See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/216213018

Toxicology of Nanoparticles: A Historical Perspective

Article in Nanotoxicology · March 2007

DOI: 10.1080/17435390701314761

CITATIONS READS
894 3,909

3 authors, including:

Vicki Stone Ken Donaldson

Heriot-Watt University The University of Edinburgh
226 PUBLICATIONS 27,927 CITATIONS 369 PUBLICATIONS 45,350 CITATIONS

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

REFNANO View project

Mechanism of cell death (apoptosis) in mesothelioma cells View project

All content following this page was uploaded by Vicki Stone on 10 September 2014.

The user has requested enhancement of the downloaded file.

Nanotoxicology, March 2007; 1(1): 2
25

Toxicology of nanoparticles: A historical perspective

GÜNTER OBERDÖRSTER1, VICKI STONE2, & KEN DONALDSON3

1
University of Rochester, School of Medicine & Dentistry, Department of Environmental Medicine, Rochester, New York,
USA, 2Napier University, School of Life Sciences, Biomedicine Research Group, Edinburgh, and 3MRC/University of
Edinburgh, Centre for Inflammation Research, Edinburgh, UK

Abstract
The rapid expansion of nanotechnology promises to have great benefits for society, yet there is increasing concern that
human and environmental exposure to engineered nanomaterials may result in significant adverse effects. That is why the
field of nanotoxicology
dealing with effects and potential risks of particulate structures B100 nm in size
has
emerged, growing significantly over the past decade from long-standing foundations of well established knowledge on the
toxicology of fibrous and non-fibrous particles and the interactions of viruses with cells. This review places nanoparticles
in the context of conventional particle toxicology and so includes references to other types of particles, such as silica and
asbestos, which have been extensively studied and can provide useful lessons relevant to newly engineered nanoparticles
(NP). Discoveries of nanoparticle-specific concepts of toxicology related to their small size and large specific surface area
go back to the early parts of the past century, although the distinctive biological effects and kinetics of NP were not
recognized until the last decade of the past century. Today, the propensity of NP to cross cell barriers, enter cells and
interact with subcellular structures is well established, as is the induction of oxidative stress as a major mechanism of
nanoparticle effects. In addition to the significance of small size and surface area of NP, uncovering the impact of many
other physico-chemical characteristics
in particular NP surface properties
for initiating effects in the mammalian
organism and the environment is now an active area of research. The article aims to cover hazards relevant to humans,
provides an introduction to some of the newly emerging literature on fate and behavior of NP in the environment, as well
as describing their ecotoxicology in a variety of species. Major milestones in the research leading to our present
understanding of nanotoxicology and the potential risks of NP to humans and the environment are summarized. These
risks are likely to be different for different nanomaterials, ranging from perceived and very low for most, to real and very
high for some. There are many questions that remain to be addressed, and we foresee for the future a continuing
extended research in nanotoxicology. A full understanding of the hazard of NP will make a major contribution to the risk
assessment that is so urgently needed to ensure that products that utilize NP are made safely, are exploited to their full
potential and then disposed of safely.

Keywords: Nanotoxicology, nanoparticle, oxidative stress, fullerene, human and environmental health

Introduction lation, such as those with a pre-existing or genetic

Continuing advances in nanotechnology promise to
be of great benefit for many applications, including
their exploitation within structural engineering,
electronics, optics, consumer products, alternative
energy, soil and water remediation, or for medicinal
uses as therapeutic, diagnostic or drug delivery
devices. Advances in nanotechnology in all of these
areas are likely to significantly benefit society and the
economy; this is reflected in the global multi-billion
dollar investment in nanotechnology. Despite bright
outlooks for the future of nanotechnology, there is
increasing concern that human exposure to some
types of engineered nanoparticles, inadvertently or
intentionally, may lead to significant adverse health
effects. In particular, susceptible parts of the popu-
disease, may be more affected. This reflects a lesson
learned from epidemiological studies on the effects
of ambient air particulate matter, where ultrafine-
particles are considered to play a role in mediating
the adverse effects in patients with cardiovascular
and respiratory diseases (EPA 2004). In addition,
there is concern of environmental contamination
and associated effects on ecosystems, which could
have significant societal implications. Realizing these
consumer fears and perceived risks, and in order to
prevent unforeseen toxic disasters, concerned reg-
ulatory agencies, funding agencies and industries
world-wide have begun to provide funds and invite
applications for evaluating toxicological profiles of
engineered nanoparticles. These particles range in
size from 1
100 nm, the same sizes covered by

Correspondence: Günter Oberdörster, University of Rochester, Dept. of Environmental Medicine, Medical Center Box 850, 575 Elmwood
Avenue, Rochester, NY 14642 USA. Tel: 1 585 275 3804. Fax: 1 585 256 2631. E-mail: gunter_oberdorster@urmc.rochester.edu

ISSN 1743-5390 print/ISSN 1743-5404 online # 2007 Informa UK Ltd.

DOI: 10.1080/17435390701314761

ambient particles which have been termed ‘ultrafine’
particles by toxicologists. The sources of these are
many-fold, both natural and anthropogenic, as
shown by some examples listed in Table I, together
with engineered nanoparticles. Ultrafine particles
and engineered nanoparticles have several physical
characteristics in common, but there are also sig-
nificant differences, yet properties of high impor-
tance for toxicological effects are common to both
(Figure 1). Both consist of primary particles below
100 nm in size, which provides a high surface area
per unit mass and potentially high surface reactivity.
Both tend to form agglomerates when suspended in
gases or liquids, dependent on concentration and the
chemistry of particle surface and the suspension
medium. Most of the ambient ultrafine particles are
generated as heterogeneous aggregates of primary
particles (e.g., combustion soot particles), whereas
most of the engineered nanoparticles are generated
as more monodispersed individual particles (e.g.,
quantum dots). Determination of the state of
agglomeration and aggregation is an essential part
of nanoparticle characterization (Figure 1). Knowl-
edge gained from toxicological studies with ultrafine
particles represents a valuable database for the
toxicology of engineered nanoparticles. Research in
this field is expanding rapidly, as evidenced by a
search of website accessible publications for ‘toxicity
or health effects’ combined with ‘ultrafine or nano-
particles’. An exponential increase over the past 5
years becomes apparent. (Figure 2) This article gives
an overview about some key historical developments
in biomedical/toxicological research leading to the
recognition of the special biological properties of
nano-sized particles, and to today’s focused research
to assess the potential risks and safety of nanomater-
ials. Specific mechanisms underlying ultrafine/nano-
particle effects will be mentioned but not discussed
in detail.
Table I. Ultrafine/nanoparticles (B100 nm): Natural and Anthropogenic Sources.
Anthropogenic
Natural Unintentional
gas to particle conversions internal combustion engines
forest fires power plants
volcanoes (hot lava) incinerators
viruses airplane jets
biogenic magnetite: metal fumes
magnetotactic bacteria (smelting, welding, etc. )
protoctists, mollusks polymer fumes
arthropods , fish, birds other fumes
human brain, meteorite? heated surfaces
ferritin (12.5 nm) frying, broiling, grilling
microparticles (B100 nm) electric motors
(activated cells)
Toxicology of nanoparticles: A historical perspective 3
Definition of terms:
The terms ‘ultrafine’ and ‘nano’ particles are differ-
ently defined with regard to their size: For toxicol-
ogists and material scientists, both are particles
B100 nm, although there are obvious differences
between them in that engineered nanomaterials
provide novel phenomena, properties and functions
at the nanoscale enabling us to measure, control and
manipulate matter in order to change those proper-
ties and functions (Roco 2003a, 2003b). For atmo-
spheric scientists, the nucleation mode (B10 nm) of
the atmospheric multimodal size distribution is often
referred to as nanoparticle. On the other hand,
combustion engineers and pharmacists define all
particles below ‘micro’ (1000 nm 1 mm) as nano-
particles. In addition the special role of combustion
in producing harmful carbon-centred nanoparticles
(Avakian et al. 2002) typified by diesel exhaust
particles (DEP) has produced a category of combus-
tion-derived nanoparticles that represents the ultra-
fine component derived from combustion and also
other sources of combustion derived particles of
small size (Donaldson et al. 2005). This may lead to
some confusion when searching for published work
on ‘nanoparticles’, in particular when information
on size characteristics is not always given in a
publication. Because of this difficulty in determining
the particle size used in many pharmaceutical
studies, no attempts are made in this review to
evaluate such particles in this paper. For the purpose
of this article, the toxicological/nanomaterials defini-
tion of particle sizes B100 nm will be used.
Particle toxicology and the roots of
nanotoxicology
The roots of the current science of nanotoxicology
can be traced to a variety of origins (Donaldson et al.
2004; Oberdörster et al. 2005). Different strands of
Intentional
engineered nanoparticles
(controlled size and shape,
designed for functionality)
metals, semiconductors, metal oxides
carbon, polymers
nano-spheres, -wires,
-needles, -tubes, -shells,
-rings, -platelets;
untreated, coated
(nanotechnology applied to many
products: cosmetics, medical, fabrics,
electronics, optics, displays, etc .)
4 G. Oberdörster et al.

Primary Particle

Agglomerated Primary Particles (Agglomerates)

(Engineered Nanoparticles)

Aggregated Primary Particles (Aggregates)

(Ambient ultrafine particles)

Agglomerated Aggregates

Figure 1. ‘Primary nanoparticles can agglomerate (coagulate) to form clusters, but may also aggregate (fuse; sinter) during generation and
then in turn can agglomerate additionally. Some mass-produced engineered nanoparticles (e.g., carbon black; TiO2) are generated partly as
aggregates and subsequently agglomerate (Bansal et al. 1993; Beaucage et al. 1999; IARC 2006).

understanding have come together, from the accu- would be unthinkable without this history and it is
mulation of knowledge during the development of within this historical context that we place nanotox-
modern particle toxicology, that enabled the current icology in this article.
paradigm to be developed. Thus the behaviour of
nanoparticles can be understood within a perspec-
Virology as precursor
tive that has embraced the toxicology of metal
fumes, radionuclides, nuisance dusts, rat lung over- In some sense, nanoparticles may resemble viruses
load, the toxicology of silica, asbestos and synthetic (18
500 nm structures), not only in size but also with
vitreous fibers and more recently from the toxicology respect to interactions with cells (Oberdörster et al.
of air pollution particles (PM10 particles below 2005). A most recent article by Douglas and Young
10 mm). In addition virology and other sciences have (2006) discusses in detail ‘a new expanded role [of
contributed to our understanding (Figure 3). These viruses] as nanoplatforms with applications in mate-
are alluded to in various sections of this document. rials science and medicine’. Indeed, accumulated
The current paradigm of nanoparticle toxicology knowledge gleaned from virology research over the

200
Number of Citations

150

100

50

0
1990 1995 2000 2005

Greater UFP surface Increasing awareness of Q-dot and

UFP O x i d a t iv e
inflammatory area is related potential for nanoparticles fullerene
hypothesis stress as
potential and to toxicity; to induce adverse effects in penetration
for Effects of major
i nt e rs t i ti a l greater o f s k in t o
air pollution mechanism of humans and environment;
translocation of t ox ic i ty o f epidermis
ultrafine engineered particles UFP effects pulmonary toxicity of
carbon NT; mitochondrial and dermis
particles (UFP) nanoparticles
(NP) and nuclear acc es s of NP

Figure 2. PubMed January 2007: Number of publications/year searching for ‘ultrafine’ or ‘nanoparticles’ and ‘toxicity’ or ‘health effects’.

