Review
title
Electroporation as an Efficient Physical
Enhancer for Skin Drug Delivery
José Juan Escobar-Chávez, PhD, Dalia Bonilla-Martínez, MSc,
Martha Angélica Villegas-González, MSc, and Alma Luisa Revilla-Vázquez, PhD
Transdermal drug delivery offers an attractive alternative
to the conventional drug delivery methods of oral admin-
istration and injection. However, the stratum corneum
acts as a barrier that limits the penetration of substances
through the skin. Application of high-voltage pulses to the
skin increases its permeability (electroporation) and
enables the delivery of various substances into and through
the skin. The application of electroporation to the skin has
been shown to increase transdermal drug delivery.
Moreover, electroporation, used alone or in combination
with other enhancement methods, expands the range of
drugs (small to macromolecules, lipophilic or hydrophilic,
charged or neutral molecules) that can be delivered trans-
dermally. The efficacy of transport depends on the electri-
cal parameters and the physicochemical properties of
T ransdermal drug delivery offers several advan-
tages over conventional routes.1,2 It avoids the
first-pass metabolism and the gastrointestinal tract.
Transdermal delivery has the potential for sustained
and controlled drug release. Moreover, it is a nonin-
vasive mode of drug delivery with no trauma or risk
From the División de Estudios de Posgrado (Tecnología Farmacéutica),
Facultad de Estudios Superiores Cuautitlán-Universidad Nacional
Autónoma de México, Cuautitlán Izcalli, Estado de México, México
(Dr Escobar-Chávez); División de Ciencias Químicas, Sección de Química
Analítica, Facultad de Estudios Superiores Cuautitlán-Universidad
Nacional Autónoma de México, Cuautitlán Izcalli, Estado de México,
México (Dr Escobar-Chávez, Dr Bonilla-Martínez, Dr Villegas-González);
and Laboratorio de Desarrollo de Métodos Analíticos-Facultad de
Estudios Superiores Cuautitlán-Universidad Nacional Autónoma de
México, Cuautitlán Izcalli, Estado de México, México (Dr Revilla-
Vázquez). Submitted for publication April 11, 2009; revised version
accepted July 13, 2009. Address for correspondence: José Juan
Escobar-Chávez, PhD, División de Estudios de Posgrado (Tecnología
Farmacéutica), Facultad de Estudios Superiores Cuautitlán-Universidad
Nacional Autónoma de México, Cuautitlán Izcalli, Estado de México,
México 54740; e-mail: josejuanescobarchavez@gmail.com.
DOI: 10.1177/0091270009344984
1262  •  J Clin Pharmacol 2009;49:1262-1283
drugs. The in vivo application of high-voltage pulses is
well tolerated, but muscle contractions are usually
induced. The electrode and patch design is an important
issue to reduce the discomfort of the electrical treatment
in humans. This review presents the main findings in the
field of electroporation—namely, transdermal drug deliv-
ery. Particular attention is paid to proposed enhancement
mechanisms and trends in the field of topical and trans-
dermal delivery.
Keywords: Electroporation; skin; physical enhancers;
transdermal drug delivery
Journal of Clinical Pharmacology, 2009;49:1262-1283
© 2009 the American College of Clinical Pharmacology
of infection. Patient compliance may be improved
by this user-friendly method. Despite the advantages
of transdermal delivery, only a small percentage of
drugs can be delivered transdermally because of the
barrier properties of the skin: only small, potent
lipophilic drugs can be delivered at therapeutic rates
by passive diffusion.3 Moreover, transport of most
drugs across the skin is very slow, and lag times to
reach steady-state fluxes are in hours. Achievement
of a therapeutically effective drug level is therefore
difficult without enhancing skin permeation.
In the past 30 years, numerous methods of over-
coming the skin barrier have been described, but they
can be broadly divided into 2 main categories defined
as either passive or active methods. Passive methods
for enhancing transdermal drug delivery include the
use of vehicles such as ointments, creams, gels, and
“passive” patch technology. More recently, such dos-
age forms have been developed and/or modified to
enhance the driving force of drug diffusion (thermo-
dynamic activity) and/or increase the permeability of
the skin. Such approaches include the use of penetra-
tion enhancers,4 supersaturated systems,5 prodrugs
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY

or metabolic approach,6,7 liposomes, and other

vesicles.8-11 However, the amount of drug that can be
delivered using these methods is still limited because
the barrier properties of the skin are not fundamen-
tally changed. On the other hand, active methods for
enhancing transdermal drug delivery involve the use
of external energy to act as a driving force and/or act
to reduce the barrier nature of the stratum corneum to
enhance permeation of drug molecules into the skin.12
Recent progress in these technologies has occurred as
a result of advances in precision engineering (bioen-
gineering), computing, chemical engineering, and
material sciences, all of which have helped to achieve
the creation of miniature, powerful devices that can
generate the required clinical response. The use of
active enhancement methods has gained importance
because of the advent of biotechnology in the latter
half of the 20th century, which has led to the genera-
tion of therapeutically active, large molecular weight
(>500 Da) polar and hydrophilic molecules, mostly
peptides and proteins.12 However, gastrointestinal
enzymes often cause degradation of such molecules,
and hence there is a need to demonstrate efficient
Figure 1.  Schematic representation of the skin layers.
delivery of these molecules by alternative administra-
tion routes. Passive methods of skin delivery are
incapable of enhancing permeation of such large sol- that enables the body to interact more intimately
utes, which has led to studies involving the use of with its environment. Essentially, the skin consists
alternative active strategies such as iontophoresis,13,14 of 4 layers: the stratum corneum (SC), which is the
sonophoresis,15 and electroporation.16 outer layer of the skin (nonviable epidermis) and
The scope of this article is, first, to detail the elec- forms the rate-controlling barrier for diffusion for
troporation technique; second, to describe the mecha- almost all compounds. It is composed of dead-
nisms by which electroporation enhances transdermal flattened, keratin-rich cells, the corneocytes. These
transport; and third, to review the increasing elec- dense cells are surrounded by a complex mixture of
troporation enhancement literature to assess the intercellular lipids—namely, ceramides, free fatty
potential for practical transdermal applications in acids, cholesterol, and cholesterol sulfate. Their
drug delivery and noninvasive clinical chemistry. most important feature is that they are structured as
A number of excellent reviews that have been ordered bilayer arrays.23 The predominant diffu-
published contain detailed discussions concerning sional path for a molecule crossing the SC appears to
many aspects of electroporation.17-19 The present be intercellular.24-26 The other layers are the remain-
review shows an updated overview of the use of elec- ing layers of the epidermis (viable epidermis), the
troporation in the pharmaceutical field, specifically dermis, and the subcutaneous tissues (Figure 1).
in the area of topical and transdermal drugs. This There are also several associated appendages: hair
focus is justified because of the magnitude of the follicles, sweat ducts, apocrine glands, and nails.
experimental data available with the use of this tech- In a general context, the skin’s functions may be
nique. The use of electroporation in experimental classified as protective, homeostasis-maintaining
medicine and pharmaceutical sciences has a long functions, or sensing.27,28
history. The importance of the protective and homeostatic
role of the skin is illustrated in one context by its
THE SKIN barrier property. This allows survival in an environ-
ment of variable temperature and water content and
The skin is the largest organ of the body,20-22 account- the presence of environmental dangers, such as
ing for more than 10% of body mass and the one chemicals, bacteria, allergens, fungi, and radiation.

Review 1263
 ESCOBAR-CHÁVEZ ET AL
Figure 2.  Processes of percutaneous absorption and transdermal
delivery.
In a second context, the skin is a major organ for
maintaining the body’s homeostasis, especially in
terms of its composition, heat regulation, blood pres-
sure control, and excretory roles.28 Third, the skin is
a major sensory organ in terms of sensing environ-
mental influences, such as heat, pressure, pain,
allergens, and microorganism entry. Finally, the skin
is an organ in a continuous state of regeneration and
repair. To perform each of these functions, the
skin must be tough, robust, and flexible, with an
effective communication between each of its intrin-
sic components.
Many agents are applied to the skin either delib-
erately or accidentally, with either beneficial or del-
eterious outcomes. The main interest in dermal
absorption assessment is related to (a) local effects in
dermatology (eg, corticosteroids for dermatitis), (b)
transport through the skin seeking a systemic effect
(eg, nicotine patches, hormonal drug patches), (c)
surface effects (eg, sunscreens, cosmetics, and anti-
infectives),29 (d) targeting of deeper tissues (eg, non-
steroidal anti-inflammatory agents),30-39 and (e)
unwanted absorption (eg, solvents in the workplace,
pesticides, or allergens).40,41 Figure 2 summarizes the
process of percutaneous absorption.
The skin became popular as a potential site for
systemic drug delivery, on one hand, because of the
possibility of avoiding the problems of stomach
emptying, pH effects, enzyme deactivation associ-
ated with gastrointestinal passage, and hepatic first-
pass metabolism and, on the other hand, because of
its capability to enable input control.
ELECTROPORATION
The electroporation method for transdermal drug
delivery (TDD) consists of applying high-voltage
pulses to skin, then creating new aqueous pores in
1264  •  J Clin Pharmacol 2009;49:1262-1283
Figure 3.  Diagram representing the in vivo experimental elec-
troporation. (A) A sampling chamber glues on the skin surface of
a rat. Ag/AgCl electrodes are placed in the sampling chamber and
secured on the skin surface (B, C), and then they are connected to
porator (D).
the skin to increase molecular permeability.42-46
Figure 3 shows a schematic representation of in vivo
animal electroporation. It has the potential to
enhance macromolecular permeability across skin
for TDD. As to the method, the voltage applied to
skin and new pathways created in skin have raised
important factors for a drug’s permeability flux.45
The drug’s molecule can pass not only through
preexisting aqueous pathways within the lipid
lamellae-corneocytes region of the SC but also
through new aqueous pores created by electropora-
tion of multilamellar lipid bilayers separating cor-
neocytes. The voltage applied to skin plays 2 roles
in creating new pathways for drug permeability
through skin and raising the force driving the mole-
cule motion to pass skin.
The technique of electroporation is normally used
on the unilamellar phospholipid bilayers of cell
membranes. However, it has been demonstrated that
electroporation of skin is feasible, even though the
SC contains multilamellar, intercellular lipid bilay-
ers with few phospholipids and no living cells.47
The electrical behavior of the human epidermal
membrane as a function of the magnitude and dura-
tion of applied voltage closely parallels the electrical
breakdown: recovery of bilayer membranes seen
during electroporation.48 The approximately 100
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY

