CHAPTER

Nanoparticles for
transdermal drug delivery
system (TDDS)
,{,{
7 ,{,{
Keishiro Tomoda* , Kimiko Makino*
*
Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki Noda,
Chiba 278-8510, Japan
{
Center for Drug Delivery Research, Faculty of Pharmaceutical Sciences, Tokyo University
of Science, 2641 Yamazaki Noda, Chiba 278-8510, Japan
{
Center for Physical Pharmaceutics, Tokyo University of Science, 2641 Yamazaki Noda,
Chiba 278-8510, Japan

CHAPTER CONTENTS
7.1 Introduction ....................................................................................................131
7.2 Nanoparticles For Transdermal Drug Delivery ....................................................134
7.3 Combination of Nanoparticle System and IP ......................................................134
7.4 Improved Nanoparticles for Iontophoretic Transdermal Drug Delivery .................140
7.5 Conclusions ....................................................................................................145
References ............................................................................................................145

7.1 INTRODUCTION
Drug delivery system (DDS) is one form of drug formulation research to develop
innovative formulation and application methods for improving drug specificity by
modifying applicable material to a therapeutic agent. Conventional administration
routes such as intravenous or oral administration cannot deliver enough drug mole-
cules to the target site, whereas almost all drugs are excreted or accumulate in unspe-
cific sites where the drug expresses side effects. Therefore induction of spatial
control is needed to deliver therapeutic agents to specific target sites to increase drug
efficiency as well as to decrease side effects. This concept is also applied to cosmetic
and therapeutic agents as transdermal drug delivery systems (TDDSs). TDDS is ben-
eficial for systemic delivery and local delivery of therapeutic agents with simple
administration, available outside medical institutions, decreased burden to patients
caused by intravenous administration, escape from the first pass effect by liver, deliv-
ery of therapeutic drugs with controlled ratio and ease of stopping administration [1].
Scopolamine formulation for the treatment of motion sickness, nitroglycerin
Colloid and Interface Science in Pharmaceutical Research and Development. http://dx.doi.org/10.1016/B978-0-444-62614-1.00007-7
© 2014 Elsevier B.V. All rights reserved.
131
132 CHAPTER 7 Nanoparticles for transdermal drug delivery system (TDDS)

formulation for the treatment of angina pectoris, nicotine formulation for the treat-
ment of smoking cessation and fentanyl formulation for the treatment of pain have
been developed as transdermal formulations [2]. Recently, rivastigmine transdermal
patch was noted for the treatment of Alzheimer’s disease [3]. Transdermal formula-
tions currently approved by the U.S. Food and Drug Administration (FDA) are
shown in Table 7.1 [4]. Development of transdermal delivery formulation, however,
has a critical problem to exert the efficiency of therapeutic agents. Skin has stratum

Table 7.1 Transdermal formulations approved by U.S. FDA

Approval
Drug Company Dosage form year

Scopolamine/Transderm Scop® Novartis Film, extended 31

release December
1979
Nitroglycerin/Nitrol® Rorer Film, extended 19 January
release 1983
Clonidine/Catapres-TTS® Boehringer Film, extended 10 October
Ingelheim release 1984
Estradio/Estraderm® Novartis Film, extended 10
release September
1986
Fentany/Duragesic® Ortho Film, extended 7 August
McNeil release 1990
Janssen
Nicotine/Habitrol® Novartis Film, extended 27
release November
1991
Nicotine/NicoDerm CQ® Sanofi Film, extended 7
aventis release November
1991
Nicotine/ProStep® Aveva Film, extended 28 January
release 1992
Testosterone/Testosterone® Alza Gel 12 October
1993
Testosterone/Androderm® Wastson Film, extended 29
Labs release September
1995
Epinephrine/lidocaine Iomed Systern; 21
hydrochloride/Iontocaine® iontophoresis December
1995
Estradio/norethindrone acetate/ Novartis Film, extended 7 August
CombiPatcj® release 1998
Lidocaine hydrochloride/ Teikoku Patch 19 March
Lidoderm® Pharma 1999
USA

Continued
 7.1 Introduction 133

Table 7.1 Transdermal formulations approved by U.S. FDA—cont’d

Approval
Drug Company Dosage form year
Ethinyl estradiol/norelgestromin/ Ortho Film, extended 20
Ortho Evra® McNeil release November
Janssen 2001
Oxybutynin/Oxytrol® Watson Film, extended 26 February
Labs release 2003
Selegiline/Emsam® Somerset Film, extended 27 February
release 2006
Methyphenidate/Daytrana® Shire DOT matrix 6 April 2006
Epinephrine/lidocaine Vyteris System; 6 May 2004
hydrochloride/LidoSite Topical iontophoresis
System®
Fentanyl hydrochloride/Ionsys® Alza System; 22 May
iontophoresis 2006
Rotigotine/Neupro® Schwarz Film, extended 9 May 2007
Pharma release
Rivastigmine/Exelon® Novartis Film, extended 6 July 2007
release
Lidocaine hydrochloride/Zingo® Anesiva Powder Ject 16 August
2007
Diclofenac epolamine/Flector® Inst Hydrogel 31 January
Biocherm 2007

