Nanotoxicology, March 2010; 4(1): 15–41

Zinc oxide nanoparticles in modern sunscreens: An analysis of potential

exposure and hazard

MEGAN J. OSMOND, & MAXINE J. MCCALL

CSIRO Future Manufacturing Flagship, North Ryde, NSW, Australia

(Received 22 September 2009; accepted 20 November 2009)

Abstract
Sunscreens containing metal oxide nanoparticles appear transparent on the skin and provide excellent protection against
sunburn caused by UV radiation. While it is likely that nanoparticles remain on the surface of the skin of healthy adult humans,
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13

and thus are considered safe for use in sunscreens, there has been no comprehensive assessment of the impact on human health
from exposure to the metal oxide nanoparticles destined for use in sunscreens, either in the workplace during the
manufacturing process, in long-term use across a range of skin conditions, or upon release into the broader environment,
either accidentally or consequent of normal sunscreen use. In this review, we focus on zinc oxide nanoparticles destined for use
in modern sunscreens, and discuss the potential for human exposure and the health hazard at each stage of their manufacture
and use. We highlight where there is a need for further research.

Keywords: Nanotoxicology, nanoparticle, zinc oxide, sunscreen

Introduction impact at the terrestrial level, although the proportion

For personal use only.

of shorter wavelengths reaching the earth’s surface

Prolonged exposure to the sun may cause painful sun-
burn. Research has established that over-exposure to
ultraviolet (UV) radiation can lead to inflammation,
immunosuppression, premature photo-ageing, DNA
damage, and photocarcinogenesis (Marrot and
Meunier 2008). In addition to direct cellular damage,
exposure to UV radiation can generate reactive oxygen
species (ROS) in skin cells, and lead to an impairment of
endogenous anti-oxidant defence systems (Darr and
Fridovich 1994; Sander et al. 2004; Sakurai et al. 2005).
Protection from over-exposure to UV radiation is
encouraged and the use of sunscreen as part of a
protection strategy is recommended (e.g., http://
www.cancer.org.au/Healthprofessionals/PositionState-
ments/sunsmart.htm). However, the rise and fall in the
use of para-aminobenzoic acid (PABA) (Wolf et al.
2001) exemplifies the potential duality of sun-protection
ingredients: they can attenuate the damaging effects of
over-exposure to UV radiation but may also elicit a range
of side-effects that are not necessarily appreciated during
the first generation of use.
UV wavelengths are broadly categorized by wave-
length into three groups (UVA, UVB and UVC).
UVC (200–290 nm) is completely absorbed by the
ozone layer and therefore has negligible biological
may alter with the depletion of stratospheric ozone
attributed to anthropogenic activities (Frederick
and Snell 1988; Madronich et al. 1998). UVB
(290–320 nm) is incompletely attenuated by the
ozone layer such that the terrestrial UV spectrum
currently contains approximately 6% of wavelengths
in this range (Diffey 2002). The remaining 94%
comprise UVA wavelengths (320–400 nm), sub-
categorized into UVAI (340–400 nm) and UVAII
(320–340 nm) (Diffey 2002; Matts 2006).
Penetration of UV radiation into human skin
decreases rapidly with increasing epidermal depth
(Bruls et al. 1984). As nucleic acids and proteins
most efficiently absorb wavelengths within the UVB
(and UVC) range, with relatively low absorbance at
wavelengths greater than 300 nm, UVB was initially
thought to be the primary damaging component of
terrestrial UV radiation. However, more recently, it
has been appreciated that wavelengths in the UVA
region penetrate deeper into the epidermis than UVB,
inducing oxidative stress linked to indirect DNA
damage and long-term photo-ageing (Forestier 2008;
Marrot and Meunier 2008). It is now accepted that
sunscreens must provide broad spectrum UV
protection (Nash et al. 2006; Forestier 2008).

Correspondence: Megan Osmond, CSIRO Future Manufacturing Flagship, PO Box 184, North Ryde, NSW, 2113, Australia. E-mail: megan.osmond@csiro.au

ISSN 1743-5390 print/ISSN 1743-5404 online
2010 Informa UK Ltd.

DOI: 10.3109/17435390903502028
 16 M.J. Osmond & M.J. McCall
There are two major classes of active ingredients
used in modern sunscreens: ‘chemical’ and ‘physical’.
Active ‘chemical’ ingredients typically are molecules
with an aromatic ring structure bearing two functional
groups, one of which acts as an electron donor and the
other as an electron acceptor following the absorption
of UV radiation (Wolf et al. 2001; Maier and Korting
2005). Active chemical ingredients include octyl-
methoxycinnamate, butyl-methoxydibenzoylmethane
and 3-(4-methylbenzylidene) camphor.
Active ‘physical’ ingredients typically are particles
of the metal oxides, such as zinc oxide (ZnO) and
titanium dioxide (TiO2) having intrinsic UV absorb-
ing properties. Sunscreens containing physical
blockers of UV have been shown to be highly effective
in protecting cells against UV-induced DNA damage
(Cayrol et al. 1999). In normal pigment size ranges
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
(150–300 nm for TiO2 and 200–400 nm for ZnO)
these particles reflect and scatter light, making the
sunscreens appear white. However, as particle sizes
decrease to smaller sub-micron (nano) dimensions
(typically between 20 and 150 nm for TiO2 and
40–100 nm for ZnO) they absorb and scatter UV
radiation, and largely absorb visible wavelengths
(Wolf et al. 2001), making the sunscreens appear
transparent on skin and thus more aesthetically pleas-
ing. In addition to facilitating consumer acceptance,
For personal use only.
nanosized metal oxide particles may also offer greater
UV protection compared to their microsized counter-
parts, at least in the UVB region (Popov et al. 2005).
A recent review of 308 sunscreen products revealed
that 3.6% contained physical blockers alone for their
active ingredients, 54.8% contained only chemical
UV-absorbers, whilst the remaining 41.6% contained
a mixture of both (Wahie et al. 2007) suggesting that
approximately half of the commercially-available
sunscreens in this study contained metal oxide par-
ticles as at least one of their active ingredients. Of
these, TiO2 was contained in ~ 45%, ZnO < 5%, but
the portion of this percentage containing nanoparti-
cles was not reported.
Currently, metal oxide nanoparticles have not been
comprehensively assessed in regard to potential
effects on human health, from exposure (accidental
or otherwise) in the workplace during nanoparticle
production or exposure through use in commercial
products (e.g., TiO2 or ZnO sunscreens), or for their
effects on ecosystems if released into the environ-
ment. It has been reported that some nanoparticles
such as TiO2 and carbon black, when inhaled by rats
or instilled in their lungs, can elicit a greater inflam-
matory response on an equivalent mass basis than fine
or bulk particles of the same material (Baggs et al.
1997; Donaldson et al. 2000; Oberdörster 2000).
However, the risk of adverse impact on health from
any material depends both on the biological toxicity
associated with the intrinsic properties of the material,
and on the degree of exposure of an organism to it.
The hazard posed may change during the life cycle of
the material from its time of manufacture through to
its demise or transformation into other forms.
Zinc oxide shows exceptional properties as an
active physical ingredient in sunscreens. It absorbs
UV more effectively than TiO2 over a broad range,
particularly in the UVA region (Mitchnick et al. 1999;
Pinnell et al. 2000), and has consequently been used
as the sole active ingredient in some broad-spectrum
sunscreens. In this review, we focus on the use of ZnO
nanoparticles destined for use in modern sunscreens.
We assess the potential risk at each stage, from pro-
duction to use of product and disposal (Figure 1). An
effort has been made to identify the gaps in current
knowledge and consequent research priorities at each
stage.
Workplace exposure
Hazard identification and potential for exposure
Controlled chemical reactions such as gas-phase or
solution-phase syntheses can be used to manufacture
ZnO nanoparticles from precursors (Perl 1993;
Kleinwechter et al. 2002; Viswanathan and Gupta
2003; Bauermann et al. 2006; Duffy et al. 2007).
Alternatively, ZnO nanoparticles can be prepared
using mechanical methods such as high energy attri-
tion milling. Where destined for incorporation into
sunscreens, they may be prepared as powders or
suspensions. Although these processes typically take
place within closed systems, inherent in their overall
production, including post-production processing
and packaging, is the possibility that ZnO nanoparti-
cles may be released incidentally as ‘dust’ or as aerosol
droplets into the ambient air. Once in the air, they
may circulate for some time before settling on work-
place surfaces or on workers themselves. Therefore,
there may be the potential for exposure of workers via
inhalation and ingestion (by swallowing airborne
nanoparticles), as well as possible dermal contact
(Figure 1, left).
Workplace variables that may influence the poten-
tial for exposure during manufacture include the
emission source, air flow dynamics including move-
ment, current and ventilation rate, and worker mobil-
ity (Brouwer et al. 2004). Particle mobility and a
changing size distribution over time may also be
important factors in exposure estimates, as airborne
nanoparticles are unlikely to remain as single particles
and may form homogenous or heterogeneous
 Analysis of ZnO for sunscreens 17
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

Figure 1. Schematic showing potential exposure to zinc oxide nanoparticles destined for use in sunscreens, from their manufacture in the
workplace and their transport to be incorporated in sunscreen, through to their use by consumers. Possible points of exposure to humans and
the environment are highlighted in red, with unregulated exposure indicated as potential hot-spots.

agglomerates with themselves or other particles
(Brouwer et al. 2004; Maynard and Kuempel 2005;
Tsai et al. 2008). Particle size and composition can
influence the efficiency of deposition in the respiratory
tract as well as the biological impact, particularly as
the lung environment may favor the stabilization of
agglomerates (Kendall et al. 2002).
Given that even small amounts of nanoparticulate
material contain enormous numbers of individual
particles, low-level chronic exposure may occur in
the workplace via inhalation or ingestion, or by other
means such as dermal absorption including the eyes.
The exposure of those directly involved in the
manufacturing process may be mitigated by imple-
menting a hierarchy of controls including process
engineering for handling and storage, and the
provision and use of adequate personal protective
equipment (PPE). Depending on the levels of control
implemented, other personnel such as administrative
staff, cleaners and trades people may be exposed
through circulating or settled nanoparticles.
Likelihood of exposure
Assessment of the likelihood and degree of workplace
exposure has been limited by a lack of robust
techniques and procedures for obtaining relevant
dose metrics for inhalation. The heightened pro-
inflammatory effects of nanoparticles in general
have most often been attributed to their greater sur-
face area relative to the same mass of larger particles
(Oberdörster 2000; Hohr et al. 2002; Hext et al. 2005;
Monteiller et al. 2007), and the importance of surface
properties (Tabata and Ikada 1988; Fubini 1997),
particle size (Limbach et al. 2005), and shape at point
of contact (Champion and Mitragotri 2006) have also
been emphasized. Although instruments are available
 18 M.J. Osmond & M.J. McCall
to characterize nanoparticulate aerosols on the basis
of mass, particle number or surface area, their effec-
tiveness may be limited as the use of static samplers
may not accurately reflect personal exposure
(Brouwer et al. 2004). In addition, these instruments
cannot quantify specific nanoparticles against a gen-
eral, high background, and so current identification of
exposure to specific nanoparticles relies on the col-
lection of airborne samples on filters that are subse-
quently analyzed by electron microscopy. Therefore,
until there are technological advances in measuring
personal exposure to specific nanoparticles in the
workplace, it is difficult to readily estimate the specific
likelihood and degree.
Information on the likelihood of workplace exposure
to nanoparticulate ZnO destined for sunscreen use
during the production phase is not publically available
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
but other studies using different nanoparticles may
provide some insight. A recent workplace study
(Tsai et al. 2008) was carried out during the produc-
tion of polymer-nanoalumina nanocomposites by twin
screw extrusion (TSE), a commercial method by which
nanoparticles and polymer melts are compounded.
Release of airborne nanoparticles during the com-
pounding process was assessed using a Fast Mobility
Particle Sizer (FMPSTM) spectrometer, placed at var-
ious points around the workplace during production.
For personal use only.
Workplace controls were not specifically described but
the presence of an exhaust system attached to the TSE
instrument and a laboratory ventilation system were
noted. Elevated levels of nanoparticles in the ambient
air at the source of production and in the breathing
zone were reported (Tsai et al. 2008).
An earlier workplace study (Maynard et al. 2004)
assessed the generation of aerosol particles during
post-production handling of single-walled carbon
nanotubes (SWCNTs) across four facilities. Particle
handling was carried out in specially-constructed
clean air enclosures and air was monitored using a
portable condensation particle counter and a size-
discriminating optical particle counter. Filter samples
were also taken and cotton gloves worn by the workers
were used to assess dermal exposure. During the
manufacturing process, when it was noted that work-
ers took care to move with slow deliberation, airborne
nanoparticles were present at only low levels. These
levels increased during clean-up and when workers
entered or left the work area (Maynard et al. 2004),
indicating that increased circulation at these points
may have disturbed surface-deposited nanoparticles.
The study also reported that detectable levels of
SWCNTs deposited on workplace surfaces were
transferred to gloves worn by workers. It should
also be noted that due to their surface chemistries,
some CNTs can be particularly prone to forming
bundles rather than dispersing, and therefore extra-
polation of levels of airborne particles from a model
using CNTs may underestimate levels for more dust-
like nanoparticles such as ZnO.
Potential health effects of workplace exposure
Inhalation. Depending on size, deposition of
inhaled particles can occur in different areas of the
respiratory tract, with theoretical modeling of
nanoparticles suggesting preferential deposition of
particles ~ 10–100 nm in the alveolar and tracheo-
bronchial regions, and even smaller nanoparticles
in the nasopharynx region (Oberdörster 2001;
Maynard and Kuempel 2005); however, deposition
efficiency in the latter region increases when
expressed as per unit surface area, as the nasopharynx
has a smaller surface area than the alveolar region
(Oberdörster 2001). In the former, alveolar macro-
phages generally phagocytose particles and migrate
via the mucociliary escalator to the tracheobronchial
region, where they can be either coughed up or
swallowed. Lung lining fluid, containing antioxidants,
lipids, mucins and proteins, provides further protec-
tion against inhaled particles (Kendall et al. 2002). If
these mechanisms fail, particles can enter the inter-
stitium where they are no longer available for clear-
ance via normal pathways. Their presence can then
chronically stimulate cells, leading to inflammation,
fibrosis and disease (Donaldson et al. 1998). Studies
with TiO2 and carbon black nanoparticles suggest that
phagocytosis of these types of nanoparticles may be
sub-optimal (Churg et al. 1998; Renwick et al. 2001;
Geiser et al. 2008) and that nanosized particles may
show long-term retention in the bronchial airways
(Kreyling et al. 2006).
Studies with animal models have suggested that
single exposures to ZnO nanoparticles as freshly gen-
erated aerosols (60 nm) or in suspension via intra-
tracheal instillation (50–70 nm) can produce at least a
transient inflammatory response (Gordon et al. 1992;
Sayes et al. 2007) (Table Ia). In a guinea pig model,
repeated inhalation (3 h/day 6 day) of the industry-
relevant dose of 5 mg/m3 of freshly generated ultrafine
ZnO (50 nm) caused lung lesions characterized
by infiltration of inflammatory cells, interstitial
thickening, and diminished lung capacity (Lam et al.
1985), although this response was not observed in a
previous single-exposure study (Lam et al. 1982).
However, interspecies differences in the response to
inhaled particles in general (Gordon et al. 1992;
Bermudez et al. 2002; Bermudez et al. 2004;
Hext et al. 2005) can make it difficult to reliably
extrapolate the results of these studies to humans.

