[Dermato-Endocrinology 1:4, 197-206; July/August 2009]; ©2009 Landes Bioscience

Review

Nanoparticles and their interactions with the dermal barrier

Marc Schneider,1,* Frank Stracke,2 Steffi Hansen3 and Ulrich F. Schaefer3
1Universität des Saarlandes; Pharmazeutische Nanotechnologie; Saarbrücken, Germany; 2Fraunhofer Institut für Biomedizinische Technik; Functional Optics Group;
St. Ingbert, Germany; 3Universität des Saarlandes; Biopharmazie und Pharmazeutische Technologie; Saarbrücken, Germany

Abbreviations: MPA, 3-mercaptopropionic acid; TMAOH, tetramethylammonium hydroxide; AOT, sodium bis(2-ethylhexyl) sulfo-
succinate; Baa-Lys-(FITC)-NLS, phenylalanine-based fullerene [bucky amino acid] (fluorescein isothiocyanate) nuclear localisation
sequence; Cit, citrate; SEM, scanning electron microscopy; TEM, transmission electron microscopy; AuNP, gold nanoparticles; AgNP,
silver nanoparticles
Key words: nanoparticles, human skin, skin models, particle penetration, particle permeation, follicular targeting

The dermal application of drugs is promising due to the
ease of application. In this context nano-scale carrier systems
were already evaluated in several studies with respect to the skin
interaction and the impact on drug penetration. At the same
time the upcoming production of engineered nano-scale mate-
rials requires a thorough safety evaluation. Drug delivery as
well as risk assessment depends crucially on the ability of such
carriers to overcome the skin barrier and reach deeper tissue
layers. Therefore, the interaction of nanoparticles with skin and
especially skin models is an intriguing field. However, the data
obtained do not show a clear image on the effect of nano-carriers.
Especially the penetration of such particles is an open and
controversially discussed topic. The literature reports different
results mainly on pig or murine skin showing strong penetration
(pig and mouse) or the opposite. Looking only at the sizes of the
particles also no conclusive picture can be obtained. Nevertheless,
size is regarded to play an important role for skin penetra-
tion. Furthermore, the state of the skin influences penetration
(hydration) and the mechanical stress is of outmost importance.
Introduction
Any kind of organism always faced nanoscale environmental
compounds interacting with their exterior barrier but a significant
interest in these interactions came up not until the broad ascent
of artificial nanoparticulate compounds.1 The great potential
for future advanced applications in areas such as energy, elec-
tronics, automotive, chemistry and life sciences led to a rapidly
growing number of nanoparticulate systems, with respect to mate-
rial composition, size, shape and formulation. Nanomedicines,
*Correspondence to: Marc Schneider; Saarland University; Pharmaceutical
Nanotechnology; Campus A4 1; Saarbrücken 66123 Germany; Tel.:
+49.681.302.2438; Fax: +49.681.302.4677; Email: marc.schneider@
mx.uni-saarland.de
Submitted: 07/10/09; Accepted: 07/13/09
Previously published online as a Dermato-Endocrinology E-publication:
http://www.landesbioscience.com/journals/dermatoendocrinology/article/9501
i­ncorporating drug delivery, diagnostics, implants, cancer therapy,
to name just a few, pose an important and appealing area for
beneficial application. Regarding final applicability it is essential to
investigate the various biological aspects of nanoparticle exposure
to the human organism. On one hand health hazards are to be
identified and assessed, on the other hand medical and pharma-
ceutical potentials may be discovered and exploited. The behavior
of nanoparticulate substances in biosystems and their physiological
effects can, up to now, neither be extrapolated straightforwardly
from bulk properties nor can they be predicted from molecular
properties of the constituents. The first step of any interaction
between an organism and any compound is the uptake of the
compound from the environment. Several pathways of absorption
exist for human (and most animal) organisms of which the skin is
the most obvious and easiest to reach.
The present review focus on skin and its barrier as well as sink
function to artificial nanoparticulate compounds. Different classes
of nanoparticulate material were studied to gain knowledge on
the interaction and possible impact on skin covering bio-resistant
particles (e.g., metal oxide, carbon-based) and biodegradable parti-
cles (e.g., liposomes, polymeric particles). Furthermore, mainly
healthy skin is treated herein. Injured and inflamed skin can be
entered even by particles larger than the nanometer scale due to
the loss of its barrier function.1 This is also the case for chemically
induced irritations of the skin, e.g., by non-polar solvents and
strong alkalines. However, particle penetration in combination
with deliberately damaged skin or skin under mechanical stress is
available and will be discussed.
Pathways for Skin Absorption
Regarding the absorption of any kind of material there can
be considered two general pathways: along the skin append-
ages or through the stratum corneum and the underlying layers.
Even though the appendages present only a small portion of
the surface they are considered to contribute and might even be
addressed for a directed delivery because of the depth they reach.
Especially the hair follicles seem to be a promising target regarding

