?:
solution).
Fig.
tration
PLGA
IM
4. Effects
of
iontophoresis
and
nanoparticle
system
on
changes
in IM
concen-
in plasma
(mean
± S.D.,
n = 3) (?:
application
of
iontophoresis
on
IM-loaded
nanoparticle
suspension,
?:
IM-loaded
PLGA
nanoparticle
suspension,
and
?:
solution).

Page 5

Author's personal copy
K. Tomoda
et al.
/ Colloids
and
Surfaces
B:
Biointerfaces
92 (2012) 50–
54
53
Fig.
5. Microscopic
image
of
HE
stained
cross
section
of skin.
combination
ticles,
of
iontophoresis
with
liposome,
polymeric
nanopar-
solid
lipid
nanoparticles
[24–28].
3.3. Transdermal
PLGA
delivery
route
of
IM and
coumarin-6-loaded
nanoparticles
Particles
through
with
this
100
ticles.
released
ered
image
observed,
coumarin-6-loaded
sis,
was
was
hand,
applied
was
IM
twice
(Fig.
in
(Figs.
fore
smaller
than
3 ?m
are
reported
to penetrate
into
skin
transaccessory
pathway
[29–33].
Also,
PLGA
nanoparticles
the
diameter
of
200
nm are
reported
to reach
dermis
[20]. In
study,
we
have
used
PLGA
nanoparticles
with
the
diameter
of
nm.
Coumarin-6
was
selected
as
a trace
marker
of
the
nanopar-
Coumarin-6
is highly
stable
in the
nanoparticles
and
hardly
from
the
nanoparticles
[23,34]. Therefore
it was
consid-
to be
an effective
trace
marker
of
the
nanoparticles.
From
the
of
HE
stained
sample,
follicles,
epidermis
and
dermis
were
as
shown
in Fig.
5.
After
2 h permeation
study
of
IM and
PLGA
nanoparticles
by applying
iontophore-
accumulation
of
IM and
coumarin-6-loaded
PLGA
nanoparticles
observed
in follicles
and
epidermis,
although
the
fluorescence
hardly
detected
in dermis,
as
shown
in Fig.
6.
On
the
other
when
IM and
coumarin-6-loaded
PLGA
nanoparticles
were
on
the
skin
without
iontophoresis,
almost
no fluorescence
detected,
as
shown
in Fig.
7.
concentration
in skin
with
applying
iontophoresis
was
about
higher
than
that
without
applying
iontophoresis
at 2 h
2).
The
difference
of
fluorescence
intensity,
however,
detected
skin
between
with
and
without
iontophoresis
was
significant
6 and
7).
Coumarin-6
is
more
hydrophobic
than
IM,
there-
interaction
between
coumarin-6
and
PLGA
nanoparticles
is
Fig.
permeability
6. Confocal
laser
scanning
microscope
images
of
cross
sections
of
the
skin
after
study
using
iontophoresis.
Fig.
permeability
7.
Confocal
laser
scanning
microscope
images
of
cross
sections
of
the
skin
after
study
without
using
iontophoresis.
higher
tion
as
loaded
to
effectively
sis
hydrophobic
with
higher
of
ological
IM-loaded
for
From
6-loaded
epidermis,
We also
IM.
nanoparticle
was
was
the
than
that
between
IM and
PLGA
nanoparticles.
The
loca-
of
fluorescence
is regarded
as
the
location
of
nanoparticles,
mentioned
above.
From
these,
IM seems
to be
released
from
IM-
PLGA
nanoparticles
accumulated
in epidermis
and
delivered
blood
stream
and
muscle.
As
discussed
in Section
3.2, IM was
delivered
to blood
and
muscle
by
applying
iontophore-
(Figs.
3 and
4).
The
results
correspond
to our
hypothesis.
Skin
is
and
intercellular
space
between
corneocytes
is filled
lipids.
IM is hydrophobic
agent
and
affinity
of IM for
skin
is
than
that
for
physiological
saline.
Therefore
the
solubility
IM in epidermis
in vivo
should
be
different
from
that
in physi-
saline
in vitro.
We
have
shown
IM was
not
released
from
PLGA
nanoparticles
resuspended
in physiological
saline
8 h
in a previous
paper
[23].
these
results,
it was
indicated
that
IM and
coumarin-
PLGA
nanoparticles
were
accumulated
in follicles
and
and
IM seemed
to be
released
from
the
nanoparticles.
studied
the
effect
of
particle
size
on
the
permeability
of
The
results
showed
that
permeability
of IM was
not
affected
by
size
between
100
and
400
nm (data
are
not
shown).
It
considered
that
the
main
route
of
permeation
of
nanoparticles
follicle,
therefore
the
permeability
of IM was
not
affected
by
size
of
nanoparticles.
4.
Conclusions
The
combined
use
of
drug-loaded
nanoparticles
and
ion-
tophoresis
riers
to
delivery
could
offer
many
additional
benefits.
Charged
nanocar-
can
be
delivered
by
iontophoresis.
IM was
effectively
delivered
blood,
which
means
this
technique
can
be applied
to systemic
of drug
through
skin.
Acknowledgement
This
work
was
supported
by
Program
for
Development
of Strate-
gic
(2010).
Research
Center
in Private
Universities
supported
by
MEXT
References
[1] M.R.
[2] R.H.
[3]
Prausnitz,
R.
Langer,
Nat.
Biotechnol.
26 (11)
(2008)
1261–1268.
Guy,
Pharm.
Res.
13 (12)
(1996)
1765–1769.
G.T.
Grossberg,
C.
Sadowsky,
J.T.
Olin,
Int.
J. Clin.
Pract.
(2010)
1–10.