Fibre Toxicology
T he role of par ticle s hape
an d b i o p e r s is t e nc e
Silica
T he ro le of pa r ticle s urface
reactivit y and its capacity
for modification
NAN OT OX IC OL OG Y
Metal Fume Fever
The role of small size and
systemic effects ( fever)
V i r o lo g y
The role of viruses as models for interactions of
nanostructures with cells and as
nanoplatforms in medicine
Figure 3. Foundations of Nanotoxicology.
last 100 years might be considered a valuable basis for
today’s research on the interactions of nanoparticles
with cells and the living organism. Beijerinck in 1898
suggested a ‘contagium vivum fluidum ’ (virus) as cause
of the tobacco mosaic (TM) disease in plants. Only
with the invention of the electron microscope in the
1930s was it possible to visualize viruses as nano-sized
structures for the first time (Kausche et al. 1939;
Ruska et al. 1940). Stanley (1935) and Bawden and
Pirie (1937) characterized the TM virus as a physi-
cochemical entity (crystalline protein nucleic acid
complex), and Bodian and Howe (1941) described a
nose to brain translocation of 30 nm poliovirus via
the olfactory neuron in chimpanzees. Eventually, in
the 1950s and 1960s with the advance of transmission
electron microscopy (TEM) and X-ray diffraction, it
was possible to determine the fine structure of
viruses. Since then, there is accumulating informa-
tion on viral mechanisms for entering cells and their
interactions with subcellular structures, as described
recently in an excellent review by Smith and Helenius
(2004), including viral entry into cells via clathrin-
mediated or caveolae-mediated pathways, their in-
tracellular endosomal and caveosomal movement
and entry into the cell-nucleus via 39 nm nuclear
pore complex (Panté & Kann 2002). Indeed, viral
capsids are considered a valuable tool or carrier
(Trojan horses) for targeted delivery of drug mole-
cules for pharmaceutical and materials applications
(Douglas & Young 2006), and the propensity of
engineered nanoparticles to endocytose and transcy-
tose appears to be due to much the same mechanisms.
Pre-1990
With regard to non-viral nano-sized particles, the
realization that the very small size of particles can be
associated with distinct and special biological effects
Toxicology of nanoparticles: A historical perspective 5
Particle Overload
T he r o l e o f d o s e a nd
dosemetric
Radionuclide
Toxicology
T h e role o f par tic le
chemistry and biokinetics
PM10
The role of small size and
systemic effects
( c ar d i o v a s c u l a r )
Deposition Studies and Models
The role of particle size, airway geometry
and breathing parameters for
d epo sition in respira to ry tract
did not emerge until later in the 20th century, in
1990. Before that, the pioneering work of Drinker et
al. (1927a, 1927b) has shown induction of acute
respiratory distress (metal [zinc] fume fever), during
occupational exposures of workers in zinc smelting
operations. At this time it was unknown that metal
fumes consist of ultrafine particles, which can
rapidly agglomerate to form larger particles. The
highly acute inflammatory potential in the respira-
tory tract of another ultrafine particle, fumed silica,
was reported by Gardner in 1938 and later studied
in more detail in experimental animals by Jötten and
Klosterkötter (1952). Again, the ultrafine size of the
particles was not considered as an important attri-
bute that might contribute to their effect; even the
1966 International Commission on Radiological
Protection (ICRP) lung particle deposition model
considered the mechanism of diffusional deposition
only down to particle sizes of 0.1 mm. Anthropogenic
athmospheric nuclei mode particles (ultrafine parti-
cles) from emissions of internal combustion engines
have been described and characterized by Whitby
et al. (1975), thereby raising attention of atmo-
spheric scientists and aerosol physicists to this class
of particles. Subsequently, the 1994 ICRP model
included detailed information about ultrafine parti-
cle deposition in all regions of the respiratory tract
(Figure 4). This was based in large part on the
ground-breaking experimental work in humans on
deposition of inhaled ultrafine particles in the lower
respiratory tract by Willie Stahlhofen’s group in
Germany (Schiller et al. 1986) and David Swift’s
studies in the human upper respiratory tract in the
U.S. (Swift et al. 1992, 1994). These deposition
studies coincided with the realization that ultrafine/
nano-sized particles have unique toxicological prop-
erties as described later in this paper. For respiratory
toxicologists, these deposition studies are of great
6 G. Oberdörster et al.
1 .0
0.8
Deposition Fraction
0.6
0.4
0.2
0.0
0.0001
Mechanisms
Deposition
Figure 4. The fractional deposition and mechanism of deposition of inhaled particles of different sizes in each region of the human
respiratory tract based on the ICRP Model (1994) assuming nose breathing.
importance, showing that deposition of inhaled
ultrafine particles (B100 nm) is apparent in all
regions of the respiratory tract, but that it is size-
dependent: Very high in the upper respiratory tract
for sizes below 10 nm, high also in the tracheobron-
chial region for 5
10 nm particles, and for 20 nm
particles in the alveolar region (Figure 3).
In addition to total regional deposition, site-
specific dosimetry, as it applies to particles, can be
traced to the studies of Brody et al. (1984, 1985) and
Warheit et al. (1984) who emphasized the extent of
deposition of asbestos fibers of small aerodynamic
diameter in the centriacinar region (terminal airways
and proximal alveoli). This focused attention on the
characteristics of this highly fragile part of the lungs
and the relative importance of the slow clearance
mechanism (phagocytic cells of the immune system)
in this region compared to that of the mucociliary
escalator of the larger airways, in contributing to the
accumulation of local dose. The high dose encoun-
tered in the centriacinar region is well documented
in pathology studies showing black dust accumula-
tion in the terminal and respiratory bronchiolar walls
of coal miners at autopsy (Churg & Green 1999).
With regard to the recognition of the toxicological
potential of ultrafine particles, studies on cell uptake
and translocation are of prime interest since this is a
most distinctive difference of these particles com-
pared to larger particles. Although research by
Bodian and Howe (1941) focused on neuronal
pathways of infection of poliovirus (as it turned out
having a size of a 30 nm particle) and discovered the
olfactory translocation route for this virus from the
nose to the brain in chimpanzees, they could not, of
course, foresee the potential importance it would
T O T AL
Nasal Pharyngeal
Laryngeal
Alveolar
Tracheobronchial
0 .001 0.01 0.1 1 10 100
Diameter (mm)
I m p a ct ion
Diffu sion
Sed im en t a t ion
have six decades later for solid nanoparticles. Even
the documentation 30 years later by DeLorenzo
(1970) of 50 nm silver-coated gold particles moving
from the nasal olfactory mucosa via the olfactory
nerve to the olfactory bulb in squirrel monkeys did
not find wide recognition by toxicologists. This
finding was not recognized as being of significance
for in vivo disposition of inhaled nano-sized parti-
cles, and instead being considered as an experimen-
tal tool in using nanogold to visualize cellular
structures. Similarly, Katz et al. (1984) and Adams
and Bray (1983) used latex and polystyrene particles
(20
500 nm) to demonstrate retrograde and ante-
rograde movement of these particles in the axons of
neurons. Czerniawska (1970) also used nanogold
particles to demonstrate perineuronal translocation
from submucosal injection sites of the olfactory
mucosa in rabbits to the cerebrospinal fluid space
bathing the brain. Again, no indication of its
potential toxicological significance was given.
In 1971, however, Berry et al. suggested that their
observation of 30 nm intratracheally-instilled gold
particles crossing the alveolar-capillary barrier and
accumulating in blood platelets in rats should be of
relevance for transporting the smallest air pollutant
particles, such as tobacco smoke, to distant organs.
They concluded that this ‘might predispose to
platelet aggregation with formation of microthrombi
atheromatous plaques,’ a hypothesis that has been
posited lately for inhaled ultrafine particles and is
now tested experimentally in the field of health
effects of particulate air pollution as part of a larger
mechanistic scheme (Figure 5). Since then, a
number of investigators in the post-1990s area
(see below) have shown in experimental animals
 Toxicology of nanoparticles: A historical perspective 7

Particle translocation
M ed i a t o r s N P A e ro s o l

Inhalation

Central nervous Neuronal Re s p i ra t o ry t r ac t Clearance Ga s t r o in t e s t i n a l

system transport deposition tr act
Translocation via
in t ers t itiu m
Lung Blood
i n fl a m ma t i on Circulation *
Humoral
mediators

Autonomic nervous system Blood vessel dysfunction E xt ra pu l mo n ar y

c o n tr o l o f h e a rt r a te & systemic inflammation organs
variability and CV function (e. g. heart and liver)

Cardiovascular Health Effects

(e.g. thrombus formation; MI)

Figure 5. The potential consequences of exposure to NP via inhalation. Modifying factors may include age, gender, disease state, genetics
and exposure to co-pollutants. *Note that once NP gain access to the CV system by any route of entry, the consequences are likely to be
comparable.

that, indeed, translocation of nanoparticles across activities of very small amounts of translocated
the alveolar capillary barrier can occur in both particles.
directions, although it is dependent on size and In the 1980s, rodent inhalation studies with
surface chemistry of the particles (Table II). Trans- ultrafine zinc-oxide particles by Mary Amdur’s
location rates, though, appear to be low. Recent group were performed. Characterization of the
human studies performed with inhaled 99mTc-la- particle size by TEM showed the primary particle
beled ultrafine ( 100 nm) particles and detection size to be between 40 and 50 nm, and the agglom-
by g-camera imaging are more controversial, show- erated particles induced significant pulmonary in-
ing either significant translocation to blood and liver flammatory responses with acute onset at exposures
(Nemmar et al. 2002) or none (Brown et al. 2002; in the mg/m3 range (Amdur et al. 1982; McCarthy
Mills et al. 2006); this discrepancy is very likely due et al. 1982). Although these were the first toxicolo-
to uncertainties of the stability of the radioactive gical inhalation studies using size characterized
label in vivo caused by variation in the oxidation of ultrafine particles, a lack of a comparison to larger
the Technetium during operation of the Technegas particles did not allow conclusions to be drawn
generator
a sealed system (the carbon particles are about the importance of particle size for eliciting an
not detected, only the label can be traced)
and by adverse response in a target organ. Later, additional
the limits of g-camera to detect the low radio- animal and clinical human exposure studies with

Table II. Particle size and surface chemistry-related alveolar-capillary translocation.

Particle size (nm) Type Translocation Localization/Effect Reference

5
20 nm Gold, albumin coated yes via caveolae Mehta et al. , 2004
8 nm Gold, albumin coated yes via ‘‘vesicles’’ Konig et al., 1993
8 nm Gold, albumin coated yes via caveolae Heckel et al ., 2004
18 nm Iridium yesa Extrapulm. org. Kreyling et al , 2003
30 nm Gold yes Platelet? Berry et al., 1977
35 nm Carbon yes Liver Oberdörster et al ., 2002
60 nm Polystyrene, pos charge ?b Thrombus, early Nemmar et al., 2002
Silva et al ., 2005
60 nm Polystyrene, neg charge ?b No thrombus Nemmar et al., 2002
80 nm Iridium yesa Extrapulm. org. Kreyling et al ., 2002
240 nm Polystyrene, lecithin yes Monocyte Kato et al., 2003
240 nm Polystyrene, uncoated no Kato et al ., 2003
400 nm Polystyrene no Nemmar et al ., 2004
a
minimal; bindirect evidence
Surface coating (chemistry) charge, size govern translocation.
8 G. Oberdörster et al.
these zinc-oxide particles were performed by this
group, resulting in acute pulmonary and systemic
symptoms of zinc fume fever (Gordon et al. 1992).
The fever-inducing effects of the inhaled ZnO
nanoparticles may indicate the potential of these
small particles to produce extra-pulmonary/systemic
effects because the ultrafine particles themselves
may have translocated across the alveolo-capillary
barrier to other sites in the body as mentioned
above. However, given that ZnO fume particles have
a very short retention halftime in the lung on the
order of 6
10 hours (Oberdörster et al. 1979;
Teague & Raabe 1980; Gordon et al. 1992) due to
dissolution in the acidic milieu of the phagolysosome
in alveolar macrophages, a direct systemic ZnO
particle effect should not be assumed. Other possi-
bilities exist, such as the role of soluble components
of the particles (in this case ionic zinc) becoming
blood-borne and affecting distant sites as well as
effects of inflammatory mediators from the lung
passing systemically to other sites. In the latter
regard temperature homeostasis is controlled by
the brain and involves a number of pyrogenic
cytokines such as interleukin-1 (IL-1), tumor ne-
crosis factor (TNF), interleukin-6 (IL-6). Thus, an
obvious concept emerging from these studies is that
local conditions at the portal-of-entry have to be
carefully evaluated with regard to their impact on
particle biopersistence and subsequent fate as nano-
particle or as dissolved constituent. For example, a
low biopersistence of nano Zn-oxide and nano Mn-
oxide in the alveolar region (alveolar macrophages)
of the lung can be contrasted with a neutral pH in
the upper respiratory tract (Lundbong et al. 1985;
Johnson 1994).