multilamellar bilayers of the SC need about 100-V

pulses for electroporation, or about 1 V per bilayer.49
Although iontophoresis involves the use of relatively
low transdermal voltages (<100 V), electroporation of
skin takes place at high transdermal voltages (~100 V
or more). There is considerable indirect evidence
that high-voltage pulses cause changes in the skin
structure.50,51 The use of electroporation in conjunc-
tion with iontophoresis can expand the scope of
transdermal delivery to larger molecules such as
therapeutic proteins and oligonucleotides. Electropo­
ration acts on the skin with some driving force on the
drug during a pulse.47 It has been shown that applica-
tion of continuous low voltage resulted in a calcein
flux 3 orders of magnitude smaller than pulsing at
high voltage under electrophoretically equivalent
conditions, suggesting that structural changes induced
in the skin by pulsing contribute more significantly
than the direct electrophoretic force acting on the
drug.47 However, iontophoresis will have secondary
effects on the skin, just as electroporation also applies
direct electromotive force on the drug during the brief
pulse period. This may be particularly true for the Figure 4.  Schematic drawing showing skin and subcutaneous
thin cell lining of the sweat ducts, which might tissue being clamped between a pair of plate electrodes. During
be electroporated with low voltages as in iontophore- the pulse application, charges build up across the stratum cor-
sis. Besides the model compounds calcein, sulforho- neum (SC).
damine, and caffeine, other drugs that have been
investigated for transdermal delivery by electropora-
tion include fentanyl,52 metoprolol,53 flurbiprofen,54 voltage drop of an applied electric pulse to the skin
cyclosporin,55 heparin,56 oligonucleotides,57,58 and develops across the SC. This voltage distribution
genes.59 Results with many of these drugs have been favors local electric breakdown (electroporation) of
less dramatic than those seen with model compounds. the SC, which is precisely the barrier targeted to be
Although studies with model compounds have permeabilized to facilitate drug transport.60
given excellent mechanistic insights, the magnitude The application of electroporation to transdermal
of flux enhancement observed may not happen for all delivery is a relatively recent development. If the
drugs; thus, each drug needs to be studied as a sepa- voltage of the applied pulses exceeds a voltage
rate case. threshold at 75 to 100 V (equivalent to the break-
down threshold of 8-10 lipid bilayers in the SC),
MECHANISMS OF ACTION microchannels or “local transport regions” are
created through the breakdown sites of the SC60
Mammalian skin has 2 layers, epidermis and dermis. (Figure 4). Transdermal transport of calcein was
The epidermis is a stratified squamous keratinizing enhanced by electroporation by 2 orders of magni-
epithelium (see Figure 1). The uppermost stratum of tude, compared with that by diffusion and iontopho-
the epidermis is the SC, which consists of about 20 resis.61 Subsequently, many small-molecule drugs
layers of flattened, enucleate, keratin-filled corneo- have been successfully delivered through the skin
cytes surrounded by lamellae of an average of 8 lipid by electroporation. Transport efficiency for small
bilayers.1 The SC forms the major barrier to most (molecular weight ≤1000) charged molecules (such
water-soluble and many hydrophobic drugs and as protoporphyrin IX), using the same polarity
contributes the major portion of the electric resis- pulses, was more than an order of magnitude higher
tance of the skin. The mechanism of electroporation than that for uncharged molecules (such as proto-
depicts the transient electric breakdown site to be porphyrin IX methyl ester) or charged molecules
the most resistive component of the electric circuit. with opposite polarity pulses. The results indicated
In transdermal electroporation, the predominant that, besides passive diffusion through electropores,

Review 1265
 ESCOBAR-CHÁVEZ ET AL
electrophoretic force of the pulses also contributes
to the electroporation-enhanced transport of these
charged molecules.62 Transport occurred during the
10 to 30 minutes after pulse application was found
to be equal to or higher than that which occurred
during pulses, indicating that passive diffusion
through electropores was also an important trans-
port mechanism.63 Auxiliary current was applied
during the recovery period of the skin to further pro-
mote drug transport by electrophoretic flow.64 Yet
the transport of uncharged or weakly charged mole-
cules was also enhanced by a current of opposite
polarity, indicating the action of electroosmotic
force.65 This is especially important for delivering
larger and uncharged molecules.
Mechanisms of Molecular Transport
Molecular transport through transiently permeabi-
lized skin by electroporation results from different
mechanisms at different times. Enhanced diffusion,
during and after pulses, and electrically driven
transport during pulses (ie, electrophoretic move-
ment and very slight electroosmosis) are the main
mechanisms of transport. The contribution of elec-
trophoresis and diffusion depends on the physico-
chemical properties of the molecule.
Electrophoretic Movement
During high-voltage pulses, the main driving force
for transport of charged molecules is electropho­
resis.47,52,53,66 Evidence for this major contribution of
electrophoresis is the drop in the drug transport with
reverse electrode polarity opposing electrophoresis.
Diffusion
Molecular transport through skin highly perm­
eabilized by electroporation is also due to enhanced
passive diffusion. Although much higher skin per-
meability is achieved during the pulse, prolonged
permeabilization and thereby transport occur after
pulsing, lasting for hours in the in vitro studies.
Evidence for the contribution of enhanced postpulse
diffusion arises from the increased transport seen
(1) with reverse electrode polarity, (2) with neutral
molecules, and (3) when the drug is added after
application of the pulses.53,66,67
Electroosmosis
In contrast with iontophoresis, the contribution of elec-
troosmosis during high-voltage pulses is low. The short
time of current application (few seconds) limits the
role of electroosmosis in drug transport by skin
1266  •  J Clin Pharmacol 2009;49:1262-1283
electroporation. Further evidence comes from the similar
anodic and cathodic transport of neutral molecules.67
ADVANTAGES AND DISADVANTAGES OF
ELECTROPORATION
The advantage common to all physical methods,
including electroporation (excluding methods using
particle carriers), is that the transport mechanism
does not depend on the uptake functions of the cell;
therefore, it can be applied equally well to all cell
types and at all stages of the cell cycle. The process is,
by itself, biochemically and biologically nontoxic.
All physical methods, including electroporation,
involve transient damage of the cell membrane in
order for the gene and drug to be transported into
cells. Therefore, collateral damage resulting in some
cell death is inevitable. This is sometimes referred to
as “toxicity” of the physical treatment. All physical
methods, including electroporation, require instru-
ments that seem less convenient than simple injec-
tion of naked DNA or with carrier particles.68-73
In addition to the benefits of avoiding the hepatic
first-pass effect and higher patient compliance, the
additional advantages and disadvantages that the
electroporation technique offers can be summarized
as follows in Table I.
APPLICATIONS OF ELECTROPORATION
Skin electroporation could be particularly appropri-
ate for topical delivery for the following reasons. (1)
Skin electroporation temporarily permeabilizes the
barrier to drug permeation and therefore could
broaden topical delivery to drugs not suitable for
delivery by passive diffusion (ie, hydrophilic,
charged, and/or large molecular mass drugs).18,51,67,78
(2) The application of high-voltage pulses can
also enhance the permeability of viable cells under-
lying the SC as demonstrated in vivo by electro-
chemotherapy (ie, the electropermeabilization of
tumors to bleomycin using high-voltage pulses) or
DNA transfection.79,80
The unique and promising release of drugs by
electroporation renders it an attractive candidate as
a physical enhancer to administer drugs throughout
the skin81-108 and for other uses as gene therapy109-111
and wound healing,112 as well as to develop large,
permeable regions in the SC113 and to reseal the epi-
dermis after electroporation.114,115
This is emphasized in Table II, which summarizes
the research on electroporation uses in the transdermal
administration of drugs.
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY

Table I  Advantages and Disadvantages of Using Electroporation as a Physical Penetration Enhancer

Advantages Disadvantages

Enhanced drug penetration (of selected drugs) over Cell damage: If the pulses are of the wrong length or
passive transport   intensity, some pores may become too large or fail to
  close after membrane discharge, causing cell damage
  or rupture77
Allows strict control of transdermal penetration rates The transport of material into and out of the cell during
  the time of electropermeability is relatively
  nonspecific77
Versatility: electroporation is effective nearly with all
cells and species types74
Efficiency: a large majority of cells take in the target
DNA or molecule75
Permits rapid termination of drug delivery through
termination of electroporation
The procedure may be performed with intact tissue76
Less anxiety provoking or painful than injection
In many cases, greater patient satisfaction
Not immunologically sensitizing

Analgesic and Anti-Inflammatory Drugs
The transdermal transport of cyclodextrin (CD) across
the porcine epidermis by electroporation was studied
by Murthy et al.81 Electroporation increased the per-
meation of h-cyclodextrin (BCD) and hydroxy propyl
h-cyclodextrin (HPCD) by several orders of magni-
tude, relative to passive transport. The presence of
BCD and HPCD enhanced the total transport of the
test permeants piroxicam and carboxyfluorescein
(CF), respectively, from both permeant solutions and
suspensions. BCD enhanced the fraction of piroxicam
transported across the epidermis into the receiver
compartment medium. This was most likely due to
the prolonged postpulse permeability state of the epi-
dermis. The fraction of CF retained in the epidermis
was increased by HPCD. The in vivo delivery of CF by
electroporation in mice demonstrated the potential of
HPCD for sustaining the transdermal absorption rate
of hydrophilic molecules.
The aim of the study of Huang et al82 was to assess
the effects of iontophoresis and electroporation on
transdermal delivery of nalbuphine (NA) and its 2
novel prodrugs, nalbuphine benzoate (NAB) and
sebacoyl dinalbuphine ester (SDN), from solutions
as well as from hydrogels. Hydroxypropyl cellulose
(HPC) and carboxymethyl cellulose (CMC) were
used in hydrogel formulations to evaluate their
feasibility for delivery of NA and its prodrugs.
Application of iontophoresis or electroporation sig-
nificantly enhanced the in vitro permeation of NA
and its prodrugs. The enhancement effect was more
pronounced after applying iontophoresis. The com-
bination of 2 electrically assisted methods enhanced
the delivery of NA; however, no such enhancement
was observed for the permeation of NAB and SDN.
Their results demonstrated that lipophilicity and
molecular size, as well as hydrogel compositions,
had significant effects on skin permeation of NA,
NAB, and SDN via passive diffusion or under the
electric field.
Sung et al83 also assessed the effects of electropora-
tion on transdermal permeation of NA and its prod-
rugs. The permeation characteristics were investigated
under various electrical factors and skin barriers to
elucidate the mechanisms involved in transdermal
delivery of NA and its prodrugs by skin electropo-
ration. The in vitro permeation studies were per-
formed using side-by-side diffusion cells. The various
electrical factors investigated were pulse voltage,
pulse duration, and pulse number; the different skin
barriers studied were intact hairless mouse skin,
SC-stripped skin, delipid skin, and furry Wistar rat
skin. Application of electroporation significantly
enhanced transdermal permeation of NA and its
prodrugs. The enhancement ratios were highest for
NA, and the 4 prodrugs showed the similar permea-
bility after electroporation. The permeation amounts
of NA and its prodrugs may be increased by applica-
tion of higher pulse voltage, pulse duration, and
pulse number. Various kinetics and mechanisms were
observed for the permeation of the hydrophilic NA

Review 1267

Table II  Research on Uses of Electroporation to Administer Different Drugs Through the Skin
Research Outcome Author
The transdermal transport of cyclo-
dextrins (CDs) across porcine epi-
dermis by electroporation
To assess the effects of iontophore-
sis and electroporation on trans-
dermal delivery of nalbuphine
(NA) and its 2 novel prodrugs:
nalbuphine benzoate (NAB) and
sebacoyl dinalbuphine ester
(SDN) from solutions as well as
from hydrogels
Effects of electroporation on trans-
dermal permeation of NA and its
prodrugs
Effect of iontophoresis combined
with treatment of other physical
enhancement methods such as
electroporation, low-frequency
ultrasound, and erbium:YAG
(yttrium-aluminum-garnet) laser
on the transdermal delivery of
sodium nonivamide acetate (SNA)
Antidiuretics
The use of macromolecules as
chemical enhancers of mannitol
transdermal transport by skin
electroporation
Antiviral drugs
Transdermal delivery of interferon
alpha-2b (IFNα2b) in hairless rats
through aqueous microchannels
created in the skin and enhanced
by iontophoresis
Examination of parameters influ-
encing electroporative transder-
mal delivery of terazosin
hydrochloride (THCl) to hairless
rat skin
To understand the mechanism of
terazosin delivery as a prelimi-
nary indicator of safety
1268  •  J Clin Pharmacol 2009;49:1262-1283
ESCOBAR-CHÁVEZ ET AL
Analgesic and Anti-Inflammatory Drugs
Electroporation increased the permeation of h-cyclodextrin Murthy et al81
(BCD) and hydroxy propyl h-cyclodextrin (HPCD) by sev-
eral orders of magnitude, relative to passive transport.
The presence of BCD and HPCD enhanced the total
transport of the test permeants piroxicam and
carboxyfluorescein (CF).
The combination of 2 electrically assisted methods Huang et al82
enhanced the delivery of NA; however, no such enhance-
ment was observed for the permeation of NAB and SDN.
The permeation amounts of NA and its prodrugs may be Sung et al83
increased by application of higher pulse voltage, pulse
duration, and pulse number.
Pulsing of high voltages followed by iontophoresis did not Fang et al84
result in increased transport over iontophoresis alone.
However, electroporation shortened the onset of transder-
mal iontophoretic delivery of SNA.
Skin electroporation increased transdermal mannitol deliv- Vanbever et al85
ery by approximately 2 orders of magnitude. The addi-
tion of macromolecules further increased transport up to
5-fold.
In vivo delivery of IFNα2b was demonstrated through Badkar et al86
micropores created in the outer layer of the skin.
Iontophoresis enhanced delivery through microporated
skin in hairless rats.
It was found that voltage, pulse length (t), and number of Sharma et al87
pulses were the 3 most important parameters influencing
transdermal delivery of THCl.
Using shorter pulses and large-area electrodes is a safer Sharma et al88
technique than large pulses and small-area electrodes
when electroporation is used to enhance skin’s permea-
bility for terazosin hydrochloride.
(continued)
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY

Table II  (continued)

Analgesic and Anti-Inflammatory Drugs

Research Outcome Author

Beta-blocker agents
The effect of electroporation on ion- The iontophoretic transport of timolol was decreased by Denet et al89
tophoretic transport of 2 beta- electroporation because the high accumulation of the
blockers, timolol and atenolol lipophilic cation timolol in the stratum corneum (SC)
resulted in a decrease of electroosmosis. In contrast,
electroosmosis was not affected by atenolol, and the
iontophoretic transport of atenolol was increased by
electroporation.
Anticancer drugs
The influence of iontophoresis and A combination of electroporation pretreatment and subse- Fang et al90
other physical enhancement quent iontophoresis resulted in a higher permeation of
methods such as electroporation 5-FU than either technique alone.
and erbium:YAG laser on the skin
permeation of 5 fluorouracil
(5-FU)
To present an overview of electro- Electrochemotherapy is a new, clinically acknowledged Sersa et al91
chemotherapy, via cell membrane method for the treatment of cutaneous and subcutaneous
permeabilizing electric pulses tumors.
Insulin
Study of transdermal transport of In vivo noninvasive insulin delivery to therapeutic levels Murthy et al92
insulin and extraction of intersti- and glucose extraction may be achieved by combining
tial glucose under anodal ionto- electroporation with anionic lipids and electroosmosis.
phoresis following electroporation
in the presence of 1,2 dimyris-
toylphophatidylserine (DMPS)
Evaluation of the synergistic effect Percutaneous absorption of insulin is synergistically Tokumoto
of electroporation (EP) and ionto- enhanced by a combined use of EP and IP. et al93
phoresis (IP) on the in vivo percu-
taneous absorption of human
insulin in rats
Hormones
Investigation of electronically facili- The flux of hPTH (1-34) with the electroporation pulses of Medi and
tated transdermal delivery of 100 and 300 V, followed by iontophoresis at 0.2 mA/cm2, Singh94
human parathyroid hormone was 10- and 5-fold higher, respectively.
(1-34), hPTH (1-34)
In vitro electrically assisted delivery A combination of electroporation and iontophoresis Chang et al95
by iontophoresis and/or elec- resulted in higher transdermal permeation than either
troporation of calcium-regulating one technique alone.
hormones, salmon calcitonin
(sCT) and parathyroid hormone
(1-34) (PTH), through human
epidermis
Transdermal penetration of estradiol Combination of electroporation and iontophoresis did not Essa et al96
through human epidermis from markedly improve estradiol penetration for ultradeform-
phosphatidylcholine (PC)–based able vesicles.
liposomes and saturated aqueous
estradiol solution
(continued)

Review 1269

Research Outcome Author
Photosensitizers
Electric pulses to increase transder-
mal transport of d-aminolevulinic
acid (ALA), a precursor to the
photosensitizer protoporphyrin IX
(PpIX)
Optimize and enhance the in vitro
skin permeation of 5-ALA by 2
resurfacing techniques:
erbium:YAG laser and micro-
dermabrasion
Oligonucleotids
Topical delivery in the skin of 39
end-modified phosphodiester
oligonucleotides using
electroporation
A needle-free method based on
transcutaneous electroporation is
described for delivering peptide
vaccines
Adaptation of the electroporation
and electrofusion technology in 2
fronts of cancer research and
treatment
Tea polyphenols
To assess the effects of electropora-
tion, iontophoresis, and their
combination on the transdermal
delivery of tea catechins across
porcine skin
Stimulants
Investigate the effect of electrotreat-
ment on skin permeability by
measuring the cumulative deliv-
ery of caffeine and sodium ascor-
byl phosphate (NAP)
Dextrans
To test factors affecting the
potency of anionic lipid transport
enhancers
The substantial synergic effects of
CaCl2 and EP on in vitro skin per-
meation of calcein and fluorescein
isothiocyanate (FITC) dextrans
1270  •  J Clin Pharmacol 2009;49:1262-1283
ESCOBAR-CHÁVEZ ET AL
Table II  (continued)
Analgesic and Anti-Inflammatory Drugs
A greater than 2-fold enhancement of PpIX production Johnson et al97
with electroporative delivery was seen versus that
obtained with passive delivery.
Application of iontophoresis or electroporation alone also Fang et al98
increased the ALA permeation by approximately 15-fold
and 2-fold, respectively. The incorporation of iontophore-
sis or electroporation with the resurfacing techniques
caused a profound synergistic effect on ALA permeation.
Electroporation increased the topical delivery of the 39 Regnier et al99
end-modified phosphodiesters by 2 orders of magnitude
compared to passive diffusion.
The efficacy of peptide delivery was comparable to that of Zhao et al100
intradermal injection with Freund’s complete adjuvant.
The pro-photosensitizer drug delta-amino levulinic acid Hui101
and the anticancer drug methotrexate have been deliv-
ered transcutaneously by electroporation. These studies
hold promise for the treatment of cancers in humans.
A synergistic effect was detected for (+)-catechin but not Fang et al102
for (–)-epicatechin after application of electroporation
followed by iontophoresis.
Electrotreatment increased the amount of caffeine and Marrat et al103
NAP in the skin. Enhancement factors (EFs) for NAP of
7.2 and 14.9 were observed following 20 minutes of
electrotreatment.
The use of proper lipid enhancers greatly extends the Sen et al104
upper size limit of transportable chemicals.
Little release of calcein was observed from SC lipid with- Tokudome and
out EP, whereas higher releases were observed after EP Sugibayashi105
with or without NaCl or CaCl2.
(continued)
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY

Table II  (continued)

Analgesic and Anti-Inflammatory Drugs
Research Outcome Author

To study the influence of sodium
dodecyl sulfate (SDS) on transder-
mal transport of diffusants by
electroporation
Folic acid antagonists
Application of electroporation to
increase the transdermal transport
of methotrexate (MTX)
A microelectrode array to minimize
the pain sensation of electropora-
tion for enhancing transdermal
drug delivery of methotrexate
Other uses
Gene transfer
Optimization of electroporation
parameters for transgene expres-
sion with minimal tissue damage
with a novel electrode
Local injection of a plasmid before
electroporation
Optimization of parameters to
obtain reproducible, high-level
gene transfer to the mouse skin
Wound healing
Application of plasmid DNA
expression vectors directly into
the wound with electroporation
To develop large permeable regions in the SC
A model of pore formation between
adjacent corneocytes
To reseal epidermis after electroporation
The resealing of porcine epidermis
after electroporation
Application of high-voltage pulsing
or iontophoresis to human, hair-
less rat, or black rat snake skin
It appears that the presence of SDS during electroporation Murthy et al106
helps in achieving the desired transport with less
electrical exposure dose.
Electroporation of MTX with an anion lipid enhancer Wong et al107
under a mild hyperthermic environment provided a
significant transdermal delivery.
In vivo transdermal electroporation using a microelectrode Wong et al108
array with 180 pulses of 150 V, 0.2 ms at 1 Hz, followed
by a 30-minute methotrexate occlusion increased more
than 4-fold the systemic methotrexate level in mice.
The highest transgene expression and efficiency of Heller et al109
individual cell transformation with minimal damage was
produced with eight 150-ms pulses at a field strength of
100 V/cm.
Electrotransfer is a promising alternative for drug and gene Préat110
delivery.
With the optimized conditions, gene transfer mediated by Chesnoy and
intradermal injection of naked DNA was comparable in Huang111
efficiency to electroporation.
Electroporation-assisted transfection with DNA plasmid Ferguson et al112
expression vectors for growth factors will be an effective
modality for enhancing cutaneous wound healing.
The model predicts that the initial radius of the first Pliquett and
aqueous pathway is approximately 5 nm for a transdermal Weaver113
voltage of 60 V at room temperature.
Both poloxamer 188 and phosphatidylcholines were Burgess et al114
effective in resealing in terms of electric conductance
and transport.
Electroporation caused large molecular transport for all Chen et al115
3 types of skin.

and lipophilic nalbuphine enanthate through differ-
ent skin barriers by applying electroporation. This
study demonstrated that electroporation may enhance
and control transdermal permeation of NA and its
prodrugs.
The effect of iontophoresis combined with treat-
ment of other physical enhancement methods such
as electroporation, low-frequency ultrasound, and
erbium:YAG (yttrium-aluminum-garnet) laser on the
transdermal delivery of sodium nonivamide acetate