For example, particle overload can occur in some
mammals leading to compromised macrophage-
mediated particle clearance and mobility, and sub-
sequent development of inflammation, fibrosis and
cancer (Morrow 1988; Oberdörster et al. 1994;
Nikula et al. 1997; Tran et al. 2000), but the rele-
vance of particle overload in humans has been ques-
tioned (Borm et al. 2004).
Metal fume fever, which is characterized by short-
term pulmonary and systemic alterations in humans,
is caused by the inhalation of freshly generated metal
oxide aerosol (Kelleher et al. 2000) and may be a
more relevant model for workplace exposure to ZnO
nanoparticles. A single 10–30 min inhalation of a high
dose of nanoparticulate ZnO aerosol containing
170 nm aggregates (from 8–40 nm primary particles)
was sufficient to induce increases in levels of the
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
inflammatory cytokines interleukin (IL)-6, IL-8,
and tumor necrosis factor (TNF)-a in broncho-
alveolar lavage (BAL) fluid within three hours of
exposure in humans (Kuschner et al. 1997).
A more likely scenario of workplace exposure,
however, might be chronic, low-level exposure. The
industry threshold limit for human inhalation of ZnO
is 5 mg/m3 for up to 8 h, but it has been shown that
less than half this exposure can induce an adverse
response in humans (Gordon et al. 1992; Fine et al.
For personal use only.
1997). However, humans appear to develop tolerance
to inhaled ZnO fumes (Fine et al. 2000), with no
known long-term effects of metal fume fever, and
acquired tolerance to inhaled ZnO has also been
shown in mice (Wesselkamper et al. 2001a).
One study comparing the effects of ZnO particle
size on inflammogenicity reported that there were no
differences in the response of healthy adults to inhaled
nanoparticulate (40 nm) or larger (291 nm) ZnO
particles (Beckett et al. 2005), although it should
be noted that the dose used was below the threshold
to detect a biological difference between inhaled ZnO
and filtered air and therefore might not be conclusive
as to potential differences between nano-ZnO and
larger ZnO particles.
Although ZnO has only low solubility in water, pH
can influence solubility (Environmental Protection
Agency [EPA] 1995) and therefore a further consid-
eration is the possibility that ZnO nanoparticles may
dissolve in the more acidic environment of the lung
lining fluid, potentially leading to transient increases
in the local Zn2+ concentration. The likelihood that
this occurs and its possible biological impact have not
yet been investigated.
Ingestion. Only one study has been published on the
effects of ingestion of ZnO nanoparticles (Table Ib),
Analysis of ZnO for sunscreens 19
with a companion study addressing zinc nano-
particles. Two weeks after 5 g/kg body weight zinc
(not ZnO) nanoparticles (58 nm) and microparticles
(1.08 mm) were gastrointestinally administered to
four-week-old mice, more severe anemia and renal
damage was observed in animals exposed to the
nanoparticles compared to the microscale particles.
Histological examination of two mice that died in the
first week showed intestinal obstruction caused by
aggregation of zinc nanoparticles in the intestine
(Wang et al. 2006), although serum biomarkers of
inflammation generally did not differ appreciably
between the two groups. In a follow up study also
in mice, treatment with 1–5 g/kg body weight 20 nm
and 120 nm ZnO particles showed relatively little
toxicity, although some pathological damage was
detected in the stomach, liver, heart and spleen
(Wang et al. 2008). It is important to note that the
doses used in both of these studies were higher than
those to which most people are likely to be exposed
and, when taken together, the studies showed little
data consistent with increased oral toxicity of ZnO
nanoparticles.
Dermal absorption. To our knowledge there have been
no studies investigating the hazard potential or fate of
ZnO nanoparticle dust that may settle on exposed
skin from the air or from physical contact with other
surfaces. Some studies are available that have assessed
nanoparticles suspended in emulsions or sunscreen
formulations, but these will be considered in the
Product use section.
Potential for translocation
Lung and airways. A range of structural and immune
cells comprise the barriers of the respiratory system
(Suzuki et al. 2008). Due to their small size, nano-
particles may retard or evade phagocytosis by macro-
phages thus giving them a greater opportunity to
penetrate into the pulmonary interstitium (Churg
et al. 1998; Oberdörster 2001; Renwick et al. 2001;
Borm and Kreyling 2004; Geiser et al. 2008;
Lynch and Dawson 2008). Nanoparticles may also
form complexes with proteins such that the fate of
the nanoparticle may be determined by that of the
protein (Kreyling et al. 2006; Semmler et al. 2004).
The biological impact of this ‘protein corona’
may alter depending on particle size and surface
characteristics as both of these have been shown to
influence the constituents of the corona in human
plasma using a range of polystyrene nanoparticles
(Lundqvist et al. 2008).
 Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

20

Table I. A summary of published data relating to potential hazards of zinc oxide nanoparticles.

Particle surface
Authors Particle size (nm) area Surface coating Administration vehicle Experimental regime Results

(a) Inhalation
In vitro
Brunner et al. 19–36 57 m2/g NR1 Nanoparticles Human mesothelioma and Cell viability was reduced after
(2006) suspended in deionized rodent fibroblast cells exposure to 3.75 ppm and was
water then diluted in cell incubated with 1–30 ppm ZnO. ~ 100% at 15 ppm. Toxicity was
culture media. attributed to ZnO solubility.
M.J. Osmond & M.J. McCall

Gojova et al. 100–200 length; 21 m2/g NR Nanoparticles Human aortic endothelial cells Doses at or above 10ug/ml initiated
(2007) 20–70 width suspended in cell exposed to ZnO nanoparticle a dose-dependent inflammatory
culture medium. concentrations ranging from response and decrease in cell
0.001–50 mg/ml in culture viability. Internalization of ZnO
media for 4 h, with cell viability nanoparticles within intracellular
and inflammatory biomarkers vesicles was detected.
measured at 4 h and 24 h.
Gopalan et al. 40–70 NR NR Nanoparticles Primary human sperm cells and DNA damage occurred in sperm
(2009) suspended in DMSO, lymphocytes incubated with cells and lymphocytes compared to
filtered through a 11.5–92.3 mg/ml ZnO. a negative control, but ZnO was
0.2 mm syringe filter Incubation with the particles only weakly photogenotoxic.
and diluted in PBS. was performed in the dark, with
pre- or simultaneous irradiation
with UV light.
Jeng and 50–70 15–25 m2/g NR Nanoparticles Mouse neuroblastoma cells Cell viability decreased at a
Swanson (2006) suspended in deionized incubated for 48 h with threshold dose of 50 mg/ml.
water then added to cell 10–100 mg/ml ZnO particles in
culture media. culture media.
In vivo2
Beckett et al. 40 & 291 NR NR Freshly generated ZnO Inhaltion exposure of human No differences were observed
(2005) vapor diluted in air. subjects to air containing between filtered air and either ZnO
500 mg3 nano- or microparticles sample. The concentration used
ZnO for 2 h with physiological was below the threshold required
endpoints assessed 24 h post- for an acute response.
exposure.
 Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

Table I (Continued)

Particle surface
Authors Particle size (nm) area Surface coating Administration vehicle Experimental regime Results

Fine et al. (1997) 300 (secondary NR NR Freshly generated ZnO Inhalation exposure of human
aggregates from vapor diluted in filtered subjects to air containing 0, 2.5
primary air. and 5 mg/m3 ZnO for 2 h on
nanoparticles) separate days. Venous blood
was withdrawn 3 and 6 h
post-exposure and subjects
recorded symptoms 0, 3, 6
and 9 h post-exposure.
Fine et al. (2000) 300 (secondary NR NR Freshly generated ZnO Inhalation exposure of naïve
aggregates from vapor diluted in filtered and chronically exposed
primary air. human subjects to air
nanoparticles) containing 5 mg/m3 ZnO
for 2 h on 1 or 3 days.
Bronchoalveolar lavage was
performed 20 h post-exposure.
Venous blood was withdrawn
3 and 6 h after each exposure.
Subjects also recorded
symptoms 0, 3, 6 and 9 h
post-final exposure.
Gordon et al. 60 (primary) NR NR Freshly generated ZnO Inhalation exposure of guinea
(1992) vapor diluted in filtered pigs, rats and rabbits to air
air. containing 2.5 or 5 mg/m3 ZnO
particles for 3 h. Lungs were
lavaged 0, 4 or 24 h after
treatment. Inhalation exposure
of human subjects to air
containing
5 mg/m3 ZnO particles for 2 h
with lung function assessed
immediately afterwards and
recorded symptoms 24 h
post-exposure.
Kuschner et al. 170 (secondary NR NR Freshly generated ZnO Inhalation exposure of human
(1997) aggregates from vapor diluted in air. subjects to air containing an
8–40 nm primary average of 33 mg/m3 for 10–30
nanoparticles) min. Bronchoalveolar lavage
was performed 3 or 20 h
post-exposure.
Symptoms of metal fume fever and
plasma levels of IL-6 were elevated
post-inhalation.
Inflammatory biomarkers and
symptoms of metal fume fever
initially increased in naïve subjects
but tolerance was induced by
repeat exposures. Chronically
exposed subjects reported fewer
symptoms than naïve subjects, but
still had elevated levels of plasma
IL-6 following exposure.
Dose-dependent elevation of
inflammatory biomarkers as well as
increased ZnO particle retention
was shown in rats and guinea pigs
but not in rabbits.
Human subjects reported
symptoms of metal fume fever
within 4–8 h of exposure although
no differences in pulmonary
function were detected
immediately post-exposure.
Biomarkers for pulmonary
inflammation were elevated in
subjects 3 h post-exposure.
Analysis of ZnO for sunscreens
21
 Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

22

Table I (Continued)

Particle surface
Authors Particle size (nm) area Surface coating Administration vehicle Experimental regime Results

Lam et al. (1982) 50 NR NR Freshly generated ZnO Inhalation exposure of guinea Most parameters of lung function
vapor diluted in air. pigs to air containing 7.8 mg/ assessed were not significantly
m3 ZnO particles alone, or influenced by single 3 h exposure
0.8–6 mg/m3 ZnO in to ZnO alone but were impaired in
combination with SiO2 for 3 h. animals treated with the ZnO in
Lung function was assessed combination with SiO2 in a
within 2 h of treatment. dose-dependent manner.
Lam et al. (1985) 50 NR NR Freshly generated ZnO Inhalation exposure of guinea Lung function was decreased and
vapor diluted in air. pigs to air containing 5 mg/m3 markers for pulmonary
M.J. Osmond & M.J. McCall

ZnO particles 3 h/day for inflammation increased following

6 days. Lung function and treatment and persisted up to 72 h
morphology were assessed at 1, post-inhalation.
24, 48 or 72 h post-inhalation.
Sayes et al. 50–70 12.1 NR Powder suspended in Intratracheal instillation of 1 or ZnO fine and nanoparticles caused
(2007) Mili-Q water, PBS or 5 mg/kg fine or nano-ZnO in a transient inflammatory response
cell culture media rats with lung inflammatory in vivo in rats. Results of
biomarkers assessed 24 h, comparative in vitro studies were
1 wk, 1 month or 3 months not always predictive of the in vivo
post-exposure. response.
Wesselkamper 300 (secondary NR NR Freshly generated ZnO Inhalation exposure of mice to Inflammatory biomarkers in the
et al. (2001a) aggregates from vapor diluted in filtered air containing 1 mg/m3 ZnO lung decreased with repeated
primary air. particles for 3 h/d for 1, 3 or 5 exposures, however total protein
nanoparticles) successive days. A subset of levels and lung pathology were
mice was exposed for 5 higher with increasing exposure,
successive days, followed by and an increase in metallothionein
re-exposure after 5 d rest. mRNA was also observed.
Bronchoalveolar lavage and Acquired tolerance was lost when
morphological assessment exposure was not on consecutive
performed 24 h post-exposure. days.
Wesselkamper 300 (secondary NR NR Freshly generated ZnO Inhalation exposure of Acquired pulmonary tolerance to
et al. (2001b) aggregates from vapor diluted in filtered 11 inbred mouse strains to inhaled ZnO fumes has a genetic
primary air. 1 mg/m2 ZnO for 3 h/day for component in mice.
nanoparticles) 1 or 5 consecutive days.
Bronchoalveolar lavage was
performed 24 h post-exposure.
 Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

Table I (Continued)

Particle surface
Authors Particle size (nm) area Surface coating Administration vehicle Experimental regime Results

(b) Ingestion
In vitro
No data available
In vivo
Wang et al. 20 & 120 4.4  105 & 9.1 NR Powder suspended in Nanoparticle suspension Little difference in toxicity between
(2008)  104 cm2/g 1% sodium carboxy administered 20 and 120 nm ZnO particles was
methyl cellulose. gastrointestianally into mice at observed, but a lower dose of the
1-, 2-, 3-, 4-, and 20 nm particles induced greater
5-g/kg body weight. pathology than higher doses.
Liver, spleen heart, pancreas and
bone were identified as target
organs for ZnO particles.
(c) Dermal
In vitro
Dufour et al. 100 (average) NR Uncoated Stock solutions of ZnO Chinese hamster ovary cells Cells were more sensitive to ZnO-
(2006) powder added to cell incubated with ZnO under dark induced clastogenic effects with
culture medium. conditions, or pre- or pre- or simultaneous UV
simultaneously irradiated with irradiation, but ZnO itself was not
UV light then assessed for strongly photo-clastogenic.
DNA damage.
Hidaka et al. NR NR NR ZnO particles extracted 1 mg/ml extracted particles ZnO particles extracted from
(2006) from sunscreens and incubated in vitro with plasmid sunscreens were found to be highly
suspended in deionized DNA in the presence of UV photoactive and caused rapid DNA
water. light and assessed for DNA damage under in vitro conditions.
strand breakage.
Rampaul et al. 14–27 NR Uncoated or dimethicone ZnO particles washed Photobleaching of methylene ZnO particles extracted from
(2007) from sunscreen or blue by ZnO extracted particles commercially available sunscreens
obtained directly from was assessed under light and showed photocatalytic activity
the manufacturer. dark conditions. comparable to uncoated TiO2.
Analysis of ZnO for sunscreens
23
 Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