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 Nanoparticles and skin

Figure 1. Color-coded simulations of three diffusing compounds in a 3D stratum corneum model, calculated for different diffusivity ratios of the com-
pounds between corneocytes (bricks) and in the intercellular space (mortar). (Top view; red indicates high concentration). For nanoparticulate compounds
a strictly intercellular route (right case) will be due. (Courtesy of M. Heisig, IWR, Ruprecht-Karls-Universität Heidelberg, Germany).77

­ anoparticulate carrier systems.2,3 The invasion of a substance

n
into a skin appendage is not yet an absorption process itself.
Compounds inside an appendage are still on the outside of the
body by definition. Nevertheless accumulation in such structures
may lead to faster and more efficient uptake due to altered barrier
morphology in the appendages what will be discussed in detail in
a separate paragraph.
Generally, the SC is assumed to be the main barrier for absorp-
tion. The hydrophilicity of the absorbent is hereby crucial. The
absorption across the stratum corneum offers two possible path-
ways which are obvious: through the corneocytes (bricks) or along
the intercellular spaces along the lipid matrix (mortar) (Fig. 1). The
latter pathway seems to be most suited for penetration offering
channel-like structures providing higher diffusivity although the
pathway is much longer (Fig. 2).4
The full understanding of the processes being responsible for
the penetration process is not yet obtained. This holds true even
more for particle penetration. For molecular species, approaches
relying on a close interaction of theoretic modeling approaches Figure 2. Sketch of the morphology of human stratum corneum: The
based on experimentally extracted data is necessary and in stratum corneum is composed out of rigid corneocytes (CC), dead cells
densely packed with keratinous filaments in a matrix of connecting
process.5-7 proteins and confined by the cornified envelope (CE), separated by the
Overall, a second barrier functionality is achieved by composing intercellular space (IS) filled with lipid compounds (fatty acids, ceramides
layers of different polar character. The SC can be considered to etc.,) and aqueous films to form lamellar lipid bilayers (LLB) (published with
be a lipoidic compartment whereas the underlying living tissue permission from ref. 4).
is a more aqueous environment. Hence lipophilic molecules can
distribute more easily in the SC and penetration is facilitated.
Furthermore, the absorbent needs to fit into the intercellular space
and need to move along the lipid phase or the aqueous phase, These two aspects of penetration and permeation, the size of
respectively, restricting the space available and therefore the size of an absorbent as well as is partition coefficient, are expressed in the
penetrating species influences the absorption behavior. Diffusion formula of Potts and Guy who found a phenomenological expres-
into the living tissue, however, is in favor of polar molecules and sion based on molecular properties to describe the absorption9
objects restricting the invasion of too lipophilic compounds.8 As
a consequence only substances with partition coefficients (logP)   (6)
between 1 and 3 are well suited for skin absorption (Fig. 3).