corneum in the outermost layer, which has a thick layer and high hydrophobicity;
therefore, the stratum corneum stands as a physical barrier for penetration of sub-
stances. The epidermal layer exists below the stratum corneum. The epidermal layer
has hydrophilicity and numerous types of enzymes. Therefore, the epidermal layer
stands as a chemical barrier. Many techniques were evaluated to enhance the perme-
ability of drug molecules. For instance, occlusive dressing technique, which hydrates
stratum corneum, is the simplest method [5]. As chemical modifications, penetration
enhancers [6], dimethylsulfoxide (DMSO) or Azone [7, 8], N-methyl-2-pyrrolidone
(NMP) [9], oleic acid [8, 10], ethanol or propylene glycol, menthol, limonene or 1,8-
cineole [9, 11, 12] have been developed. As physical modifications, iontophoresis
(IP), which promotes skin permeability using electropotential energy [13–15]; elec-
troporation, by producing small pores on the surface of stratum corneum by adding
pulse voltage [16]; sonophoresis, using the effect of cavitation of ultrasonic wave
[17]; and microneedle, by producing new permeation routes [18], have been studied.
The combination of physical and chemical enhancement techniques has been studied
[19, 20]. The use of both chemical enhancers and physical enhancers, however, needs
high dose or high potency, which induces irritation, causes damage and reduces the
skin barrier function. Therefore, it is desirable to deliver therapeutic agents across the
134 CHAPTER 7 Nanoparticles for transdermal drug delivery system (TDDS)

stratum corneum keeping the normal skin barrier function without the aid of chem-
ical or physical enhancers. For this purpose, drug-loaded nanoparticles have obtained
increasing attention.

7.2 NANOPARTICLES FOR TRANSDERMAL DRUG DELIVERY

Both chemical modification and physical modification have advantages and disad-
vantages for application of transdermal formulation. In recent years extensive studies
have been carried out on the delivery of therapeutic agents and enhancement of
absorption through skin, maintaining the homeostasis of skin. Nano-sized colloidal
particles, such as micelles, liposomes, and solid-lipid, polymeric and inorganic nano-
particles have been developed as carriers to enhance percutaneous absorption of ther-
apeutic agents [21–28]. Encapsulation of therapeutic agents into colloidal carriers
enables the agents not only to enhance the permeability without skin denaturation
but also to protect the agents from degradation and to control the release ratio of
the agents from the colloidal carrier [23].
The penetration pathways of substances into and through the skin are separated to
intracellular pathways, intercellular pathways and trans-appendicular pathways [21].
Trans-appendicular pathways include hair follicles and sweat glands. Intercellular
pathways are considered to be the most probable way, but substances whose molec-
ular weights are more than 500 and ionic substances accumulate in appendicular
organs because these substances seldom pass through the stratum corneum [29].
The main penetration pathway of nanoparticles is reported to be the trans-
appendicular pathway [30]. Whereas the effect of trans-appendicular pathways on
the penetration rate of substances was considered to be negligible, because appen-
dicular organs exist only 0.1% in cutaneous surfaces, they are revaluated for their
ability to reach deep into thick stratum corneum, as they have many capillary vessel
and neurocyte distributions near their roots and function as a reservoir of substances
themselves [31]. Submicron-sized particles are reported to penetrate through folli-
cles more effectively than micro-sized particles [32]. The effects of particle size
of gold nanoparticles on the permeability of the nanoparticles through rat skin have
been evaluated. The result confirmed that gold nanoparticles <100 nm effectively
permeate through rat skin ex vivo (Figure 7.1) [33]. From these insights, nanoparti-
cles <100 nm in size are considered to be beneficial for transdermal drug delivery.

7.3 COMBINATION OF NANOPARTICLE SYSTEM AND IP

Colloidal nanoparticles have charges and/or a stabiliser layer on the surfaces to keep
the stability in the medium by electrostatic repulsion or steric hindrance of surface-
modified stabiliser between particles. IP was applied to the colloidal nanoparticles
for transdermal delivery by utilising the charge property. IP is normally applied
for ionic and water-soluble agents. Although non-ionic agents are also reported to
 7.3 Combination of nanoparticle system and IP 135

10

Amounts of gold permeated through

8

rat skin (µg)

6

0
0 1 2 3 4 6 10 24
Time (h)
: 15 nm : 100 nm : 200 nm

FIGURE 7.1
Amounts of gold permeated through rat skin (mean  S.D., n ¼ 3).