1990 2000
Identifying nanoparticles’ toxic potential
As mentioned in the previous section, despite several
early experimental studies with solid nano-sized
(ultrafine) particles which showed their transloca-
tion beyond the respiratory tract and also demon-
strated inflammatory pulmonary responses, it was
not appreciated that these effects were especially
strong for ultrafine particles. A first side-by-side
comparison of the pulmonary inflammatory effect
and retention of ultrafine and fine particles of TiO2
(20 nm and 250 nm) and of Al2O3 (20 nm and
500 nm) was reported in two companion publica-
tions by Ferin et al. (1990) and Oberdörster et al.
(1990). They observed a significantly greater pul-
monary inflammation and interstitial translocation
from the same mass of ultrafine than fine particles
when intratracheally instilled in rats. Based on these
results the authors suggested an increased pulmon-
ary toxicity of ultrafine particles and concluded that
‘particles of 0.02
0.03 mm in diameter penetrate
more easily than particles of ca . 0.2
0.5 mm’ and
that ‘dusts can be highly toxic to the lung if they
consist of ultrafine particles and are persistent in the
lung’ so that ‘occupational exposure standards for
‘nuisance’ dusts should be reconsidered to take into
account such possibly damaging effect of ultrafine
particles.’
Based on results of additional studies with carbon
and TiO2 particles of different sizes, this same group
reported two years later their findings that the
elicited pulmonary response ‘correlated best with
the surface area of the retained particles rather than
their mass, volume, or numbers’, and concluded that
‘the increased pulmonary toxicity of ultrafine parti-
cles is related to their larger surface area and to their
increased interstitial access’ (Oberdörster et al.
1992). They also pointed out the importance of
the particle surface area concept for ‘so-called
nanophase materials which include powders of
metals, ceramics and ceramic-based composites
being developed for new uses in the electronics
industry consisting of particles in the ultrafine size
range’ and which are ‘designed on an atomic scale,
i.e., relating atomic configuration to macroscopic
properties’ so that ‘proportionally more atoms or
molecules are located at particle surfaces’. They
concluded that ‘toxic responses to new technology
metal compounds may not be extrapolated from
known metal toxicology’ (Oberdörster & Ferin
1992).
Oxidative stress as a mechanism of action of
nanoparticles at the cellular level
As described before, the occurrence and physio-
chemical characteristics of ambient airborne ultra-
fine particles (nuclei mode) has been reported in the
1970s by Whitby et al. (1975). With this knowledge
and with increasing awareness about an association
between health effects (increased hospital admis-
sions and acute mortality due to respiratory and
cardiovascular conditions) and traffic-related parti-
culate air pollution (EPA 2004) coupled with the
above-mentioned and new experimental findings of
a greater effect of ultrafine particles, an ultrafine
particle hypothesis of air pollution particles was
formulated in the mid 1990s by Oberdörster et al.
(1995): They suggested that ‘ultrafine particles
always present in the urban atmosphere . . . play a
role in causing acute lung injury in sensitive parts of
the population’. The new studies contributing to this
hypothesis were carried out with airborne ultrafine
particles (CMD 20 nm) generated by heating

PTFE (polytetrafluoroethylene) which caused 100%
mortality in rats when inhaled for 15 min at a mass
concentration of 50 mg/m3. Concurrently, Seaton et
al. (1995) proposed a mechanistic hypothesis stating
that ‘ultrafine particles are able to provoke alveolar
inflammation, with release of mediators capable, in
susceptible individuals, of causing exacerbations of
lung disease and of increasing blood coagulability,
thus also explaining the observed increases in
cardiovascular deaths associated with urban pollu-
tion episodes.’
In the late 80s and early 90s there was a surge in
mechanistic particle research aimed at asbestos and
the synthetic vitreous fibers as substitutes. Oxidative
stress by asbestos fibers was advanced as a factor in
their ability to cause cell damage and disease as early
as 1975 (Gabor & Anca 1975). However Mossman
and Landesman (1983) and Mossman et al. (1986)
refined the oxidative stress hypothesis for asbestos
and placed it in a pathogenesis paradigm that still
prevails today and then demonstrated the role of
anti-oxidants in ameliorating asbestos effects (Moss-
man et al. 1990). Others demonstrated the ability of
anti-oxidants like glutathione to ameliorate the
harmful effects of asbestos (Quinlan et al. 1994;
Faux et al. 1994; Donaldson & Golyasnya 1995). In
mainstream molecular medicine, the link between
oxidative stress and inflammation and the role of the
oxidative stress-responsive pro-inflammatory sys-
tems, including transcription factors such as AP-1
and NF-kB grew alongside these developments
(Schreck et al. 1992; Meyer et al. 1993) and were
rapidly incorporated into a central role for oxidative
stress-driven inflammation in the pathogenesis of
asbestosis (Janssen et al. 1995; Gilmour et al. 1997).
In the meantime, oxidative stress had been identified
as a likely common pathway for the adverse effects of
other harmful particles including crystalline silica
(Kennedy et al. 1989) and coal mine dusts (Dalal et
al. 1991; Vallyathan et al. 1993).
The generalization of the ‘oxidative stress hypoth-
esis’ to nanoparticles occurred in the mid-90s when
the Edinburgh group was describing the oxidative
potential of PM10 (Gilmour et al. 1996) and had
evoked the ultrafine component as a key component
of PM10 in driving the adverse cardiovascular effects
of PM10 (Seaton et al. 1995). Using the ultrafine and
fine TiO2 used by Oberdörster and Ferin in their
earlier studies, Donaldson et al. demonstrated that
the ultrafine TiO2 had greater oxidative activity than
the larger but respirable TiO2 (Donaldson et al.
1996). The Edinburgh group went on to demon-
strate increased oxidative stressing activity of a
number of ultrafine/nanoparticles including carbon
black (Stone et al. 1998), polystyrene beads (Brown
et al. 2001) and metals (Zhang et al. 1998). This
Toxicology of nanoparticles: A historical perspective 9
group suggested a number of oxidative stress-indu-
cing mechanisms when nanoparticles interact with
cells (Figure 6).
By 1998 Donaldson et al. performed studies on
the pulmonary toxicity of ultrafine and fine particles
and suggested particle surface area as a dose metric
to assess pulmonary effects, and they in addition
pointed out that surface area ‘provides the opportu-
nity for surface chemistry to have a profound effect,
particularly free radicals or transition metal
mediated Fenton chemistry, leading to the genera-
tion of oxidants’. Thus, a number of particle
parameters such as size, surface area and surface
chemistry, especially oxidative stressing activity,
were identified in the 1990s as key dosemetric
parameters for nanoparticle toxicology. Today, the
importance of these, as well as additional particle
parameters (e.g., charge, crystallinity, coating and
biopersistence), has been well accepted as important
determinants of nanoparticle toxicity. Knowledge
about only one or two characteristics of nanoparti-
cles (size alone; surface reactivity or surface area
alone) is not sufficient to interpret their biological/
toxicological effects, so that the multitude and
interplay of all has to be considered (Oberdörster
et al. 2005; Warheit et al. 2006).
Since the initial hypotheses by Oberdörster et al.
(1995) and Seaton et al. (1995), epidemiological
studies have shown an association between number
concentrations of ambient particles (reflecting ultra-
fine particles) and cardiac and vascular health effects
in people with compromised cardiovascular systems
(Peters et al. 2004, Rückerl et al. 2006). Details of
the mechanisms underlying the Seaton hypothesis
for ultrafine and fine particle effects with respect to
induction of acute phase responses and direct access
of particles from the lung via translocation to the
blood circulation (Figure 5) are presently studied by
several groups.
Immunogenicity of nanoparticles
An important finding of immune-modulatory effects
of engineered nanoparticles in the late 1990s was
first reported by Erlanger’s group at Columbia
University. This group showed that C60 fullerenes
have antigenic properties when functionalized with
serum albumin or derivatives of lysine, thereby
acting as a hapten which can induce antibodies after
i.p. injection in mice (Chen et al. 1998). They
showed the resulting IgG response was directed at
the fullerenes. Subsequently, Erlanger et al. (2005)
found that the C60 specific monoclonal antibodies
also recognized and specifically bound to single
walled carbon nanotubes (SWCNT). They directly
imaged the antibodies on SWNT using atomic force
10 G. Oberdörster et al.

Figure 6. Hypothetical cellular interactions of NP. The diagram shows the potential effects of NP with emphasis on potential oxidative
stress induced effects and their consequences. (A) Particle-associated characteristics induce lipid peroxidation, intracellular oxidative stress
and increased cytosolic calcium ion concentration; (B) NP may be actively endocytosed via different mechanisms, including caveolae,
clathrin coated pits, or receptor mediated mechanisms. In phagocytic cells phagocytosis triggers activation of NADPH oxidase and
generation of ROS; (C) Particles and their associated metals, as well as oxidative stress, can activate the EGF receptor; (D) Oxidative stress,
receptor activation and increased calcium ions activate transcription of pro-inflammatory genes via transcription factors such as NF-kB; (E)
NP may enter the cell by passive diffusion and remain non-membrane bound from where they may enter mitochondria; (F) and disrupt
normal electron transport leading to oxidative stress. (G) Free particles may also enter the nucleus via the nuclear pore complex and interact
with the genetic material. (H) Lipid peroxide-derived products such as 4-hydroxy nonenal form DNA adducts that may lead to genotoxicity
and mutagenesis. (Note that particle and cell structures are not to scale).

microscopy. This work raises the question of poten-
tial immune responses elicited when functionalized
nanoparticles come into contact with the organism
and acting as sensitizing agents.
Post 2000
The new millennium has seen increasing numbers of
studies with engineered nanomaterials (dendrimers,
quantum dots, carbon nanotubes, metals, etc.)
addressing questions of their kinetics, effects and
underlying cellular and molecular mechanisms de-
rived from in vitro and in vivo studies. This was
largely driven by rapid growth of the number of
nanotech companies producing new nanomaterials
with novel properties and applications in diverse
fields, ranging from engineering, diverse consumer
products, energy production, environmental appli-
cations, and nanomedicine. Both positive and nega-
tive aspects were highlighted to either promote (e.g.,
‘green nanotechnology’) or bedevil (e.g., morator-
ium on nanotechnology) uses/applications of nano-
materials. A general concern for both proponents
and opponents of nanotechnology is the unknown
potential of some nanomaterials for causing harm to
human health and environment; however, whereas
advocates point to the many benefits that nanotech-
nology will bring (Roco 2003a, 2003b; The Royal
Society 2004; US Nanotechnology Initiative 2002;
European Commission 2005; Malsch & Oud 2004),
its adversaries view it as so unpredictably dangerous
that any use poses indeterminate social and environ-
mental risks and must not proceed in the absence of
well-supported risk assessment and societal under-
standing. Improved understanding of the factors
likely to make nanomaterials hazardous is essential
to solving this dilemma, and the reality is likely to be
different for different nanomaterials, ranging from
the safe and innocuous for most to the highly toxic
for some.
Indeed, a number of studies have been published
over the past five years demonstrating significant
toxicity of different nanomaterials, be they carbon
nanotubes, metal nanoparticles, fullerenes, quantum