198 Dermato-Endocrinology 2009; Vol. 1 Issue 4


Figure 3. Relation between absorption and lipophilicity. Optimal absorp-
tion conditions are for 1 ≤ log (P) ≤ 3 (P being the octanol/water partition
coefficient).
Table 1 “Best case” transport parameters of
nanoparticles over a typical stratum corneum
barrier
r/nm Kp/m s-1 JSS/s-1 m-2 with c = 1020 m-3
5 3.0 × 10-12 3.0 × 108
10 1.5 × 10-12 1.5 × 108
50 3.0 × 10-13 3.0 × 107
100 1.5 × 10-13 1.5 × 107
Calculations were based on a diffusion path length of h = 500 µm (Moghimi et al.,12 Potts et al.78), an
area fraction of lipid channels of 1% and a viscosity of 0.3 Pa s, which is estimated from the diffusion
coefficient of 5-fluoruracil (Moghimi et al.12).
Hereby D is the not restricted diffusivity of the permeant in the
membrane (skin), h represents the length of the diffusion pathway,
MW is the molecular weight (for most molecules the molecular
weight correlates well with the molecular volume), P is the octanol/
water partition coefficient (accessibility of P is better than the value
of the membrane/water coefficient Km), and Kp is the permeability
coefficient. Several attempts were made to optimize this relatively
crude phenomenological expression but only slight improvements
were achieved. Most approaches find the molecular weight to be
a strong effector and reliable measure to predict permeabilities
and maximum fluxes of solutes through the stratum corneum, as
shown by Magnusson et al.10
Empirical findings by Bos et al.11 known as the “500 Dalton
rule,” even indicate a molar mass exclusion effect for dermal
penetration, which is not predicted by the quantitative ­penetration
models of Potts and Guy and others (Potts,9 Magnusson10).
Nevertheless it is shown that diffusion of high-MW corpuscles is
drastically impeded in the stratum corneum. As nanoparticles have
molar masses on the order of 106 Dalton and more no significant
transport over an intact dermal barrier is to expect for human
beings concerning these predictions.
Assuming that particles penetration and diffusion will follow
the intercellular route a “best case” flux of particles over the
stratum corneum can be estimated by classical diffusion theory.
www.landesbioscience.com
Nanoparticles and skin
The particles need to diffuse a distance of some hundred μm (say
h’ = 500 μm) via the lipid-filled intercellular space, which has a
cross sectional area of about ε = 1% of the total surface area.12
Approximating the situation (coarse simplification) by assuming a
homogeneous membrane matrix (hence neglecting any size exclu-
sion effects at pores, adsorption at corneocyte surfaces and phase
transition steps) of diffusivity D and furthermore no limitation in
drug amount (infinite dose) on the outer surface, one may calculate
steady state flux parameters over the barrier. The diffusivity D for
spherical nanoparticles is approximated by the Stokes-Einstein
relation and data from Moghimi et al.12
  (7)
(with k: Boltzmann constant, η: dynamic viscosity, r: particle
radius, h’: diffusion distance, ε: area fraction of intercellular
spaces). Obviously, the particle flux drops with the reciprocal of r.
A modified maximum particle flux can be defined by assuming a
maximum nanoparticle density of 74% (“closest packed”) of the
reciprocal of the particle volume:
  (8)
Following these assumptions, the maximum particle flux
decreases with the fourth power of the particle radius. Some exem-
plifying results of such estimations are given in Table 1. In reality,
the first particles will arrive at the inner interface of the barrier
not until a lag time elapsed. During this lag time the steady state
conditions, i.e., the constant gradient over the barrier, develops.
This time also increases with the particle radius. Together with the
impeding effects due to the inhomogeneous, porous nature of the
stratum corneum as well as the finite dose and exposure time, this
strongly limits transport of nanoparticulate material into human
skin far below the best case estimation.
Penetration & Permeation Studies of Nanoparticulate
Materials
As it is know that nanosized carriers behave differently when
interacting with biological barriers, their potential for transdermal
drug delivery purposes was addressed in several studies. Applying
drugs in pharmaceutical formulations containing nanoparticulate
material as drug delivery devices was carefully investigated.13-23
At the same time safety issues gained more and more importance
to assess the risk when exposed unintended to these particles.24-26
Here open question for nanoparticulate potential to induce adverse
effects based on the nanoparticles properties and their interaction
mechanism are of interest. This includes the interaction with
proteins which may alter the nanoparticles behavior after adsorp-
tion to the surface as well as nanoparticles might impact on the
behavior of proteins.27 Furthermore genotoxic effects induced
either directly or indirectly (inflammatory processes-mediated)
can play a roll as well as oxidative stress due to the formation of
active oxygen species. Some nanoparticulate materials might be
Dermato-Endocrinology 199
 Nanoparticles and skin