penetrate through skin by electro-osmosis fluid, the contribution to the permeability

is minimal. Moreover the IP reportedly enables only low-molecule substances to be
permeable. On the other hand, encapsulation of substances into charged nanoparti-
cles is applicable to various substances, irrespective of molecular weight and
charges.
Indomethacin was selected as an ionic model drug, poly (lactide-co-glycolide)
(PLGA) as a carrier and polyvinyl alcohol (PVA) as a stabiliser. PVA-coated
indomethacin-loaded PLGA nanoparticles of 100 nm were prepared by emulsifica-
tion with solvent evaporation method. PLGA has terminal carboxyl group in the mol-
ecules that have de-protonation and negative charge above neutral pH [34]. The
indomethacin-loaded PLGA nanoparticles were suspended in 5 mM NaCl solution,
and the permeability of indomethacin through rat skin was studied ex vivo and
in vivo. Significantly higher amounts of indomethacin were permeated through rat
skin when indomethacin was loaded in PLGA nanoparticles than when indomethacin
was free molecules. Application of IP enhanced the permeability of indomethacin
about five times higher than that obtained by passive diffusion (Figure 7.2) [35].
Indomethacin-loaded PLGA nanoparticles were suspended in 5 mM NaCl solu-
tion. The abdominal skin of rats was shaved, and two square gauze (2
2 cm) sheets
were placed on the shaved skin spot. Indomethacin-loaded PLGA nanoparticle sus-
pension was infused on the donor side gauze, and the same volume of 5 mM NaCl
solution was infused to the receptor side. Translatability to circulation, accumulative
ability inside skin and muscle accumulation under the set area of gauze were eval-
uated. Transition to circulation, accumulation inside skin and accumulation in mus-
cle were hardly detected when indomethacin solution in 5 mM NaCl solution was
136 CHAPTER 7 Nanoparticles for transdermal drug delivery system (TDDS)

12

Amounts of indomethacin permeated

through rat skin (µg/cm2)
10

0 2 4 6 8
Time (h)
FIGURE 7.2
Effect of the combination of nanoparticle system with iontophoresis on the permeability of
indomethacin through rat skin ex vivo ( , application of iontophoresis (3 V/cm) on
indomethacin-loaded PLGA nanoparticle suspension; ○, indomethacin-loaded PLGA
•
nanoparticle suspension; □, free molecule of indomethacin) (mean  S.D., n ¼ 3).

200
Indomethacin concentration

5 300

Indomethacin concentration
Indomethacin concentration

250

in plasma (ng/mL)
in skin (×104 ng/g)

4
in muscle (ng/g)

150
200
3
100 150
2
100
50
1 50

0 100 200 300 400 500 600 0 100 200 300 400 500 600 0 100 200 300 400 500 600
Time (min) Time (min) Time (min)
(a) (b) (c)
FIGURE 7.3
Effect of the combination of nanoparticle system with iontophoresis on the permeability of
indomethacin through rat skin in vivo. (a) Amounts of indomethacin accumulation in skin.
(b) Amounts of indomethacin accumulation in muscle. (c) Indomethacin concentration in
•
plasma ( , application of iontophoresis (3 V/cm) on indomethacin-loaded PLGA nanoparticle
suspension; ○, indomethacin-loaded PLGA nanoparticle suspension; □, free molecule of
indomethacin) (mean  S.D., n ¼ 3).

applied. Meanwhile, it was detected 6 h after the experiment started when

indomethacin-loaded nanoparticle suspension was applied (Figure 7.3).
Indomethacin in molecule state seems to remain only in stratum corneum because
of the high hydrophobicity, whereas indomethacin-loaded PLGA nanoparticles
can penetrate through stratum corneum via the trans-appendicular route and transit
to circulation and muscle. When IP was applied, accumulation of indomethacin in
skin significantly increased in the initial 60 min. Subsequently, the concentration
 7.3 Combination of nanoparticle system and IP 137

of indomethacin in skin gradually decreased, and the accumulation of indomethacin

in the muscle gradually increased. Indomethacin concentration in plasma was also
apparently increased by application of IP.
To estimate the transdermal delivery route of indomethacin-loaded PLGA nano-
particles, coumarin 6 was encapsulated with indomethacin into PLGA nanoparticles
as a fluorescent trace maker. After the permeability study was carried out, a vertical
section of the skin was excised and observed by using a confocal laser scanning
microscope. The obtained images revealed that the nanoparticles mainly accumu-
lated in follicles; therefore the nanoparticles delivered indomethacin via trans-
appendicular pathway to muscle and circulation (Figure 7.4) [36].
Indomethacin is an ionic therapeutic agent. The permeability of estradiol (non-
ionic therapeutic agent) has been studied by the combination of charged nanoparticle
system and IP. Estradiol-loaded PLGA nanoparticles were suspended in 5 mM phos-
phate buffer, and the permeability of estradiol through rat skin was studied ex vivo
and in vivo. In a similar way of indomethacin, the permeability of estradiol through
rat skin ex vivo increased by encapsulation of estradiol into PLGA nanoparticles. The
enhancement effect was also accelerated by applying IP, although free molecules of
estradiol were not affected by applying IP (Figure 7.5).