Review 1271
 ESCOBAR-CHÁVEZ ET AL
(SNA) was examined by Fang et al.84 Iontophoresis
increased the transdermal flux of SNA in vitro as
compared to the passive diffusion without any
enhancement. Furthermore, iontophoresis was always
the most potent enhancement method for SNA per-
meation among the physical enhancement methods
tested. Pulsing of high voltages (electroporation)
followed by iontophoresis did not result in increased
transport over iontophoresis alone. However, elec-
troporation shortened the onset of transdermal
iontophoretic delivery of SNA. Pretreatment of low-
frequency ultrasound alone on skin did not increase
the skin permeation of SNA. The combination of ion-
tophoresis and sonophoresis increased transdermal
SNA transport more than each method by itself. The
enhancement of drug transport across shunt routes
and reduction of the threshold voltage in the presence
of an electric field may contribute to this synergistic
effect. Use of an erbium:YAG laser was a good method
for enhancing transdermal absorption of SNA because
it allows precise control of SC removal, and this abla-
tion of SC could be reversible to the original normal
status. The combination of laser treatment and ionto-
phoresis also synergized the skin permeation of SNA,
possibly due to a gradual drop in the electric resis-
tance of the skin. The results in this present study
point out that the choice of certain conditions with
suitable physical enhancement methods can induce a
synergistic effect on transdermal delivery of SNA
during iontophoresis.
Antidiuretic Drugs
Macromolecules were investigated as chemical
enhancers of transdermal transport by skin electropo-
ration for Vanbever et al.85 Although unable to enhance
passive or iontophoretic transport, macromolecules
are proposed to enhance electroporation-assisted
delivery by stabilizing the increased permeability
caused by high-voltage pulses. Skin electroporation
increased transdermal mannitol delivery by approxi-
mately 2 orders of magnitude. The addition of macro-
molecules further increased transport up to 5-fold.
Macromolecules present during pulsing enhanced
mannitol transport after pulsing for hours, apparently
by a macromolecule-skin interaction. No enhance-
ment was observed during passive diffusion or low-
voltage iontophoresis, suggesting that macromolecules
interact specifically with transport pathways created
at high voltage. Although all macromolecules studied
enhanced transport, those with greater charge and size
were more effective. This study demonstrates that
1272  •  J Clin Pharmacol 2009;49:1262-1283
macromolecules can be used as transdermal transport
enhancers uniquely suited to skin electroporation.
Antiviral Drugs
Badkar et al86 demonstrated transdermal delivery of
interferon alpha-2b (IFNα2b) in hairless rats through
aqueous microchannels (micropores) created in the
skin and enhanced by iontophoresis.
The Altea Therapeutics PassPort System was con-
figured to form an array of micropores (2.0 cm2; 72
micropores/cm2) on the rat abdomen. The transder-
mal patch (Iomed TransQ1-GS-hydrogel) was satu-
rated with an IFNα2b solution and applied. Delivery
was evaluated with and without cathodic iontophore-
sis (0.1 mA/cm2). Intravenous delivery was performed
to support pharmacokinetic calculations. IFNα2b was
not delivered through intact skin by itself (passive
delivery) or during iontophoresis. However, passive
delivery through micropores was achieved in vivo in
rats. A dose of 397 ng was delivered over 6 hours,
with steady-state serum concentrations reaching
a plateau at 1 hour postpatch application. These
levels dropped rapidly after patch removal and
returned to baseline within 2 hours of patch removal.
Iontophoresis-enhanced delivery through micropores
resulted in a 2-fold increase in the dose delivered in
the hairless rat. In vivo delivery of IFNα2b was dem-
onstrated through micropores created in the outer
layer of the skin. Iontophoresis enhanced delivery
through microporated skin in hairless rats.
The use of electroporation pulses as a physical
means of enhancing the permeability of skin to
deliver drugs is in the early stages of development. In
this article, a systematic study examining the para­
meters influencing electroporative transdermal deliv-
ery of terazosin hydrochloride to hairless rat skin is
reported by Sharma et al.87 It was found that voltage,
pulse length (t), and number of pulses were the 3
most important parameters, in that order. For creating
a significant enhancement in drug delivery to the
skin, without causing any apparent change in its
external appearance, it was necessary to deliver 5 or
more exponentially decaying electroporation pulses,
at 88 ± 2.5 V (voltage across the skin), with a decay
time constant of 20 ms. Electrodes with larger area
could attain the same voltages across the skin with a
much lower applied voltage and possessed other
advantages with regard to performance of the drug
delivery system.
In another article, Sharma et al88 described a study
in which the reversibility of the electroporation
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY
technique is evaluated with in vitro methods. The
skin’s reversal from an enhanced permeation mode as
a result of electroporation to the base level was used as
an index to understand the mechanism of drug deliv-
ery and also as a preliminary indicator of safety.
Maximum delivery of the model drug, terazosin hydro-
chloride, occurred during the pulsing. Electroporative
delivery with a wire electrode (small-area electrode,
0.56 cm2) using 20 pulses at 88 V and pulse length of
20 ms did not cause any damage to the skin. Increasing
the pulse length to 60 ms, while keeping the rest of the
parameters fixed, caused a visible change in the exter-
nal appearance of the skin. However, with the use of a
spiral electrode (large-area electrode, 2.74 cm2) at a
60-ms pulse length, there was minimal damage to the
skin. This may be attributed to the more uniform flow
of current over the whole skin area.
These findings indicate that using shorter pulses
and large-area electrodes is a safer technique than
large pulses and small-area electrodes when elec-
troporation is used to enhance skin’s permeability
for drug delivery.
Beta-Blocker Agents
Denet et al89 studied the effect of electroporation on
iontophoretic transport of 2 beta-blockers, timolol
(lipophilic) and atenolol (hydrophilic), to have a bet-
ter understanding of the mechanism of combination.
The transdermal delivery of these beta-blockers
through human SC was studied in 3-compartment
diffusion cells. The transport of mannitol was evalu-
ated to assess the electroosmotic flow. The ionto­
phoretic transport of timolol was decreased by
electroporation because the high accumulation of
the lipophilic cation timolol in the SC resulted in a
decrease of electroosmosis. In contrast, electroosmo-
sis was not affected by atenolol, and the iontophoretic
transport of atenolol was increased by electropora-
tion. Using 2 different beta-blockers, the authors
showed that lipophilicity and positive charges affect
the electrotransport of drugs. Understanding the effect
of the physicochemical properties of the drug, as well
as the electrical parameters, is thus essential for the
optimization of transdermal drug delivery by a com-
bination of electroporation and iontophoresis.
Anticancer Drugs
The influence of iontophoresis and other physical
enhancement methods such as electroporation
and erbium:YAG laser on the skin permeation of
Review 1273
5-fluorouracil (5-FU) was examined by Fang et al.90
Iontophoresis increased the in vitro transdermal
transport of both the anionic and nonionic forms of
5-FU. A combination of electroporation pretreat-
ment and subsequent iontophoresis resulted in a
higher permeation of 5-FU than either technique
alone. It appeared that electroporation treatment
exerted a disruptive influence on the SC. The SC
layers in the skin were partly ablated by the laser,
resulting in a great enhancement effect on the skin
permeation of 5-FU. Application of iontophoresis
further increased the drug permeation across laser-
pretreated skin. The laser was consistently the most
potent technique to enhance 5-FU delivery among
the physical enhancement methods examined in
this study.
Electrochemotherapy is a local drug delivery
approach aimed at treatment with palliative intent
of cutaneous and subcutaneous tumor nodules of
different histologies. Electrochemotherapy, via cell
membrane permeabilizing electric pulses, poten-
tiates the cytotoxicity of nonpermeant or poorly
permeant anticancer drugs with high intrinsic cyto-
toxicity, such as bleomycin or cisplatin, at the site of
electric pulse application. An overview of preclini-
cal and clinical studies was presented, and the treat-
ment procedure was further critically evaluated by
Sersa et al.91 In clinical studies, electrochemotherapy
has proved to be a highly efficient and safe approach
for treating cutaneous and subcutaneous tumor nod-
ules. The treatment response for various tumors
(predominantly melanoma) was approximately 75%
complete and 10% partial response of the treated
nodules. Electrochemotherapy is a new, clinically
acknowledged method for the treatment of cutane-
ous and subcutaneous tumors. Its advantages are
high effectiveness on tumors with different histolo-
gies, simple application, minimal side effects, and
the possibility of effective repetitive treatment.
Insulin
Transdermal transport of insulin and extraction of
interstitial glucose under anodal iontophoresis (elec-
troosmosis) following electroporation in the presence
of 1,2-dimyristoylphophatidylserine (DMPS) was
studied by Murthy et al.92 An earlier study showed
that DMPS increased the transport of insulin across
porcine epidermis under electroporation by ~4-fold.
It was suggested that DMPS increased the lifetime
of electropores in the epidermis, resulting in an
enhanced transport of permeants. When electro­osmosis
 ESCOBAR-CHÁVEZ ET AL
was applied across the epidermis follow­ing elec-
troporation with DMPS, the enhancement of
insulin transport was ~18-fold over electroporation
alone. When the same strategy was applied to extract
interstitial glucose, the enhancement was ~23-fold
over electroporation alone. Real-time transdermal
insulin transport kinetics was measured using fluo-
rescein isothiocyanate (FITC)–labeled insulin and a
custom-made vertical diffusion apparatus that had a
fluorescence cuvette as the receiver compartment.
Insulin transport by electroporation alone showed a
nonlinear kinetics that is most likely due to the
resealing of the electropores with time. The transport
kinetics was more linear when electroporation was
carried out in the presence of DMPS, confirming ear-
lier studies that suggested that DMPS stabilizes trans-
port paths formed by electroporation. The data suggest
that in vivo, noninvasive insulin delivery to thera-
peutic levels and glucose extraction may be achieved
by combining electroporation with anionic lipids and
electroosmosis.
Tokumoto et al93 evaluated the synergistic effect of
electroporation (EP) and iontophoresis (IP) on the in
vivo percutaneous absorption of human insulin in
rats. Passive diffusion and IP alone (0.4 mA/cm2)
resulted in almost no skin permeation of insulin at
pH 7, whereas EP treatment (150 or 300 V, 10 ms, and
10 pulses) resulted in a high plasma level of insulin,
and the combined use of EP and IP led to a further
increase of the plasma level of insulin compared
with that measured after EP alone. Interestingly, a
much higher plasma level was observed when the pH
of the insulin solution at 7 was increased to 10. The
nonassociation ratio of insulin was significantly
higher at pH 10 than at pH 7. Insulin monomers and
dimers were observed in addition to the normal form
of insulin, hexamer, albeit in low percentages, at pH
10, whereas most of the insulin was in the hexamer
form at pH 7. To confirm the influence of the aggrega-
tion properties of insulin, the commercially available
human insulin analog insulin lispro was then evalu-
ated. Its skin permeation was found to be extremely
high compared to that of conventional human insu-
lin without increasing the solution pH. Marked
decreases in blood glucose levels reflecting the
increases in the plasma concentration of insulin
were also observed after EP/IP treatment. The pres-
ent study suggests that percutaneous absorption of
insulin is synergistically enhanced by a combined
use of EP and IP and that altering the aggregation
properties of insulin is important to enhance the per-
cutaneous absorption of insulin by IP and/or EP.
1274  •  J Clin Pharmacol 2009;49:1262-1283
Hormones
Electronically facilitated transdermal delivery of
human parathyroid hormone (1-34), hPTH (1-34),
was investigated in vitro by Medi and Singh94 using
dermatomed porcine skin. The effect of iontopho-