activated by light and result in photo-

sensitation.27
As already mentioned above, particle
penetration is most likely along two
possible routes: the intercellular route,
following the lipid channels between
the corneocytes to deeper skin layers
and the appendage route (hair follicles,
sweat glands). Both ways have shown
considerable interaction with nano-
particulate formulations.2,14,15,28
In this paragraph an overview of
penetration and permeation studies
on nanoparticulate compounds into
human skin and its models (mainly
different animal skins) will be given,
with an emphasis on skin models and Figure 4. PLGA nanoparticles in human skin furrows 5 hours after administration. (a) A two photon
micrograph at z = 15 μm subsurface depth showing superficial keratin fluorescence and clearly resolved
penetration conditions.
single particles. (b) A pseudo-color overlay of two photon and confocal images at z = 28 μm revealing
This overview will be subdivided the release of a drug dummy from the particles and its cutaneous uptake. The two photon channel (green)
according to the particles’ chemical shows keratinous layers and single particles, the 488 nm-excited confocal channel (blue) addresses the
composition, which concomitantly fluorescein-labeled particles solely and the 543 nm-excited confocal channel (red) exclusively shows the
includes a classification according to dummy compound (Texas Red) (published with permission from ref. 4).
application foci of the studies: risk
assessment for the metal and mineral nanoparticles, pharmaceu-
tical applications for organic nanoscale compounds.
Polymeric particles. For drug delivery purposes, the most
promising and mostly applied carrier technology is based on
polymeric materials. Especially on biomaterials that offer an
intrinsic biocompatibility and biodegradability. Hence, there are
several studies on drug penetration encapsulated in polymer-based
particles that show a marked difference between the conventional
and nanoparticulate formulations.13-17,29 A crucial question for
the investigation of nanoparticulate drug delivery carriers is the
site of drug release from the particles, i.e., does the release occur
in suspension or on the skin surface leaving the carrier particles
outside or do the particles penetrate the skin to release the drug
within the tissue? However, when it comes to particle penetration
into the skin there is only limited information available. FITC-
dextran particles of different sizes (up to 4 μm) were investigated
with and without mechanical stress.30 A clear cut-off with respect
to the size could be determined (≤1 μm) but only for skin under
mechanical stress. Nevertheless, these data are quite surprising Figure 5. Scheme of mechanism of ultradeformable vesicle transport
through a stratum corneum pore. The vesicle will penetrate into the pore if
regarding the size of the penetrating species. Kohli and Alpar the force from the osmotic pressure exceeds the deformation resistance of
16
also found latex particles (up to 500 nm) to penetrate while the vesicle (published with permission from ref. 4).
applying mechanical stress to the barrier. Shim and co-workers
applied polymer-based particles of 40 and 130 nm onto the skin of
guinea-pigs and detected clear differences between the penetrated two photon and confocal laser scanning microscopy (CLSM).18
drug amounts. However, working with hairless animals did not The simultaneous investigation of particle movement and load
show any penetration into the skin clearly indicating that the release was enabled by multimodal fluorescence strategy: the
hair follicles might be an important pathway for skin invasion of particles were covalently labeled with fluorescein and physically
particulate materials.29 loaded with Texas Red as a drug dummy for release. Furthermore
On the other hand, passive penetration (without mechanical dermal structures were visualized by keratinous autofluorescence.
stress) into the skin of polymer particles (d ~ 300 nm) seemed The excitation wavelengths were chosen to allow to separately
not to occur for human skin within 6 hours after application but investigating each of the fluorescent compounds. The covalently
release a drug dummy for cutaneous absorption was observed using labeled particles were only distributed in the skin wrinkles and