dots, etc. (Lam et al. 2004; Warheit et al. 2004;
Sayes et al. 2004; Hoshino et al. 2004; Derfus et al.
2004; Kirchner et al. 2005; Shvedova et al. 2005).
However, for a full understanding of risk, exposure
and toxicokinetic data are both missing, as these are
required to combine with hazard data to begin to
understand risk; such extra studies are urgently
needed.
On the other hand, many experimental studies
have evaluated the potential of nanomaterials to be
used for therapeutic or diagnostic purposes (Vicent
& Duncan 2006). Such studies suggest that the
properties of nanoparticles make them useful for
beneficial medical applications by allowing the
ability to cross barriers (Gulyaev et al. 1999) as
well as access to specific tissue and intracellular
locations. Furthermore, some of the properties
known to be responsible for toxicity, can also be
exploited in a controlled manner for therapeutic
purposes, e.g., in the killing of pathogens (silver
nanoparticles [Baker et al. 2005]) or tumour cells
(haematite [Ito et al. 2004]). While the potential for
uncontrolled adverse effects may slow advances in
nanomedicine, it has been possible to develop
polymer therapeutics to market with improved
drug efficacy and reduced side effects for che-
motherapeutic agents (Vicent & Duncan 2006).
Improved understanding of the risk factors related
to nanomaterials in the body will aid the future
development and exploitation of a wider variety of
nanomedicines.
There has been an upsurge of recent reviews
which have summarized our present understanding
of ultrafine/nanoparticle toxicology (Donaldson et
al. 2002; Borm & Kreyling, 2004; Hoet et al. 2004;
Kreyling et al. 2004; Biswas & Wu 2005; Donaldson
et al. 2005; Oberdörster et al. 2005; Donaldson et al.
2006; Lam et al. 2006; Nel et al. 2006). These
reviews discuss to different degrees fundamental
concepts of the toxicology of nano-sized particles
or are also focused on very specific aspects of one
group of nanoparticles, and the reader is referred to
these articles for more information on mechanistic
and other aspects. It should be noted, however, that
toxicological information on nanomaterials needs to
consider actual human exposure levels. Any parti-
culate material, whether nano-sized or larger, will
give rise to adverse effects at high enough doses.
Unfortunately, doses or exposure concentrations of
nanomaterials that are used in in vitro or in vivo
toxicological studies are most often extraordinarily
high, and also delivered as a bolus (extremely high
dose rate), so as to have no or limited relevancy to
real or anticipated human exposures. Despite this, a
critical discussion of dose and dose rate that were
used is mostly lacking in peer-reviewed articles.
Toxicology of nanoparticles: A historical perspective 11
Although high dose studies can provide useful
information, caution has to be exercised when
extrapolating results or mechanisms to the in vivo
situation.
Several of the above-mentioned reviews also point
out the need for toxicological testing specifically
geared towards nanomaterials, and many confer-
ences and workshops, nationally and internationally,
have been sponsored by regulatory bodies and
scientific societies to discuss this as an urgent need.
This is reflected in reports from such meetings (e.g.,
Scientific committee on emerging and newly identi-
fied health risks of The Royal Society 2004; Inter-
national Life Sciences Institute [ILSI] 2005;
Scientific Committee on Emerging and Newly
Identified Health Risks [SCENIHR] 2005; National
Nanotechnology Initiative [NNI] 2006); however, it
appears that these reports are not followed by
appropriate action, thus creating the impression
that there may be just too many of these meetings
without serious follow-up action. On the other hand,
they help to spread a greater awareness of the
potential for nanomaterials to be both beneficial
but also possibly harmful if we do not act pro-
actively to address toxicity issues in order to allow
the safe development of new nanoparticles. Such
awareness is necessary both to prevent disasters due
to premature applications without having recognized
adverse effects of nanomaterials, and to realize the
maximum potential of a safe and sustainable nano-
technology.
Nanoparticle specific effects related to their un-
ique surface and size characteristics have been
reported in increasing numbers in recent years
(Figure 2), and only some can be mentioned in
this brief historical overview. It should be pointed
out, though, that several discoveries made and
reported decades ago have only now been ‘redis-
covered’, and the significance of the earlier studies
for the toxicology of nanoparticles is now being
recognized. Examples are the translocation of nano-
gold particles across the alveolo-capillary barrier in
either direction observed 3 decades ago (Berry et al.
1977); and described again recently (Heckel et al.
2004); the neuronal translocation pathway of viruses
(nano-sized particles) and of nanogold from the nose
to the olfactory bulb (old studies: Howe & Bodian
1940; DeLorenzo 1970; new studies: Oberdörster et
al. 2004; Elder et al. 2006); or the localization of
nanogold particles in mitochondria after entry into
cells following in vivo exposure in the 1970s
(DeLorenzo 1970; Gopinath et al. 1978) and
recently demonstrated in vitro in macrophages with
ambient ultrafine particles, thereby inducing oxida-
tive stress and severe mitochondrial damage (Li et al.
2003).
12 G. Oberdörster et al.

In addition to mitochondrial access of nanoparti- The ‘concept of differential adsorption’ of blood

cles after cell endocytosis, their entry into the cell
nucleus via the nuclear pore complex (NPC) was
recently observed (Panté & Kann 2002), and found
to be limited by the functional diameter of the NPC
of 39 nm. Such nuclear access was associated with
strong in vitro cytotoxicity of 1.4 nm gold particles
(55 gold atoms, Au55) where 50% of the particles
after entering the nucleus were bound to DNA
(Tsoli et al. 2005). The authors suggest the reason
for the cytotoxicity ‘is based on the perfect combina-
tion [Au55] cluster size and major groove [of DNA]
dimension’ and suggested a molecular model of B-
DNA combined with Au55 clusters irreversibly
attached to the major grooves (see Figure 6G).
Diffusion may be a likely mechanism of uptake of
these very small particles into cells (Figure 6E) as
suggested by Geiser et al. (2005).
The significance of particle surface area as an
appropriate dosemetric for expressing dose-response
relationships has also preceded the era of purposeful
toxicological research with nanoparticles: Timbrell
(1982) evaluated the degree of fibrosis in lung
autopsy samples of asbestos miners and determined
retained fibers by their mass, number and surface
area. The miners had been exposed to different types
of asbestos, with large differences in fiber size, at
four different locations. The authors found that
retained fiber surface area correlated best with the
degree of fibrosis, indicating ‘that the fibrogenicity of
the chrysotile per unit surface area was similar to
that of amphiboles’. As discussed before, the con-
cept of particle surface area as dosemetric for the
biologic/toxicologic activity of nanoparticles has now
well been established.
Another important concept for nanomedicine and
now also for nanotoxicology has been discussed in
detail in a recent review by Müller and Keck (2004):
proteins for targeting different organs/cells by nano-
particles (Figure 7). This concept implies that for
intravenously delivered nanoparticles ‘the final de-
termining factor for organ distribution is the blood
proteins adsorbing onto the particle surface’ (Müller
& Keck 2004). The work of Kreuter demonstrating
that coating of nanoparticle surfaces with apolipo-
protein E from blood plasma makes them to breach
the blood brain barrier (uptake by endothelial cells
via the LDL receptor and transcytosis of the
particles across the barrier) is one recent example
of this concept (Kreuter 2001).
The concept of differential adsorption can be
expanded further to the lung as portal-of-entry for
inhaled nanoparticles: Coating of the particles after
depositing on the conducting airways or the alveoli
with the components of the lining fluid (proteins,
lipids) can be an important determinant of their fate.
For example, even large 240 nm polystyrene parti-
cles were found to translocate across the alveolo-
capillary barrier in the lung when coated with
lecithin, whereas uncoated particles did not (Table
II, Kato et al. 2003). Furthermore, coating with
lining fluid components is not only dependent on the
physicochemical properties of the particle surface
but may also be different for different mammalian
species, which could make it very difficult for
toxicologists to predict from results in experimental
animals the biokinetics and potential effect of
nanoparticles in humans. Assessing the extent and
implications of lung and blood protein coatings for
endocytotic and transcytotic processes of nanopar-
ticles is now an area of great interest by scientists
and of high importance. First results indicate that
both nanoparticle surface Characteristics and Size
determine the binding properties of plasma proteins
(Cedervall et al. 2007).

“Concept of Differential Adsorption”

NP physicochemical properties: Body compartment media:

• Size • Respiratory tract lining fluid
• S ur f a c e A re a • Gastrointestinal milieu
• Charge • Plasma proteins
• Surface hydrophobicity • Other mucosal membranes
• Redox activity • Intra/extracellular fluid
• Others

deter mine

Protein/lipid adsorption patterns

d et e r m i n e

Localisation in target tissues/cells

Figure 7. Concepts of differential absorption. The physicochemical properties of NP that influence adsorption of body proteins or lipids
from different body compartments and that subsequently determine the fate of NP in the body. Adapted from Muller and Heinemann
(1989).

There are numerous other examples of continuing
advances in our basic understanding about the
kinetics and effects of nanoparticles, and it appears
that it is not possible based on findings with one
specific engineered nanoparticle to derive general-
ized conclusions that all nanoparticles will follow the
same pattern of uptake and induce the same effects.
There are too many variables, not only size, but
surface characteristics (chemistry, charge, porosity,
etc.), persistence of coating and others that influence
NP effects and kinetics. For example, results of
state-of-the-art retention studies with inhaled radio-
active iridium nanoparticles showing less than 1%
translocation from lung deposits to other organs
(Kreyling et al. 2003; Semmler et al. 2004) or of
more than 10% of deposited Mn-oxide nanoparti-
cles on the nasal olfactory mucosa to the CNS (Elder
et al. 2006) may or may not be applicable to other
nanomaterials. A better understanding is needed
about the interplay of factors contributing and
affecting uptake and disposition from deposits in
the respiratory tract or from other portals of entry.
As another example, the most recent first report of
penetration of differently charged quantum dots
(hydrodynamic diameter 14
35 nm) across the stra-
tum corneum of the skin (Ryman-Rasmussen et al.
2006) shows that, indeed, these engineered nano-
particles can reach epidermis and dermis where they
might interact with cells of the immune system
(macrophages, dendritic cells) or be subjected to
specific translocation pathways via lymph and blood
vessels or even sensory nerves (Oberdörster et al.
2005). In a subsequent study this group observed the
penetration of amino acid functionalized fullerenes
(35 nm) across the stratum corneum to the stratum
granulosum of the skin, using a flexed porcine skin
model (Rouse et al. 2007). This finding confirms
earlier recommendations by Tinkle et al. (2003) that
simulating in vivo conditions of skin flexing is a
significant methodological improvement over a static
skin model in vitro. However, a better mechanistic
understanding is needed before extrapolating these
results of skin penetration to other NP.
Fullerene toxicology
C60 (Buckminster fullerene) was first discovered by
Kroto et al. (1985) and describes 60 linked carbon
atoms in a highly stable icosahedron structure,
consisting of 60 vertices and 32 faces (12 pentagonal
and 20 hexagonal). The term fullerene now applies
to molecules composed entirely of carbon that form
spheres or tubes. Some features of C60 mean that it
is potentially of great value to a number of industries
and in medicine. For example the use of C60 is being
Toxicology of nanoparticles: A historical perspective 13
investigated for use in optics and superconductors
(Da Ros et al. 1999) and for drug delivery (Vogelson
2001). Recently a number of cosmetic products such
as face creams that contain C60 nanoparticles have
entered the market (Halford 2006) . C60 particles
have already begun production on a large scale with
the opening of Mitsubishi’s first fullerene plant in
Japan which aims to produce fullerenes by the ton
(Tremblay 2003).
A number of studies have shown that the C60
fullerene has antioxidant actions. For example,
Wang et al. (1999) demonstrated that lipid soluble
and water soluble C60 derivatives prevented super-
oxide and hydroxyl radical initiated lipid peroxida-
tion to a greater extent than the natural antioxidant,
vitamin E. In rats Gharbi et al. (2005) found that
C60 was able to prevent CCl4 induced liver toxicity,
supporting the theory that C60 is an antioxidant.
Xiao et al. (2005), using electron spin resonance
(ESR) also found that chemically generated hydroxyl
radicals were quenched by PEG-modified and hy-
droxyl fullerenes. Fiorito et al. (2006) demonstrated
that C60 fullerenes and single wall carbon nanotubes
(SWNT) failed to induce the production of NO by
the J774 murine macrophage cell line. These studies
together suggest that far from being toxic, C60 and
its derivatives could actually be of beneficial health
effect.
In contrast a number of other studies have
presented data suggesting that C60 and its derivatives
actually have prooxidant and toxic effects. Sayes
et al. (2004) looked at the effects of treating three
cell lines with nano-C60 prepared in THF. The
nano-C60 was cytotoxic to all three cell types at
doses above 50 ppb, with LC50 ranging from 2
50
ppb depending on the cell type. Treatment of the
cells with nano-C60 was shown to induce membrane
damage and the production of peroxyl radicals. The
role of oxidative damage was confirmed as addition
of L-ascorbic acid prevented membrane damage. A
number of ecotoxicology studies have also used
THF as a solvent to disperse C60, but it has been
suggested that the THF may alter or enhance
toxicity of the nanoparticles (see below), and so
this data requires some careful consideration. Kamat
et al. (1998) also used rat liver microsomes to
demonstrate oxidative damage to lipids induced by
C60 following photosensitisation. The relevance of
this model is questionable since the liver tissue
would not normally be exposed to UV light.
High aspect ratio nanoparticles (HARN)
There has been considerable interest in the fibrous,
high aspect ratio nanoparticles (HARN) given the
14 G. Oberdörster et al.
previous experience with asbestos. Asbestos is a term
used to describe a range of crystalline fibrous silicate
minerals classified as serpentines, containing the
single type chrysotile asbestos, and the amphiboles
that contains crocidolite, amosite, tremolite and
anthophyllite. Of these, chrysotile (white asbestos)
has been the most-used commercially. It has been
suggested that asbestos is a nano material and hence
the first commercially-used HARN (Royal Society
and Royal Academy of Engineering 2004) since the
chrysotile asbestos fibrils are B100 nm in diameter.
Amphibole fibers can be around 100
200 nm in
diameter.
A detailed history of asbestos cannot be presented
here, but asbestos has been in use as an insulating
material since 500 BC according to archaeological
finds in Finland and the reader is referred to specific
reviews by Selikoff and Lee (1978). A full under-
standing of the asbestos hazards did not become
apparent until studies on synthetic vitreous fibers
(SVF) brought them together with asbestos within a
single ‘fiber paradigm’. This paradigm highlighted
the importance of length and biopersistence. Length
was important because, above a key length, the fibers
cannot be adequately phagocytosed by the alveolar
macrophages, preventing clearance and so the dose
accumulates. Biopersistence is also important be-
cause long fibers that are biosoluble weaken, break
and become short enough to be cleared and so do
not persist in the milieu of the lungs. Insulation
wools, rock and slag wool and chrysotile asbestos are
all low-biopersistence fibers whilst crocidolite asbes-
tos is a highly biopersistent fiber. The overall
conclusions from many studies were that the two
factors of length and biopersistence dominated the
pathogenicity of respirable mineral and vitreous
fibers. Importantly, in this context, long fibers are
greater than approximately 20 mm.
The final emergence of a paradigm for the
mechanism of pathogenicity of mineral and vitreous
fibers might therefore be thought to have set the
scene for a useful paradigm for applying to fibrous
nanomaterials. However, it is by no means certain
that fibers other then asbestos or SVF fibers adhere
to this model since the new nano-fibers are made of
a wide range of materials. These new high aspect
ratio nanomaterials are presently dominated by the
carbon nanotubes. These are currently increasing in
scale of production and are set to reach enormous
global quantities in the early part of the 21st century
(Donaldson et al. 2006). The carbon nanotubes are
comprised largely of carbon, the structure being
essentially a single seamless cylinder of graphite in
the case of a single walled carbon nanotube. It is
important to note that they can be contaminated by
other metals such as iron, which can undergo redox
cycling and generate ROS (Kagan et al. 2006;
Shvedova et al. 2005) like asbestos (Shen et al.
1995). Multiwall carbon nanotubes can be visua-
lized as stacks of single nanotubes, one inside the
other. There are a number of other HARN including
nanowires, nanorods (Chen & von Mikecz 2005),
nanobelts (Wang 2004) nanosprings (Bell et al.
2006), etc. It should be noted that nanoparticles
readily agglomerate and nanotubes form ‘nanoropes’
as the individual nanotubes twist around each other,
forming larger fibrous structures (Maynard et al.
2004).
Carbon nanotubes represent one of the fastest-
developing nanoparticle materials with production
set to increase rapidly as a consequence of the useful
properties of this material (Donaldson et al. 2006).
As mentioned above, carbon nanotubes have a
tendency to form tangles and ropes; however these
may also deagglomerate in the lungs due to the
activity of surfactant. A number of material science
studies utilize surfactants to deagglomerate nano-
tubes for imaging and characterization purposes
(Blackburn et al. 2006; Lisunova et al. 2006). This
would suggest that in the lung nanotubes have
the potential to exhibit the properties of other
high aspect ratio particles such as asbestos. Nano-
tubes are HARN and may therefore act either like
nanoparticles and/or like fibers (Donaldson et al.
2004).
For example, the pulmonary toxicity of single-
walled and multi-walled carbon nanotubes delivered
at high doses and dose rates to the lower respiratory
tract of rats and mice induced a high acute inflam-
matory response with granuloma formation and
fibrosis as late effects (Lam et al. 2004; Warheit et
al. 2004; Jia et al. 2005; Muller et al. 2005;
Shvedova et al. 2005). Inhalation studies at low
concentrations with slow build up in the lungs have
not yet been published. Nevertheless, the present
results indicate at least for high doses that carbon
nanotubes are potentially toxic to humans, and that
strict hygiene measures should be taken to limit
exposure during their manipulation (Muller et al.
2005). At the same time, in vitro studies on
fibroblasts showed that with increasing degree of
sidewall functionalization of single-walled carbon
nanotubes their toxicity decreases (Sayes et al.
2006), similar to what Colvin’s group had observed
earlier with functionalized vs. underivatized C60
fullerenes (Sayes et al. 2004).
In the present context there are two important
questions to address: (i) Does asbestos behave like a
nanoparticle?, and (ii) Do man-made HARN behave
like asbestos? These questions are addressed in detail
below.