FIGURE 7.4
Fluorescent image of vertical section of skin 6 h after the permeability study started by the
combination of nanoparticle system with iontophoresis in vivo.
138 CHAPTER 7 Nanoparticles for transdermal drug delivery system (TDDS)

permeated through rat skin (ng/cm2)

200

Amounts of estradiol
150

100

50

0 2 4 6 8
Time (h)
FIGURE 7.5
Effect of the combination of nanoparticle system with iontophoresis on the permeability of
•
estradiol through rat skin ex vivo ( , application of iontophoresis (3 V/cm) on estradiol-loaded
PLGA nanoparticle suspension; ○, estradiol-loaded PLGA nanoparticle suspension; n,
application of iontophoresis (3 V/cm) on free molecule of estradiol; □, free molecule of
estradiol suspension) (mean  S.D., n ¼ 3).

From an in vivo experiment, it was shown that estradiol concentration in muscle

significantly increased by applying estradiol-loaded PLGA nanoparticles compared
to free estradiol. The enhancement effect was accelerated by applying IP. This phe-
nomenon was significant 3 h after the permeation study started, but the values of
estradiol concentration in muscle were almost constant between 3 and 9 h. The con-
centration of estradiol in muscle seems to be saturated within 3 h; subsequently estra-
diol would penetrate to other organs and blood. Estradiol concentration in plasma
was significantly higher when estradiol-loaded PLGA nanoparticles were applied
than when the estradiol molecule itself was applied. When IP was not applied, the
estradiol concentration in plasma was almost constant between 3 and 9 h after the
permeation study started (Figure 7.6). These results suggested that estradiol first
migrated to muscle and subsequently penetrated into plasma.
Transdermal delivery route of estradiol-loaded PLGA nanoparticles was evalu-
ated by encapsulating coumarin 6. The results showed that estradiol and coumarin 6-
loaded PLGA nanoparticle accumulation was observed in stratum corneum and
barely in follicles after the permeation study by passive diffusion. When estradiol
and coumarin 6-loaded PLGA nanoparticles were applied on the skin with IP, much
stronger fluorescence of coumarin 6 was observed in follicles and stratum corneum
than that without IP (Figure 7.7). The main permeation route of estradiol and cou-
marin 6-loaded PLGA nanoparticles was follicles.
The fluorescence was, however, detected only in follicles and was not observed in
deeper skin. The data indicates that estradiol and coumarin 6-loaded PLGA nanopar-
ticles exist only in follicles. On the other hand, estradiol was detected in plasma and
 7.3 Combination of nanoparticle system and IP 139

200 10

Estradiol concentration
Estradiol concentration 8

in plasma (ng/mL)
in muscle (ng/g)
150
6
100
4

50 2

0 3 6 9 0 3 6 9
Time (h) Time (h)
(a) (b)
FIGURE 7.6
Effect of the combination of nanoparticle system with iontophoresis on the permeability of
estradiol through rat skin in vivo. (a) Amounts of indomethacin accumulation in muscle.
•
(b) Estradiol concentration in plasma ( , application of iontophoresis (3 V/cm) on
estradiol-loaded PLGA nanoparticle suspension; ○, estradiol-loaded PLGA nanoparticle
suspension; □, free molecule of estradiol in PBS) (mean  S.D., n ¼ 3).

100 mm 100 mm

(a) (b)
FIGURE 7.7
Fluorescent images of vertical section of skin after 8 h of permeability study ex vivo. (a)
Application of iontophoresis on estradiol-loaded PLGA nanoparticles. (b) Estradiol-loaded
PLGA nanoparticles.

muscle. Estradiol, a hydrophobic agent, would have higher affinity with skin than
donor phase because skin is more hydrophobic than donor phase. Estradiol seems
to be released from the nanoparticles after accumulating in follicles and to migrate
in muscle and bloodstream. IP enhanced accumulation of the nanoparticles in
follicles; therefore, accumulation of estradiol in muscle and blood would be
enhanced [37].
140 CHAPTER 7 Nanoparticles for transdermal drug delivery system (TDDS)