200 Dermato-Endocrinology 2009; Vol. 1 Issue 4

 Nanoparticles and skin
on the skin surface whereas the released Texas Red significantly
penetrated the epidermis (Fig. 4). These investigations where
carried out on excised human skin stored at -26°C. The freezing
procedure preserves the barrier properties of skin, but other rele-
vant characteristics are likely altered (e.g., pH).31,32 Furthermore
cellular autofluorescence vanishes from the viable epidermis due to
depletion of NADH and flavines.
Protein particles. These kinds of particles, although already
commercially available for oral application, are not yet applied on
skin.
Solid lipid nano particles (SLN). In the early 90th solid lipid
nanoparticles were introduced as drug carriers in the pharmaceu-
tical field. In general SLN are composed of physiological solid
lipids manufactured by a high pressure homogenization process.
Such systems applied onto skin results in higher drug perme-
ation. However, this enhanced drug absorption is not the result of
penetrating particles but of the occlusive effect as a result of surface
coverage.33 No intact penetration of the SLN is reported to our
knowledge.
Liposomes. Liposomes are composed of a closed bilayer of
phospholipids offering a hydrophobic compartment (lipid layer) as
well as a hydrophilic compartment (inner volume of the liposome).
Their advantage, with respect to pharmaceutical application as drug
carriers, is the wide variety of drugs to be incorporated as well as the
biocompatibility, inherently connected with natural phospholipids.
Hence, liposomes are the biggest group of nanoparticulate carriers
used for application in cosmetics or for therapeutic purposes.
Regarding the penetration behavior of liposomes it is still under
discussion if such objects might penetrate (intact) the skin.34-37
However, ultraflexible liposomal structures are assumed to pene-
trate successfully38,39 whereas conventional liposomes failed.40,41
As the liposomes are bigger in size (≥50 nm) than the skin
openings passive penetration is obstructed and a driving force is
required large enough to drag the liposomes through the skin.
Cevc and co-author proposed a mechanism to explain the observed
dermal penetration based on a hydration gradient driven transport
(Fig. 5).42 Thus the different activity of water on the two barrier
sides is responsible for aggregate migration.
The water activity gradient in the stratum corneum is reported to
be also responsible for the motion of the liposomes along the pores
of the intercellular space.
Metallic particles. Gold nanoparticles (AuNP) were widely
used for cellular imaging and gain again more and more attention
in recent years. Due to the ease of preparation of different sized
particles with comparatively small size distributions43 and their
good contrast for electron microscopy techniques they are as well
suited for studying the dermal penetration. There is one study
based on rat skin showing the penetration of different sized AuNP
(15 nm, 102 nm and 198 nm). Gold could be detected in an
acceptor compartment with permeation kinetics linked with the
size of the applied particles. The smallest particles were found to
aggregate by TEM in deeper skin layers whereas the larger two
particles types only reach the viable epidermis and dermis.43
Beryllium nanoparticles (up to 1 μm) were found in the stratum
corneum using tape-stripping as well as in deeper skin layers based
www.landesbioscience.com Dermato-Endocrinology
on TEM imaging.30 In contrast to this, Maghemite (γ-Fe2O3) and
iron core shell particles (d ≤ 20 nm) were shown to penetrate into
the top layer and eventually could reach the viable epidermis of
human skin.44 This study was conducted using electron dispersion
spectrometry SEM on vertical skin sections. This technique allows
chemical identification of iron particles in the tissue. In addition,
these particles were also found in the hair follicles. The authors
tried as well to investigate possible mechanism and investigated the
barrier properties (resistance) and found that the formulation vehi-
cles (solvent) has already a marked influence on the barrier that can
not be identified with the visualization tools. However, the authors
hydrated the skin deliberately before applying the nanoparticulate
formulations what might significantly alter the barrier properties as
a result of swelling. Another study investigated the in vitro penetra-
tion and permeation of silver nanoparticles (AgNP) through human
skin.45 The ­polyvinylpyrrolidone coated polydispers particles were
found to penetrate into the stratum corneum (TEM analysis) and
even permeated into an aqueous acceptor compartment (electro-
thermal atomic absorption spectroscopy). Particle sizes were
distributed between 10 and 50 nm and applied from an ethanolic/
synthetic sweat formulation. Even though particles themselves were
only imaged in the SC the analytical measurement of Ag in the
acceptor indicates permeation. Furthermore, abraded and healthy
skin were compared and showed—as expected—marked differ-
ences in permeation kinetics and permeated amounts showing
higher amounts and faster permeation onsets for the damaged skin
samples.
Mineral particles. Mineral particles are of huge relevance due
to their production and usage in large amounts. In 2003/2004
approximately 1,000 t of these materials were fabricated for their
usage in sun protection agents with sizes between 50–500 nm.25
Sun-screen-grade nanoparticles are composed of titanium dioxide
and (TiO2) and zinc oxide (ZnO).
Regarding the passive diffusion of TiO2 the EU Scientific
Committee on Cosmetics and Non-Food Products (SCCNP)46
published a paper based on studies with micro- and nano-sized
material. Herein they state that these particles remain on the skin
surface or on the outer layer of the stratum corneum and do not
penetrate into or through the living skin.47-49 Confirmation was
obtained with studies on human, porcine or murine skin50-53
for particles within a size range between 10 and 100 nm. These
data were recently confirmed by the outcome of an EU project
(NanoDerm). Here, TiO2 are only found in the top layer of the
stratum corneum and the openings of the hair follicle.54 Similar
results were obtained for ZnO.50,55 Just recently two studies on
ZnO56 and TiO2,57 were published; ZnO penetration was investi-
gated in vivo with human volunteers and located the particulated
materials only on the skin surface and their accumulation in
skin folds and/or hair follicles.56 In vitro measurements of the
penetration of TiO2 particles between 20 and 100 nm showed the
nanoparticles only in the top 3–5 layers for all skin samples used
(porcine skin, healthy human skin and human skin grafted on a
severe combined immuno-deficient mouse model).57
Carbon nanotubes (CNT)/fullerenes. Carbon materials are
considered as drug delivery systems due to their inert nature