(1) Does asbestos behave like a nanoparticle? If
asbestos were to act like a nanoparticle then,
on current understandings of nanoparticle
toxicity its surface area would be the most
accurate descriptor of its pathogenicity. In fact
it has been suggested (Timbrell 1982) that
surface area correlates well with pulmonary
fibrosis caused by asbestos and might be a
good dose metric for that endpoint. However,
the most important descriptor of fiber patho-
genicity is length and biopersistence, as de-
scribed above (Davis et al. 1986). It would be
anticipated that short fibers act like particles
and at a high enough surface area dose would
induce inflammation. Certainly the non-fibrous
particulate in some SVF samples have been
held responsible for surface-area related rat
lung overload, that complicated the interpreta-
tion of studies involving mixed exposures to
long biopersistent fibers plus large amounts of
non-fibrous particles (Brown et al. 2005).
(2) Do HARN behave like asbestos? If the same
paradigm for asbestos and vitreous fibers was
operative for HARN, then any biopersistent
HARN longer than about 20 mm might cause
fibrosis, cancer, pleural changes and mesothe-
lioma, if the fiber number is sufficient and
fibers are not phagocytizable. The current
paradigm of fiber pathogenicity does not dis-
criminate between different compositions of
long biopersistent fibers, except in-so-far as
composition determines biopersistence. There
are instances of two long biopersistent fiber
types
erionite (Wagner et al. 1985) and SiC
(Davis et al. 1996) showing special proclivity to
cause mesothelioma for reasons that were not
easily explained, since they were not especially
long or more biopersistent than amphiboles.
The chemical basis of the enhanced pathogeni-
city of these two fibers has not been elucidated.
This suggests that some fiber types may possess
surface or chemical reactivity that imparts
added pathogenicity over and above what
would be anticipated for long biopersistent
fibers. We do not know if CNT or any other
HARN could have this potential but there is
substantial ability to derivatize and change the
CNT surface and alter simple measures of
toxicity (Sayes et al. 2006). Carbon particles
would not normally be anticipated to have
specially active surfaces, however, due to their
small size, nanoparticle carbon black, com-
posed of degenerated graphitic crystallites, is
able to generate more oxidative stress in vitro
than fine carbon black (Wilson 2001). This
paradox may be explained by the complex
Toxicology of nanoparticles: A historical perspective 15
chemical events that occur in cells and tissue
fluids, where oxidations are possible in a
potentially highly catalytic environment.
The role of length in determining CNT toxicity is
yet to be investigated, although there is at least one
study purporting to study it. However, long CNT in
this case were only 820 nm and short CNT were
220 nm long. Both of these samples are less than
1 mm and would not even be classified as fibers in a
workplace, far less approach the 20 mm length range
considered ‘long’ in the current fiber paradigm. In
fact, as might be expected neither sample proved
very effective at inducing TNFa production by cells
in vitro nor causing inflammation when implanted
subcutaneously (Sato et al. 2005).
As mentioned above, chrysotile fibrils consist of
nanofibers which are of low biopersistence in lungs
(Bernstein et al. 2003a, 2003b). Carbon nanotubes
are a few nm (single-walled) or a few hundred nm
(multi-walled) in diameter and so can approach the
dimensions of asbestos. It is worth noting that singlet
CNT (and probably some asbestos fibers) may not
be visible by conventional imaging used to quantify
lung burden and so could confound attempts to
quantify the dose
if they were present in the lungs
as singlets, which is doubtful. The ‘length hypoth-
esis’ needs to be rigorously tested by using CNT that
approach 20 mm in length. The strategy for assessing
the pathogenicity of ‘long CNT’ should recognize
endpoints from classical fiber pathogenicity such as
durability, biopersistence, persistent inflammation
and cell proliferation (Warheit et al. 1995); Donald-
son et al. 2004). Chronic studies should emphasize
carcinogenic and mesothelioma endpoints in the
lungs and positive controls would include amphibole
asbestos. Pleural mesothelioma is a key endpoint to
study but is a rare tumour even in animal experi-
ments; however hamsters have been shown to be a
species especially susceptible to mesothelioma
(McConnell et al. 1999) and so hamster studies
may be considered. Since some types of CNT appear
to be flexible and have the ability to bend or curl up
to form a more compact particle that is an agglom-
erate (Maynard et al. 2004; Shvedova et al. 2005)
the rigidity of CNT may be a factor in driving
toxicity, since CNT rolled into a ball could be more
easily cleared by phagocytic cells and may not be like
rigid asbestos fibers at all. In the latter case the
overall length is less important in producing fru-
strated phagocytosis, a key process in fiber patho-
genicity. It is possible that nanotubes may prove
especially fibrogenic as large granulomas have been
described in rodents exposed by instillation (Lam
et al. 2004; Warheit et al. 2004). However, these
were instillation studies where the nanotubes may
16 G. Oberdörster et al.
well have agglomerated into large tangles, producing
an abnormal response and so an inhalation study is
urgently required to determine the pathogenicity by
physiological exposure routes and CNT aerosols
resembling workplace conditions.
Interactions of nanoparticles with the
environment
The awareness of a potential negative impact of
nanomaterials on the environment has also become
central to the discussion of nanomaterial safety: ‘ . . .
a growing public debate is emerging on whether the
environmental and societal costs of nanotechnology
outweigh its many benefits’ (Colvin 2003). The
diverse array of products containing nanoparticles
inspires the certainty that nanoparticles will be
released into the environment in significant quan-
tities, presently with unknown consequences. In fact,
this is not a problem of the future, currently released
nanoparticles include those used in consumer pro-
ducts that are washed into waste water and disposed
of via land-fill sites, as well as nanoparticles that are
intentionally released, such as those used for the
remediation of polluted land (e.g., zerovalent iron
NP [Zhang 2003]). Tratnjek et al. (2006) recently
reported that such particles were typically between
10 and 100 nm, although in laboratory conditions
they often agglomerate into larger particles. At the
current time the impact of particle agglomeration on
toxicity is not understood; many of the toxicology
studies in rodents and cell lines have used agglom-
erated and aggregated rather than mono-dispersed
particles, and such studies indicate that even in the
agglomerated forms the smaller particles are more
potent in many respects (see previous sections).
When considering the potential risk associated
with nanoparticle release into the environment, we
need to consider both exposure and hazard. For
substances purposefully released into the environ-
ment, such as the zerovalent iron described above,
estimations of exposure concentrations for various
organisms can be obtained. However, for substances
that are unintentionally released, exposure assess-
ment is extremely difficult, not least because of the
large number of currently unidentified release
sources, including both domestic and industrial.
There is a tendency within our article to use the
generic term ‘nanoparticle’ when referring to parti-
cles of less than 100 nm in at least one dimension.
But as described previously, the array of nanoparti-
cles currently available and under development is
immense. When assessing exposure of organisms to
nanoparticles in the environment, it is likely that
initial studies will focus on nanoparticles of a specific
chemical composition, but over time it will be
essential to estimate or determine the combination
or total exposure. This does not assume that all
nanoparticles are of equal risk or hazard, but that a
risk assessment must be generated that is based upon
a combination of the different types of nanoparticles
most prevalent in the environment.
Once nanoparticles are released into the environ-
ment there is a requirement to understand their fate
and transport, in order to determine in which
environments the particles are most likely to occur
or even accumulate, and hence which organisms are
most likely to be exposed. Currently a search of any
publication database, for the terms nanoparticles,
environment and fate or transport, yields very few
publications. There is however a large collection of
publications relating to naturally occurring colloids,
their composition and behaviour. Colloids are stable
suspensions of organic or inorganic particles with
diameters between 1 and 1000 nm (although precise
definitions vary between sources), and hence such
research is incredibly relevant to the environmental
fate and behaviour of nanoparticles. Colloids have
previously been studied because of their role in the
transport of adsorbed contaminants in subsurface
environments (McCarthy & Zachara 1989). In 1989
McCarthy called ‘attention to the potentially critical
but poorly understood role of colloids in facilitating
contaminant transport’. This complements the sug-
gestions that nanoparticles may adsorb endogenous
molecules in the human body as described in
previous sections (Müller & Keck 2004) (Figure
7). This would suggest, that lessons relating to the
behaviour of nanoparticles in different environments
must be communicated widely in order to allow
extrapolation to other situations and hence a rapid
advancement of experimental design and under-
standing.
A full history of the science of environmental
colloids is beyond the scope of this article, but is
reviewed elsewhere by Kanti and Khilar (2006). To
quote from Kanti:
Until some two decades ago, it was believed that
only the soil liquid and gaseous phases were
mobile and could facilitate the transport of
chemicals and nutrients in subsurface flow. It is
now generally accepted that also part of the soil
solid phase is mobile under different geochemical
conditions and that mobile organic and inorganic
soil particles, colloidal in nature may facilitate or
retard the contaminant transport.
A large proportion of the naturally occurring
colloid particles are composed of clay minerals,
oxides of Fe and Al, silica and/or natural organic
matter (Kanti & Khilar 2006). Elimelech (1994)

investigated the effect of particle size on the kinetics
of particle deposition onto surfaces, including other
particles, both theoretically and experimentally. This
study used positively charged latex particles and
negatively charged glass beads ranging from 0.08