retic current density, electroporative pulse voltages,
and also electroporation followed by iontophoresis
was investigated on the in vitro percutaneous absorp-
tion of hPTH (1-34). Iontophoresis at 0.5 mA/cm2
current density significantly enhanced the flux
of hPTH (1-34) in comparison to passive flux.
Electroporation pulses of 100, 200, and 300 V signi­
ficantly increased the flux of hPTH (1-34) in com-
parison with the passive as well as iontophoretic
flux at 0.5 mA/cm2. The electroporative flux of
hPTH (1-34) was found to vary linearly with the
pulse amplitude. The principal barrier of the skin,
SC, was found perturbed following the pulses, as
evident by light microscopy studies. The application
of electroporation pulses followed by iontophoresis
further increased the flux by several fold. The flux of
hPTH (1-34) with the electroporation pulses of 100
and 300 V followed by iontophoresis at 0.2 mA/cm2
was 10- and 5-fold higher, respectively, in compari-
son to the flux with corresponding pulses alone.
This shows the synergistic effect of iontophoresis in
combination with electroporation on skin permea-
bility of hPTH (1-34). The results indicate the pos-
sibility of designing controlled transdermal delivery
systems for hPTH (1-34) using electroporation fol-
lowed by iontophoresis.
Electrically assisted delivery by iontophoresis
and/or electroporation was used in vitro to deliver
the calcium-regulating hormones, salmon calcitonin
(sCT) and parathyroid hormone (1-34) (PTH), through
human epidermis by Chang et al.95 Such delivery
could be useful for the chronic treatment of post-
menopausal osteoporosis and other clinical indica-
tions as a superior alternative to parenteral delivery.
The sCT (50 mg/mL) or PTH (1-34) (100 mg/mL)
formulation was prepared in citrate buffer (pH 4.0
or 5.0, respectively). Epidermis separated from
human cadaver skin was used. Iontophoresis was
applied using a constant current power source
and electroporation with an exponential pulse gen-
erator. A combination of electroporation and ionto-
phoresis resulted in higher transdermal permeation
than either one technique alone. Electroporation
also shortened the lag time of iontophoretic trans-
dermal delivery of salmon calcitonin. Pulsing at
lower voltages followed by iontophoresis did not
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY
result in increased transport (over iontophoresis
alone), perhaps because the transdermal voltage was
very low. The transdermal transport of salmon calci-
tonin by pulsing with 15 pulses (1 ppm) of 500 V
(200 ms) followed by iontophoresis led to a quick
input and high flux. The average transdermal volt-
age was only about 50 V for a 500-V study.
Essa et al96 investigated transdermal penetration
of a model lipophilic drug (estradiol) through human
epidermis from phosphatidylcholine (PC)–based
liposomes and saturated aqueous estradiol solu-
tion (control). Representative examples of cholate-
containing ultradeformable (Transfersomes), nonrigid
(pure PC), and membrane-stabilized (cholesterol-
containing) vesicles were used. Transdermal pene-
tration studies involved occluded passive penetration
for 12 hours and cathodic iontophoresis (0.8 mA/
cm2) for 8 hours for all systems. Combined elec-
troporation (5 pulses, 100 V, 100 ms, 1-minute spac-
ing) and iontophoresis (0.8 mA/cm2, for 2 hours)
was also employed for ultradeformable vesicles and
control. Estradiol penetration parameters from dif-
ferent formulations were compared. All vesicles had
essentially the same particle size, with ultradeform-
able liposomes showing the highest negative zeta
potential (–29 mV). Occluded passive penetration
improved estradiol skin penetration from liposomes
relative to control. Iontophoretic studies revealed
the superiority of ultradeformable vesicles regarding
drug skin penetration and deposition compared to
traditional liposomes. Combination of electropora-
tion and iontophoresis did not markedly improve
estradiol penetration for ultradeformable vesicles.
The combination results implied repair of the skin
barrier due to the penetration-retarding effect of PC
monomers released from liposomes.
Photosensitizers
Selectivity of photodynamic therapy can be improved
with localized photosensitizer delivery, but topical
administration is restricted by poor diffusion across
the SC. Johnson et al97 used electric pulses to
increase transdermal transport of d-aminolevulinic
acid (ALA), a precursor to the photosensitizer proto-
porphyrin IX (PpIX). ALA-filled electrodes were
attached to the surface of excised porcine skin or the
dorsal surface of mice. Pulses were administered
and, in some in vivo cases, a continuous DC poten-
tial (6 V) was concomitantly applied. For in vitro
14C ALA penetration, 10-mm layers parallel to the
SC were assayed by liquid scintillation analysis, and
Review 1275
10-mm cross sections were examined autoradio-
graphically. As the electrical dose increased, there
was an increase in penetration depth. In vivo deliv-
ery was assayed by measuring the fluorescence of
PpIX in skin samples. A greater than 2-fold enhance-
ment of PpIX production with electroporative deliv-
ery was seen versus that obtained with passive
delivery. Superimposition of a DC potential resulted
in a nearly 3-fold enhancement of PpIX production
versus passive delivery. Levels were higher than the
sum of PpIX detected after pulse-alone and DC-alone
delivery. Electroporation and electrophoresis are
likely factors in electrically enhanced delivery.
5-Aminolaevulinic acid (ALA) is used as a pro-
toporphyrin IX-precursor for the photodynamic
therapy of superficial skin cancer and cutaneous
metastases of internal malignancies. However, the
permeability of hydrophilic ALA across the skin is
very low. The objective of Fang et al98 was to opti-
mize and enhance the in vitro skin permeation of
ALA by 2 resurfacing techniques: erbium:YAG
laser and microdermabrasion. Light microscopic
changes in pig skin caused by these techniques
were also compared. The electrically assisted
methods, iontophoresis and electroporation, were
also used to facilitate ALA permeation across laser-
or microdermabrasion-treated skin. Among the
modalities tested in this study, the erbium:YAG laser
showed the greatest enhancement of ALA permeation.
The laser fluence was found to play an important
role in controlling the drug flux, producing enhance-
ment ratios from 4-fold to 246-fold relative to the
control. The skin permeation of ALA across micro-
dermabrasion-treated skin was approximately 5- to
15-fold higher than that across intact skin. Both the
ablated effect of the SC and ALA flux was proportio­
nal to the treatment duration of microdermabrasion.
The application of iontophoresis or electroporation
alone also increased the ALA permeation by approx-
imately 15-fold and 2-fold, respectively. The incor-
poration of iontophoresis or electroporation with the
resurfacing techniques caused a profound synergis-
tic effect on ALA permeation. This basic study has
encouraged the further investigation of ALA perme-
ation by laser or microdermabrasion.
Oligonucleotids
The feasibility of topical delivery in the skin of 39
end-modified phosphodiester oligonucleotides using
electroporation was investigated by Regnier et al.99
Experiments were performed in vitro, using hairless
 ESCOBAR-CHÁVEZ ET AL
rat skin. Five pulses of (200 V, 450 ms) were applied.
The 39 end modifications of the 15-mer oligonucle-
otide were (1) 39-aminohexyl; (2) biotin, with a trieth-
yleneglycol arm; (3) methylphosphonate links
between nucleotides 13, 14, and 15; and (4) 2-O-
methyl nucleotides at 13, 14, and 15 positions. All the
modifications were efficient to protect the oligonucle-
otides against degradation in the skin. Electroporation
increased the topical delivery of the 39 end-modified
phosphodiesters by 2 orders of magnitude compared
to passive diffusion, without significant differences
between the derivatives. Oligonucleotide concentra-
tions in the range of 1 mm could be achieved in the
viable skin. The delivery of a phosphorothioate con-
gener was lower than phosphodiester delivery due to
the interaction of phosphorothioate with the SC.
Consequently, 39 end-protected phosphodiesters
could be an interesting alternative to phosphorothio-
ate oligonucleotides for topical treatment of cutane-
ous diseases.
A needle-free method based on transcutaneous
electroporation is described for delivering peptide
vaccines by Zhao et al.100 The Kb-binding OVA-
peptide SIINFEKL was used as an example to induce
the peptide-specific cytotoxic T lymphocyte (CTL)
response in mice. A saturated anionic lipid was
added during electroporation, and postpulse elec-
troosmosis was applied to enhance the vaccine deliv-
ery. Electroporation was found to stimulate the exodus
of Langerhans cells (LC) from the skin. The peptide
transported into and through murine skin was mea-
sured using a Franz diffusion apparatus. Most peptide
was retained in the skin rather than passing through
the skin in the process. The peptide was delivered to
the dorsal skin of mice by in vivo electroporation. An
electroporation-transportable oligonucleotide with a
CpG motif was used as adjuvant. The efficacy of pep-
tide delivery was comparable to intradermal injection
with Freund’s complete adjuvant. Peptide-specific
CTL response to the vaccine delivered by needle-free
electroporation/electroosmosis was equivalent to that
delivered by intradermal injection, as determined by
production of the peptide-specific interferon gamma
(IFN-γ) in the ELISPOT assay.
Electroporation and the associated phenomenon
of electrofusion have been widely adapted as tools to
a broad range of biomedical research and therapy. In
their study, Hui101 summarized the adaptation of the
electroporation and electrofusion technology in 2
fronts of cancer research and treatment. The first
was genetic manipulation of hematopoietic cells for
the purpose of cancer treatment. High-efficiency
transfection methods have been developed to transfect
1276  •  J Clin Pharmacol 2009;49:1262-1283
natural killer (NK) cells, peripheral blood stem cells,
and bone marrow–derived dendritic cells. Hybrids
of tumor cells and bone marrow–derived dendritic
cells have been formed by electrofusion
for the purpose of tumor vaccines. The second front
was the use of transcutaneous electroporation to
deliver anticancer drugs and vaccines across the
skin. Methods to extend the upper molecular weight
limit of transcutaneous electroporation have been
developed. The pro-photosensitizer drug delta-amino
levulinic acid, the anticancer drug methotrexate,
and peptide vaccines designed for cancer prevention
and immunotherapy have been delivered transcuta-
neously by electroporation. These studies hold
promise for the treatment of cancers in humans.
Tea Polyphenols
Tea polyphenols, including (+)-catechin, (–)-epicat-
echin, and (–)-epigallocatechin-3-gallate (EGCG),
have been shown to possess potent antioxidant and
chemopreventive activities. The aim of the study by
Fang et al102 was to assess the effects of electropora-
tion, iontophoresis, and their combination on the
transdermal delivery of tea catechins across porcine
skin. The permeation characteristics were investi-
gated using various analogs of catechins, pH values,
and modes of electroporation and iontophoresis.
The mechanisms by which these catechins were
transported via the skin were elucidated by examin-
ing the electric conductivity, transepidermal water
loss (TEWL), and fusion of SC lipid liposomes
(SCLL). The isomers, (+)-catechin and (–)-epicate-
chin, showed different behaviors of skin permeation
and local skin deposition with the electrically
assisted methods. The results suggest evidence of
selective skin absorption of (–)-epicatechin over
(+)-catechin. A synergistic effect was detected for
(+)-catechin but not for (–)-epicatechin after applica-
tion of electroporation followed by iontophoresis.
The presence of a gallic acid ester in the structure of
EGCG significantly increased the skin uptake of cat-
echins. However, a negligible amount of or no EGCG
molecules permeated across the skin. The mecha-
nisms involved in the enhancement of electropora-
tion may be the skin reservoir effect and an increase
in skin permeability. The TEWL profiles suggest that
in addition to the force of electrorepulsion, the skin
hydration effect and structural alterations may also
have contributed to the enhancement by iontophore-
sis. Electroporation did not influence the skin bar-
rier function, although the skin permeability
increased according to the SCLL fusion study.
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY
Stimulants
Electrically assisted transport can increase the rate
and extent of delivery; moreover, it also enables the
administration of polar and charged molecules into
the skin. The objective of Marrat et al103 was to inves-