201
 Nanoparticles and skin
and their volume to be filled with drugs.58 With respect to the
interaction of these materials with skin several studies are under-
taken looking at the toxic impact of these materials on skin cells
(keratinocytes).59-61 To our knowledge only one work investigated
the penetration behavior of carbon materials, i.e., fullerenes into
skin.62 Rouse et al. coupled a peptide with a fullerene and added
a fluorescent marker (3.5–400 nm) to facilitate the observation of
the complex. No penetration of the complex in skin was found as
long as the skin was untreated. Simulating the mechanical stress
of the skin (such as walking)—flexing of the skin—resulted in
penetration of the particulate material into skin. In addition, the
penetration was dependent on the time the mechanical stress was
applied; flexing need to be going on for at least 60 min to allow
penetration. This may indicate that the structure of the skin is
altered applying mechanical stress.
Semiconductor nanocrystals. Semiconductor nanocrystals, also
known as Quantum dots (QD) are widely used for non-invasive
imaging purposes. The QD offer several advantageous properties
that offer superior detection and experimental control compared to
many other nanoparticulate materials. For all penetration processes
the size of the permeant is considered to be one of the main factors.
QD are, without additional surface coating available in a very small
size range; below 10 nm. Besides the size-dependent fluorescent
emission, the QD show typically small size-distributions and
favorable optical properties. The inherent fluorescence guarantees
a label-free—not altering the chemical composition—material.63
Furthermore, the powerful and convenient fluorescent methods
can be non-invasively applied. Typically, most of the experimental
data is based on confocal microscopy, sometimes supported with
TEM images and quantitative analysis.64
As a result they are applied many-fold in several areas and are
used as well for understanding of the interaction of nano-sized
materials with skin. There are several publications dealing with QD
and the interaction with skin. An important aspect is that none of
the present publications deals with human skin but with murine
skin64,65 (mouse, rat) and pig skin.21,64 As the different skin types
do differ and no information on particle penetration exists care
should be taken to generalize the different data.
In a recent work, healthy skin was compared with damaged
skin regarding the QD penetration. Skin was damaged applying
tape-stripping to remove the stratum corneum or a sand paper
was used to physically damage the skin and facilitate penetration
because of a missing or ruptured main barrier. However, no clear
penetration was observed.64 Rat skin without stratum corneum as
well as healthy rat skin did not show any permeability for the types
of QD used in contrast to the results obtained for pig skin with
the same particles.21 Only the abraded skin showed some penetra-
tion following the authors although this trend is not obvious.64
Surprisingly, following earlier results, even flexing did not facilitate
the penetration of the QD into the skin.
In contrast to these data obtained for rat skin, for porcine
and mice strong penetration was reported.21,65 The findings in
these two papers are really surprising and unexpected and with
respect to other data on particle penetration the reliability might
be questioned. Ryman-Rasmussen et al. applied two types of
202 Dermato-Endocrinology
QD—spherical and ellipsoidal—with different surface coatings
(PEG-amine-, carboxy- and PEG-functionalized) from a borate
buffer (pH = 9) onto porcine skin and basically found that all
particles do penetrate into skin after 24 hours. Neither the size nor
the surface modification had any influence on the QD absorption.
Following the penetration of molecules (Potts-Guy equation) this
is really surprising for particulate material. Chu and co-workers
applied QD in vivo as well as in vitro on mouse skin (which differs
from human skin morphology). With fluorescence methods they
found penetration in the skin and in addition analyzing the animal
organs with inductive coupled plasma atomic emission spectrom-
etry (ICP.AES), the QD were detected in lung, heart, liver and
the kidneys. Comparable high levels of the QD were kept over
one week. Not proven is the presence of particles in these organs
because ICP only detects ionic species not particulate material.
Just recently, another study identified a further condition
destroying/reducing the barrier function of skin: ultraviolet radia-
tion (UVR).66 Carboxylated QD applied on mouse skin in a
glycerol vehicle showed increased penetration if the skin is exposed
to UV radiation (290–400 nm) compared to low level of pene-
trated QD without illumination. However the overall penetration
was low in both cases. The authors could even demonstrate the
possible pathway of the intruding QD along the intercellular space
using silver enhanced TEM imaging.
Follicular Penetration
The role of the appendages in skin absorption was long
considered negligible due to their low surface coverage of only
0.1%. Recently in the case of the follicle orifices this number was
corrected to be higher than previously assumed.67 Evaluation of
the follicles of several anatomical sites (lateral forehead, upper arm,
forearm, thorax, back, thigh, calf ) showed that every body region
features its own hair follicle characteristics. A maximum surface
coverage was found at the forehead (1.28%) and a minimum on
the forearm (0.09%). The available infundibular volume which
determines the potential follicular reservoir was found to be
highest at the forehead and the calf region and similarly to the
estimated reservoir of the stratum corneum.67 However, so far no
direct correlation to regional differences in skin absorption of small
molecules has been investigated with respect to the hair follicles.
For small molecules the trans-follicular pathway is usually
ignored in view of the dominance of especially the lipid pathway,
although the matter is little understood. Nonetheless, for several
compounds (e.g., caffeine, hydrocortisone, testosterone68-70) the
follicular route seems to promote permeability at the beginning of
skin absorption while it is overruled by the other higher capacity
pathways at later times in the transport process.68
In the context of the absorption of nanoparticles the follicles
have received much higher attention. It is commonly agreed that
in non-damaged skin the hair follicles play a central role in the
penetration of solid nanoparticles.2 Being invaginations of the
epidermis hair follicles may reach deep into the subcutaneous
fat (depending for example on the state of development of the
follicle). Nonetheless, an efficient barrier, which is similar to the