7.4 IMPROVED NANOPARTICLES FOR IONTOPHORETIC

TRANSDERMAL DRUG DELIVERY
The combination of charged nanoparticles system with IP is beneficial for transder-
mal drug delivery. The commonly used preparation method of polymeric nanopar-
ticles needs stabiliser. The stabiliser adsorbs on the surface of polymeric
nanoparticles and then generates an electric shielding effect of the charged nanopar-
ticles. That causes diminution of the enhancement effect of IP in the case of trans-
dermal delivery of nanoparticles. Indomethacin-loaded PLGA nanoparticles were
prepared by using a combination of anti-solvent diffusion (nanoprecipitation)
method [38] with selective solvation phenomenon [39, 40]. The anti-solvent diffu-
sion method is based on interfacial deposition of polymer following displacement of
polymer-dissolved solvent (Marangoni effect) [38]. The procedure is very simple,
mixing polymer-dissolved solvent with non-solvent (solvent needs miscible nature
to non-solvent). Selective solvation phenomenon is based on polymer-solvent–
non-solvent interaction. When the polymer has polar groups, selective solvation
occurs if Lewis base (electron pair donor) is selected as a solvent and Lewis acid
(electron pair acceptor) is selected as a non-solvent. Electrostatic interaction between
polymer and solvent should be stronger than that between solvent and non-solvent.
When the conditions are proper, polymer nanoparticles are separated out and are cov-
ered with solvent shell by selective solvation phenomenon. These conditions form
the polymer molecules in the selective solvation shell to be further solvated by
non-solvent molecules and then form a complete stabilisation chain [40]. If the
applied solvent and non-solvent are strong Lewis base and acid, respectively, smaller
and more stable polymeric nanoparticles can be obtained [40]. NMP, a strong Lewis
base, as a solvent and distilled water, a strong Lewis acid in this condition, as a non-
solvent were selected [41]. Indomethacin and PLGA were dissolved in NMP, and the
solvent solution was dropped in distilled water. Indomethacin-loaded PLGA nano-
particles were separated out spontaneously by Marangoni effect and kept in stable
condition by preferential solvation phenomenon (Figure 7.8). The indomethacin-
loaded PLGA nanoparticles prepared by anti-solvent diffusion with preferential sol-
vation method are referred to as non-coated nanoparticles. Alternatively, the
indomethacin-loaded PLGA nanoparticles prepared by emulsification with solvent
evaporation (the nanoparticles are covered with PVA) are referred as PVA-coated
nanoparticles.
Non-coated and PVA-coated indomethacin-loaded PLGA nanoparticles were
prepared at an average size of 100 nm. Table 7.2 shows the physicochemical prop-
erties of non-coated and PVA-coated nanoparticles.
Electrophoretic mobilities, the migration rate of colloids per unit electrical field
(mm/s cm/V), of non-coated nanoparticles calculated by Ohshima’s equation [42]
showed more negative values than those of PVA-coated nanoparticles in phosphate
buffer (pH 7.4; Figure 7.9).
Electrophoretic mobility can be regarded as the indicator of sensibility of colloids
against applied electrical fields; therefore, non-coated nanoparticles are supposed to
7.4 Improved nanoparticles for iontophoretic transdermal drug delivery 141

Dissolved
substances
(indomethacin and PLGA)

Anti-solvent diffusion

Preferential solvation
Solvent
(NMP) Molecular interaction
PLGA NMP NMP H2O

Anti-solvent Stable suspension

(H2O) : NMP molecule

FIGURE 7.8
Schematic representation of the method using anti-solvent diffusion with preferential
salvation.

Table 7.2 Properties of non-coated nanoparticles and PVA-coated

nanoparticles
Non-coated PVA-coated
nanoparticles nanoparticles

Particle diameter (nm) 100.00  0.92 102.07  1.36

Polydispersity index 0.130  0.013 0.087  0.025
Indomethacin content in 4.16  0.05 4.25  0.09
nanoparticles (w/w%)
Indomethacin encapsulation 51.1  4.00 36.4  2.97
efficiency (w/w%)
Recovery of nanoparticles (w/w%) 71.1  5.81 51.1  3.36
Electrophoretic mobility at I ¼ 5 mM 10.23  0.96 0.15  0.05
(mm/s cm/V)
Electrophoretic mobility at 5.89  0.24 0.10  0.00
I ¼ 154 mM (mm/s cm/V)
Particle softness parameter (1/l) (nm) 3.21 9.31
Particle surface charge density (mM) 47.57 0.23

have more sensibility against IP. Particle softness parameter and surface charge num-
ber density were also determined by a curve-fitting procedure reported previously
[43, 44]. Particle softness parameter of non-coated nanoparticles was about three
times lower than that of PVA-coated nanoparticles, meaning that non-coated nano-
particles were more rigid than PVA-coated nanoparticles because non-coated nano-
particles do not have a surface polymer layer (PVA). Particle surface negative charge
142 CHAPTER 7 Nanoparticles for transdermal drug delivery system (TDDS)

Ionic strength (M) Ionic strength (M)

12
14
16
02
04
06
08
10
02
04
06
08
10
12
14
16

0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0 0

Electrophoretic mobility
Electrophoretic mobility
–2
–0.05

(µm/s·cm/V)
(µm/s·cm/V)
–4

–6 –0.10

–8
–0.15
–10

–12 –0.20
(a) (b)
FIGURE 7.9
Electrophoretic mobility of indomethacin-loaded PLGA nanoparticles prepared by (a) anti-
solvent diffusion with preferential solvation and (b) emulsification with solvent evaporation at
in phosphate buffer with various ionic strengths at 32  C (mean  S.D., n ¼ 3).