24

Table I (Continued)

Particle surface
Authors Particle size (nm) area Surface coating Administration vehicle Experimental regime Results

Yang et al. 20 NR NR Powder suspended in Primary mouse embryo ZnO nanoparticles exerted
(2008) FBS then diluted in cell fibroblast cells incubated with comparatively more cytotoxic and
culture medium. ZnO nanoparticles, carbon oxidative stress on cells than other
black, single-walled carbon nanoparticles measured, but was
nanotubes, and silicon dioxide less genotoxic than single-walled
nanoparticles. carbon nanotubes. Cytotoxicic
effects were attributed to particle
M.J. Osmond & M.J. McCall

composition while genotoxicity

was attributed to particle shape.
Ex vivo
Cross et al. 15–30 NR Polymethylsilsesquioxane 60% ZnO in caprylic Sunscreen applied to full- Negligible penetration through the
(2007) capric triglyceride or thickness human abdominal stratum corneum.
20% ZnO in caprylic skin sections mounted on
capric triglyceride diffusion cells for
incorporated into 24 h. Also viewed by electron
sunscreen formulation. microscopy.
Dussert et al. 117 NR NR 60% dispersion of Sunscreen applied to human No penetration through stratum
(1997) zinc oxide in mineral abdominal skin and viewed by corneum.
oil/triglyceride electron microscopy.
incorporated into
sunscreen formulation.
Gamer et al. 80, with 90% NR Uncoated 10.3% wt ZnO in Emulsion applied to full- Negligible penetration through the
(2006) < 160 nm oil/water emulsion. thickness porcine abdominal stratum corneum.
skin samples mounted on
diffusion cells for
24 h. Skin stripping was also
performed.
Kuo et al. (2009) 10 NR Uncoated Nanoparticles Single application of ZnO nanoparticles detected
suspended in solutions nanoparticle solution onto 20–30 mm from the skin surface
of PBS, oleic acid, hairless mouse dorsal skin when applied to skin samples in the
ethanol or oleic samples mounted on diffusion presence of oleic acid and/or
acid-ethanol. cells and viewed by ethanol but did not penetrate
multiphoton microscopy. through stratum corneum in
control solution (PBS).
 Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

Table I (Continued)

Particle surface
Authors Particle size (nm) area Surface coating Administration vehicle Experimental regime Results

In vivo
Zvyagin et al. 26–30 NR NR Commercially available One application to forearm, ZnO nanoparticles settled into skin
(2008) sun protective cheek, shoulder and feet of four folds and hair follicles but did not
formulation. subjects of different ethnic penetrate through the stratum
background. Penetration corneum.
assessed by multiphoton
microscopy and SEM/EDX. Ex
vivo human skin samples also
assessed.
Roberts et al. 26–30 NR Polymethylsilsesquioxane Commercially available One application of formulation ZnO nanoparticles found in skin
(2008) suspension of coated rubbed into forearms of 60 folds but no penetration into viable
ZnO nanoparticles in human volunteers of different epidermis.
caprylic/capric age, race and sex for 5 min.
triglycerides. Penetration assessed by
multiphoton microscopy and
SEM/EDX.
Gulson et al. 20 & > 100 NR Uncoated 18% wt 68ZnO in Sunscreens containing Not yet available.
(2009) sunscreen formulation. traceable 68ZnO applied to
human skin twice daily for five
days with exposure to water and
UV radiation.
(d) Environment
Adams et al. 480 nm–4 mm NR NR Powder suspended in E. coli and B. subtilis incubated B. subtilis more sensitive to ZnO
(2006a) (in suspension) Milli-Q water and with 10–5000 ppm overnight than E. coli, with negligible
diluted in liquid under light or dark conditions difference in the presence of
bacterial media. then plated on agar and sunlight compared to dark
colonies counted. incubation for B. subtilis, but more
pronounced light-induced growth
inhibition observed for E. coli.
Adams et al. 480 nm–4 mm NR NR Powder suspended in E. coli or B. subtilis incubated Nanoparticles formed micron-
(2006b) (in suspension) Milli-Q water and with 10–5000 ppm nano- or sized aggregates in suspension and
diluted in bacterial bulk-sized in sunlight or dark were not significantly more toxic to
media. conditions. Cultures were gram-positive or gram-negative
plated on agar and colonies bacteria than particles with a
counted. primary size in the micron-range.
D. magna exposed to B subtilis was more sensitive to ZnO
0.2–1 ppm ZnO in than E. coli. Toxicity was observed
spring water. for B. subtilis under both light and
Analysis of ZnO for sunscreens

dark conditions but E. coli was less

sensitive under dark conditions
25
 Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

26

Table I (Continued)

Particle surface
Authors Particle size (nm) area Surface coating Administration vehicle Experimental regime Results

than light. All concentrations used

showed toxicity towards D. magna.
Aruoja et al. 30 NR NR Powder suspended in OECD 201 algal growth Bulk and nano ZnO showed similar
(2009) algal medium. inhibition test guidelines toxicity to P. subcapitata and
followed to assess toxicity of toxicity was attributed largely to
ZnO nanoparticles to dissolved zinc ions.
P. subcapitata.
M.J. Osmond & M.J. McCall

Brayner et al. 8–14 NR NR Particles ± various Bacteriological tests on E. coli Concentration dependent
(2006) dispersing agents on solid agar plates with varying bactericidal activity observed, with
suspended in bacterial concentrations of ZnO internalisation of particles and
medium. nanoparticles. disruption of cellular membranes.
Franklin et al. 30 NR NR Stock solutions P. subcapitata incubated with a Nanoparticles were no more toxic
(2007) prepared in algal test range of ZnO concentrations in to P. subcapitata than bulk ZnO or
medium. algal media and cell counts dissolved ZnCl2. Toxicity
obtained. attributed to dissolved zinc ions.
Heinlaan et al. 50–70 NR NR Powder suspended in V. fischeri, D. magna and Toxicity and growth inhibition was
(2008) Milli-Q water and T. platyurus incubated with detected for all ZnO preparations,
diluted in test media. bulk and nano-sized ZnO. with negligible difference between
bulk and nano ZnO, and zinc ions
Toxicity was attributed to
dissolved zinc ions.
Huang et al. 60 NR Polyvinyl alcohol Particles suspended in S. agalactiae and S. aureus Concentration-dependent
(2008) aqueous ethylene glycol incubated with ZnO bactericidal activity and damage to
media and diluted in nanoparticles in solid and cellular membranes.
bacterial medium. liquid media.
Jiang et al. (2009) 1448 (secondary NR NR Powder suspended in Bacteria incubated with ZnO in ZnO nanoparticles were more toxic
aggregates from filter-sterilized liquid media then plated on to gram positive and negative
primary 20 nm deionized water agar and colonies counted. bacteria compared to bulk ZnO,
nanoparticles) then diluted in 1 g/l with toxicity attributed largely to
NaCl. particle exposure rather than
dissolved zinc ions.
Jones et al. 50–70 NR NR Powder suspended in Gram-positive and negative Toxicity observed towards gram
(2008) sterile double-distilled bacteria incubated with ZnO in negative and gram positive
water. liquid media then plated on bacteria, with greater growth
agar and colonies counted. inhibition observed with
decreasing particle size.
 Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.

Table I (Continued)

Particle surface
Authors Particle size (nm) area Surface coating Administration vehicle Experimental regime Results
2
Lin and Xing 20 58 m /g NR Powder suspended in Seeds from six higher plant Exposure to ZnO nanoparticles
(2007) deionized water. species incubated in decreased seed germination and
nanoparticle or control root elongation in a number of
suspensions then germinated higher plant species.
on filter paper under dark
incubation conditions.
Reddy et al. 13 NR NR Powder suspended in E. coli and S. aureus grown on Concentration-dependent toxicity
(2007) liquid or solid bacterial solid media containing ZnO to gram negative and gram positive
media. nanoparticles or incubated in bacteria at lower doses than
liquid media containing 0–10 primary human T lymphocytes.
mM ZnO nanoparticles.
Yamamoto 100–800 26–0.85 m2/g NR Powder suspended in Bacteria incubated with ZnO Toxicity increased with decreasing
(2001) saline. nanoparticles across a range of particle size and increasing
concentrations. concentration.
Zhu et al. (2008) 180 (in NR NR Powder suspended in Zebrafish 96-h embryo-larval Concentration dependent toxicity
suspension, Milli-Q water. bioassay was used to assess largely attributed to dissolved zinc
from 20 nm toxicity of ZnO to D. rerio with ions, but residual toxicity was
nanoparticles) toxicological endpoints attributed to nanoparticles
assessed within 96 h exposure. themselves.
Zhu et al. (2009) 20 ‡ 90 m2/g Uncoated Powder suspended in Developing zebrafish incubated ZnO nanoparticles in water settle
zebrafish culture with settled aggregates of ZnO out of water column into
medium. nanoparticles for 96 h followed aggregates within 48 h. Dose
by assessment of developmental dependent toxicity to zebrafish
endpoints. attributed to combination of intact
ZnO nanoparticles and dissolved
Zn2+. Toxicity of nanoparticles was
mitigated in the presence of
formulated sediment.
1
NR, Not reported.
2
Only a subset of published literature associated with metal fume fever has been included here.
Analysis of ZnO for sunscreens
27
 28 M.J. Osmond & M.J. McCall
Particle retention and response in animals may
not necessarily be predictive of that in humans
(Nikula et al. 1997; Nemmar et al. 2004) and the
majority of translocation studies performed in animals
have not specifically investigated ZnO. Nevertheless,
a number of these studies have indicated that other
types of inhaled nanoparticles such as carbon, man-
ganese oxide and silver can translocate relatively
swiftly from the lung to extrapulmonary organs, the
central nervous system, and lung-associated lymph
nodes (Takenaka et al. 2001; Oberdörster et al. 2002;
Elder et al. 2006). TiO2 and gold nanoparticles have
also been shown to penetrate into red blood cells
in vitro in a size-dependent manner (Rothen-
Rutischauser et al. 2006), with the subsequent
biodistribution of gold nanoparticles after intravenous
administration also showing size-dependence (de
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
Jong et al. 2008). Additionally, nanoparticles can
permeate the blood-brain barrier depending on sur-
face characteristics and charge, as well as concentra-
tion (Kreuter 2004; Lockman et al. 2004). In contrast
to these studies, a much lower proportion of systemic
uptake and translocation to extrapulmonary organs
of insoluble iridium nanoparticles was reported
in rats, reaching a maximum after one week and
steadily clearing thereafter (Kreyling et al. 2002;
Semmler et al. 2004).
For personal use only.
In humans, particle translocation from lungs after
inhalation has not specifically been investigated using
ZnO nanoparticles. In studies addressing other types
of nanoparticles, inhaled radioactive carbon nanopar-
ticles were reported to rapidly translocate from the
lung into systemic circulation (Nemmar et al. 2002),
although the stability of the label in biological fluids
has been questioned making it difficult to distinguish
between translocation of free label and particle-bound
label (Wiebert et al. 2006b). Later studies, using more
stable radio-labeled carbonaceous particles in healthy
or asthmatic humans, showed little evidence of trans-
location from the lungs to systemic circulation
(Brown et al. 2002; Wiebert et al. 2006a, 2006b).
However, even if the rate of uptake and/or transloca-
tion is low and subject to subsequent clearance, over
periods of chronic and sustained exposure particle
accumulation may occur (Kreyling et al. 2006).
Gastrointestinal tract. Orally-ingested, solid micro-
particles that do not dissolve can pass between
the epithelial cells of the digestive tract and translocate
throughout the body via the lymph or circu-
latory systems, eventually being eliminated in urine
(Volkheimer 1974). Some types of nanoparticles can
pass through the gastrointestinal mucosa and enter the
bloodstream, where they may undergo translocation to
secondary organs dependent on size and surface
characteristics (Hussain et al. 2001; reviewed by
Hagens et al. 2007). However, data showing the
specific translocation of ZnO nanoparticles from the
gastrointestinal tract are not available although ZnO
nanoparticles have been shown to have only low
solubility in artificial gastric fluid (Wang et al. 2008).
Is more research required?
A range of epidemiological data exists for the health
effects and biomarkers for human exposure to ultra-
fine combustion particles via air pollution (Lewtas
2007), which may provide some insight into the
possible effects of inhaled manufactured nanoparti-
cles in general. However, we can find scant informa-
tion relevant to potential workplace hazards from
inhalation, dermal exposure to dust, or oral ingestion
posed by ZnO nanoparticles manufactured for
sunscreens (Table Ia, Ib). The number of studies
investigating the possible translocation of nanoparti-
cles in general from lungs or the gastrointestinal tract
into systemic circulation is also small and is non-
existent for ZnO nanoparticles. While it is known
that inhalation of ZnO aerosols containing nanopar-
ticles can lead to adverse health effects in humans,
specific research into the potential translocation and
secondary health effects of ZnO nanoparticles is lack-
ing. Currently, one must extrapolate from data
obtained for other kinds of nanoparticles, which is
problematic given not all nanoparticles exhibit the
same behaviors and toxicity. Further research is
required to clarify if translocation occurs, and whether
is likely to be only at low levels and subject to clear-
ance mechanisms. Specifically, research priorities
should include: (i) whether workers manufacturing
or handling ZnO nanoparticles are at any risk from
accidental or incidental exposure in the workplace;
(ii) whether inhaled ZnO nanoparticles in the pulmo-
nary or gastrointestinal systems remain as particles or
dissolve in biological fluids; (iii) whether ZnO nano-
particles in the body translocate from their primary
site of entry; and (iv) whether they are subject to
adequate clearance mechanisms in the body.
Transport and storage of ZnO nanoparticles
prepared for sunscreens
Hazard identification and potential for exposure
After production and packaging, ZnO nanoparticles
typically are transported from the site of manufacture
to the site of their formulation into sunscreen. At this
 Analysis of ZnO for sunscreens 29

stage they may be in the form of a powder or dispersed
in an oil or aqueous medium depending on their
surface coating. Assuming the nanoparticles are
enclosed within appropriate packaging, their transport
and storage would likely involve only low risk of
personal exposure or release into the environment,
apart from the potential risk of accidental spillage
(Figure 1, left). It is difficult to assess the possible
differences in hazard potential when transporting
nanoparticulate forms of ZnO compared to non-
nano due to a lack of experience and knowledge.
During sunscreen formulation (Figure 1, centre),
the hazard and potential exposure for workers from
accidental or incidental release of ZnO nanoparticles
as powder or nanoparticulate aerosol are similar to
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.
those encountered during the manufacture of the
nanoparticles, and they will not be reiterated here.
Product use
Hazard identification and potential for exposure
Percutaneous absorption of topically applied sub-
stances can be divided into two stages: penetration
through the epidermal layers and absorption into the
blood stream (Harvell and Maibach 1994). The func-
tion of the epidermis (Figure 2) is to minimize water
loss whilst maintaining a barrier against external
organisms and particles (Goldstein and Abramovits