2.51 mm diameter, and demonstrated that as particle
size increased deposition into a packed bed column
increased. This would suggest that in the environ-
ment larger particles are less mobile within porous
media such as soil, due to their propensity to deposit
or adsorb onto surfaces, and hence become trapped
(Elimelech 1994). More recently, Zhuang et al.
(2005) studied amphiphillic (both hydrophilic and
hydrophobic) colloids due to their prevalence in the
environment in the form of ‘microorganisms, man-
ufactured nanoparticles, proteins and natural occur-
ring mineral colloids with organic matter coatings’,
and found that as particle size increased from 20

420 nm, the deposition or retention within a porous
media decreased, reaching a minimum value at
around 100 nm. Above 100 nm retention within
the porous media increased again, suggesting that
mobility of amphiphillic nanoparticles within a
porous media is not linearly related to particle size
at this lower size range (Figure 8). Interestingly, this
deposition behaviour of nano-sized colloidal struc-
tures in soil is to some extent similar to the
deposition behaviour of inhaled particles in the
respiratory tract (Figure 4), which for the smallest
particles is governed by diffusion. This becomes less
efficient with increasing size such that particles
between 100 to 300 nm experience least deposi-
tion and remain airborne, whereas larger particles
Matrix Deposition Rate
0 100 200 300 400
Particle Diameter (nm)
Figure 8. Deposition rates of colloidal particles in a model
simulating movement in soil. As particle size increases from
20 nm, colloid deposition rate decreases to reach a minimum at
100 nm, but then subsequently increases at greater particle sizes.
Due to their lower propensity to deposit, colloid particles around
100 nm are the most mobile within a porous media. Colloid
deposition rate varies with saturation of flow conditions, being
higher at low saturation and lower at 100% saturation (Drawn
after Zhuang et al. 2005).
Toxicology of nanoparticles: A historical perspective 17
show increased deposition due to impaction and
sedimentation (Figure 4).
The techniques, models and knowledge currently
available in relation to colloid behaviour, are all
applicable to engineered nanoparticles and will
provide a useful knowledge base on which to
investigate such environmental issues. For example,
atomic force microscopy (AFM) has been used to
characterize freshwater natural aquatic colloids
(Lead et al. 2005), allowing identification of fibrils
10 nm in diameter and 100 nm or more in length, as
well as spherical colloids 30
50 nm in diameter.
Techniques such as flow filed-flow fractionation (FI
FFF) and size exclusion chromatography (SEC)
coupled to UV-absorbance (UVA) and inductively
coupled plasma mass spectrometry (ICP-MS) have
been used to investigate colloidal binding of con-
taminants such as U and Ni in water extracts of
sediments collected from rivers (Jackson et al. 2005).
Despite the current lack of knowledge relating to
which environments, and hence which organisms,
are most likely to be exposed to nanoparticles, the
toxicological impact of nanoparticles on a range of
organisms has started to emerge. Such studies
already span the breadth of microorganisms, plants,
invertebrates and vertebrates, although relatively few
publications are available within each category.
One of the first such studies was published by
Oberdörster (2004) and involved treating large
mouth bass fish with nC60 (0.5 and 1 ppm for 48
h). The exposed fish were found to exhibit signs of
lipid peroxidation in the brain which the authors
suggested could be due to NP transport to the brain
as previously reported in mammals (DeLorenzo
1970), along with the lack of neural antioxidant
defence mechanisms that occur naturally in neural
tissue. However, the nC60 was prepared initially in
an organic solvent tetrahydrofuran (THF) in order
to aid solubilisation, before evapouration to remove
the solvent. These studies are interesting, but cau-
tion is required since THF is a neurotoxin; addi-
tionally Brant et al. (2005), have demonstrated that
even after filtration and evaporation, THF remains
trapped between nC60 agglomerates. Since THF is
not an environmentally relevant solvent or vehicle,
special consideration will need to be given to the way
in which particles are prepared in order to generate
an environmentally relevant protocol. For these
reasons, particle preparation is likely to form a major
component of future research, because until
standardised, well characterised, but relevant
and validated protocols are in place, it will be
difficult to interpret ecotoxicology studies, but also
to allow routine testing and comparison between
laboratories.
18 G. Oberdörster et al.
In order to investigate the role of THF in fullerene
toxicity, Zhu et al. (2006) in a subsequent study
exposed adult fat head minnow to nC60 (0.5 ppm)
prepared either in THF or by stirring in water for
weeks. The nC60 prepared in THF induced 100%
mortality within 6
18 h of exposure. In contrast, the
nC60 generated by water stirring had no impact on
lethality over the same time period. Sub-lethal
effects of the water-stirred nC60 still occurred and
included lipid peroxidation in the gill and increased
expression in the liver of the biotransformation
isoenzyme family, CYP2. Again, this clearly demon-
strates the potential impact of the particle prepara-
tion technique on the results of such studies.
Some studies have also been published that have
investigated the effects of nanoparticles on aquatic
invertebrates. One of the first studies in this area was
by Lovern and Klaper (2006), and used both nC60
and TiO2 (Degussa P25, 25 nm diameter) to treat
Daphnia magna (water fleas). Again the study used
THF to disperse the particles, but this technique
was also compared with sonication in medium for 30
min. The TiO2 and the nC60 particles were both
found to be more potent at killing the organisms
when prepared in THF than when prepared by
sonication. For example, the LC50 of TiO2 prepared
in THF was 5.5 ppm, but for sonicated TiO2
mortality never exceeded 11%, even at concentra-
tions up to 500 ppm. In this study, the nC60 was
generally more potent at inducing lethality than the
TiO2. The question remains as to whether the
particles prepared in THF were more toxic because
they were better dispersed or because of THF
induced toxicity. Similar results were obtained in
the above mentioned study by Zhu et al. (2006)
using THF or water stirring solubilization of nC60 to
expose Daphnia magna .
Recent studies by this group have avoided the
potential for both THF and sonication to enhance
nC60 toxicity, by preparing the particles by stirring in
water alone (Oberdörster et al. 2006). They con-
firmed a lower toxicity in freshwater invertebrates
(Daphnia magna and Hyalella azteca as well as a
Figure 9. Daphnia magna exposed to (a) 20 nm fluorescent carboxylated polystyrene beads (2.64 mg/l for 1 h), (b) 14 nm carbon black
(1 mg/l for 48 h) or (c) 25 nm TiO2 particles (1 mg/l for 48 h). Pictures provided by P. Rosenkranz and T. Fernandes.
marine harpacticoid) of nC60 solubilized by stirring
compared to earlier findings in different aquatic
species with THF solubilized nC60. However, spe-
cies differences in toxicity may also play a role.
The lethality results of the studies published by
Lovern and Klaper (2006) and Zhu et al. (2006)
both indicate that in the acute tests conducted on
Daphnia magna, the lethality of the nanoparticles
tested is relatively low. However, the sublethal effects
of nanoparticles on these species have not yet been
studied. Preliminary results from the group of
V. Stone (Rosenkranz et al. 2006) indicating accu-
mulation within the gastrointestinal tract and other
body compartments, suggests that such research will
be needed in order to determine effects on beha-
viour, feeding and reproduction as well as accumu-
lation within the food chain. In addition, the
particles adhere to the exoskeleton surfaces of the
exposed organisms, suggesting multiple routes of
exposure and absorption (Figure 9). Similar obser-
vations have been reported by Oberdörster et al.
(2006), after exposure of Daphnia magna to nano-
iron particles. The effects of any nanoparticles on
other aquatic or terrestrial invertebrates have not as
yet been published.
Oberdörster et al. (2006) investigated whether
gene array technology might act as a useful tool to
screen nanoparticle toxicity. They treated fathead
minnow for 5 days with 50 ppm nano-iron particles
(70 nm diameter) composed of an iron oxide shell
with an elemental iron core. Brain, liver, gonads,
gills and other tissues showed that very few genes
were changed in the exposed animals suggesting at
the dose and in the conditions employed these
particles were not effective at inducing toxicity in
the fathead minnow. However, before such conclu-
sions can be made further studies are required in
these fish to determine the effects of different
particle concentrations as well as comparison with
concentrations expected in the environment.
With respect to the effects of nanoparticles on
plants, very little work has been published. Yang
and Watts (2005) have recently investigated the

phytotoxicity of 13 nm aluminium nanoparticles on
the root growth from seeds of five different plant
species. The studies found that at relatively high
concentrations (2 mg ml 1), Al nanoparticles in-
hibited root growth, while larger Al particles of 200