stratum corneum in the upper part of the follicle and features tight
2009; Vol. 1 Issue 4
 Nanoparticles and skin
junctions in the lower part of the follicle71 prevents particles from
easily invading into the living cells.72 Instead, once entered into the
follicle openings particles will be stored there until being cleared by
hair growth or sebum production. Clearance is much slower than
for example for the stratum corneum as inside the follicles particles
are to a certain degree protected from natural desquamation, textile
contact and washing. This makes the follicles an excellent storage
site that can be used for drug release and offering a shortcut to the
viable skin layers and the systemic circulation.
Lademann and co-workers reported that only follicles containing
growing hair or showing sebum production took up particles.73
Others, “non-active” follicles, were clogged by cellular debris and
needed re-opening by washing or light peeling. This was kind
of surprising as the direction of hair growth and sebum flow are
opposed to the invasion of particles into the follicles suggesting an
“active” uptake mechanism transporting nanoparticles deep into
the follicle.2 Instead, experiments on excised pig ear skin with and
without massage suggested that this is a purely mechanical process.
The natural movement of the hair together with the scaly struc-
ture of the hair cuticula works like a gear pump which transports
particles having an appropriate size into the follicle. The optimum
particle size was in the range of 400–700 nm73 and coincided with
the size of the scales of the cuticula. In addition material properties
did not seem to influence the particle uptake or penetration depth.
However, also particles of other sizes were found to penetrate into
human follicles such as polystyrene particles of 20 nm14 as well as
iron-based particles.44
Several methods were proposed for the investigation of the
follicular pathway. Animal studies with hairless and hair bearing
species are not appropriate as even hairless species do have follicles.
Some assumptions to that rule are newborn hairless rats that
develop follicles only within a few days after birth.70 Also, follicle
free skin can be artificially created by scarring. However there is a
common consensus that murine skin is not an adequate model for
the skin absorption in humans. Rat skin for instance has a density
of 289 ± 21 hair follicles/cm2 in contrast to human abdominal
skin and pig skin from the back showing a density of 11 ± 1
hair follicles/cm2 only.74 Therefore some effort has been taken
to develop suitable alternative methods that are based on human
skin. Barry and coworkers developed the “SC sandwich” which is
composed of an epidermal and a stratum corneum membrane.75
Due to the random distribution of hair follicles on the skin surface
there is only negligible chance for a direct connection across both
membranes.75 Therefore, if the shunts contribute significantly
to the permeation transport through the sandwich will be much
reduced compared to what would be expected from simple increase
in path length.
Otberg et al. chose a more work intensive approach by closing
each hair follicle opening with varnish wax under microscopic
observation.68 At the control skin site the varnish wax was applied
directly adjacent to each follicle in order to compensate for the
area reduction at the test site. The same group also suggested a
differential stripping technique for selective sampling of the folli-
cular content.76 In a first step tape-stripping removes substances
remaining on the skin surface or having penetrated into the
www.landesbioscience.com Dermato-Endocrinology
stratum corneum. Substances (or particles) inside the follicles are
protected and are sampled in a second step by a cyanoacrylate-
biopsy that removes the content of the hair follicles.
Conclusion
First of all, one can state that the penetration of particulate
materials into skin is a very complex issue. Ab initio predictions
are therefore not (yet) a practical approach to describe the penetra-
tion processes. For this reason, up to now only experimental data
for particle penetration is available and will be indispensible also
in the future. However, the experiments can not be transferred
straightforwardly form the non-particulate situation, but has to
take into account the particulate nature of the analyte and not
only the material. Furthermore, practical and ethical consider-
ations limit the freedom of the experimental design. In addition,
the choice of the experimental parameters as well as the following
interpretation of the data obtained strongly depends on the field
of the research and the particular problem investigated. Therefore,
it is of outmost importance when looking at the available data to
take the experimental settings into account. Furthermore, care
has to be taken when transferring data from one model system to
another and hence far-reaching conclusions are made for instance
regarding safety hazards.
Looking at Table 2, problematic data might be the results on
the penetration and permeation of Quantum dots. For mouse skin
as well as for porcine skin penetration was reported whereas on rat
skin only for abraded skin permeation into deeper skin layers was
observed. In any case, the data available yields a heterogeneous
picture.
In contrast, for human skin only 2 publications report passive
permeation of particles beyond the stratum corneum. In both
cases small primary particles are used even though at least two of
the particle types were described to form bigger agglomerates. As
a consequence only small amounts were found in the viable part
of the skin.
Another important issue is the mechanical stress applied to the
skin when investigating the penetration behavior. There seems to
be a tremendous influence on the kind of mechanical stimulus as
well as on the time that stimulation is present. Particles as big as
2 μm were reported to penetrate into human skin.30 In contrast
to that for much smaller particles such as the surface-modified
fullerenes flexation of more than 60 min was necessary to improve
penetration into deeper layers of porcine skin.62 However, it is not
clear if the possible agglomerates (~250 nm) penetrate or if the
primary particles penetrate.
As an overall consequence a standardized operation procedure
is not only desirable but essential to be able to investigate and
eventually predict the behavior of nano-scale materials with skin.
A well established protocol need to be defined to make the results
comparable with each other. Slight differences in the treatment of
the skin samples or the set-up might already contribute to altered
penetration behavior. Pretreatment, way of application, cleaning/
rinsing procedures, hydration state, and detection methods will
influence the results and need to be carefully controlled.
203
 Nanoparticles and skin