Page 6

Author's personal copy
54
K.
Tomoda
et al.
/ Colloids
and
Surfaces
B:
Biointerfaces
92 (2012) 50–
54
[4]
C.R.
Higuchi,
346–352.

Behl,

G.L.

Flynn,
Ho,

T.

Kurihara,
Pierson,

N.
Invest.

Harper,

W.

Smith,
75

W.I.

N.F.H.

C.L.

J.
Dermatol.

(1980)
[5]
[6] J.W.
[7]
[8]
99
[9] A.
299–306.
[10]
[11] B.
[12]
Pharm.
[13]
362–367.
[14] B.
(2007)
[15]
(1993) 10504–10508.
[16] H.
291–297.
[17]
922–925.
[18]
Sci.
A.C.
Williams,
B.W.
Barry,
Adv.
Drug
Deliv.
Rev.
56 (2004)
603–618.
Wiechers,
R.A.
de
Zeeuw,
Drug
Des.
Deliv.
6 (2)
(1990)
87–100.
M.
Goodman,
B.W.
Barry,
Int.
J. Pharm.
57 (1)
(1989)
29–40.
T.
Ghafourian,
P.
Zandasrar,
H.
Hamishekar,
A.
Nokhodchi,
J. Control.
Release
(2004)
113–125.
Naik,
L.A.R.M.
Pechtold,
R.O.
Potts,
R.H.
Guy,
J. Control.
Release
37
(1995)
B.J.
Drakulic,
I.O.
Juranic,
S.
Eric,
M.
Zloh,
Int.
J. Pharm.
(363)
(2008)
40–49.
Sapra,
S.
Jain,
A.K.
Tiwary,
AAPS
J. 10
(1)
(2008)
120–132.
T.
Masada,
W.I.
Higuchi,
V.
Srinivasan,
U.
Rohr,
J. Fox,
C.
Behl,
S.
Pons,
Int.
J.
49 (1)
(1989)
57–62.
B.
Mudry,
R.H.
Guy,
M.B.
Delgado-Charro,
J. Control.
Release
111
(2006)
Mudry,
P.A.
Carrupt,
R.H.
Guy,
M.B.
Delgado-Charro,
J. Control.
Release
122
165–172.
M.R.
Praunitz,
V.G.
Bose,
R.
Langer,
J.C.
Weaver,
Proc.
Natl.
Acad.
Sci.
U.S.A.
90
Ueda,
K.
Sugibayashi,
Y.
Morimoto,
J. Control.
Release
37
(1995)
S.
Henry,
D.V.
McAllister,
M.G.
Allen,
M.R.
Prausnitz,
J. Pharm.
Sci.
87 (8)
(1998)
F. Bounoure,
M.L.
Skiba,
M.
Besnard,
P.
Arnaud,
E. Mallet,
M.
Skiba,
J. Dermatol.
52 (2008)
170–177.
[19]
[20]
S.
Mitragotri,
Pharm.
Res.
17
(11)
(2004)
1354–1359.
H.
Kawashima,
Tsujimoto,