300 nm diameter had no significant effect. The
effects of the nanoparticles on root growth were
decreased by the addition of an antioxidant, di-
methyl sulphoxide, suggesting that oxidative stress
may play a role in the effects of the nanoparticles on
root elongation.
The literature relating to the effects of nanoparti-
cles on microorganisms is much more extensive and
diverse than for plants, invertebrates and vertebrates.
The toxicity of nanoparticle to microorganisms has
been widely studied in relation to the development of
antimicrobial agents and devices for use in the
environment, in industry and in medical devices.
Such applications and research has also lead to the
use of nanoparticles as antimicrobial agents in
consumer products such as underwear.
It is important to realize that many of the
applications that employ nanoparticles for antibac-
terial purposes use materials that are already known
to possess antibacterial properties, for example TiO2
and silver. The nanoparticle nature and associated
larger specific surface area of the material simply
enhances the antibacterial activity.
Nanoparticles of TiO2 have been studied and used
extensively as an antimicrobial agent. Following the
initial discovery of photo-induced water cleavage on
TiO2 electrodes (Fujishima & Honda 1972), TiO2
photocatalysis has been found to generate various
active oxygen species, such as hydroxyl radical and
hydrogen peroxide by reductive reactions or oxida-
tive reactions under light (Sunada et al. 1998). The
reactive oxygen species formed by the UV-induced
photocatalysis then induce antimicrobial effects by
killing bacteria, but also by destroying endotoxin,
thereby preventing endotoxin induced biological
responses (Sunada et al. 1998). The nanoparticle
form of TiO2 is now being exploited due to the
relatively larger surface area over which the photo-
chemical reactions can occur (Mills & Hunte 1997)
for applications in water and air purification.
Composite materials containing different TiO2
and other nanomaterials are also being developed
for antimicrobial applications. For example, Lee
(2005) investigated the ability of TiO2-coated
multi-walled nanotubes to kill bacterial endospores.
They found that the composite irradiated with UV
light was more effective than either UV light alone or
UV irradiated TiO2 particles. However, the compo-
site of nanotubes and TiO2 appeared to cause
endospore aggregation so that some of the spores
were protected and survived.
Toxicology of nanoparticles: A historical perspective 19
The antimicrobial properties of silver-coated sur-
faces and impregnated materials have long been
recognized, with strong biocidal effects being ob-
served in 16 species of bacteria, including Escherichia
Coli in the 1970s (Spadaro et al. 1974). This
recognition has led to the use of metallic silver in a
diverse array of applications including dental resin
composites and coatings for medical devices. Coat-
ings for medical devices have, however, proven
disappointing, due to inactivation caused by subse-
quent coating with blood plasma. Impregnation of
the material has been suggested to improve the
outcome, but silver metal or silver salts cannot be
incorporated into biomaterials and so studies have
now begun to investigate incorporation of silver
nanoparticles in medical devices (Furno et al.
2004). Again due to the recognition that a nanopar-
ticulate form of silver would increase the surface area
for reactivity, Aymonier et al. 2002 developed
hybrids of silver nanoparticles with hyperbranched
macromolecules and demonstrated their use as
effective antimicrobial surface coatings.
Monodispersed silver nanoparticles (12 nm) have
been shown to induce lethality in E. Coli (Sondi &
Salopek-Sondi 2004). Electron microscopy of the
bacteria suggests that the mechanism of death
involves the generation of pits in the cell wall, and
the accumulation of nanoparticles in the cell mem-
brane. Other metal oxides also developed for anti-
microbial purposes include nano-magnesium oxide
and iron oxide. Huang et al. (2005) have demon-
strated that for nano-MgO, bactericidal efficacy
increases with decreasing particle size. Iron oxide
nanoparticles are already being used, not for biome-
dical purposes, but for environmental remediation
purposes in some countries, including the USA.
Remediation involves the release of tonne quantities
of the nanoparticulate into a contaminated land site.
Zerovalent iron nanoparticles have been demon-
strated by Zhang (2003) to result in the removal of
oxygen from groundwaters. This is an important
deleterious effect and supports the suggestion that
the environmental risk resulting from nanoparticle
release needs to be assessed.
As mentioned previously, all of the materials
described above have antimicrobial activity regard-
less of the particle size, but this activity is enhanced
when delivered in a nanoparticulate form. One
material which is not inherently antimicrobial is
carbon, however C60 fullerenes, prepared by stirring
in water, have recently been found to form agglom-
erates 25
500 nm in diameter that, at low concen-
trations ( ]0.4 mg l 1), inhibit the growth of E. coli
and Bacillus subtilis (Fortner et al. 2005).
With respect to the antimicrobial effects of nano-
particles, an important emerging concept is that
20 G. Oberdörster et al.
substances with biocidal properties have these prop-
erties enhanced when presented in a nanoparticulate
form. However, there is insufficient evidence to
suggest that all nanoparticles have antimicrobial
effects, or in fact that all nanoparticles are toxic to
any organism encountered in an exposed environ-
ment. Research into the ecotoxicology of nanopar-
ticles is in its infancy compared to the field of
toxicology, and so much more work will be required
before any generalised statements can be made
regarding nanoparticle ecotoxicology.
Outlook
Undoubtedly, there will be a steady stream of new
findings to advance our understanding of mechan-
isms related to the toxicology and ecotoxicology of
nanomaterials, some will show significant adverse
effects of certain materials
raising new concerns
about human and environmental health
and others
portraying a more benign nature of other different
materials. At present, we lack a model to predict
hazard or safety just based on the physicochemical
characteristics of new nanomaterials that can be
used for risk assessment or for safe product design.
Likewise, there is little information about human
exposure (both level and duration of exposure) of
nanomaterials, and since risk is a function of hazard
and exposure, knowledge of both hazard and ex-
posure are necessary to determine a risk. Experi-
mental findings of high as well as lack of toxicity
(hazard) often are based on flawed study designs and
need to be viewed critically. Regulatory agencies,
professional societies, NGOs and industry are in-
volved in developing and validating standard guide-
lines for toxicological and ecotoxicological testing
strategies of nanomaterials to pre-empt and avoid
untoward surprises from intentional or unintentional
exposures to nanomaterials (e.g., SCENIHR 2004).
Special attention should also be given to sensitive
parts of the population, i.e., people with compro-
mised organ systems or specific genetic disorders.
These will predictably be more at risk, a lesson
learned from population studies of ambient particu-
late air pollution. Thus, including animal models of
compromised human conditions in future nanotox-
icology research should be an important goal to
cover this aspect of increased susceptibility, as was
shown, for example, by an increased translocation of
nanoparticles across tight junctions of the alveolo-
capillary barrier in inflammatory conditions (Heckel
et al. 2004).
There currently exists a battery of validated
toxicity and ecotoxicity tests suitable for chemicals,
however much work is required to optimize these for
nanomaterials that are often poorly understood in
terms of their behaviour in living organisms and in
soil and water. Improved understanding of the
kinetics of nanomaterials in the mammalian organ-
ism and the environment is essential in the develop-
ment of realistic and reliable hazard assessment.
However, studies should not be limited to optimisa-
tion of validated tests. Instead we also require an
understanding of the mechanisms of uptake, toxicity
and the potential for bioaccumulation/bioconcentra-
tion in different species. Much work is required to
close the gap regarding our current understanding of
nanomaterial behaviour in human models versus the
environment, but information from human models
can be useful in informing study design in other
phyla.
There have been many conferences, meetings and
workshops organized by national and international
organizations to discuss these and other issues, with
as many calls for developing testing strategies, with
only few proposals, followed though by far less
action. Many claims have been made on one hand
about the potential benefits of nanotechnology and
on the other the potential risks. Only through the
development of a sound evidence base in all aspects
of hazard and exposure assessment can we hope to
generate an informed and reliable risk assessment.
Quantitative risk assessment of nanomaterials
should be an important goal of the next chapter in
the history of nanotoxicology; this has now started.
Acknowledgements
Support of G.O. is acknowledged through AFOSR
MURI Grant (No. FA9550-04-1-0430), NSF
SGER Grant (No. BES-0427262) and EPA PM
Center Grant (No. RD-83241501). Support of K.D.
is via The Colt Foundation.
References
Adams RJ, Bray D. 1983. Rapid transport of foreign particles
microinjected into crab axons. Nature 303:718
720.
Amdur MO, McCarthy JF, Gill MW. 1982. Respiratory response
of Guinea pigs to zinc oxide fume. Am Ind Hyg Assoc J
43:897
889.
Avakian MD, Dellinger B, Fiedler H, Gullet B, Koshland C,
Marklund S, Oberdörster G, Safe S, Sarofim A, Smith KR,
Schwartz D, Suk WA. 2002. The origin, fate, and health effects
of combustion by-products: A research framework. Environ
Health Perspect 110(11):1155
1162.
Aymonier C, Schlotterbeck U, Antonietti L, Zacharias P, Tho-
mann R, Tiller JC, Mecking S. 2002. Hybrids of silver
nanoparticles with amphiphilic hyperbranched macromolecules
exhibiting antimicrobial properties. Chem Commun 24:3018

3019.
Baker C, Pradhan A, Pakstis L, Pochan DJ, Shah SI. 2005.
Synthesis and antibacterial properties of silver nanoparticles. J
Nanosci Nanotechnol 5:244
249.
Bansal RC, Wang M-J, Donnet J-B. 1993. Carbon black: Science
and technology. 2nd ed. New York: Marcel Dekker. 482 pp.

Bawden FC, Pirie NW. 1937. The isolation and some properties
of liquid crystalline substances from Solanaceous plants
infected with three strains of tobacco mosaic virus. Proceedings
of the Royal Society of London, Series B, Biological Sciences,
Vol. 832 (3 August 1937). pp 274
320.
Beaucage G, Rane S, Schaefer DW, Long G, Fischer D. 1999.
Morphology of polyethylene-carbon black composites. J Poly-
mer Sci: Part B: Polymer Phys 37:1105
1119.
Beijerinck MJ. 1898. Concerning a contagium vivum fluidum as
cause of the spot disease of tobacco leaves. Verhandelingen der
Koninkyke akademie Wettenschapppen te Amsterdam 65:3

21. [Phytopathological Classics Number 7, 1942. St Paul,
Minnesota: American Phytopathological Society Press].
Bell DJ, Dong L, Nelson BJ, Golling M, Zhang L, Grutzmacher
D. 2006. Fabrication and characterization of three-dimensional
InGaAs/GaAs nanosprings. Nano Lett 6(4):725
729.
Bernstein DM, Chevalier J, Smith P. 2003a. Comparison of
Calidria chrysotile asbestos to pure tremolite: Inhalation
biopersistence and histopathology following short-term expo-
sure. Inhal Toxicol 15(14):1387
1419.
Bernstein DM, Rogers R, Smith P. 2003b. The biopersistence of
Canadian chrysotile asbestos following inhalation. Inhal Tox-
icol 15(13):1247
1274.
Berry JP, Arnoux B, Stanislas G, Galle P, Chretien J. 1977. A
microanalytic study of particles transport across the alveoli:
Role of blood platelets. Biomed 27:354
357.
Blackburn JL, Engtrakul C, McDonald TJ, Dillon AC, Heben
MJ. 2006. Effects of surfactant and boron doping on the BWF
feature in the Raman spectrum of single-wall carbon nanotube
aqueous dispersions. J Phys Chem B Condens Matter Mater
Surf Interfaces Biophys 110:25551
25558.
Biswas P, Wu C-Y. 2005. Nanoparticles and the environment. J
Air Waste Manage Assoc 55:708
746.
Bodian D, Howe HA. 1941. Experimental studies on intraneural
spread of poliomyelitis virus. In: Bordley JI, editor. Bulletin of
the Johns Hopkins Hospital. Baltimore: The Johns Hopkins
Press. pp 248
267.
Borm PA, Kreyling W. 2004. Toxicological hazards of inhaled
nanoparticles
potential implication for drug delivery. J
Nanosci Nanotechnol 4(5):521
531.
Brant J, Lecoanet H, Hotze M, Wiesner M. 2005. Comparison of
electrokinetic properties of colloidal fullerenes (n-C60) formed
using two procedures. Environ Sci Technol 39:6343
6351.
Brody AR, Warheit DB, Chang LY, Roe MW, George G, Hill LH.
1984. Initial deposition pattern of inhaled minerals and
consequent pathogenic events at the alveolar level. Ann NY
Acad Sci 428:108
120.
Brody AR, Hill LH, Warheit DB. 1985. Induction of early alveolar
injury by inhaled asbestos and silica. Federation Proc 44:2596

2601.
Brown RC, Bellmann B, Muhle H, Davis JM, Maxim LD. 2005.
Survey of the biological effects of refractory ceramic fibres:
Overload and its possible consequences. Ann Occup Hyg
49(4):295
307.
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 DM, Wilson MR, MacNee W, Stone V, Donaldson K.
2001. Size-dependent proinflammatory effects of ultrafine
polystyrene particles: A role for surface area and oxidative
stress in the enhanced activity of ultrafines. Toxicol Appl
Pharmacol 175:191
199.
Cedervall T, Lynch I, Lindman S, Tulin E, Nilsson H, Dawson
KA, Linse S. 2007. Novel methods to quantify binding rates
and affinities of proteins to nanoparticles: Effects of nanopar-
ticle composition and size. PNAS 104:2050
2055.
Toxicology of nanoparticles: A historical perspective 21
Chen B-X, Wilson SR, Das M, Coughlin DJ, Erlanger BF. 1998.
Antigenicity of fullerenes. Antibodies specific for fullerenes and
their characteristics. PNAS USA 95:10809
10813.
Chen M, von Mikecz A. 2005. Formation of nucleoplasmic
protein aggregates impairs nuclear function in response to
SiO2 nanoparticles. Experim Cell Res 305(1):51
62.
Churg A, Green F. 1999. Pathology of occupational lung disease.
2nd ed. Baltimore: Williams and Wilkins.
Czerniawska A. 1970. Experimental investigations on the pene-
tration of 198Au from nasal mucous membrane into cerebrosp-
inal fluid. Acta Otolaryng 70:58
61.
Colvin VL. 2003. The potential environmental impact of en-
gineered nanomaterials. Nature Biotechnol 21:1166
1170.
Dalal NS, Jafari B, Petersen M, Green FY, Vallyathan V. 1991.
Presence of stable coal radicals in autopsied coal-miners lungs
and its possible correlation to coal-workers pneumoconiosis.
Arch Environ Health 46:366
372.
Dalal NS, Newman J, Pack D, Leonard S, Vallyathan V. 1995.
Hydroxyl radical generation by coal mine dust: possible
implication to coal workers’ pneumoconiosis (CWP). Free
Radic Biol Med 18(1):11
20.
Da Ros T, Prato M. 1999. Medicinal chemistry with fullerenes
and fullerene derivatives. Chemical Communications 663

669.
Davis JM, Addison J, Bolton RE, Donaldson K, Jones AD, Smith
T. 1986. The pathogenicity of long versus short fibre samples of
amosite asbestos administered to rats by inhalation and
intraperitoneal injection. Br J Exp Pathol 67(3):415
430.
Davis JM, Brown DM, Cullen RT, Donaldson K, Jones A, Miller
A, McIntosh BG, Searl A. 1996. A comparison of methods for
determining and predicting the pathogenicity of mineral fibres.
Inhal Toxicol 8:747
770.
DeLorenzo A. 1970. The olfactory neuron and the blood-brain
barrier. In: Wolstenholme G, Knight J, editors. Taste and smell
in vertebrates. London: J. & A. Churchill. p 151
176.
Derfus AM, Chan WCW, Bhatia SN. 2004. Probing the cyto-
toxicity of semiconductor quantum dots. Nano Letters
4(1):11
18.
Donaldson K, Golyasnya N. 2004. Cytogenetic and pathogenic
effects of long and short amosite asbestos. J Pathol 177:303

307.
Donaldson K, Beswick PH, Gilmour PS. 1996. Free radical
activity associated with the surface of particles: A unifying
factor in determining biological activity? Toxicol Lett 88(1