Stratum corneum
Stratum lucidum

Epidermis
Stratum granulosum
Stratum spinosum

Stratum basale
Hair shaft

Sebaceous gland

Bulge
Dermis

Matrix cells

Melanocytes

Dermal papilla

Blood vessels
Sub cutaneous

Connective tissue
Adipose tissue

Figure 2. Schematic of the structure of human skin. The epidermis consists of five thin layers, with the outmost layer, the stratum corneum,
largely providing the vital barrier function of the skin. Hair follicles are embedded in the skin, well into the viable dermis. The outer sheath of
the hair follicle is continuous with the epidermis. Blood vessels feed the skin cells as well as the hair follicle.
 30 M.J. Osmond & M.J. McCall
2003; Elias 2005; Brown et al. 2006). The outermost
layer, the stratum corneum, presents the most formi-
dable penetration barrier (Benson 2005) and is a
relatively impermeable two-compartment system of
10–15 layers of keratinocyte-rich corneocyte cells
embedded within a lipid-rich extracellular matrix
(Elias 1983; Benson 2005). This tissue is a self-
renewing layer, with cells moving up from the deeper
layers into the upper layers of the epidermis, differ-
entiating and losing transcriptional and metabolic
activity, eventually reaching the stratum corneum as
non-viable, protein-dense corneocytes (Goldstein and
Abramovits 2003). ZnO nanoparticles contact this
surface layer during normal use of sunscreens
containing them (Figure 1, right). However, the
effects of different skin ages, skin types, or health
conditions on the potential for deep penetration of
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
ZnO nanoparticles from sunscreens have not yet been
established, and so it is not known if they can reach
the deeper living tissues under some conditions, and,
if so, what the impact of that penetration might be.
A number of characteristics of the stratum corneum
can alter across a lifetime (Cerimele et al. 1990;
Hermanns-Le et al. 2004; Waller and Maibach
2005). The degree to which innate barrier function
may become impaired with age is equivocal as data on
this subject are contradictory (Cerimele et al. 1990;
For personal use only.
Harvell and Maibach 1994) but it is recognized that
barrier integrity can be more easily disrupted in the
aged compared to the young, and that recovery occurs
more slowly (Ghadially et al. 1995). Skin penetrability
can also show large intra- and inter-individual vari-
ability (Hostynek 2003). For example, human skin
may be more susceptible to penetration in areas of the
body where it is thinner, or at sites of flexing, such as
joints (Tinkle et al. 2003; Rouse et al. 2007). Con-
sequently, barrier disruption can be influenced by
anatomical location (Breternitz et al. 2007). Stratum
corneum pH is also integral to normal skin-barrier
function (Hachem et al. 2003) and can vary in an age
(Fluhr et al. 2004; Choi et al. 2007), gender and site-
specific manner (Wagner et al. 2003). The presence of
skin pathologies such as atopic dermatitis can also
impair barrier function (Bouwstra and Ponec 2006).
Additionally, exposure of skin to a threshold UV dose
may cause a delayed, temporary decrease in its barrier
function (Haratake et al. 1997; Gelis et al. 2002; Jiang
et al. 2007; Mortensen et al. 2008) and alterations in
humidity and hydration status can also lead to a
temporary decline in skin barrier function (Zhai
and Maibach 2001; Sato et al. 2002; Benson 2005;
Elias 2005). Many of these factors are likely to be
relevant to the assessment of skin penetration by metal
oxide particles, as consumers use sunscreens out-
doors and often in aqueous environments.
Titanium dioxide nanoparticles (Lademann et al.
1999; Nanoderm 2007) and ZnO nanoparticles
(Roberts et al. 2008; Zvyagin et al. 2008) in sun-
screens have been shown to settle into skin furrows
and hair follicles, and it has been assumed that the
majority in the latter locations are most likely
displaced by sebum production and hair growth
(Nohynek et al. 2007). Interestingly, though, hair
follicles are being investigated as a target site for
delivery of drugs (Lademann et al. 2005; Vogt et
al. 2005). Although the outer root sheath of a hair
follicle is continuous with the epidermis, very few
differentiated corneocytes are present in the lower
follicular infundibulum and consequently the
epidermis is relatively permeable at this point (Vogt
et al. 2005). Vogt et al. (2006) showed that 40 nm
fluorescent beads, but not larger particles, penetrated
deeply into the hair follicle, although they were not
detected below the base of the follicle in the dermis
(Vogt et al. 2006). The penetrability of a hair follicle
can be influenced by sebum production and/or hair
growth (Lademann et al. 2001) as well as exfoliation
(Otberg et al. 2004). When considering the potential
routes of entry of ZnO nanoparticles through the
epidermis, it is important to consider typical human
behaviors prior to the application of sunscreens. For
example, skin on the face or legs may be exfoliated,
shaved or waxed just prior to sunscreen application. If
hair follicles are a site of deposition of nanoparticles
and are additionally vulnerable to penetration under
some circumstances, then they may provide a route of
entry for nanoparticles in sunscreens to reach living
skin cells.
It is also relevant to consider the potential impact of
incidental ingestion of low levels of sunscreen from
hand to mouth transfer, and also from the use of ZnO
nanoparticles in sun-protective lip balms. These
exposures may make the gastrointestinal tract a
secondary route of exposure to ZnO nanoparticles
in sunscreen, albeit most likely at very low levels
(Figure 1, right).
Likelihood of exposure
Particles of ZnO in sunscreen are not expected to
penetrate the stratum corneum (Rostan et al. 2002)
and the majority of studies on samples of healthy skin
conducted to date support this view (Nohynek et al.
2007; Environmental Working Group [EWG] 2009)
(Table Ic). In humans, the percutaneous absorption
of zinc from ZnO (size not given), following 72-h
topical application of ZnO emulsions or ointments
onto ex vivo skin samples, was shown to be less than
2% of the applied dose, although still detectable

irrespective of the vehicle (Pirot et al. 1996). Within a
3-h period, only a low, statistically-insignificant
increase in serum zinc levels was observed in healthy
male subjects on whom 100 g 40% ZnO ointments
(size not given) were applied (Derry et al. 1983). Near
nanosized (~117 nm) ZnO particles applied to ex vivo
skin samples from healthy adults remained on top of
the stratum corneum (Dussert et al. 1997). Consis-
tent with this, penetration into tissue was not detected
in experiments with diffusion cells or by electron
microscopy following a single 24-h exposure of
ex vivo human skin samples to sunscreen containing
26–30 nm ZnO (Cross et al. 2007). Similarly, within a
24-h time-frame, ZnO nanoparticles did not pene-
trate past the first few layers of full-thickness ex vivo
porcine skin samples (Gamer et al. 2006). Under in
vivo conditions using multiphoton microscopy,
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
Zvyagin et al. (2008) and Roberts et al. (2008)
showed that ZnO nanoparticles accumulated into
skin folds and hair follicles when applied to the
forearms of human volunteers, but did not
penetrate the stratum corneum (Roberts et al.
2008; Zvyagin et al. 2008).
In contrast, quantum dots with a range of physio-
chemical characteristics penetrated through ex vivo
porcine skin mounted on flow-through diffusion cells
following 8- or 24-h exposure (Ryman-Rasmussen
For personal use only.
et al. 2006), and maghemite and iron nanoparticles
were detected in the stratum granulosum and viable
epidermis of full-thickness human skin samples also
mounted on diffusion cells (Baroli et al. 2007),
showing that nanoparticle penetration can occur
under some circumstances. ZnO nanoparticles were
also shown to pass through the stratum corneum
when they were applied hairless mouse skin mounted
on diffusion cells (Kuo et al. 2009), but the use of this
model which is known to be far more penetrable than
human skin (Scott et al. 1986; van Ravenzwaay and
Leibold 2004; Scientific Committee on Consumer
Products [SCCP] 2006), particularly in the presence
of chemical enhancers (oleic acid and/or ethanol),
may make the relevance of these findings to dermal
penetration of ZnO nanoparticles during normal sun-
screen use somewhat questionable.
Although Roberts et al (2008) used a relatively
large number of subjects (60) of different age, race
and gender, the conclusions from all the studies
described here are limited by the use of a single or
few ZnO applications, small application areas and
short exposure periods. From this perspective, it is
interesting that TiO2 nanoparticles (4 nm) were
shown to penetrate through ear skin of pigs when
applied for 30 consecutive days compared with no
penetration after only 24 h using ex vivo skin samples
mounted on diffusion cells, a common technique
Analysis of ZnO for sunscreens 31
used in ZnO (and TiO2) nanoparticle penetration
studies to date (Wu et al. 2009). The authors also
reported a list of biological effects in hairless mice
after 60 day exposure to topically applied TiO2
including decreased body weight, skin damage and
organ damage (Wu et al. 2009). This study highlights
the importance of realistic exposure periods in asses-
sing the long-term use of nanoparticles, such as ZnO
in sunscreen. For example, it is possible that a build-
up of a secondary diffusion barrier due to the for-
mation of stable bonds between electrophilic metals
and proteins of the skin may retard dermal penetra-
tion by as much as days (Hostynek 2003). Currently
there is no information on whether similar interac-
tions occur between ZnO nanoparticles and skin
surface proteins, and whether such interactions
may influence the results of penetration studies
that only address short-term exposures, such as those
that are available to date.
Some early studies have shown that the state of the
skin, as well as the delivery vehicle, can influence the
percutaneous absorption of ZnO microparticles
(Agren 1991) and suggest that these factors may
influence the absorption of ZnO nanoparticles, too.
In impaired skin models, serum zinc increased after
application of ZnO (size not given) to wounded rat
(Hallmans 1978) and human (Hallmans 1977a) skin,
while the human study also showed elevated levels of
zinc in urine. These studies did not differentiate
between zinc ions released from ZnO and ZnO par-
ticles, per se. Nevertheless, they serve to illustrate that
skin can be penetrated by zinc, and that the potential
for such penetration can be influenced by the state of
the skin to which the ZnO is applied. More recently,
UV exposure has been implicated in enhanced pen-
etration of quantum dot nanoparticles through mildly
sunburned murine skin into the dermis in vivo
(Mortensen et al. 2008), albeit still at a very low level.
A recent study has been conducted in which sun-
screen containing traceable ZnO particles was applied
twice daily for five consecutive days to humans
undergoing normal activities at a Sydney beach
(Gulson et al. 2009). The ZnO was enriched to
> 99.9% with a stable zinc isotope, 68Zn, with the
intention that measurements of altered isotopic
ratios of zinc in blood and urine samples by multi-
collector ICP-MS (inductively-coupled plasma mass-
spectrometry) would distinguish any zinc dermally
absorbed from sunscreen from that naturally present
in the body. Two sunscreens, one with 68ZnO nano-
particles (20 nm) and the other with 68ZnO particles
(> 100 nm), were used to determine if dermal absorp-
tion of ZnO was dependent on particle size. The data
from this study, when available, may provide informa-
tion on potential dermal absorption of uncoated ZnO
 32 M.J. Osmond & M.J. McCall
particles in a specific sunscreen formulation, under
conditions of normal use by humans.
Potential health effects of dermal exposure
An initial concern over the use of metal oxide particles
in sunscreens was that ZnO and TiO2 are semi-
conductors. UV absorption by semi-conductors can
excite electrons from the valence band to the con-
duction band, leaving a ‘hole’ in the former. Most
electrons return to the valence band, releasing excess
energy as heat, but ~ 10% do not return and can
generate free radicals on the surface of the metal oxide
in the presence of water (Wolf et al. 2001). The
inherent photocatalytic activity of ZnO increases
with decreasing particle size (Park and Kang 1997;
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
Casey et al. 2006), and also can be influenced by
particle morphology and the method of preparation
(Wang et al. 2007). Coating or doping are techniques
widely used to reduce the semiconductor activity of
metal oxide particles for sunscreens. Consistent with
this, microsized ZnO particles, both coated and
uncoated, were reported to be both photo-stable
and non-photocatalytic (Mitchnick et al. 1999), and
an absence of photo-genotoxicity in studies with
Chinese hamster ovary cells has also been reported
For personal use only.
(Dufour et al. 2006).
Zinc oxide particles extracted from commercially-
available sunscreens have been shown to be photo-
catalytically active in vitro, but were not investigated
for biological impact in vivo (Rampaul et al. 2007).
Similarly, TiO2 and ZnO nanoparticles present in a
number of commercially-available sunscreens were
recently identified as initiators of the accelerated
weathering of surface coatings on roofs manufactured
by an Australian company (Barker and Branch 2008).
The incidental contact with the roofs during instal-
lation by workers who used these sunscreens trans-
ferred nanoparticles in sufficient amounts to
photocatalytically initiate the free-radical driven deg-
radation of the roof coating. In this case, biological
impact was not assessed. In one study, however, ZnO
particles isolated from commercially-available sun-
screen were shown to induce DNA damage in vitro
after UV irradiation, producing hydroxyl radicals
that attacked the DNA sugar-phosphate chain
(Hidaka et al. 2006). In contrast, microsized ZnO
particles in sunscreen were found to protect UVB-
irradiated murine keratinocyte cells from cytotoxic
stress, DNA damage, disintegration of the cell mem-
brane and oxidative stress, provided the sunscreen
was applied thickly enough to screen at least 90% of
UVB radiation; otherwise an increase of intracellular
ROS was detected (Hayashi et al. 2001).
Even if ZnO nanoparticles in sunscreens do not
penetrate into living tissues, there remains the possi-
bility that dissolved Zn2+ may. Zinc is the second-
most abundant, essential trace-metal after iron
(St. Croix et al. 2005) and is ubiquitous in the
body, being present in all organs, tissues and fluids
(Rostan et al. 2002). Zinc-binding proteins form the
largest class of proteins in multicellular animals
(Nyborg and Peersen 2004), and constitute a large
proportion of the proteins hypothesized to be involved
in the regulation of gene expression (Tupler et al.
2001). Consequently, physiological levels of zinc are
critical for normal gene expression (Nyborg and
Peersen 2004) and intracellular free zinc is generally
found only in picomolar concentrations (Peck and
Ray 1971; Simons 1991), with its cellular distribution
tightly regulated (Maret 1995; St. Croix et al. 2005).
Within the physiological range, zinc is an antioxidant
(Rostan et al. 2002; Prasad et al. 2004) but, should
intracellular levels shift too far in either direction
outside this range, zinc can become harmful to the
cell (Krones et al. 2005; St. Croix et al. 2005;
Wiseman et al. 2006). Disruptions to cellular zinc
homeostasis have been linked in in vitro systems to
oxidative stress, mitochondrial dysfunction, and loss
of cellular viability (Maret 1995; Sensi et al. 1999;
Dineley et al. 2003), and excess zinc can also act as a
neurotoxin (Frederickson et al. 2005). Zinc-ion
release from ZnO nanoparticles is more rapid than
the release from ZnO microparticles (Yang and Xie
2006). The potential for disruption of intracellular
levels of zinc from daily applications of sunscreens
containing ZnO nanoparticles over long terms should
be evaluated, given we know that zinc released from
ZnO can undergo percutaneous absorption.
Potential for translocation
Early studies showed that 65Zn, from 65ZnO particles
in tape applied to the skin of rats, showed a rapid
accumulation in pancreas, kidney, liver, serum, hair
and bone, and was excreted in urine and faeces
(Hallmans 1977b; Hallmans et al. 1978). However,
intact ZnO nanoparticles have not yet been shown to
penetrate through healthy human skin and the poten-
tial translocation of ZnO nanoparticles arising from
dermal absorption has not been investigated.
Is more research required?
At this point it is difficult to conclude whether dermal
exposure to ZnO nanoparticles in sunscreen poses
a hazard from internal exposure to free ZnO