Table 2 Overview on particle penetration experiments performed with different types of skin and different
particles

Particle type Ø of core (nm) Ø in vehicle (nm) Skin type Treatment Permeation Reference
QD PEG 6 x 9c 40* porcine - yes‡ Zhang et al. 2008,1
QD-COOH 4.6a 14* “ - yes‡ Ryman-Rasmussen et al.
2006,2
6 x 12b 18* “ - yes‡ “
QD-NH2 4.6a 14* “ - yes‡ “
6 x 12b 18* “ - yes‡ “
QD-PEG 4.6a 14* “ - yes‡ “
6 x 12b 18* “ - yes‡ “
Carbon-Baa-Lys-(FITC)-NLS 3.5a 40–250§ “ full thickness skin flexed yes‡ Rouse et al. 2007,3
(60–90 min)
Carbon-Baa-Lys-(FITC)-NLS 3.5a 40–250§ “ - no “
ZnO 80a <160§ “ - no Gamer et al. 2006,4
TiO2 30 x 60 x 10# “ - no “
TiO2 20§ “ - no Gontier et al. 2008,5
PS-COOH 20a* porcine ear - no Alvarez-Roman et al. 2004,6
“ 200a* “ - no “
QD-COOH 4.6a* 14 rat - no Zhang et al. 2008,7
“ “ “ “ flexed no “
“ “ “ “ tape-stripped no “
“ “ “ “ abraded yes “
QD-COOH 6 x 12b* 18 “ - no Zhang et al. 2008,7
“ “ “ “ flexed no “
“ “ “ “ tape-stripped no “
“ “ “ “ abraded yes “
AuNP-Cit 15a “ - yes† Sonavane et al. 2008,8
“ 102a “ - yes† “
“ 198a “ - yes† “
QD-MPA 4.1a* mouse - yes† Chu et al. 2007,9
QD-COOH 29a glycerol “ - yes‡ Mortensen et al. 2008,10
“ 30a glycerol “ UV irradiation yes‡ “
ZnO 15–40 30* Human heat separated no Cross et al. 2007,11
ZnO 15–40 20–30* “ in vivo no Zvyagin et al. 2008,12
TiO2 26–30 20–30* “ in vivo no Roberts et al. 2008,13
TiO2 20§ “ no Gontier et al. 2008,5
PLGA 290a “ - no Stracke et al. 2006,14
FITC-dextran 500a “ - no Tinkle et al. 2003,15
“ 1000a “ - no “
“ 2000a “ - no “
“ 4000a “ - no “
FITC-dextran 500a “ flexed yes‡ “
“ 1000a “ “ yes‡ “
“ 2000a “ “ no “
“ 4000a “ “ no “
γ-Fe2O3-TMAOH 6 50# “ full thickness skin yes‡ Baroli et al. 2007,16
Fe-AOT 5 5–10–80# “ “ yes‡ “
AgNP-PVP 25# “ - yes† Larese et al. 2009,17
aspherical, belliptical, cnail-shaped, *monodispers, #polydispers, §aggregates, †in acceptor compartment or in tissues for in vivo experiments, ‡deeper skin layers (dermis and viable epidermis). MPA, 3-mercapto-

propionic acid; TMAOH, tetramethylammonium hydroxide; AOT, sodium bis(2-ethylhexyl) sulfosuccinate; Baa-Lys-(FITC)-NLS, phenylalanine-based fullerene [Bucky amino acid] (Fluorescein isothiocyanate) nuclear
localisation sequence; Cit, Citrate.

204 Dermato-Endocrinology 2009; Vol. 1 Issue 4

 Nanoparticles and skin
Nevertheless, the growing number of nanoparticle occurrence
in manufacturing and application need a better understanding of
the relevant mechanisms of nanoparticulate penetration into skin,
not least for an adequate risk assessment.
An issue not being studied so far is the influence of the nano-
particulate formulations on biochemical processes in the skin. This
might be an intriguing field for future investigations.
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