: NMP molecule shell : PVA molecule

: Indomethacin

(a) (b)
FIGURE 7.10
Conceptual design of indomethacin-loaded PLGA nanoparticles prepared by (a) anti-solvent
diffusion with preferential solvation and (b) emulsification with solvent evaporation.

number density of non-coated nanoparticles was about 200 times higher than that of
PVA-coated nanoparticles (Table 7.2). PLGA molecules have negative charges at
neutral pH because of their ionised terminal carboxyl groups [34]. Anti-solvent dif-
fusion with preferential solvation method enables preparation of nanoparticles while
maintaining high charge density derived from the terminal group of PLGA molecule
(Figure 7.10).
Permeability of indomethacin through rat skin by using non-coated and PVA-
coated nanoparticles with or without IP were evaluated ex vivo and in vivo [45].
In the case of passive diffusion, the permeability of indomethacin through rat skin
7.4 Improved nanoparticles for iontophoretic transdermal drug delivery 143

permeated through rat skin (µg/cm2)

35

Amounts of indomethacin
30
25 *
20
15
**
10
*
5

0 2 4 6 8
Time (h)
FIGURE 7.11
Amounts of indomethacin permeated through rat skin (mean  S.D., n ¼ 3). ○, Application of
iontophoresis on non-coated indomethacin-loaded PLGA nanoparticles suspension; , non-
coated indomethacin-loaded PLGA nanoparticles suspension; □, application of iontophoresis
•
on coated indomethacin-loaded PLGA nanoparticles suspension; n, PVA-coated
indomethacin-loaded PLGA nanoparticles suspension (*p < 0.005, **p < 0.05).

of non-coated nanoparticles was lower than that of PVA-coated nanoparticles. On the

other hand, when IP was applied, the permeability of indomethacin through rat
skin significantly increased, especially in the case of non-coated nanoparticles
(Figure 7.11).
Non-coated nanoparticles seem to be more hydrophobic than PVA-coated nano-
particles because non-coated nanoparticles do not have a coating layer of PVA [35,
42]. That causes high affinity towards the stratum corneum. Therefore non-coated
nanoparticles accumulated higher in the stratum corneum. However, the non-coated
nanoparticles were difficult to pass through the stratum corneum because of the high
hydrophobicity. Non-coated nanoparticles can be affected by IP more effectively
than PVA-coated nanoparticles (Table 7.2) and could more effectively pass through
the stratum corneum by applying IP. Accumulation amounts of indomethacin were
significantly higher when non-coated nanoparticles were used than when PVA-
coated nanoparticles were used (Figure 7.12).
It was suggested that IP increases the transfer of indomethacin from skin to recep-
tor phase. Procedure of transdermal permeation of therapeutic agents consists of six
parts as follows: (1) diffusion of agents in the substrate, (2) partition of agents from
substrate to stratum corneum, (3) diffusion of agents in the stratum corneum, (4) par-
tition of agents from stratum corneum to epidermis and dermis, (5) diffusion of
agents in the epidermis and dermis and (6) immigration of agents from epidermis
and dermis to circulation (circulation replaces receptor phase in ex vivo experiment).
In this procedure, parts 2–4 would be enhanced by using non-coated nanoparticles
because hydrophobicity was higher than PVA-coated nanoparticles. Also, parts 1,
5 and 6 would be enhanced by applying IP. These combined effects seem to enhance
144 CHAPTER 7 Nanoparticles for transdermal drug delivery system (TDDS)

250

Indomethacin amounts inside skin

after permeation study (µg)
200

150

100

50

0
(a) (b) (c) (d)
FIGURE 7.12
Cumulative amounts of indomethacin inside skin after 8 h of permeation study (mean  S.D.,
n ¼ 3). (a) Non-coated nanoparticles applied with iontophoresis. (b) Non-coated
nanoparticles without iontophoresis. (c) PVA-coated nanoparticles applied with
iontophoresis. (d) PVA-coated nanoparticles without iontophoresis.
Indomethacin concentration

4 Indomethacin concentration
0.5
in plasma (µg/mL)
0.4
in muscle (µg/g)

3
0.3 *
2 ** *
0.2
1 **
0.1

0 1 2 3 4 5 6 0 1 2 3 4 5 6
Time (h) Time (h)
(a) (b)
FIGURE 7.13
Effect of the combination of nanoparticle system with iontophoresis on the permeability of
indomethacin through rat skin in vivo. (a) Amounts of indomethacin accumulation in muscle.
(b) Indomethacin concentration in plasma (○, application of iontophoresis on non-coated
indomethacin-loaded PLGA nanoparticles suspension; , non-coated indomethacin-loaded
PLGA nanoparticles suspension; □, application of iontophoresis on PVA-coated
•
indomethacin-loaded PLGA nanoparticles suspension; n, PVA-coated indomethacin-loaded
PLGA nanoparticles suspension (*p < 0.0005, **p < 0.005, ***p < 0.01).