nanoparticles. It is probable that the nanoparticles do
not penetrate through the stratum corneum to living
tissue in healthy, intact skin after a limited number of
exposures (Table Ic). However, it is not clear whether
this situation might alter over a lifetime’s use of
sunscreen, when the influences of repeated exposure,
aging, skin health, waxing, shaving, and penetration
enhancers such as UV radiation or skin hydration are
taken into consideration. Specifically, research should
be directed at two areas: (i) To determine if zinc
detected in vivo from the application of ZnO particles
to skin is in the form of dissolved Zn2+ cations or solid
nanoparticles; and (ii) to determine the effects on
health from long-term exposure to sunscreens con-
taining nanoparticles of ZnO, and particularly the
effects of free-radical generation on the surface of
the skin or possibly in hair follicles.
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
Transplacental and childhood exposure
There are critical periods in utero and during child-
hood when exposure to certain compounds can
affect reproductive development and/or capacity
(Makri et al. 2004; Pryor et al. 2000). While the
degree of exposure is critical, the timing of exposure
may also have profound impacts.
For personal use only.
Low molecular weight, lipid-soluble, non-ionized
substances are most able to diffuse through the pla-
centa or into mammary glands from maternal circu-
lation (Makri et al. 2004). Solid, orally-ingested
microparticles have been shown to rapidly translocate
into the milk of lactating women as well as transpla-
centally (Volkheimer 1974) but studies conducted
with nanoparticles are few in number. In one exper-
iment, foetal lethality and developmental abnormal-
ities in mice were associated with maternal
interperitoneal injection of carbon(60) fullerenes,
suggesting transplacental transfer of the nanoparticles
via maternal blood flow (Tsuchiya et al. 1996). This
contrasts with another study where, although mouse
embryonic cells were shown to internalize polysty-
rene-based nanoparticles in vitro, there were no asso-
ciated developmental abnormalities or toxicities
in vivo (Bosman et al. 2005). Therefore, it is possible
that developmental effects associated with certain
nanoparticles may not be representative of all nano-
particles (Hagens et al. 2007) and initially they should
be assessed on a case-by-case basis.
If nanoparticles can translocate through the pla-
centa or into mammary glands, the possible disrup-
tions to the developing foetus or neonate and their
impact on adult-onset pathologies bear serious con-
sideration. The possibility that pregnant females com-
pared to non-pregnant females may also exhibit a
Analysis of ZnO for sunscreens 33
differential response to nanoparticles should also be
considered. Pregnant BALB/c mice that received
controlled doses of intra-nasally administered suspen-
sions of TiO2 nanoparticles or diesel-exhaust particles
during pregnancy showed an elevated inflammatory
response to both compared to non-pregnant controls.
Additionally, the observed responses correlated with
microarray analyses revealing altered patterns of gene
expression for a number of pathways involved in the
inflammatory response and immune regulation, cell
proliferation and metabolism. Furthermore, the
in utero exposure potentiated offspring to diminished
lung function (Fedulov et al. 2008). These data
suggest that pregnancy may alter the response to
nanoparticles even if they are considered inert in
the non-pregnant state, and that this may impact
foetal development.
Children may be more susceptible to adverse effects
from exposure to potentially hazardous materials due
to a variety of factors, including potentially immature
detoxification systems, as well as differences in behav-
ior and metabolism (Selevan et al. 2000). Children
have a greater ratio of surface area to volume than
adults – approximately 2.7–1.3 times higher in new-
borns and children, respectively (Selevan et al. 2000).
Consequently, although lower mass amounts of
sunscreen may be applied, their effective exposures
may be greater, particularly if multiple applications
are administered over the course of each day.
Currently there are no studies on the long-term
effects of in utero or childhood exposure to ZnO
nanoparticles in sunscreens, and therefore this is an
area of research that should be addressed.
Ecological impact of product use and disposal
Hazard identification and potential for exposure
In Australia, the intentional disposal of ZnO nano-
particles is covered by existing regulations for disposal
of inorganic zinc salts, which are known to have
biocidal activity (Bruschi and Thomas 2006). How-
ever, the normal use of sunscreens and other per-
sonal-care products leads to uncontrolled release of
their ingredients to the natural environment (for
example, by swimming or wet-weather run-off) and
to water-treatment and sewage systems (for example
from showering) (Daughton and Ternes 1999). The
release to sea-water of four commonly-used chemical
UV absorbers in sunscreens has been observed to
indirectly cause rapid and complete bleaching of
hard corals, even at extremely low concentrations,
by affecting symbiotic algae that nourish reef-building
coral species; the chemicals induce a lytic viral cycle in
 34 M.J. Osmond & M.J. McCall
symbiotic zooxanthellae with latent infections, leading
to infection of neighboring coral communities
(Danovaro et al. 2008). Similarly, the bioaccumula-
tion of up to 2 mg/kg lipid comprised of six different
chemical sunscreen components was identified in fish
found in several German lakes used for recreational
swimming during summer periods, even though
the concentration in the water was much lower
(Daughton and Ternes 1999). This suggests that
although sunscreen ingredients may enter ecosystems
only at very low levels and may not necessarily elicit an
acute response, a continual influx may exert more
subtle, long-term effects (Daughton and Ternes 1999).
It is likely that there will be environmental exposure
to ZnO nanoparticles from human use of sunscreens
but the fate and toxicity of nanoparticles in the envi-
ronment is expected to be influenced by the degree of
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
aggregation, stability, transport, sedimentation,
adsorption of environmental molecules and bio-
uptake, as well as potential dissolution. The specific
behavior of ZnO nanoparticles washed from
sunscreens in the relevant environments has not yet
been addressed.
Potential effects of environmental exposure
For personal use only.
A small number of studies have been published within
the last few years addressing the impact of ZnO
nanoparticles (not in sunscreen) on the environment
(Table Id). ZnO has been shown to possess
antimicrobial activity (Sawai 2003; Reddy et al.
2007) that increases with decreasing particle size
and concomitant increase in specific surface area
(Yamamoto 2001; Jones et al. 2008), and which
appears to arise from the generation of ROS from
the particle surfaces (Yamamoto 2001; Yamamoto
et al. 2004). TEM micrographs of bacteria incubated
with nano-sized ZnO have also shown nanoparticles
located inside and outside of the cell, with associated
membrane damage (Brayner et al. 2006; Huang et al.
2008). However the antimicrobial effect of ZnO
diminished markedly when bacteria were exposed
under dark conditions, suggesting that the anti-
bacterial effect might be enhanced by visible light
(Jones et al. 2008). This is consistent with photoca-
talytically-activated ROS toxicity. In contrast,
Adams et al. reported that the toxicity of ZnO to
bacteria was largely size- and light-independent
(Adams et al. 2006a, 2006b). Moreover, incubation
with 3–10 mM nanosized ZnO caused 100% growth
inhibition of Escherichia coli, while lower concentra-
tions between 1 and 1.5 mM supported only a slight
increase in colony growth (Brayner et al. 2006),
suggesting a dose-dependent effect.
When assessing the potential ecotoxicity of ZnO
nanoparticles, the dynamics of the particles in solu-
tion should be fully characterized, as toxicity may be
ascribed erroneously to the nanoparticle itself when in
fact it may be due to Zn2+ released from the nano-
particle. ZnO is insoluble in water (Perl 1993) but
appears to be at least partially soluble in some bio-
logical fluids. For example, Franklin et al. (2007)
showed that ZnO nanoparticles rapidly aggregated
and then dissolved in a neutral freshwater algal
medium. Nano-sized ZnO was shown to be no
more toxic than bulk ZnO and dissolved ZnCl2 to
the freshwater algae Pseudokirchneriella subcapitata,
and the toxicity exhibited by all three could be attrib-
uted to released Zn2+ (Franklin et al. 2007). This
result was confirmed in another laboratory using the
same organism (Aruoja et al. 2009). The importance
of characterizing particle solubility was also demon-
strated by Brunner et al. (2006), who showed that the
partially-soluble nanoparticles ZnO and Fe2O3 were
more cytotoxic to rodent and human cell cultures
than the insoluble particles Ca3(PO4)2 and TiO2
(Brunner et al. 2006). Consistent with this, suspen-
sions of bulk and nanoparticulate ZnO, as well as
dissolved ZnSO4, were toxic to bacteria and crusta-
ceans, and this toxicity was shown to be largely due to
solubilized zinc rather than the ZnO particles
themselves (Heinlaan et al. 2008).
In contrast, a recent study demonstrated that a
solution containing ~ 180 nm aggregates of ZnO
nanoparticles, filtered through 100 nm acrodisc
syringe filters to exclude the nanoparticles, exhibited
toxicity to zebrafish, again indicative of released Zn2+-
mediated toxicity. However, the filtered solution con-
taining only the dissolved zinc ions was less toxic than
the original nanoparticle suspension suggesting that
intact nanoparticles may still enhance overall toxicity
(Zhu et al. 2008). A follow-up study also suggested
that the toxicity of ZnO nanoparticles to developing
zebrafish could be attributed to a combination of
intact ZnO nanoparticles and released Zn2+. Inter-
estingly, this study also reported that when the assay
was conducted in the presence of sediment, the tox-
icity of the nanoparticles was largely mitigated
(Zhu et al. 2009). Similarly, although dissolved
Zn2+ ions were shown to contribute to some toxicity
against certain bacterial strains, toxicity was largely
attributed to exposure to ZnO particles, with nano-
particles exerting a more toxic effect than bulk par-
ticles (Jiang et al. 2009). The impact of chemical
stability of metallic nanoparticles on cellular toxicity
has recently been reviewed (Auffan et al. 2009).
Zinc oxide nanoparticles may exhibit some toxicity
to plants, based on limited studies. Exposure to
suspensions of ZnO nanoparticles inhibited seed