tigate the effect of electrotreatment on skin permeabil-
ity by measuring the cumulative delivery of caffeine
and sodium ascorbyl phosphate. Furthermore, confo-
cal microscopy was used to visualize the effect of
electrotreatment on the penetration of calcein. Porcine
ear skin was used for the in vitro permeation studies,
which involved application of either the caffeine or
sodium ascorbyl phosphate (NAP)–containing gels
using the roll-on supplied with the electrotreatment
device. Electrotreatment increased the amount of caf-
feine and NAP in the skin. Enhancement factors (EFs)
for NAP of 7.2 and 14.9 were observed following 20
minutes of electrotreatment and either immediate
sampling or a further 60 minutes of passive diffusion
compared with passive diffusion for either 20 or 80
minutes. The effect on caffeine permeation was less
significant (EF = 2.1 for a 20-minute electrotreatment
compared with passive diffusion for 20 minutes). The
confocal microscopy images showed that electrotreat-
ment significantly increased calcein permeation;
fluorescence was observed deep into the viable
epidermis-reaching depths of up to 60 to 80 microns.
They have shown that electrotreatment increases skin
permeability and the cumulative delivery of cosmeti-
cals into the skin.
Dextrans
Sen et al104 found that anionic phospholipids, but
not cationic or neutral phospholipids, enhance the
transdermal transport of molecules by electropora-
tion. DMPS, phosphatidylserine from bovine brain
(brain-PS), dioleoylphosphatidylserine (DOPS), and
dioleoylphosphatidylglycerol (DOPG) were used to
test factors affecting the potency of anionic lipid
transport enhancers. DMPS with saturated acyl
chains was found to be a much more potent trans-
port enhancer than those with unsaturated acyl
chains (DOPS and DOPG). Saturated DMPS was also
more effective in delaying resistance recovery after
pulsing and had a greater affinity in the epidermis
after pulsing. Using fluorescent carboxyl fluorescein
and FITC-labeled dextrans as test water-soluble mol-
ecules for transport, as well as rhodamine-labeled
phospholipids to track anionic phospholipids, they
found, by conventional and confocal fluorescence
Review 1277
microscopy, that transport of water-soluble mole-
cules was localized in local transport spots or regions
(LTRs) created by the electroporation pulses. Anionic
phospholipids, especially DMPS, were located at the
center of the LTRs and spanned the entire thickness
of the SC. They deduced that, after being driven into
the epidermis by negative electric pulses, saturated
anionic phospholipids mix and are retained better
by the SC lipids. Anionic lipids prefer loose layers
or vesicular rather than multilamellar forms, thereby
prolonging the structural recovery of SC lipids to the
native multilamellar form. In the presence of 1 mg/
mL DMPS in the transport milieu, the flux of FITC-
Dextran-4k was enhanced by 80-fold and reached
175 µg/cm2/min. Thus, the use of proper lipid
enhancers greatly extends the upper size limit of
transportable chemicals. Understanding the mecha-
nism of lipid enhancers enables one to rationally
design better enhancers for transdermal drug and
vaccine delivery by electroporation.
After reporting the substantial synergic effects of
CaCl2 and EP on in vitro skin permeation of calcein
and FITC dextrans, Tokudome and Sugibayashi105
investigated the mechanisms for these effects by
considering changes in lamellar structure and bar-
rier recovery time of the biggest skin barrier, the SC,
by this combined treatment. The change in skin
lamellar structure was evaluated by lipid mobility in
the SC using attenuated total reflection Fourier
transform infrared (ATR-FTIR), calcein release from
SCLL, in vitro skin permeation of calcein, and
TEWL.
The C-H stretching band of skin lipids produced
with EP was blue-shifted when compared to that with-
out EP. Asymmetric C-H stretching was highest with
EP in CaCl2 solution. Little release of calcein was
observed from SCL without EP, whereas higher
releases were observed after EP with or without
NaCl or CaCl2. In particular, high calcein release
(>20%) was observed over 60 minutes with EP in
CaCl2 solution. The in vitro permeation study of cal-
cein was conducted through excised hairless rat skin
that was pretreated with EP before skin excision.
Permeation rate was highest in skin excised immedi-
ately after in vivo EP, and this rate decreased with
time after EP treatment. TEWL recovered to control
levels within 2 hours after EP in distilled water or
NaCl solution, whereas high TEWL was maintained
after EP in CaCl2 solution. These results suggest
that at least lamellar destruction of SC must be
related to the enhanced skin permeation of drugs by
the combination of CaCl2 and EP. On the other hand,
 ESCOBAR-CHÁVEZ ET AL
a prolonged enhancing effect on the skin permeation
of calcein by this combination may be due to a
high lamellar destruction and/or delayed barrier
repair of SC.
The objective of the experiment of Murthy et al106
was to study the influence of sodium dodecyl sulfate
(SDS) on transdermal transport of diffusants by elec-
troporation. The resistance of porcine epidermis in
contact with SDS solution (0.2% w/v) dropped by
40% within 24 hours. SDS improved the efficiency
of transdermal delivery of glucose and dextrans of
molecular weight (MW) 4 kDa (FD4K) and 10 kDa
(FD10K) by electroporation. However, the transport
of dextran MW 35 kDa (FD35K) was not influenced
significantly. Pretreatment of epidermis with SDS
solution reduced its electroporation threshold from
80 to 60 V. It appears that the presence of SDS during
electroporation helps to achieve the desired trans-
port with less electrical exposure dose. SDS enhanced
the transdermal delivery of molecules by electropo-
ration most likely by facilitating the barrier disrup-
tion during pulse application and also by prolonging
the lifetime of electropores created by the pulse.
Folic Acid Antagonists
The topical administration of methotrexate (MTX)
for the treatment of psoriasis and neoplastic diseases
is restricted by the poor diffusion of MTX across the
SC. Wong et al107 applied electroporation to increase
the transdermal transport of MTX. Electrodes were
placed either side-by-side on the surface of excised
full-thickness pig skin or on a piece of skin clamped
between compartments of a vertical diffusion cham-
ber. Sixty rectangular electric pulses at 120 V, 1 ms,
and 1 Hz were applied across the skin. MTX was left
on the skin surface for an additional 10 minutes to
take advantage of diffusion through electropores.
Cumulative drug transport was measured by radio-
active tracing, using [3H]-methotrexate, from punch
biopsy samples taken from under the cathode.
Using side-by-side electrodes, treatment with the
pulses alone resulted in a 2.5-fold increase; adding
anionic lipid enhancers to the pulses resulted in a 4.4-
fold enhancement compared with passive diffusion.
Concurrent iontophoresis for the 11-minute time
period made a no significant contribution. To reduce
tissue resistance, we used 40°C hyperthermia in a ver-
tical diffusion chamber; transport was increased
11-fold to 53 µg/cm2 (flux 290 µg/cm2 h). MTX pene-
tration profiles indicated that more than half of the
MTX was confined to the epidermis and papillary
dermis. The tissue concentration in this superficial
1278  •  J Clin Pharmacol 2009;49:1262-1283
reactive unit was 1.7 mmol/L. Electropo­ration of MTX
with an anion lipid enhancer under a mild hyperther-
mic environment provided a significant transdermal
delivery within a short application time. The method
may be an effective means of drug delivery for treating
psoriasis or other MTX-sensitive disorders and avoids
potential systemic toxicity.
In another study, Wong et al108 designed a micro-
electrode array to minimize the pain sensation of
electroporation for enhancing transdermal drug
delivery. The influence of the size of the electrode-
skin contact area and of the distance between elec-
trodes on the pain sensation was tested on human
volunteers. The pain level decreased with the dimen-
sion of electrode-skin contact area and with inter-
electrode distance. When both reached about 0.5
mm, the pain level was not perceptible even at the
threshold of transdermal electroporation level of 60
electric pulses at 150 V, 1 ms at 1 to 10 Hz. The elec-
tric thresholds for effective drug delivery, using
toluidine blue O as a marker on mouse skin, were
found to be the same for microelectrode arrays as for
larger electrodes and wider interelectrode distances.
In vivo transdermal electroporation using a micro-
electrode array with 180 pulses of 150 V, 0.2 ms at 1
Hz, followed by a 30-minute methotrexate occlusion
increased more than 4-fold the systemic methotrex-
ate level in mice. The results demonstrated the
potential of painless delivery of significant amounts
of chemotherapeutic agents through skin with the
new electrode arrays in a clinical setting.
Other Uses
Gene Transfer
The easy accessibility of skin makes it an excellent
target for gene transfer protocols. To take advantage
of skin as a target for gene transfer, it is important to
establish an efficient and reproducible delivery
system. Electroporation is an established technique
for enhancing plasmid delivery to many tissues in
vivo. A critical component of this technique is the
electrode configuration. Electroporation parameters
were optimized by Heller et al109 for transgene
expression with minimal tissue damage with a novel
electrode. The highest transgene expression and effi-
ciency of individual cell transformation with mini-
mal damage was produced with eight 150-ms pulses
at a field strength of 100 V/cm. This electrode design
offers the potential for easier and more reproducible
electrically mediated cutaneous plasmid delivery
than the simple electrodes currently commercially
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY
available. This electrode can be a valuable tool in
determining the applicability of electrically medi-
ated cutaneous gene transfer.
The use of a low-intensity current (iontophoresis)
and high-voltage pulses (electroporation that per-
meabilizes lipid bilayers) has a potential for the
administration of conventional and biotechnology-
produced drugs. Iontophoresis and electroporation
enhance transdermal delivery of drugs, including
peptides and oligonucleotides. Electrochemotherapy
(ie, combination of a systemic or local delivery of a
nonpermeant cytostatic drug with electroporation)
kills tumor cells locally. Recently, Préat110 has shown
that the local injection of a plasmid before electropo-
ration significantly increases gene transfection. Hence,
electrotransfer is a promising alternative for drug
and gene delivery.
Intradermal injection of naked DNA results in
gene transfer to skin cells, but the efficiency of this
gene transfer method is relatively low and variable.
Chesnoy and Huang111 have systematically opti-
mized several parameters to obtain reproducible,
high-level gene transfer to the mouse skin. Older
mice (~7 weeks) showed a significant decrease in
gene expression compared with younger mice (4-5
weeks old). The composition of the solvent vehicle
(electrolyte vs nonelectrolyte) strongly affected gene
expression in the skin. A higher level of gene expres-
sion was achieved when naked DNA was dissolved
in isotonic phosphate-buffered saline solution com-
pared with isotonic dextrose solution. Finally, trans-
fection efficiency in older mice was greatly improved
by increasing the ionic strength of the solvent vehi-
cle. The improved transfection efficiency was due to
an enhanced DNA uptake by the skin cells. Gene
transfer was most evident in the subdermal smooth
muscle cells and epidermal cells. With the opti-
mized conditions, gene transfer mediated by intrad-
ermal injection of naked DNA was comparable in
efficiency to electroporation. However, cellular dis-
tributions of the gene transfer of the 2 methods were
different.
Wound Healing
The major goal of wound-healing biology is to
determine how a wound can be induced to repair
damaged tissue faster and more efficiently. Enhance­
ment of dermal and epidermal regeneration is an
extremely important goal for the treatment of many
different types of wounds. Exogenous application of
growth factors to the wound site has been shown to
have potential to improve wound healing. Frequent
Review 1279
applications of large amounts of growth factor have
been required. Gene therapy has the potential to
produce growth factors deep within the wound,
where they can be effective as well as able to
constantly replenish growth factor that is destroyed
by peptidases. Ferguson et al112 have shown that
application of plasmid DNA expression vectors
directly into the wound is an inefficient modality.
Electroporation, the application of an electrical field