the permeability of indomethacin through rat skin significantly by the combination

of non-coated nanoparticles with IP. The effect of the nanoparticle system and IP on
transition to circulation of indomethacin and accumulation in muscle through rat skin
in vivo is shown in Figure 7.13.
Transition to circulation of indomethacin by using non-coated nanoparticles and
IP was about five times higher than that by using PVA-coated nanoparticles and IP.
 References 145

Effectiveness of combination of nanoparticle system with IP for the delivery of drugs

through the skin have reported the combination of IP with liposome, unilamellar ves-
icles, polymeric nanoparticles and solid-lipid nanoparticles [46–49]. Hydrophobicity
of the nanoparticles has substantial contribution for accumulation of drugs to skin
and muscle through skin. Also, high-charged nanoparticles are more suitable for ion-
tophoretic drug delivery.

7.5 CONCLUSIONS
Nanoparticles have been gaining attention for a transdermal drug delivery carrier.
Indomethacin and estradiol permeability through rat skin were enhanced by encap-
sulating into PLGA nanoparticles. Application of IP to nanoparticle system enhanced
accumulation of the nanoparticles into follicles. High-charged indomethacin-loaded
PLGA nanoparticles were prepared by anti-solvent diffusion with preferential solva-
tion method. The nanoparticles showed high electrophoretic mobility. Highly
charged indomethacin-loaded PLGA nanoparticles showed greater permeability of
indomethacin through rat skin than PVA-coated indomethacin-loaded PLGA nano-
particles when IP was applied. The highly charged indomethacin-loaded PLGA
showed greater accumulation inside skin than PVA-coated indomethacin-loaded
PLGA nanoparticles because highly charged nanoparticles have higher hydrophobic-
ity than PVA-coated indomethacin-loaded PLGA nanoparticles. These results sug-
gested that highly charged nanoparticles have great potential for iontophoretic
TDDS.

REFERENCES
[1] M.R. Prausnitz, R. Langer, Nat. Biotechnol. 26 (11) (2008) 1261–1268.
[2] R.H. Guy, Pharm. Res. 13 (12) (1996) 1765–1769.
[3] G.T. Grossberg, C. Sadowsky, J.T. Olin, Int. J. Clin. Pract. 64 (5) (2010) 651–660.
[4] FDA website http://www.accessdata.fda.gov/Scripts/cder/Drugsatfda.
[5] C.R. Behl, G.L. Flynn, T. Kurihara, N. Harper, W. Smith, W.I. Higuchi, et al., J. Invest.
Dermatol. 75 (1980) 346–352.
[6] A.C. Williams, B.W. Barry, Adv. Drug Deliv. Rev. 56 (5) (2004) 603–618.
[7] J.W. Wiechers, R.A. Zeeuw, Drug Des. Deliv. 6 (2) (1990) 87–100.
[8] M. Goodman, B.W. Barry, Int. J. Pharm. 57 (1) (1989) 29–40.
[9] T. Ghafourian, P. Zandasrar, H. Hamishekar, A. Nokhodchi, J. Control. Release
99 (2004) 113–125.
[10] A. Naik, L.A.R. Pechtold, R.O. Potts, R.H. Guy, J. Control. Release 37 (3) (1995)
299–306.
[11] B.J. Drakulic, I.O. Juranic, S. Eric, M. Zloh, Int. J. Pharm. 363 (2008) 40–49.
[12] B. Sapra, S. Jain, A.K. Tiwary, AAPS J. 10 (1) (2008) 120–132.
[13] T. Masada, W.I. Higuchi, V. Srinivasan, U. Rohr, J. Fox, C. Behl, et al., Int. J. Pharm.
49 (1) (1989) 57–62.
146 CHAPTER 7 Nanoparticles for transdermal drug delivery system (TDDS)