germination of corn and virtually terminated the root
growth of radish, rape, ryegrass, lettuce, corn and
cucumber (Lin and Xing 2007) with a lower IC50
than previously reported for some other plant
species using non-nano ZnO (Paschke et al. 2006;
Paschke et al. 2000). Importantly, the phytotoxicity of
the filtered ZnO nanoparticle solution containing
Zn2+ was shown to be insignificant, suggesting that
the observed phytotoxicity was due to the exposure to
intact nanoparticles rather than released Zn2+ (Lin
and Xing 2007).
Is more research required?
The paucity of information in Table Id suggests a lack
of public research targeted specifically to determine
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
the impact of ZnO nanoparticles from sunscreens in
the environment, although powdered ZnO has
received some attention. Studies so far have tended
to focus on isolated organisms, rather than holistic
studies to address the effects on symbiotic organisms
or potential bioaccumulation progressing up the food
chain. An analysis of the impact of ZnO nanoparticles
on an ecosystem might be appropriate, where possi-
ble, in order to detect potentially subtle, synergistic
effects that might not otherwise be revealed under
For personal use only.
isolated experimental conditions. In addition, studies
are needed on the state of ZnO particles released to
the environment from swimmers using sunscreen, as
these particles are likely to be coated with ingredients
from the sunscreen formulation, and may behave
differently from pristine ZnO particles in both model
and natural environments. Indeed, studies on the
movement and fate of ZnO nanoparticles released
to the environment should precede any comprehen-
sive toxicity studies so that the aggregation state, and
altered surface coatings, may be identified; and then
relevant forms of the nanoparticles may be tested in
ecotoxicity studies to obtain useful data.
Conclusions
ZnO nanoparticles show the broadest UV protection
of all the active ingredients currently available in
commercial sunscreens (Pinnell et al. 2000), and it
is important to bear this in mind when considering the
potential risks associated with their use. Their incor-
poration in sunscreens provides broad protection
against the known carcinogenic, immunosuppressant
and photo-ageing effects of UVA and UVB. There-
fore, avoidance of sunscreens containing ZnO nano-
particles on the basis of unknown or perceived hazards
would appear to be counterproductive, particularly
when considered against the potential for adverse
Analysis of ZnO for sunscreens 35
effects arising from the long-term use of some other
active sunscreen ingredients (Schlumpf et al. 2001).
Nonetheless, there is clearly a dearth of research into
the specific risks to human health and the environ-
ment associated with ZnO nanoparticles destined for
use in sunscreens. Recently, the Environmental
Working Group released a statement in favor of using
sunscreens containing ZnO and TiO2 nanoparticles
on the basis of current evidence, although it was noted
that the impact of compromised skin had not been
completely addressed by existing studies and that
there were “significant unaddressed safety concerns
for workers handling nano materials” (EWG 2009).
The report also noted that insufficient environmental
assessments are available for regulators to adequately
control potential hazard associated with mainstream
use of these materials.
In summary, future research efforts should be
expended to obtain information on and an under-
standing of: (i) Inhalation exposure for workers and
other personnel during the manufacturing phase; (ii)
the impact of less than healthy skin and long-term use
on the skin-penetrability of ZnO nanoparticles in
sunscreen as well as their potential translocation;
(iii) the potential for free radical activity from ZnO
nanoparticles lodged in hair follicles; (iv) the transport
and fate of ZnO nanoparticles after discharge into the
environment arising from manufacturing and con-
sumer use; and (v) their ultimate effects on ecosys-
tems. In order to facilitate comparison between
different studies, research should be conducted
together with a thorough characterization of the phys-
ical and chemical properties of the ZnO nanoparticles
studied, as several metrics in addition to size may
influence their behavior. Furthermore, the potential
risk for any detrimental effects on foetal development
arising from maternal use of sunscreens containing
ZnO nanoparticles should be ascertained.
Acknowledgements
The authors gratefully acknowledge Brendan Carney
(Department of Infrastructure, Transport, Regional
Development & Local Government Australia) for
providing advice regarding the possible OH&S issues
surrounding the transport of ZnO nanoparticles in
Australia. We would also like to thank Glenn McLeod
for preparing the diagrams used in this manuscript,
and Phil Casey (CSIRO) for his critical reading,
suggestions and comments.
Declaration of interest: Work on this manuscript
was supported by the Nanosafety Theme in the CSIRO
Future Manufacturing Flagship, which receives its
 36 M.J. Osmond & M.J. McCall
funding from the Australian Government. The authors
report no conflicts of interest. The authors alone are
responsible for the content and writing of the paper.
References
Adams L, Lyon D, McIntosh A, Alvarez P. 2006a. Comparative
toxicity of nano-scale TiO2, SiO2 and ZnO water suspensions.
Wat Sci Tech 54:327–334.
Adams LK, Lyon DY, Alvarez PJJ. 2006b. Comparative eco-
toxicity of nanoscale TiO2, SiO2, and ZnO water suspensions.
Water Res 40:3527–3532.
Agren MS. 1991. Influence of two vehicles for zinc oxide on zinc
absorption through intact skin and wounds. Acta Derm Venereol
71:153–156.
Aruoja V, Dubourguier HC, Kasemets K, Kahru A. 2009. Toxicity
of nanoparticles of CuO, ZnO and TiO2 to microalgae
Pseudokirchneriella subcapitata. Sci Total Environ 407:
1461–1468.
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
Auffan M, Rose J, Wiesner MR, Bottero JY. 2009. Chemical
stability of metallic nanoparticles: A parameter controlling
their potential cellular toxicity in vitro. Environ Pollut
157:1127–1133.
Baggs RB, Ferin J, Oberdörster G. 1997. Regression of pulmonary
lesions produced by inhaled titanium dioxide in rats. Vet Pathol
34:592–597.
Barker P, Branch A. 2008. The interaction of modern sunscreen
formulations with surface coatings. Prog Org Coat 62:
313–320.
Baroli B, Ennas MG, Loffredo F, Isola M, Pinna R, Lopez-
Quintela MA. 2007. Penetration of metallic nanoparticles in
For personal use only.
human full-thickness skin. Soc Invest Dermatol 127:1701–1712.
Bauermann LP, Bill J, Aldinger F. 2006. Bio-friendly synthesis of
ZnO nanoparticles in aqueous solution at near-neutral pH and
low temperature. J Phys Chem B 110:5182–5185.
Beckett W, Chalupa D, Pauly-Brown A, Speers D, Stewart J,
Frampton M, Utell M, Huang L, Cox C, Zareba W, et al.
2005. Comparing inhaled ultrafine versus fine zinc oxide parti-
cles in healthy adults. Am J Respir Crit Care Med 171:
1129–1135.
Benson HA. 2005. Transdermal drug delivery: Penetration
enhancement techniques. Curr Drug Deliv 2:2–33.
Bermudez E, Mangum JB, Asgharian B, Wong BA, Reverdy EE,
Janszen DB, Hext PM, Warheit DB, Everitt JI. 2002. Long-term
pulmonary responses of three laboratory rodent species to sub-
chronic inhalation of pigmentary titanium dioxide particles.
Toxicol Sci 70:86–97.
Bermudez E, Mangum JB, Wong BA, Asgharian B, Hext PM,
Warheit DB, Everitt JI. 2004. Pulmonary responses of mice, rats,
and hamsters to subchronic inhalation of ultrafine titanium
dioxide particles. Toxicol Sci 77:347–357.
Borm PJ, Kreyling W. 2004. Toxicological hazards of inhaled
nanoparticles – potential implications for drug delivery. J
Nanosci Nanotechnol 4:521–531.
Borm PJA, Schins RPF, Albrecht C. 2004. Inhaled particles and
lung cancer, Part B: Paradigms and risk assessment. Int J Cancer
110:3–14.
Bosman SJ, Nieto SP, Patton WC, Jacobson JD, Corselli JU,
Chan PJ. 2005. Development of mammalian embryos exposed
to mixed-size nanoparticles. Clin Exp Obstet Gynecol
32:222–224.
Bouwstra JA, Ponec M. 2006. The skin barrier in healthy and
diseased state. Biochim Biophys Acta 1758:2080–2095.
Brayner R, Ferrari-lliou R, Brivois N, Djediat S, Benedetti MF,
Fievet F. 2006. Toxicological impact studies based on
Escherichia coli bacteria in ultrafine ZnO nanoparticles colloidal
medium. Nano Lett 6:866–870.
Breternitz M, Flach M, Prassler J, Elsner P, Fluhr JW. 2007. Acute
barrier disruption by adhesive tapes is influenced by pressure,
time and anatomical location integrity and cohesion assessed by
sequential tape stripping; a randomized, controlled study. Br J
Dermatol 156:231–240.
Brouwer DH, Gijsbers JH, Lurvink MW. 2004. Personal exposure
to ultrafine particles in the workplace: Exploring sampling
techniques and strategies. Ann Occup Hyg 48:439–453.
Brown JS, Zeman KL, Bennett WD. 2002. Ultrafine particle
deposition and clearance in the healthy and obstructed lung.
Am J Respir Crit Care Med 166:1240–1247.
Brown MB, Martin GP, Jones SA, Akomeah FK. 2006. Dermal
and transdermal drug delivery systems: Current and future
prospects. Drug Deliv 13:175–187.
Bruls WA, Slaper JC, van der Leun JC, Berrins L. 1984. Trans-
mission of human epidermis and stratum corneum as a function
of thickness in the ultraviolet and visible wavelengths.
Photochem Photobiol 40:485–494.
Brunner TJ, Wick P, Manser P, Spohn P, Grass RN,
Limbach LK, Bruinink A, Stark WJ. 2006. In vitro cytotoxicity
of oxide nanoparticles: Comparison to asbestos, silica, and the
effect of particle solubility. Environ Sci Technol 40:4374–4381.
Bruschi S, Thomas S. 2006. A review of the potential occupa-
tional health & safety implications of nanotechnology. In:
Council ASaC, editor: accessed from http://www.safe-
workaustralia.gov.au/NR/rdonlyres/8FBDE049-ADB4-417D-
8771-B9DA1D6BC7E6/0/ASCCReviewOHSImplications-
Nanotechnology2006.pdf..
Casey PS, Rossouw CJ, Boskovic S, Lawrence KA, Turney TW.
2006. Incorporation of dopants into the lattice of ZnO
nanoparticles to control photoactivity. Superlattic Microst
39:97–106.
Cayrol C, Sarraute J, Tarroux R, Redoules D, Charveron M,
Gall Y. 1999. A mineral sunscreen affords genomic protection
against ultraviolet (UV)B and UVA radiation: In vitro and in situ
assays. Br J Dermatol 141:250–258.
Cerimele D, Celleno L, Serri F. 1990. Physiological changes in
ageing skin. Br J Dermatol 122:13–20.
Champion JA, Mitragotri S. 2006. Role of target geometry in
phagocytosis. PNAS 103:4930–4934.
Choi EH, Man MQ, Xu P, Xin S, Liu Z, Crumrine DA, Jiang YJ,
Fluhr JW, Feingold KR, Elias PM, et al. 2007. Stratum corneum
acidification is impaired in moderately aged human and murine
skin. J Invest Dermatol 127:2847–2856.
Churg A, Stevens B, Wright JL. 1998. Comparison of the uptake of
fine and ultrafine TiO2 in a tracheal explant system. Am J Physiol
274:L81–86.
Cross SE, Innes B, Roberts MS, Tsuzuki T, Robertson TA,
McCormick P. 2007. Human skin penetration of sunscreen
nanoparticles: In-vitro assessment of a novel micronized zinc
oxide formulation. Skin Pharmacol Physiol 20:148–154.
Danovaro R, Bongiorni L, Corinaldesi C, Giovannelli D,
Damiani E, Astolfi P, Greci L, Pusceddu A. 2008. Sunscreens
cause coral bleaching by promoting viral infections. Environ
Health Perspect 116:441–447.
Darr D, Fridovich I. 1994. Free radicals in cutaneous biology.
J Invest Dermatol 102:671–675.
Daughton CG, Ternes TA. 1999. Pharmaceuticals and personal
care products in the environment: Agents of subtle change?
Environ Health Perspect 107:907–938.
de Jong WH, Hagens WI, Krystek P, Burger MC, Sips AJ,
Geertsma RE. 2008. Particle size-dependent organ distribution
of gold nanoparticles after intravenous administration.
Biomaterials 29:1912–1919.