across cells to permeabilize the cell membrane, has led
us to explore the possibility of using the technique
to enhance transfection efficiency. They have identi-
fied electroporation parameters that improve the
efficiency of DNA transfection in cutaneous wounds,
and they have shown that electroporation itself
does not impair wound healing. They are now on
the threshold of exploring whether electroporation-
assisted transfection with DNA plasmid expression
vectors for growth factors will be an effective modal-
ity for enhancing cutaneous wound healing.
To Develop Large Permeable Regions
in the Stratum Corneum
The main barrier to transdermal drug delivery in
human skin is the SC. Pulsed electric fields (PEFs) of
sufficient amplitude can create new aqueous path-
ways across this barrier and enhance drug delivery
through the skin. Pliquett and Weaver113 described a
model of pore formation between adjacent corneo-
cytes that predicts the following sequence of events:
(1) the PEF rapidly charges the SC near the electrode
until the transepidermal potential difference is large
enough to drive water into a small region of the SC,
creating new aqueous pathways. (2) PEFs then drive
a high-current density through this newly created
electropore to generate Joule heating that warms the
pore perimeter. (3) This temperature rise at the
perimeter increases the probability of further elec-
troporation there as the local sphingolipids reach
their phase transition temperature. (4) This heat-
generated wave of further electroporation propagates
outward until the surface area of the pore becomes
so large that the reduced current density no longer
generates sufficient heat to reach the phase transi-
tion temperature of the sphingolipids. (5) Cooling
and partial recovery occur after the field pulse.
This process yields large, high-permeability
regions in the SC at which molecules can more read-
ily cross this skin barrier. A model is presented for
this process that predicts that the initial radius of
the first aqueous pathway is approximately 5 nm for
a transdermal voltage of 60 V at room temperature.
 ESCOBAR-CHÁVEZ ET AL
To Reseal Epidermis After Electroporation
The resealing of porcine epidermis after electropora-
tion was investigated by Burgess et al.114 Porcine
epidermis was subjected to electroporation (30
pulses at 100 V, 1 ms, and 1 Hz) in a vertical diffu-
sion apparatus, in the presence of 2 mg/mL dimyris-
toylphosphatidylserine, to produce a long-lasting
permeable state. Resealing treatments include incu-
bation in 0.0625 to 0.25 mM poloxamer 188 (P188)
or incorporation of phosphatidylcholines (PC) and/
or cationic lipids with additional pulses. The recov-
ery of electric resistance of the epidermis samples
after electroporation with or without resealing treat-
ments was monitored. The transports of carboxyflu-
orescein and glucose were measured during the
recovery process. Both P188 and PC were effective
in resealing in terms of electric conductance and
transport, with P188 reacting more rapidly and com-
pletely. P188-mediated lipid exchange between SC
lipid particles was measured by fluorescence reso-
nance energy transfer (FRET). Lipid reorganization
facilitated by P188 and PC is suggested to be a major
resealing mechanism of electroporation damage.
High-voltage pulsing of human skin (approxi-
mately 100 V across the skin, 1-ms pulses) has been
hypothesized to cause electroporation of the SC and
large fluxes of drugs and other molecules across the
skin, through newly created aqueous pathways. In
contrast, iontophoresis (<0.5 mA per cm2, <1 V
across the skin) has long been used in transdermal
drug delivery and is believed to involve preexisting
pathways associated with hair follicles and sweat
ducts. Either high-voltage pulsing or iontophoresis
was applied to human, hairless rat, or black rat
snake skin by Chen et al.115 Hairless rat skin contains
more hair follicles than human skin, and snake skin
does not contain any hair follicles. All 3 types of
skin had comparable electrical resistances at low
voltages; however, the iontophoretic transport of
charged fluorescent molecules was significant for
human and hairless rat skin, but no transport occu­
rred across snake skin, indicating that hair follicles
and sweat ducts play a major role in iontophoresis.
Electroporation caused large molecular transport for
all 3 types of skin and involved spontaneously form-
ing localized transport regions not associated with
appendages. These experiments thus provide further
support for the hypothesis that high-voltage pulsing
causes electroporation in the SC and that this trans-
port mechanism is fundamentally different from
iontophoresis.
1280  •  J Clin Pharmacol 2009;49:1262-1283
CONCLUSIONS
It should be evident from this review that electropo-
ration holds a lot of promise for the future of trans-
dermal drug delivery. Electroporation is an efficient
method for enhancing transdermal drug delivery in
vitro and in vivo and expands the range of com-
pounds delivered transdermally. It could be a prom-
ising alternative as a noninvasive delivery of
macromolecules (up to at least 40 kDa) and fast and/
or pulsatile transdermal delivery.
The combined use of electroporation with other
physical enhancers such as iontophoresis is likely to
yield useful and interesting data, which will inten-
sify the efforts to more fully explore electropora-
tion as a means of transdermal drug delivery.
The reader should realize that electrically assisted
delivery of drugs involves several chemical, bio-
chemical, and physiological processes, and there is
overlap in the mechanisms involved in transport via
electroporation.
In summary, electroporation is one of the physic-
ochemical methods for gene and drug delivery. It is
superior in some aspects but also has several draw-
backs. In some cases, as attested by the voluminous
literature, it is still the preferred method. New devel-
opments are expected in the future to make this
method even more versatile. Pulse protocol and
electrode design need to be optimized to reduce the
main adverse effect (ie, muscle contraction).
Financial disclosure: José Juan Escobar-Chávez acknowledges
the PROFIP/UNAM grant and PAPITT 209709.
REFERENCES
1. Elias PM. Epidermal lipids, barrier function and desquamation.
J Invest Dermatol. 1983;80:44-49.
2. Cleary GW. Transdermal delivery systems: a medical rationale.
In: Shah VP, Maibach HI, eds. Topical Drug Bioavailability,
Bioequivalence and Penetration. New York: Plenum; 1993:17.
3. Scheuplein RJ, Blank IH. Permeability of the skin. Physiol Rev.
1971;51:702-747.
4. Williams AC, Barry BW. Penetration enhancers. Adv Drug Deliv
Rev. 2004;56:603-618.
5. Pellet M, Raghavan SL, Hadgraft J, Davis AF. The application of
supersaturated systems to percutaneous drug delivery. In: Guy
RH, Hadgraft J, eds. Transdermal Drug Delivery. New York:
Marcel Dekker; 2003:305.
6. Tsai JC, Guy RH, Thornfeldt CR, et al. Metabolic approaches to
enhance transdermal drug delivery: 1. Effect of lipid synthesis
inhibitors. J Pharm Sci. 1998;85:643-648.
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY
7. Elias PM, Feingold KR, Tsai J, Thornfeldt C, Menon G.
Metabolic approach to transdermal drug delivery. In: Guy RH,
Hadgraft J, eds. Transdermal Drug Delivery. New York: Marcel
Dekker; 2003:285.
8. Schreier H, Bouwstra J. Liposomes and niosomes as topical
drug carriers: dermal and transdermal drug delivery. J Control
Release. 1994;30:1-15.
9. Cevc G. Transfersomes, liposomes and other lipid suspensions
on the skin: permeation enhancement, vesicle penetration, and
transdermal drug delivery. Crit Rev Ther Drug Carrier Syst.
1996;13:257-388.
10. Cevc G. Transferosomes: innovative transdermal drug carriers.
In: Rathbone MJ, Hadgraft J, Roberts MS, eds. Modified Release
Drug Delivery Technology. New York: Marcel Dekker; 2003:533.
11. Godin B, Touitou E. Ethosomes: new prospects in transdermal
delivery. Crit Rev Ther Drug Carrier Syst. 2003;20:63-102.
12. Brown MB, Traynor MJ, Martin GP, Akomeah FK. Transdermal
drug delivery systems: skin perturbation devices. In: Jain KK, ed.
Drug Delivery Systems. Vol. 37. Totowa, NJ: Humana; 2008:
119-139.
13. Escobar-Chávez JJ, Merino-Sanjuán V, López-Cervantes M,
et al. The use of iontophoresis in the administration of nicotine
and new non-nicotine drugs through the skin for smoking cessation.
Curr Drug Discov Technol. 2009 Sep 1. [Epub ahead of print]
14. Merino V, Micó-Albiñana T, Nácher A, Díez-Sales O, Herráez
M, Merino-Sanjuán M. Enhancement of nortriptyline penetration
through human epidermis: influence of chemical enhancers and
iontophoresis. J Pharm Pharmacol. 2008;60:415-420.
15. El-Kamel AH, Al-Fagih IM, Alsarra IA. Effect of sonophoresis
and chemical enhancers on testosterone transdermal delivery
from solid lipid microparticles: an in vitro study. Curr Drug Deliv.
2008;5:20-26.
16. Zhang L, Widera G, Rabussay D. Enhancement of the effective-
ness of electroporation-augmented cutaneous DNA vaccination
by a particulate adjuvant. Bioelectrochemistry. 2004;63:369-373.
17. Banga AK, Bose S, Ghosh TK. Iontophoresis and electropora-
tion: comparisons and contrasts. Int J Pharm. 1999;179:1-19.
18. Prausnitz MR. A practical assessment of transdermal drug
delivery by skin electroporation. Adv Drug Deliv Rev. 1999;
35:61-76.
19. Vanbever R, Préat V. In vivo efficacy and safety of skin elec-
troporation. Adv Drug Deliv Rev. 1999;35:77-88.
20. Forslind BA. Domain mosaic model of skin barrier. Acta Derm
Venereol. 1994;74:1-6.
21. Potts RO, Francoeur ML. The influence of stratum corneum
morphology on water permeability. J Invest Dermatol. 1991;96:
495-499.
22. Potts RO, Guy RH. Predicting skin permeability. Pharm Res.
1992;9:663-669.
23. Ellias PM. Epidermal barrier function: intercellular lamellar
lipid structures, origin, composition and metabolism. J Control
Release. 1991;15:199-208.
24. Barry BW. Dermatological formulations: percutaneous absorp-
tion. In: Swarbick J, ed. Drugs and the Pharmaceutical Sciences.
New York: Marcel Dekker; 1983:202.
25. Hadgraft J. Skin, the final frontier. Int J Pharm. 2001;224
(1-2):1-18.
26. Guy RH, Hadgraft J. Transdermal Drug Delivery. New York:
Marcel Dekker; 2003.
Review 1281
27. Walters KA, Roberts MS. Dermatological and Transdermal
Formulations. New York: Marcel Dekker; 2002.
28. Nevill AM. The need to scale for differences in body size and
mass: and explanation of Klieber’s 0.75 mass exponent. Am
Physiol Soc. 1994;77:2870-2873.
29. Olvera-Martínez BI, Cazares-Delgadillo J, Calderilla-Fajardo
SB, Villalobos-García R, Ganem-Quintanar A, Quintanar-Guerrero
D. Preparation of polymeric nanocapsules containing octyl
methoxycinnamate by the emulsification-diffusion technique:
penetration across the stratum corneum. J Pharm Sci. 2005;94:
1552-1559.
30. Escobar-Chávez JJ, Quintanar-Guerrero D, Ganem-Quintanar
A. In vivo skin permeation of sodium naproxen formulated in
PF-127 gels: Effect of Azone and Transcutol. Drug Develop Ind
Pharm. 2005;31:447-454.
31. Escobar-Chávez JJ, López-Cervantes M, Naïk A, Kalia YN,
Quintanar-Guerrero D, Ganem-Quintanar A. Applications of the
thermoreversible Pluronic F-127 gels in pharmaceutical formula-
tions. J Pharm Pharmaceut Sci. 2006;9:339-358.
32. Miyazaki S, Yokouchi Ch, Nakamura T, Hashiguchi N,
Hou W-M, Takada M. Pluronic F-127 gels as a novel vehicle for
rectal administration of indomethacin. Chem Pharm Bull. 1986;
34:1801-1808.
33. Chi S-Ch, Do K, Tan HK, Chun HW. Anti-inflammatory and
analgesic transdermal gel. US patent number 5,527,832, 1996.
34. Fang JY, Leu YL, Wang YY, Tsai YH. In vitro topical applica-
tion and in vivo pharmacodynamic evaluation of nonivamide
hydrogels using Wistar rat as an animal model. Eur J Pharm Sci.
2002;15:417-423.
35. Shin SC, Cho CW, Oh IJ. Effects of non ionic surfactants as
permeation enhancers towards piroxicam from the poloxamer gel
through rat skins. Int J Pharm. 2001;222:199-203.
36. Liaw J, Lin Y-Ch. Evaluation of poly(ethylene oxide)-
poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) gels
as a release vehicle for percutaneous fentanyl. J Control Release.
2000;68:273-282.
37. Wang YY, Hong CT, Chiu WT, Fang JY. In vitro and in vivo
evaluations of topically applied capsaicin and nonivamide from
hydrogels. Int J Pharm. 2001;224:1-2.
38. Kattan El AF, Asbill CS, Kim N, Michniak BB. Effect of formu-
lation variables on the percutaneous permeation of ketoprofen
from gel formulations. Drug Deliv. 2000;7(3):147-153.
39. Curdy C, Kalia YN, Naïk A, Guy RH. Piroxicam delivery into
human stratum corneum in vivo: iontophoresis versus passive
diffusion. J Control Release. 2001;76:73-79.
40. Mattorano DA, Kupper LL, Nylander-French LA. Estimating
dermal exposure to jet fuel (naphthalene) using adhesive tape
strip samples. Ann Occup Hyg. 2004;48:139-146.
41. Chao Y-Ch, Nylander-French LA. Determination of keratin