[14] B. Mudry, R.H. Guy, M.B. Delgado-Charro, J. Control. Release 111 (2006) 362–367.
[15] B. Mudry, P.A. Carrupt, R.H. Guy, M.B. Delgado-Charro, J. Control. Release 122 (2007)
165–172.
[16] M.R. Praunitz, V.G. Bose, R. Langer, J.C. Weaver, Proc. Natl. Acad. Sci. U. S. A.
90 (1993) 10504–10508.
[17] H. Ueda, K. Sugibayashi, Y. Morimoto, J. Control. Release 37 (1995) 291–297.
[18] S. Henry, D.V. McAllister, M.G. Allen, M.R. Prausnitz, J. Pharm. Sci. 87 (8) (1998)
922–925.
[19] F. Bounoure, M.L. Skiba, M. Besnard, P. Arnaud, E. Mallet, M. Skiba, J. Dermatol. Sci.
52 (2008) 170–177.
[20] S. Mitragotri, Pharm. Res. 17 (11) (2000) 1354–1359.
[21] G. Cevc, U. Vierl, J. Control. Release 141 (2010) 277–299.
[22] H. Tsujimoto, K. Hara, C.C. Haung, T. Yokoyama, H. Yamamoto, H. Takeuchi, et al., J.
Soc. Powder Technol. 41 (2004) 867–875.
[23] N. Miyazaki, A. Takahashi, W. Kubo, J. Pharm. Pharm. Sci. 6 (2) (2003) 238–245.
[24] A. Azeem, S. Talegaonkar, L.M. Negi, F.J. Ahmad, R.K. Khar, Z. Iqbal, Int. J. Pharm.
422 (2012) 436–444.
[25] S.K.H. Khalil, G.S. El-Feky, S.T. El-Banna, W.A. Khalil, Carbohydr. Polym. 90 (2012)
1244–1253.
[26] S. Hama, K. Takahashi, Y. Inai, K. Shiota, R. Sakamoto, A. Yamada, et al., J. Pharm. Sci.
101 (8) (2012) 2909–2916.
[27] R.M. Hathout, A.H. Elshafeey, Eur. J. Pharm. Biopharm. 82 (2012) 230–240.
[28] W.I. Choi, J.H. Lee, J.Y. Kim, J.C. Kim, Y.H. Kim, G. Tae, J. Control. Release
157 (2012) 272–278.
[29] J.D. Bos, M.M.H.M. Meinardi, Exp. Dermatol. 9 (2000) 165–169.
[30] T.W. Prow, J.E. Grice, L.L. Lin, R. Faye, M. Butler, W. Becker, et al., Adv. Drug Deliv.
Rev. 63 (2011) 470–491.
[31] J. Lademann, H. Richter, A. Teichmann, N. Otberg, U. Blume-Peytavi, J. Luengo, et al.,
Eur. J. Pharm. Biopharm. 66 (2007) 159–164.
[32] R. Toll, U. Jacobi, H. Richter, J. Lademann, H. Schaefer, U. Blume-Peytavi, J. Invest.
Dermatol. 123 (2004) 168–176.
[33] G. Sonavane, K. Tomoda, A. Sano, H. Ohshima, H. Terada, K. Makino, Colloids Surf. B:
Biointerfaces 65 (2008) 1–10.
[34] S.K. Sahoo, J. Panyam, S. Prabha, V. Labhasetwar, J. Control. Release 82 (1) (2002)
105–114.
[35] K. Tomoda, H. Terashima, K. Suzuki, T. Inagi, H. Terada, K. Makino, Colloids Surf. B:
Biointerfaces 88 (2011) 706–710.
[36] K. Tomoda, H. Terashima, K. Suzuki, T. Inagi, H. Terada, K. Makino, Colloids Surf. B:
Biointerfaces 92 (2012) 50–54.
[37] K. Tomoda, A. Watanabe, K. Suzuki, T. Inagi, H. Terada, K. Makino, Colloids Surf. B:
Biointerfaces 97 (2012) 84–89.
[38] H. Fessi, F. Puisieux, J.P. Devissaguet, N. Ammoury, S. Benita, Int. J. Pharm. 55 (1989)
R1–R4.
[39] J.Y. Xiong, X.Y. Liu, P.D. Sawant, S.B. Chen, T.S. Chung, K.P. Pramoda, J. Chem. Phys.
121 (24) (2004) 12626–12631.
[40] J.Y. Xiong, X.Y. Liu, S.B. Chen, T.S. Chung, J. Phys. Chem. 109 (2005) 13877–13882.
 References 147

[41] K. Tomoda, N. Yabuki, H. Terada, K. Makino, Colloids Surf. A: Physicochem. Eng.

Asp., in revision.
[42] H. Ohshima, J. Colloid Interface Sci. 163 (1994) 474–483.
[43] K. Makino, M. Ikekita, T. Kondo, S. Tanuma, H. Ohshima, Colloid Polym. Sci.
272 (1994) 487–492.
[44] K. Makino, F. Fukai, S. Hirata, H. Ohshima, Colloids Surf. B: Biointerfaces 7 (1996)
235–238.
[45] K. Tomoda, N. Yabuki, H. Terada, K. Makino, Colloid Polym. Sci., in submission.
[46] S. Lee, J. Lee, Y.W. Choi, Biol. Pharm. Bull. 39 (2) (2007) 393–396.
[47] H. Chen, H. Zhu, J. Zheng, D. Mou, J. Wan, J. Zhang, et al., J. Control. Release 139 (1)
(2009) 63–72.
[48] J.Y. Fang, K.C. Sung, H.H. Lin, C.L. Fang, J. Control. Release 60 (1) (1999) 1–10.
[49] W. Liu, M. Hu, W. Liu, C. Xue, H. Xu, X.L. Yang, Int. J. Pharm. 364 (1) (2008) 135–141.