Derry JE, McLean WM, Freeman JB. 1983. A study of the
percutaneous absorption from topically applied zinc oxide
ointment. J Parenter Enteral Nutr 7:131–135.
Diffey BL. 2002. Human exposure to solar ultraviolet radiation. J
Cosmet Dermatol 1:124–130.
Dineley KE, Votyakova TV, Reynolds IJ. 2003. Zinc inhibition of
cellular energy production: Implications for mitochondria and
neurodegeneration. J Neurochem 85:563–570.
Donaldson K, Li XY, MacNee W. 1998. Ultrafine (nanometre)
particle mediated lung injury. J. Aerosol Sci 29:553–560.
Donaldson K, Stone V, Gilmour PS, Brown DM, MacNee W.
2000. Ultrafine particles: Mechanisms of lung injury. Roy Soc
London Phil Tr A 358:2741–2749.
Duffy GM, Pillai SC, McCormack DE. 2007. A novel processing
route for the production of nanoparticulate zinc oxide using an
isophthalate precursor. Smart Mater Struct 16:1379–1381.
Dufour EK, Kumaravel T, Nohynek GJ, Kirkland D, Toutain H.
2006. Clastogenicity, photo-clastogenicity or pseudo-photo-
clastogenicity: Genotoxic effects of zinc oxide in the dark, in
pre-irradiated or simultaneously irradiated Chinese hamster
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
ovary cells. Mut Res 607:215–224.
Dussert A-S, Gooris E, Hemmerle J. 1997. Characterization of
the mineral content of a physical sunscreen emulsion and its
distribution onto human stratum corneum. Int J Cos Sci 19:
119–129.
Elder A, Gelein R, Silva V, Feikert T, Opanashuk L, Carter J,
Potter R, Maynard A, Ito Y, Finkelstein J, et al. 2006. Trans-
location of inhaled ultrafine manganese oxide particles to the
central nervous system. Environ Health Perspect 114:
1172–1178.
Elias PM. 1983. Epidermal lipids, barrier function, and desqua-
mation. J Invest Dermatol 80:44S–49S.
For personal use only.
Elias PM. 2005. Stratum corneum defensive functions: An
integrated view. J Invest Dermatol 125:183–200.
EPA. 1995. Zinc oxide; Toxic chemical release reporting;
Community right-to know. Federal Register 60:47334–47337.
Environmental Working Group (EWG). 2009. Nanotechnology &
sunscreens. Accessed from the website: http://www.ewg.org/
cosmetics/report/sunscreen09/investigation/Nanotechnology-
Sunscreens.
Fedulov AV, Leme A, Yang Z, Dahl M, Lim R, Marian TJ,
Kobzik L. 2008. Pulmonary exposure to particles during preg-
nancy causes increased neonatal asthma susceptibility. Am J
Respir Cell Mol Biol 38:57–67.
Fine JM, Gordon T, Chen LC, Kinney P, Falcone G, Beckett W.
1997. Metal fume fever: Characterization of clinical and plasma
IL-6 responses in controlled human exposures to zinc oxide
fume at and below the threshold limit value. J Occup Environ
Med 39:722–726.
Fine JM, Gordon T, Chen LC, Kinney P, Falcone G, Sparer J,
Beckett WS. 2000. Characterization of clinical tolerance to
inhaled zinc oxide in naive subjects and sheet metal workers.
J Occup Environ Med 42:1085–1091.
Fluhr JW, Behne MJ, Brown BE, Moskowitz DG, Selden C,
Mao-Qiang M, Mauro TM, Elias PM, Feingold KR. 2004.
Stratum corneum acidification in neonatal skin: Secretory
phospholipase A2 and the sodium/hydrogen antiporter-1 acidify
neonatal rat stratum corneum. J Invest Dermatol 122:320–329.
Forestier S. 2008. Rationale for sunscreen development. J Am Acad
Dermatol 58:S133–1389.
Franklin NM, Rogers NJ, Apte SC, Batley GE, Gadd GE,
Casey PS. 2007. Comparative toxicity of nanoparticulate
ZnO, bulk ZnO, and ZnCl2 to a freshwater microalga
(Pseudokirchneriella subcapitata): The importance of particle
solubility. Environ Sci Technol 41:8484–8490.
Analysis of ZnO for sunscreens 37
Frederick JE, Snell HE. 1988. Ultraviolet radiation levels during the
Antarctic spring. Science 241:438–440.
Frederickson CP, Koh JY, Bush AI. 2005. The neurobiology of zinc
in health and disease. Nat Rev Neurosci 6:449–462.
Fubini B. 1997. Surface reactivity in the pathogenic response to
particulates. Environ Health Perspect 105:1013–1020.
Gamer AO, Leibold E, van Ravenzwaay B. 2006. The in vitro
absorption of microfine zinc oxide and titanium dioxide through
porcine skin. Toxicol in Vitro 20:301–307.
Geiser M, Casaulta M, Kupferschmid B, Schulz H, Semmler-
Behnke M, Kreyling W. 2008. The role of macrophages in
the clearance of inhaled ultrafine titanium dioxie particles.
Am J Respir Cell Mol Biol 38:371–376.
Gelis C, Mavon A, Delverdier M, Paillous N, Vicendo P. 2002.
Modifications of in vitro skin penetration under solar irradiation:
Evaluation of flow-through diffusion cells. Photochem Photobiol
75:598–604.
Ghadially R, Brown BE, Sequeira-Marin SM, Feingold KR,
Elias PM. 1995. The aged epidermal permeability barrier.
J Clin Invest 95:2281–2290.
Gojova A, Guo B, Kota RS, Rutledge JC, Kennedy IM, Barakat AI.
2007. Induction of inflammation in vascular endothelial cells by
metal oxide nanoparticles: Effect of particle composition.
Environ Health Perspect 115:403–409.
Goldstein AM, Abramovits W. 2003. Ceramides and the stratum
corneum: Structure, function, and new methods to promote
repair. Int J Dermatol 42:256–259.
Gopalan R, Osman I, Amani A, Matas Md, Anderson D. 2009. The
effect of zinc oxide and titanium dioxide nanoparticles in the
comet assay with UVA photoactivation of human sperm and
lymphocytes. Nanotox 3:33–39.
Gordon T, Chen LC, Fine JM, Schlesinger RB, Su WY,
Kimmel TA, Amdur MO. 1992. Pulmonary effects of inhaled
zinc oxide in human subjects, guinea pigs, rats, and rabbits. Am
Ind Hyg Assoc J 53:503–509.
Gulson B, Gomez L, Casey P, Korsch M, Greenoak G, Trotter J,
McCulloch M, McCall MJ. 2009. Are zinc oxide particles in
sunscreens absorbed through the skin? 4th International Con-
ference on Nanotechnology – Occupational and Environmental
Health, Helsinki: page 85 of programme.
Hachem J-P, Crumrine D, Fluhr J, Brown BE, Feingold KR,
Elias PM. 2003. pH directly regulates epidermal permeability
barrier homeostasis, and stratum corneum integrity/cohesion.
J Invest Dermatol 121:345–353.
Hagens WI, Oomen AG, de Jong WH, Cassee FR, Sips AJ. 2007.
What do we (need to) know about the kinetic properties
of nanoparticles in the body? Regul Toxicol Pharmacol
49:217–229.
Hallmans G. 1977a. Treatment of burns with zinc-tape. A study of
local absorption of zinc in humans. Scand J Plast Reconstr Surg
11:155–161.
Hallmans G. 1977b. Zinc resorption from zinc-tape during wound
healing. An experimental study in rats. Scand J Plast Reconstr
Surg 11:27–32.
Hallmans G. 1978. Local absorption of zinc from wounds treated
with various zinc-compounds. Acta Derm Venereol 58:
251–256.
Hallmans G, Nilsson AM, Sjostrom R. 1978. Absorption, retention
and excretion of 65Zn in rats with wounds treated with 65Zn-
labelled zinc tape. Scand J Plast Reconstr Surg 12:111–117.
Haratake A, Uchinda Y, Schmuth M, Tanno O, Yasuda R,
Epstein JH, Elias PM, Holleran WM. 1997. UVB-induced
alterations in permeability barrier function: Roles for epidermal
hyperproliferation and thymocyte-mediated response. J Invest
Dermatol 108:769–775.
 38 M.J. Osmond & M.J. McCall
Harvell JD, Maibach HI. 1994. Percutaneous absorption and
inflammation in aged skin: A review. J Am Acad Dermatol
31:1015–1021.
Hayashi S, Takeshita H, Nagao N, Nikaido O, Miwa N. 2001. The
relationship between UVB screening and cytoprotection by
microcorpuscular ZnO or ascorbate against DNA photodamage
and membrane injuries in keratinocytes by oxidative stress.
J Photochem Photobiol B 64:27–35.
Heinlaan M, Ivask A, Blinova I, Dubourguier HC, Kahru A. 2008.
Toxicity of nanosized and bulk ZnO, CuO and TiO2 to bacteria
Vibrio fischeri and crustaceans Daphnia magna and
Thamnocephalus platyurus. Chemosphere 71:1308–1316.
Hermanns-Le T, Uhoda I, Smitz S, Pierard GE. 2004. Skin tensile
properties revisited during ageing. Where now, what next?
J Cosmet Dermatol 3:35–40.
Hext PM, Tomenson JA, Thompson P. 2005. Titanium dioxide:
Inhalation toxicology and epidemiology. Ann Occup Hyg
49:461–472.
Hidaka H, Kobayashi H, Koike T, Sato T, Serpone N. 2006. DNA
damage photoinduced by cosmetic pigments and sunscreen
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
agents under solar exposure and artificial UV illumination.
J Oleo Sci 55:249–261.
Hohr D, Steinfartz Y, Schins RPF, Knaapen AM, Martra G,
Fubini B, Borm PJA. 2002. The surface area rather than the
surface coating determines the acute inflammatory response after
instillation of fine and ultrafine TiO2 in the rat. Int J Hyg Environ
Health 205:239–244.
Hostynek JJ. 2003. Factors determining percutaneous metal
absorption. Food Chem Toxicol 41:327–345.
Huang Z, Zheng X, Yan D, Yin G, Liao X, Kang Y, Yao Y,
Huang D, Hao B. 2008. Toxicological effect of ZnO
nanoparticles based on bacteria. Langmuir 24:4140–4144.
For personal use only.
Hussain N, Jaitley V, Florence A. 2001. Recent advances in the
understanding of uptake of microparticulates across the gastro-
intestinal lymphatics. Adv Drug Deliv Rev 50:107–142.
Jeng HA, Swanson J. 2006. Toxicity of metal oxide nano-
particles in mammalian cells. J Environ Sci Health A
41:2699–2711.
Jiang SJ, Chu AW, Lu ZF, Pan MH, Che DF, Zhou X. 2007.
Ultraviolet B-induced alterations of the skin barrier and epider-
mal calcium gradient. Exp Dermatol 16:985–992.
Jiang W, Mashayekhi H, Xing B. 2009. Bacterial toxicity compar-
ison between nano- and micro-scaled oxide particles. Environ
Pollut 157:1619–1625.
Jones N, Ray B, Ranjit KT, Manna AC. 2008. Antibacterial activity
of ZnO nanoparticle suspensions on a broad spectrum of micro-
organisms. FEMS Microbiol Lett 279:71–76.
Kelleher P, Pacheco K, Newman LS. 2000. Inorganic dust pneu-
monias: The metal-related parenchymal disorders. Environ
Health Perspect 108:685–696.
Kendall M, Tetley TD, Wigzell E, Hutton B, Nieuwenhuijsen M,
Luckham P. 2002. Lung lining liquid modified PM2.5 in favor of
particle aggregation: A protective mechanism. Am J Physiol
Lung Cell Mol Physiol 282:L109–114.
Kleinwechter H, Janzen C, Knipping J, Wiggers H, Roth P. 2002.
Formation and properties of ZnO nano-particles from gas phase
synthesis processes. J Mater Sci 37:4349–4360.
Kreuter J. 2004. Influence of the surface properties on nanoparticle-
mediated transport of drugs to the brain. J Nanosci Nanotechnol
4:484–488.
Kreyling W, Semmler-Behnke M, Moller W. 2006. Ultrafine
particle-lung interactions: Does size matter? J Aerosol Med
19:74–83.
Kreyling WG, Semmler M, Erbe F, Mayer P, Tanenaka S,
Schulz H, Oberdörster G, Ziesenis A. 2002. Translocation of
ultrafine insoluble iridium particles from lung epithelium to
extrapulmonary organs is size dependent but very low. J Toxicol
Environ Health A 65:1513–1530.
Krones CJ, Klosterhalfen B, Butz N, Hoelzl F, Junge K, Stumpf M,
Peiper C, Klinge U, Schumpelick V. 2005. Effect of zinc pre-
treatment on pulmonary endothelial cells in vitro and pulmonary
function in a porcine model of endotoxemia. J Surg Res
123:251–256.
Kuo T-R, Wu C-L, Hsu C-T, Lo W, Chiang S-J, Lin S-J,
Dong C-Y, Chen C-C. 2009. Chemical enhancer induced
changes in the mechanisms of transdermal delivery of zinc oxide
nanoparticles. Biomaterials 30:3002–3008.
Kuschner WG, D’Alessandro A, Wong H, Blanc PD. 1997. Early
pulmonary cytokine responses to zinc oxide fume inhalation.
Environ Res 75:7–11.
Lademann J, Otberg N, Jacobi U, Hoffman RM, Blume-Peytavi U.
2005. Follicular penetration and targeting. J Invest Dermatol
Symp Proc 10:301–303.
Lademann J, Otberg N, Richter H, Weigmann HJ, Lindemann U,
Schaefer H, Sterry W. 2001. Investigation of follicular penetra-
tion of topically applied substances. Skin Pharmacol Appl Skin
Physiol 14:17–22.
Lademann J, Weigmann H, Rickmeyer C, Barthelmes H,
Schaefer H, Mueller G, Sterry W. 1999. Penetration of titanium
dioxide microparticles in a sunscreen formulation into the horny
layer and the follicular orifice. Skin Pharmacol Appl Skin Physiol
12:247–256.
Lam HF, Conner MW, Rogers AE, Fitzgerald S, Amdur MO.
1985. Functional and morphologic changes in the lungs of
guinea pigs exposed to freshly generated ultrafine zinc oxide.
Toxicol Appl Pharmacol 78:29–38.
Lam HF, Peisch R, Amdur MO. 1982. Changes in lung volumes
and diffusing capacity in guinea pigs exposed to a combination of
sulfur dioxide and submicron zinc oxide mixed in a humidified
furnace. Toxicol Appl Pharmacol 66:427–433.
Lewtas J. 2007. Air pollution combustion emissions: Characteri-
zation of causative agents and mechanisms associated with
cancer, reproductive, and cardiovascular effects. Mut Res
636:95–133.
Limbach LK, Li Y, Grass RN, Brunner TJ, Hintermann MA,
Muller M, Gunther D, Stark WJ. 2005. Oxide nanoparticle
uptake in human lung fibroblasts: Effects of particle size,
agglomeration, and diffusion at low concentrations. Environ
Sci Technol 39:9370–9376.
Lin D, Xing B. 2007. Phytotoxicity of nanoparticles: Inhibition
of seed germination and root growth. Environ Pollut 150:
243–250.
Lockman PR, Koziara JM, Mumper RJ, Allen DD. 2004. Nano-
particle surface charges alter blood-brain barrier integrity and
permeability. J Drug Target 12:635–641.
Lundqvist M, Stigler J, Elia G, Lynch I, Cedervall T, Dawson KA.
2008. Nanoparticle size and surface properties determine the
protein corona with possible implications for biological impacts.
PNAS 105:14265–14270.
Lynch I, Dawson KA. 2008. Protein-nanoparticle interactions.
NanoToday 3:40–47.
Madronich S, McKenzie RL, Bjorn LO, Caldwell MM. 1998.
Changes in biologically active ultraviolet radiation reaching
the earth’s surface. J Photochem Photobiol B 46:5–19.
Maier T, Korting HC. 2005. Sunscreens – which and what for?
Skin Pharmacol Physiol 18:253–262.
Makri A, Goveia M, Balbus J, Parkin R. 2004. Children’s suscep-
tibility to chemicals: A review by developmental stage. J Toxicol
Environ Health B Crit Rev 7:417–435.
Maret W. 1995. Metallothionein/disulfide interactions, oxidative
stress, and the mobilization of cellular zinc. Neurochem Int
27:111–117.