protein in a tape-stripped skin sample from jet fuel exposed skin.
Ann Occup Hyg. 2004;48:65-73.
42. Chizmadzhev YA, Zamitsin VG, Weaver JC, Potts UO.
Mechanism of electroinduced ionic species transport through a
multilamellar lipid system. Biophysical J. 1995;68:749-765.
43. Chizmadzhev YA, Indenbom AV, Kuzmin PI, Galichenko SV,
Weaver JC, Potts RO. Electrical properties of skin at moderate
voltages: contribution of appendageal macropores. Biophys J.
1998;74:843-856.
44. Jiali B, Hong W, Huiping W, Jiguang G. Quantitative descrip-
tion and model of molecule transport through skin. Presented at:
 ESCOBAR-CHÁVEZ ET AL
26th Annual International Conference of the IEEE EMBS; Septem­
ber 1-5, 2004; San Francisco, California.
45. Treco DA, Selden RF. Non-viral gene therapy. Mol Med Today.
1995;1:314-321.
46. Weaver JC. Electroporation theory: concepts and mechanisms.
In: Nickoloff JA, ed. Electroporation Protocols for Microorganisms.
Totowa, NJ: Humana; 1995:1.
47. Prausnitz, MR, Bose VG, Langer R, Weaver JC. Electroporation
of mammalian skin: a mechanism to enhance transdermal drug
delivery. Proc Natl Acad Sci USA. 1993;90:10504-10508.
48. Inada H, Ghanem AH, Higuchi WI. Studies on the effects of
applied voltage and duration on human epidermal membrane
alteration: recovery and the resultant effects upon iontophoresis.
Pharm Res. 1994;11:687-697.
49. Weaver JC, Chizmadzhev Y. Electroporation. In: Polte C,
Postow E, eds. Biological Effects of Electromagnetic Fields. Boca
Raton, FL: CRC Press; 1996:247.
50. Edwards DA, Prausnitz MR, Langer R, Weaver JC. Analysis of
enhanced transdermal transport by skin electroporation. J Control
Release. 1995;34:211-221.
51. Prausnitz MR. Do high-voltage pulses cause changes in skin
structure? J Control Release. 1996;40:321-326.
52. Vanbever R, Lecouturier N, Preat V. Transdermal delivery of
metoprolol by electroporation. Pharm Res. 1994;11:1657-1662.
53. Vanbever R, LeBoulenge E, Preat V. Transdermal delivery of
fentanyl by electroporation: 1. Influence of electrical factors.
Pharm Res. 1996;13:559-565.
54. Choi MJ, Maibach HI. Topical vaccination of DNA antigens:
topical delivery of DNA antigens. Skin Pharmacol Appl Skin
Physiol. 2003;16:271-282.
55. Wang S, Kara M, Krishnan TR. Topical delivery of cyclosporin
A coevaporate using electroporation technique. Drug Dev Ind
Pharm. 1997;23:657-663.
56. Prausnitz MR, Edelman ER, Gimm JA, Langer R, Weaver JC.
Transdermal delivery of heparin by skin electroporation.
Biotechnology. 1995;13:1205-1209.
57. Zewert TE, Pliquett UF, Langer R, Weaver JC. Transdermal
transport of DNA antisense oligonucleotides by electroporation.
Biochem Biophys Res Commun. 1995;212:286-292.
58. Brand RM, Wahl A, Iverson PL. Effects of size and sequence
on the iontophoretic delivery of oligonucleotides. J Pharm Sci.
1998;87:49-52.
59. Zhang L, Hofmann GA. Electric pulse mediated transdermal
drug delivery. Proc Int Symp Control Release Bioact Mater.
1997;24:27-28.
60. Pliquett U, Gallo S, Hui SW, Gusbeth C, Neumann E. Local and
transient structural changes in stratum corneum at high electric
fields: contribution of Joule heating. Bioelectrochemistry. 2005;
67:37-46.
61. Fang JY, Huang Y-B, Wang H-Y, Tsai Y-H. Electrically-assisted
skin permeation of two synthetic capsaicin derivatives, sodium
nonivamide acetate and sodium nonivamide propionate, via rate-
controlling polyethylene membranes. Biol Pharm Bull. 2005;28:
1695-1701.
62. Murthy SN, Sen A, Zhao YL, Hui SW. pH influences the post-
pulse permeability state of skin after electroporation. J Control
Release. 2003;93:49-57.
1282  •  J Clin Pharmacol 2009;49:1262-1283
63. Hu Q, Liang W, Bao J, Ping Q. Enhanced transdermal delivery
of tetracaine by electroporation. Int J Pharm. 2000;202:121-124.
64. Bommannan DB, Tamada J, Leung L, Potts RO. Effect of elec-
troporation on transdermal iontophoretic delivery of luteinizing
hormone releasing hormone (LHRH) in vitro. Pharm Res. 1994;
11:1809-1814.
65. Murthy SN, Zhao Y, Hui SW, Sen A. Synergistic effect of
anionic lipid enhancer and electroosmosis for transcutaneous
delivery of insulin. Int J Pharm. 2006;326(1/2):1-6.
66. Regnier V, Préat V. Mechanisms of a phosphorothioate oligo-
nucleotide delivery by skin electroporation. Int J Pharm. 1999;
184:147-156.
67. Vanbever R, Leroy MA, Préat V. Transdermal permeation of
neutral molecules by electroporation. J Control Release. 1998;54:
243-250.
68. Li LH, McCarthy P, Hui SW. High-efficiency electrotransfec-
tion of human primary hematopoietic stem cells. FASEB J.
2001;15:586-588.
69. Yew NS, Zhao H, Przybylska M, et al. CpG-depleted plasmid
DNA vectors with enhanced safety and long-term gene expression
in vivo. Mol Ther. 2002;5:731-738.
70. Chen S, Shohet RV, Bekeredjian R, Frenkel P, Grayburn PA.
Optimization of ultrasound parameters for cardiac gene delivery
of adenoviral or plasmid deoxyribonucleic acid by ultrasound-
targeted microbubble destruction. J Am Coll Cardiol. 2003;42:
301-308.
71. Ding Z, Fach C, Sasse A, Godecke A, Schrader J. A minimally
invasive approach for efficient gene delivery to rodent hearts.
Gene Ther. 2004;11:260-265.
72. Tanaka S, Iwai M, Harada Y, et al. Targeted killing of carcino-
embryonic antigen (CEA)–producing cholangiocarcinoma cells by
polyamidoamine dendrimer–mediated transfer of an Epstein-Barr
virus (EBV)–based plasmid vector carrying the CEA promoter.
Cancer Gene Ther. 2000;7:1241-1250.
73. Keravala A, Groth AC, Jarrahian S, et al. A diversity of serine
phage integrases mediate site-specific recombination in mamma-
lian cells. Mol Genet Genomics. 2006;276:135-146.
74. Nickoloff JA. Preface. In: Nickoloff JA, ed. Electroporation
Protocols for Microorganisms. Totowa, NJ: Humana; 1995:5.
75. Miller EM, Nickoloff JA. Escherichia coli electrotransforma-
tion. In: Nickoloff JA, ed. Electroporation Protocols for
Microorganisms. Totowa, NJ: Humana; 1995:105.
76. Tetsuichiro S. In Vivo Electroporation. http://www.frontier.
kyoto-u.ac.jp/rc01/in_vivo_electroporation.html. Accessed February
15, 2003.
77. Weaver JC. Electroporation theory: concepts and mechanisms.
In: Nickoloff JA, ed. Electroporation Protocols for Microorganisms.
Totowa, NJ: Humana; 1995:1.
78. Pliquett U, Langer R, Weaver JC. Changes in the electrical
properties of human stratum corneum due to electroporation.
Biochim Biophys Acta. 1995;1239:111-121.
79. Mir L, Orlowski S, Belehradek JJ, Paolelli C. Electrochemo­
therapy: potentiation of antitumor effect of bleomycin by electric
pulses. Eur J Cancer. 1991;27:68-72.
80. Heller R, Jaroszeski R, Glass LF, et al. Phase I/II trial for the
treatment of cutaneous and subcutaneous tumor using electro-
chemotherapy. Cancer. 1996;77:964-971.
 ELECTROPORATION AS A PHYSICAL ENHANCER FOR SKIN DRUG DELIVERY
81. Murthy SN, Zhao Y-L, Sen A, Hui SW. Cyclodextrin enhanced
transdermal delivery of piroxicam and carboxyfluorescein by
electroporation. J Control Release. 2004;99:393-402.
82. Huang JF, Sung KC, Hu O Y-P, Wang JJ, Lin YH, Fang JY. The
effects of electrically assisted methods on transdermal delivery of
nalbuphine benzoate and sebacoyl dinalbuphine ester from solu-
tions and hydrogels. Int J Pharm. 2005;297:162-171.
83. Sung KC, Fang J-Y, Wang JJ, Hu O Y-P. Transdermal delivery
of nalbuphine and its prodrugs by electroporation. Eur J Pharm
Sci. 2003;18:63-70.
84. Fang J-Y, Hwang T-L, Huang Y-B, Tsai Y-H. Transdermal
iontophoresis of sodium nonivamide acetate: V. Combined effect
of physical enhancement methods. Int J Pharm. 2002;235:95-105.
85. Vanbever R, Prausnitz MR, Préat V. Macromolecules as novel
transdermal transport enhancers for skin electroporation. Pharm
Res. 1997;14:638-644.
86. Badkar AV, Smith AM, Eppstein JA, Banga AK. Transdermal
delivery of interferon alpha-2B using microporation and ionto-
phoresis in hairless rats. Pharm Res. 2007;24:1389-1395.
87. Sharma A, Kara M, Smith FR, Krishnan TR. Transdermal drug
delivery using electroporation: II. Factors influencing skin revers-
ibility in electroporative delivery of terazosin hydrochloride in
hairless rats. J Pharm Sci. 2000;89:536-544.
88. Sharma A, Kara M, Smith FR, Krishnan TR. Transdermal drug
delivery using electroporation: I. Factors influencing in vitro
delivery of terazosin hydrochloride in hairless rats. J Pharm Sci.
2000;89:528-535.
89. Denet A-N, Ucakar B, Préat V. Transdermal delivery of timolol
and atenolol using electroporation and iontophoresis in combina-
tion: a mechanistic approach. Pharm Res. 2003;20:1946-1951.
90. Fang J-Y, Hung Ch-F, Fang Y-P, Chan T-F. Transdermal ionto-
phoresis of 5-fluorouracil combined with electroporation and
laser treatment. Int J Pharm. 2004;270:241-249.
91. Sersa G, Miklavcic D, Cemazar M, Rudolf Z, Pucihar G,
Snoj M. Electrochemotherapy in treatment of tumours. Eur J Surg
Oncol. 2008;34:232-240.
92. Murthy SN, Zhao Y-L, Marlan K, Hui SW, Kazim AL, Sen A.
Lipid and electroosmosis enhanced transdermal delivery of insu-
lin by electroporation. J Pharm Sci. 2006;95:2041-2050.
93. Tokumoto S, Higo N, Sugibayashi K. Effect of electroporation
and pH on the iontophoretic transdermal delivery of human insu-
lin. Int J Pharm. 2006;326:13-19.
94. Medi BM, Singh J. Electronically facilitated transdermal
delivery of human parathyroid hormone (1-34). Int J Pharm.
2003;263:25-33.
95. Chang S-L, Hofmann GA, Zhang L, Deftos LJ, Banga AK. The
effect of electroporation on iontophoretic transdermal delivery of
calcium regulating hormones. J Control Release. 2000;66:127-133.
96. Essa EA, Bonner MC, Barry BW. Electrically assisted skin
delivery of liposomal estradiol: phospholipid as damage retar-
dant. J Control Release. 2004;95:535-546.
97. Johnson PG, Hui SW, Oseroff AR. Electrically enhanced per-
cutaneous delivery of d-aminolevulinic acid using electric pulses
and a DC potential. Photochem Photobiol. 2002;75:534-540.
98. Fang JY, Lee W-R, Shen S-C, Fang Y-P, Hu C-H. Dermatological
surgery and lasers enhancement of topical 5-aminolaevulinic
acid delivery by erbium:YAG laser and microdermabrasion: a
Review 1283
comparison with iontophoresis and electroporation. Br J Dermatol.
2004;151:132-140.
99. Regnier V, Tahiri A, André N, Lemaitrec M, Préat V.
Electroporation-mediated delivery of 3′-protected phosphodiester
oligodeoxynucleotides to the skin. J Control Release. 2000;67:
337-346.
100. Zhao YL, Murthy SN, Manjili MH, Guana LJ, Sena A, Hui SW.
Induction of cytotoxic T-lymphocytes by electroporation-enhanced
needle-free skin immunization. Vaccine. 2006;24:1282-1290.
101. Hui SW. The application of electroporation to transfect
hematopoietic cells and to deliver drugs and vaccines transcuta-
neously for cancer treatment. Technol Cancer Res Treat.
2002;1:373-384.
102. Fang JY, Hung CF, Hwang TL, Wong WW. Transdermal deliv-
ery of tea catechins by electrically assisted methods. Skin Phar­
macol Physiol. 2006;19:28-37.
103. Marrat F, Levy JL, Santi P, Kalia YN. In vitro evaluation of
electrotreatment on skin permeability. J Cosmet Dermatol. 2008;7:
105-111.
104. Sen A, Zhao Y, Zhang L, Hui SW. Enhanced transdermal
transport by electroporation using anionic lipids. J Control
Release. 2002;82:399-405.
105. Tokudome Y, Sugibayashi K. Mechanism of the synergic
effects of calcium chloride and electroporation on the in vitro
enhanced skin permeation of drugs. J Control Release. 2004;95:
267-274.
106. Murthy SN, Sen A, Hui SW. Surfactant-enhanced transdermal
delivery by electroporation. J Control Release. 2004;98:307-315.
107. Wong T-W, Zhao Y-L, Sen A, Hui SW. Pilot study of topical
delivery of methotrexate by electroporation. Br J Dermatol.
2005;152:524-530.
108. Wong TW, Chen Ch-H, Huang Ch-Ch, Lin Ch-D, Hui SW.
Painless electroporation with a new needle-free microelectrode
array to enhance transdermal drug delivery. J Control Release.
2006;110:557-565.
109. Heller LC, Jaroszeski MJ, Coppola D, McCray AN, Hickey J,
Heller R. Optimization of cutaneous electrically mediated plas-
mid DNA delivery using novel electrode. Gene Ther. 2007;14:
275-280.
110. Préat V. Drug and gene delivery using electrotransfer. Ann
Pharm Fr. 2001;59:239-244.
111. Chesnoy S, Huang L. Enhanced cutaneous gene delivery fol-
lowing intradermal injection of naked DNA in a high ionic
strength solution. Mol Ther. 2002;5:57-62.
112. Ferguson M, Byrnes C, Sun L, et al. Wound healing enhance-
ment: electroporation to address a classic problem of military
medicine. World J Surg. 2005;29:S55-S59.
113. Pliquett U, Weaver JC. Feasibility of an electrode-reservoir
device for transdermal drug delivery by noninvasive skin elec-
troporation. IEEE Trans Biomed Eng. 2007;54:536-538.
114. Burgess SE, Zhao YL, Sen A, Hui SW. Resealing of electropo-
ration of porcine epidermis using phospholipids and poloxamers.
Int J Pharm. 2007;336:269-275.
115. Chen T, Langer R, Weaver JC. Skin electroporation causes
molecular transport across the stratum corneum through local-
ized transport regions. J Investig Dermatol Symp Proc. 1998;3:
159-165.