K.
Hara,
C.C.
Haung,
T.
Yokoyama,
H.
Yamamoto,
H.
Takeuchi,
Y.
K.
Akagi,
N.
Miwa,
J. Soc.
Powder
Technol.
41
(2004)
867–875.
[21]
G.
Surf. B: Biointerfaces
[22] N.
[23]
terfaces
[24] S.
[25] H.
J.
[26] J.-Y.
[27]
[28] S. Nicoli,
214 (2001)
[29] R.
58
[30] J.
Thomas,
[31] H.
ibayashi,
[32]
(1)
[33]
[34]
(1997)
Sonavane,
K.
Tomoda,
A.
Sano,
H.
Ohshima,
H.
Terada,
K.
Makino,
Colloids
65 (2008)
1–10.
Miyazaki,
A.
Takahashi,
W.
Kubo,
J. Pharm.
Pharm.
Sci.
6 (2)
(2003)
238–245.
K.
Tomoda,
H.
Terashima,
K.
Suzuki,
T.
Inagi,
K.
Makino,
Colloids
Surf.
B: Bioin-
88 (2011)
706.
Lee,
J. Lee,
Y.W.
Choi,
Biol.
Pharm.
Bull.
30
(2)
(2007)
393–396.
Chen,
H.
Zhu,
J. Zheng,
D.
Mou,
J. Wan,
J.
Zhang,
T.
Shi,
Y.
Zhao,
H.
Xu,
X. Yang,
Control.
Release
139
(2009)
63–72.
Fang,
K.C.
Sung,
H.-H.
Lin,
C.-L.
Fang,
J.
Control.
Release
60
(1999)
1–10.
W.
Liu,
M.
Hu,
W.
Liu,
C.
Xue,
H.
Xu,
X.L.
Yang,
Int.
J. Pharm.
364
(2008)
135–141.
P. Santi,
P. Couvreur,
G.
Couarraze,
P.
Colombo,
E.
Fattal,
Int.
J. Pharm.
31–35.
Alvarez-Román,
A.
Naik,
Y.N.
Kalia,
H.
Fessi,
R.H.
Guy,
Eur.
J.
Pharm.
Biopharm.
(2004)
301–316.
Lademann,
N.
Otberg,
H.
Richter,
L.
Meyer,
H.
Audring,
A.
Teichmann,
S.
A.
Knüttel,
W.
Sterry,
Skin
Res.
Technol.
13 (2)
(2007)
119–132.
Todo,
E.
Kimura,
H.
Yasuno,
Y.
Tokudome,
F.
Hashimoto,
Y.
Ikarashi,
K.
Sug-
Biol.
Pharm.
Bull.
33 (2010)
1394–1399.
R.
Alvarez-Román,
A.
Naik,
Y.N.
Kalia,
R.H.
Guy,
H.
Fessi,
J.
Control.
Release
99
(2004)
53–62.
X.
Wu,
B.
Biatry,
C.
Cazeneuve,
R.H.
Guy,
Pharm.
Res.
26 (8)
(2009)
1995–2001.
M.P.
Desai,
V.
Labhasetwar,
E.
Walter,
R.J.
Levy,
G.L.
Amidon,
Pharm.
Res.
14
1568.