Marrot L, Meunier JR. 2008. Skin DNA photodamage and its
biological consequences. J Am Acad Dermatol 58:S139–148.
Matts PJ. 2006. Solar ultraviolet radiation: Definitions and termi-
nology. Dermatol Clin 24:1–8.
Maynard AD, Baron PA, Foley M, Shvedova AA, Kisin ER,
Castranova V. 2004. Exposure to carbon nanotube material:
Aerosol release during the handling of unrefined single-walled
carbon nanotube material. J Toxicol Environ Health A 67:87–107.
Maynard AD, Kuempel ED. 2005. Airborne nanostructured
particles and occupational health. J Nanopart Res 7:587-614.
Mitchnick MA, Fairhurst D, Pinnell SR. 1999. Microfine zinc
oxide (z-cote) as a photostable UVA/UVB sunblock agent.
J Am Acad Dermatol 40:85–89.
Monteiller C, Tran L, MacNee W, Faux SP, Jones A, Miller B,
Donaldson K. 2007. The pro-inflammatory effects of low-
toxicity low-solubility particles, nanoparticles and fine particles,
on epithelial cells in vitro: The role of surface area. Occup
Environ Med 64:609–615.
Morrow PE. 1988. Possible mechanisms to explain dust overload-
ing of the lungs. Fund Appl Toxicol 10:369–384.
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
Mortensen LJ, Oberdörster G, Pentland AP, DeLouise LA. 2008.
In vivo skin penetration of quantum dot nanoparticles in the
murine model: The effect of UVR. NanoLetters 8:2779–2787.
Nanoderm. 2007. Quality of skin as a barrier to ultra-fine particles.
Available from the website: www.uni-leipzig.de/~nanoderm/
Downloads/Nanoderm_Final_Summary.pdf.
Nash JF, Tanner PR, Matts PJ. 2006. Ultraviolet A radiation:
Testing and labeling for sunscreen products. Dermatol Clin
24:63–74.
Nemmar A, Hoet PH, Vanquickenborne B, Dinsdale D,
Thomeer M, Hoylaerts MF, Vanbilloen H, Mortelmans L,
Nemery B. 2002. Passage of inhaled particles into blood
For personal use only.
circulation in humans. Circulation 105:411–414.
Nemmar A, Hoylaerts MF, Hoet PHM, Nemery B. 2004. Possible
mechanisms of the cardiovascular effects of inhaled particles:
Systemic translocation and prothrombotic effects. Toxicol Lett
149:243–253.
Nikula KJ, Avila KJ, Griffith WC, Mauderly JL. 1997. Lung tissue
responses and sites of particle retention differ between rats and
cynomolgus monkeys exposed chronically to diesel exhaust and
coal dust. Fundam Appl Toxicol 37:37–53.
Nohynek GJ, Lademann J, Ribaud C, Roberts MS. 2007. Grey goo
on the skin? Nanotechnology, cosmetic and sunscreen safety.
Crit Rev Toxicol 37:251–277.
Nyborg JK, Peersen OB. 2004. That zincing feeling: The effects
of EDTA on the behaviours of zinc-binding transcriptional
regulators. Biochem J 381:e3–4.
Oberdörster G. 2000. Toxicology of ultrafine particles: In vivo
studies. Roy Soc London Phil Tr A 358:2719–2740.
Oberdörster G. 2001. Pulmonary effects of inhaled ultrafine par-
ticles. Int Arch Occup Environ Health 74:1–8.
Oberdörster G, Ferin J, Lehnert BE. 1994. Correlation between
particle size, in vivo particle persistence, and lung injury. Environ
Health Perspect 102:173–179.
Oberdörster G, Sharp Z, Atudorei V, Elder A, Gelein R,
Lunts A, Kreyling W, Cox C. 2002. Extrapulmonary trans-
location of ultrafine carbon particles following whole-body
inhalation exposure of rats. J Toxicol Environ Health A
65:1531–1543.
Otberg N, Richter H, Knuttel A, Schaefer H, Sterry W, Lademann J.
2004. Laser spectroscopic methods for the characterization of
open and closed follicles. Laser Phys Lett 1:46–49.
Park SB, Kang YC. 1997. Photocatalytic activity of nanometer size
ZnO particles prepared by spray pyrolysis. J Aerosol Sci 28:
S473–474.
Analysis of ZnO for sunscreens 39
Paschke MW, Perry LG, Redente EF. 2006. Zinc toxicity thre-
sholds for reclamation forb species. Wat Air Soil Pollut
140:317–330.
Paschke MW, Redente EF, Levy DB. 2000. Zinc toxicity thresholds
for important reclamation grass species of the western United
States. Environ Toxicol Chem 19:2751–2751.
Peck EJ, Ray WJJ. 1971. Metal complexes of phosphoglucomu-
tase in vivo. Alterations induced by insulin. J Biol Chem
246:1160–1167.
Perl AS. 1993. Zinc oxide. Am Ceramic Soc Bull 72:122–123.
Pinnell SR, Fairhurst D, Gillies R, Mitchnick MA, Kollias N. 2000.
Microfine zinc oxide is a superior sunscreen ingredient to
microfine titanium dioxide. Dermatol Surg 26:309–314.
Pirot F, Millet J, Kalia YN, Humbert P. 1996. In vitro study of
percutaneous absorption, cutaneous bioavailability and bio-
equivalence of zinc and copper from five topical formulations.
Skin Pharmacol 9:259–269.
Popov AP, Priezzhev AV, Lademann J, Myllylä R. 2005.
TiO 2 nanoparticles as an effective UV-B radiation skin-
protective compound in sunscreens. J Phys D: Appl Phys
38:2564–2570.
Prasad AS, Bao B, Beck FWJ, Kucuk O, Sarkar FH. 2004.
Antioxidant effect of zinc in humans. Free Radic Biol Med
37:1182–1190.
Pryor JL, Hughes C, Foster W, Hales BF, Robaire B. 2000. Critical
windows of exposure for children’s health: The reproductive
system in animals and humans. Environ Health Perspect
108:491–503.
Rampaul A, Parkin IP, Cramer LP. 2007. Damaging and protective
properties of inorganic components of sunscreens applied to
cultured human skin cells. J Photochem Photobiol A, Chem
191:138–148.
Reddy KM, Feris K, Bell J, Wingett DG, Hanley C, Punnoose A.
2007. Selective toxicity of zinc oxide nanoparticles to
prokaryotic and eukaryotic systems. Appl Phys Lett 90:
213902-1–213902-3.
Renwick LC, Donaldson K, Clouter A. 2001. Impairment of
alveolar macrophage phagocytosis by ultrafine particles. Toxicol
Appl Pharmacol 172:119–127.
Roberts M, Roberts M, Robertson T, Sanchez W, Thorling C,
Zou Y, Zhao X, Becker W, Zvyagin A. 2008. In vitro and in vivo
imaging of xenobiotic transport in human skin and in the rat
liver. Journal of Biophotonics 1:478–493.
Rostan EF, DeBuys HV, Madey DL, Pinnell SR. 2002. Evidence
supporting zinc as an important antioxidant for skin. Int J
Dermatol 41:606–611.
Rothen-Rutischauser B, Schurch S, Haenni B, Kapp N, Gehr P.
2006. Interaction of fine particles and nanoparticles with red
blood cells visualized with advanced microscopic techniques.
Environ Sci Technol 40:4353–4359.
Rouse JG, Yang J, Ryman-Rasmussen JP, Barron AR, Monteiro-
Riviere NA. 2007. Effects of mechanical flexion on the pene-
tration of fullerene amino acid-derivatized peptide nanoparticles
through skin. Nano Lett 7:155–160.
Ryman-Rasmussen JP, Riviere JE, Monteiro-Riviere NA. 2006.
Penetration of intact skin by quantum dots with diverse
physicochemical properties. Toxicol Sci 91:159–165.
Sakurai H, Yasui H, Yamada Y, Nishimura H, Shigemoto M. 2005.
Detection of reactive oxygen species in the skin of live mice and
rats exposed to UVA light: A research review on chemilumines-
cence and trials for UVA protection. Photochem Photobiol Sci
4:715–720.
Sander CS, Chang H, Hamm F, Elsner P, Thiele JJ. 2004. Role of
oxidative stress and the antioxidant network in cutaneous
carcinogenesis. Int J Dermatol 43:326–335.
 40 M.J. Osmond & M.J. McCall
Sato J, Denda M, Chang S, Elias PM, Feingold KR. 2002.
Abrupt decreases in environmental humidity induce abnor-
malities in permeability barrier homeostasis. J Invest Dermatol
119:900–904.
Sawai J. 2003. Quantitative evaluation of antibacterial activities of
metallic oxide powders (ZnO, MgO and CaO) by conductimet-
ric assay. J Microbiol Meth 54:177–182.
Sayes CM, Reed KL, Warheit DB. 2007. Assessing toxicity of fine
and nanoparticles: Comparing in vitro measurements to in vivo
pulmonary toxicity profiles. Toxicol Sci 97:163–180.
Scientific Committee on Consumer Products (SCCP). 2006. Basic
criteria for the in vitro assessment of dermal absorption of
cosmetic ingredients. Scientific Committee on Consumer
Products SCCP/0970/60:
Schlumpf M, Cotton B, Conscience M, Haller V, Steinmann B,
Lichtensteiger W. 2001. In vitro and in vivo estrogenicity of UV
screens. Environ Health Perspect 109:239–244.
Scott RC, Walker M, Dugard PH. 1986. A comparison of the in
vitro permeability properties of human and some laboratory
animal skins. Int J Cos Sci 8:189–194.
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
Selevan SG, Kimmel CA, Mendola P. 2000. Identifying critical
windows of exposure for children’s health. Environ Health
Perspect 108:451–455.
Semmler M, Seitz J, Erbe F, Mayer P, Heyder J, Oberdörster G,
Kreyling WG. 2004. Long-term clearance kinetics of inhaled
ultrafine insoluble iridium particles from the rat lung, including
transient translocation into secondary organs. Inhal Toxicol
16:453–459.
Sensi SL, Yin HZ, Carriedo SG, Rao SS, Weiss JH. 1999. Pref-
erential Zn2+ influx through Ca2+-permeable AMPA/kainate
channels triggers prolonged mitochondrial superoxide produc-
tion. PNAS 96:2414–2419.
For personal use only.
Simons TJB. 1991. Intracellular free zinc and zinc buffering in
human red blood cells. J Memb Biol 123:63–71.
St. Croix CM, Leelavanichul K, Watkins SC, Kagan VE, Pitt BR.
2005. Nitric oxide and zinc homeostasis in acute lung injury.
Proc Am Thorac Soc 2:236–242.
Suzuki T, Chow CW, Downey GP. 2008. Role of innate immune
cells and their products in lung immunopathology. Int J Biochem
Cell Biol 40:1348–1361.
Tabata Y, Ikada Y. 1988. Effect of the size and surface charge of
polymer microspheres on their phagocytosis by macrophage.
Biomaterials 9:356–362.
Takenaka S, Karg E, Roth C, Schulz H, Ziesenis A, Heinzmann U,
Schramel P, Heyder J. 2001. Pulmonary and systemic distribu-
tion of inhaled ultrafine silver particles in rats. Environ Health
Perspect 109:547–551.
Tinkle SS, Antonini JM, Rich BA, Roberts JR, Salmen R,
DePree K, Adkins EJ. 2003. Skin as a route of exposure and
sensitization in chronic beryllium disease. Environ Health
Perspect 111:1202–1208.
Tran CL, Buchanan D, Cullen RT, Searl A, Jones AD,
Donaldson K. 2000. Inhalation of poorly soluble particles. II.
Influence of particle surface area on inflammation and clearance.
Inhal Toxicol 12:1113–1126.
Tsai SJ, Ashter A, Ada E, Mead JL, Barry CF, Ellenbecker MJ.
2008. Airborne nanoparticle release associated with the com-
pounding of nanocomposites using nanoalumina as fillers. Aeros
Air Qual Res 8:160–177.
Tsuchiya T, Oguri I, Yamakoshi YN, Miyata N. 1996. Novel
harmful effects of [60]fullerene on mouse embryos in vitro
and in vivo. FEBS Lett 393:139–145.
Tupler R, Perini G, Green MG. 2001. Expressing the human
genome. Nature 409:832–833.
van Ravenzwaay B, Leibold E. 2004. The significance of in vitro rat
skin absorption studies to human risk assessment. Toxicol in
Vitro 18:219–225.
Viswanathan R, Gupta RB. 2003. Formation of zinc oxide nano-
particles in supercritical water. J Supercrit Fluids 27:187–193.
Vogt A, Combadiere B, Hadam S, Stieler KM, Lademann
J, Schaefer H, Autran B, Sterry W, Blume-Peytavi U. 2006.
40 nm, but not 750 or 1,500 nm, nanoparticles enter epidermal
CD1a+ cells after transcutaneous application on human skin.
J Invest Dermatol 126:1316–1322.
Vogt A, Mandt N, Lademann J, Schaefer H, Blume-Peytavi U.
2005. Follicular targeting – a promising tool in selective derma-
totherapy. J Invest Dermatol Symp Proc 10:252–255.
Volkheimer G. 1974. Passage of particles through the wall of the
gastrointestinal tract. Environ Health Perspect 9:215–225.
Wagner H, Kostka K-H, Lehr C-M, Schaefer UF. 2003. pH
profiles in human skin: Influence of two in vitro test
systems for drug delivery testing. Eur J Pharm Biopharm
55:57–65.
Wahie S, Lloyd JJ, Farr PM. 2007. Sunscreen ingredients and
labelling: A survey of products available in the UK. Clin Exp
Dermatol 32:359–364.
Waller JM, Maibach HI. 2005. Age and skin structure and
function, a quantitative approach (I): Blood flow, pH, thick-
ness, and ultrasound echogenicity. Skin Res Technol
11:221–235.
Wang B, Feng W, Wang M, Wang T, Gu Y, Zhu M, Ouyang H,
Shi J, Zhang F, Zhao Y, et al. 2008. Acute toxicological impact of
nano- and submicro-scaled zinc oxide powder on healthy adult
mice. J Nanopart Res 10:263–276.
Wang B, Feng WY, Wang TC, Jia G, Wang M, Shi JW, Zhang F,
Zhao YL, Chai ZF. 2006. Acute toxicity of nano- and micro-
scale zinc powder in healthy adult mice. Toxicol Lett
161:115–123.
Wang H, Xie C, Zhang W, Cai S, Yang Z, Gui Y. 2007. Com-
parison of dye degradation efficiency using ZnO powders with
various size scales. J Haz Mat 141:645–652.
Wesselkamper SC, Chen LC, Gordon T. 2001a. Development of
pulmonary tolerance in mice exposed to zinc oxide fumes.
Toxicol Sci 60:144–151.
Wesselkamper SC, Chen LC, Kleeberger SR, Gordon T. 2001b.
Genetic variability in the development of pulmonary tolerance to
inhaled pollutants in inbred mice. Am J Physiol Lung Cell Mol
Physiol 281:L1200–1209.
Wiebert P, Sanchez-Crespo A, Falk R, Philipson K, Lundin A,
Larsson S, Moller W, Kreyling W, Svartengren M. 2006a. No
significant translocation of inhaled 35-nm carbon particles to the
circulation in humans. Inhal Toxicol 18:741–747.
Wiebert P, Sanchez-Crespo A, Seitz J, Falk R, Philipson K,
Kreyling W, Moller W, Sommerer K, Larsson S,
Svartengren M. 2006b. Negligible clearance of ultrafine particles
retained in healthy and affected human lungs. Eur Respir J
28:286–290.
Wiseman DA, Wells SM, Wilham J, Hubbard M, Welker JE,
Black SM. 2006. Endothelial response to stress from exogenous
Zn2+ resembles that of NO-mediated nitrosative stress, and is
protected by MT-1 overexpression. Am J Physiol Cell Physiol
291:555–568.
Wolf R, Wolf D, Morganti P, Ruocco V. 2001. Sunscreens. Clin
Dermatol 19:452–459.
Wu J, Liu W, Xue C, Znou S, Lan F, Bi L, Xu H, Yang X,
Zeng F-D. 2009. Toxicity and penetration of TiO2 nano-
particles in hairless mice and porcine skin after subchronic
dermal exposure. Toxicol Lett 191:1–8.

Yamamoto O. 2001. Influence of particle size on the antibacterial
activity of zinc oxide. Int J Inorgan Mat 3:643–646.
Yamamoto O, Komatsu M, Sawai J, Nakagawa H. 2004. Effect of
lattice constant of zinc oxide on antibacterial characteristics.
J Mater Sci Mater Med 15:847–851.
Yang H, Liu C, Yang D, Zhang H, Xi Z. 2008. Comparative study
of cytotoxicity, oxidative stress and genotoxicity induced by four
typical nanomaterials: The role of particle size, shape and
composition. J Appl Toxicol 29:69–78.
Yang Z, Xie C. 2006. Zn2+ release from zinc and zinc oxide
particles in simulated uterine solution. Colloids Surf
B Biointerfaces 47:140–145.
Nanotoxicology Downloaded from informahealthcare.com by University of Toronto on 02/19/13
For personal use only.
Analysis of ZnO for sunscreens 41
Zhai H, Maibach HI. 2001. Effects of skin occlusion on percuta-
neous absorption: An overview. Skin Pharmacol Physiol 14:1–10.
Zhu X, Wang J, Zhang X, Chang Y, Chen Y. 2009. The impact of
ZnO nanoparticle aggregates on the embryonic development of
zebrafish (Danio rerio). Nanotechnology 20:195103.
Zhu X, Zhu L, Duan Z, Qi R, Li Y, Lang Y. 2008. Comparative
toxicity of several metal oxide nanoparticle aqueous suspensions
to zebrafish (Danio rerio) early developmental stage. J Environ Sci
Health A Tox Hazard Subst Environ Eng 43:278–284.
Zvyagin A, Zhao X, Gierden A, Sanchez W, Ross J, Roberts M.
2008. Imaging of zinc oxide nanoparticle penetration in human
skin in vitro and in vivo. Journal of Biomedical Optics 13:064031.