3):293
298.
Donaldson K, Li XY, MacNee W. 1998. Ultrafine (nanometre)
particle mediated lung injury. J Aerosol Sci 29(5/6):553
560.
Donaldson K, Brown D, Clouter A, Duffin R, MacNee W,
Renwick L, Tran L, Stone V. 2002. The pulmonary toxicology
of ultrafine particles. J Aerosol Med
Deposition Clearance &
Effects in the Lung 15(2):213
220.
Donaldson K, Stone V, Tran CL, Kreyling W, Borm PJ. 2004.
Nanotoxicology 188. Occup Environ Med 61(9):727
728.
Donaldson K, Tran L, Jimenez LA, Duffin R, Newby DE, Miles
N, MacNee W, Stone V. 2005. Combustion-derived nanopar-
ticles: A review of their toxicology following inhalation ex-
posure. Particle Fibre Toxicol 2(1):10 (on line; doi: 10.1186/
1743-8977-2-10).
Donaldson K, Aitken R, Tran L, Stone V, Duffin R, Forest G,
Alexander A. 2006. Carbon nanotubes: A review of their
properties in relation to pulmonarytoxicology and workplace
safety. Toxicological Sci 92(1):5
22.
Douglas T, Young M. 2006. Viruses: Making friends with old foes.
Science 312:873
875.
Drinker P, Thomson RM, Finn JL. 1927a. Metal fume fever: II.
Resistance acquired by inhalation of zinc oxide on two
successive days. J Ind Hyg Tox 9(3):98
105.
22 G. Oberdörster et al.
Drinker P, Thomson RM, Finn JL. 1927b. Metal fume fever: IV.
Threshold doses of zinc oxide preventive measures and the
chronic effects of repeated exposures. J Ind Hyg Tox 9:331

345.
Ehrenberg M, McGrath JL. 2005. Binding between particles and
proteins in extracts: Implications for microrheology and
toxicity. Acta Biomaterialia 1:305
315.
Elder A, Gelein R, Silva V, Feikert T, Opanashuk L, Carter J,
Potter R, Maynard A, Ito Y, Finkelstein J, Oberdörster G.
2006. Translocation of inhaled ultrafine manganese oxide
particles to the central nervous system. Environ Health
Perspect 114:1172
1178.
Elimelech M. 1994. Effect of particle size on the kinetics of
particle deposition under attractive double layer interactions. J
Colloid Interface Sci 164:190
199.
EPA. 2004. Air quality criteria for particulate matter (Vol. III)
600/P-95-001cF. Washington, DC 20460: Office of Research
and Development.
Erlanger BF, Chen B-X, Zhu M, Brus L. 2005. Binding of an
anti-fullerene IgG monoclonal antibody to single wall carbon
nanotubes. Nano Lett 1(9):465
467.
European Commission. 2005. Research needs on nanoparticles.
In: Tomellini R, de Villepin C, editors. Proceedings of work-
shop held in Brussels on 25
26 January 2005. (www.cordis.lu/
nanotechnology). 96 pp.
Faux SP, Michelangeli F, Levy LS. 1994. Calcium chelator Quin-
2 prevents crocidolite-induced DNA strand breakage in human
white blood cells. Mutat Res 311(2):209
215.
Ferin J, Oberdörster G, Penney DP, Soderholm SC, Gelein R,
Piper HC. 1990. Increased pulmonary toxicity of ultrafine
particles? I. Particle clearance, translocation, morphology. J
Aerosol Sci 21:381
384.
Fiorito S, Serafino A, Andreola F, Bernier P. 2006. Effects of
fullerenes and single-wall carbon nanotubes on murine and
human macrophages. Carbon 44(6):1100
1105.
Fortner JD, Lyon DY, Sayes CM, Boyd AM, Falkner JC, Hotze
EM, Alemany LB, Tao YJ, Guo W, Ausman KD, Colvin VL,
Hughes JB. 2005. C60 in water: nanocrystal formation and
microbial response. Environ Sci Technol 39:4307
4316.
Fujishima A, Honda K. 1972. Electrochemical photolysis of water
at a semiconductor electrode. Nature 238:37
38.
Furno F, Morley KS, Wong B, Sharp BL, Arnold PL, Howdle
SM, Bayston R, Brown PD, Winship PD, Reid HJ. 2004. Silver
nanoparticles and polymeric medical devices: A new approach
to prevention of infection? J Antimicrob Chemother 54:1019

1024.
Gabor S, Anca Z. 1975. Effect of asbestos on lipid peroxidation in
the red cells. Br J Ind Med 32(1):39
41.
Gardner LU. 1938. Experimental Pneumoconiosis. In: Lanza AJ,
editor. Silicosis and asbestosis. New York: Oxford University
Press.
Geiser M, Rothen-Rutishauser B, Kapp N, Schurch S, Kreyling
W, Schulz H, Semmler M, Im Hof V, Heyder J, Gehr P. 2005.
Ultrafine particles cross cellular membranes by nonphagocytic
mechanisms in lungs and in cultured cells. Environ Health
Perspect 113:1555
1560.
Gharbi N, Pressac M, Hadchouel M, Szwarc H, Wilson SR,
Moussa F. 2005. 60]fullerene is a powerful antioxidant in vivo
with no acute or subacute toxicity. Nano Lett 5(12):2578

2585.
Gilmour PS, Brown DM, Lindsay TG, Beswick PH, MacNee W,
Donaldson K. 1996. Adverse health-effects of PM(10) Parti-
cles
involvement of iron in generation of hydroxyl radical.
Occupat Environ Med 53:817
822.
Gilmour PS, Brown DM, Beswick PH, MacNee W, Rahman I,
Donaldson K. 1997. Free radical activity of industrial fibers:
Role of iron in oxidative stress and activation of transcription
factors. Environ Health Perspect 105:1313
1317.
Gopinath PG, Gopinath G, Kumar A. 1978. Target site of
intranasally sprayed substances and their transport across the
nasal mucosa: A new insight into the intranasal route of drug
delivery. Curr Therapeutic Res 23(5):596
607.
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(8):503
509.
Gulyaev AE, Gelperina SE, Skidan IN, Antropov AS, Kivman
GY, Kreuter J. 1999. Significant transport of doxorubicin into
the brain with polysorbate 80-coated nanoparticles. Pharm Res
16:1564
1569.
Halford B. 2006. Fullerene for the face. Chemical & Engineering
News 84:47.
Heckel K, Kiefmann R, Dorger M, Stoeckelhuber M, Goetz AE.
2004. Colloidal gold particles as a new in vivo marker of early
acute lung injury. Am J Physiol Lung Cell Mol Physiol
287:L867
878.
Hoet PHM, Bruske-Hohlfeld I, Salata OV. 2004. Nanoparticles

known and unknown health risks. J Nanobiotechnol 2(12):12

27.
Hoshino A, Fujioka K, Oku T, Suga M, Sasaki YF, Ohta T,
Yasuhara M, Suzuki K., Yamamoto K. 2004. Physicochemical
properties and cellular toxicity of nanocrystal quantum dots
depend on their surface modification. Nano Lett 4(11):2163

2169.
Howe HA, Bodian D. 1940. Portals of entry of poliomyelitis virus
in the chimpanzee. Proc Soc Exp Biol Med 43:718
721.
Huang L, Li DQ, Lin YJ, Wei M, Evans DG, Duan X. 2005.
Controllable preparation of Nano-MgO and investigation of its
bactericidal properties. J Inorg Biochem 99:986
993.
IARC. 2006. Baan R, Straif K, Grosse Y, et al. on behalf of the
WHO International Agency for Research on Cancer Mono-
graph Working Group. Carcingenicity of carbon black; titanium
dioxide, and talc. The Lancet 7:295
296.
International Commission on Radiological Protection (ICRP).
1994. Annals of the ICRP: Human respiratory tract model for
radiological protection. Smith H, editor. ICRP Publication 66.
Vol. 24, Nos. 1
3, Oxford, UK: Pergamon.
International Commission on Radiological Protection (ICRP).
1966. Deposition and retention models for internal dosimetry
of the human respiratory tract. Task Group on Lung Dynamics.
Health Phys 12:173
208.
International Life Sciences Institute (ILSI). 2005. Oberdörster et
al. and a report from the ILSI Research Foundation/Risk
Science Institute Nanomaterial Toxicity Screening Working
Group. Principles for characterizing the potential human health
effects from exposure to nanomaterials: Elements of a screening
strategy. Particle Fibre Toxicol 2005(2):8.
Ito A, Kuga Y, Honda H, Kikkawa H, Horiuchi A, Watanabe Y,
Kobayashi T. 2004. Magnetite nanoparticle-loaded anti-HER2
immunoliposomes for combination of antibody therapy with
hyperthermia. Cancer Lett 212:167
175.
Jackson BP, Ranville JF, Bertsch PM, Sowder AG. 2005.
Characterization of colloidal and humic-bound Ni and U in
the ‘‘dissolved’’ fraction of contaminated sediment extracts.
Environ Sci Technol 39:2478
2485.
Janssen YM, Barchowsky A, Treadwell M, Driscoll KE, Mossman
BT. 1995. Asbestos induces nuclear factor kappa B (NF-kappa
B) DNA-binding activity and NF-kappa B-dependent gene
expression in tracheal epithelial cells. Proc Natl Acad Sci USA
92(18):8458
8462.
Jia G, Wang H, Yan L, Wang X, Pei R, Yan T, Zhao YL, Guo X.
2005. Cytotoxicity of carbon nanomaterials: Single-wall nano-

tube, multi-wall nanotube, and fullerene. Environ Sci Technol
39:1378
1383.
Johnson NF. 1994. Phagosomal pH and glass fiber dissolution in
cultured nasal epithelial cells and alveolar macrophages: a
preliminary study. Environ. Health Perspectives 102(suppl. 5):
97
102.
Jötten KW, Klosterkötter W. 1952. Die Bedeutung der Löslichkeit
der Kieselsäure in der Lunge von Kaninchen. Arch Hyg
136:451
467.
Kagan VE, Tyurina YY, Tyurin VA, Konduru NV, Potapovich AI,
Osipov AN, Kisin ER, Schwegler-Berry D, Mercer R, Castra-
nova V, Shvedova AA. 2006. Direct and indirect effects of
single walled carbon nanotubes on RAW 264.7 macrophages:
Role of iron. Toxicol Lett 165(1):88
100.
Kamat JP, Devasagayam TPA, Priyadarsini KI, Mohan H, Mittal
JP. 1998. Oxidative damage induced by the fullerene C-60 on
photosensitization in rat liver microsomes. Chemico-Biological
Interactions 114:145
159.
Kanti ST, Khilar KC. 2006. Review on subsurface colloids and
colloid-associated contaminant transport in saturated porous
media. Adv Colloid Interface Sci 119:71
96.
Kato T, Yashiro T, Murata Y, Herbert DC, Oshikawwa K, Bando
M, Ohno S, Sugiyama Y. 2003. Evidence that exogenous
substances can be phagocytized by alveolar epithelial cells and
transported into blood capillaries. Cell Tiss Res 311:47
51.
Katz LC, Burkhalter A, Dreyer WJ. 1984. Fluorescent latex
microspheres as a retrograde neuronal marker for in vivo and in
vitro studies of visual cortex. Nature 310:498
500.
Kausche GA, Pfankuch E, Ruska H. 1939. Die Sichtbarmachung
von pflanzlichem Virus im Übermikroskop. Naturwissenschaf-
ten 27:292
299.
Kennedy TP, Dodson R, Rao NV, Hopkins C, Baser M, Tolley E,
Hoidal J. 1989. Dusts causing pneumoconiosis generate OH
and produce hemolysis by acting as Fenton catalysts. Archives
of Biochemistry and Biophysics 269:359
364.
Kirchner C, Liedl T, Kudera S, Pellegrino T, Javier AM, Gaub
HE, Stolzle S, Fertig N, Parak WJ. 2005. Cytotoxicity of
colloidal CdSe and CdSe/ZnS nanoparticles. Nano Lett
5(2):331
338.
Konig MF, Lucocq JM, Weibel ER. 1993. Demonstration of
pulmonary vascular perfusion by electron and light micro-
scropy. J Appl Physiol 75(4):1877
1883.
Kreuter J. 2001. Nanoparticulate systems for brain delivery of
drugs. Adv Drug Delivery Rev 47:65
81.
Kreyling W, Semmler M, Erbe, Mayer P, Takenaka S, Schulz H,
Oberdörster G, Ziesenis A. 2003. Translocation of ultrafine
insoluble iridium particles from lung epithelium to extrapul-
monary organs is size dependent but very low. J Toxicol
Environ Health 65A(20):1513
1530.
Kreyling WG, Semmler M, Moller W. 2004. Dosimetry and
toxicology of ultrafine particles. J Aerosol Med 17(2):140
152.
Kroto HW, Heath JR, Obrien SC, Curl RF, Smalley RE. 1985.
C-60-Buckminsterfullerene. Nature 318:162
163.
Lam CW, James JT, McCluskey R, Hunter RL. 2004. Pulmonary
toxicity of single-wall carbon nanotubes in mice 7 and 90 days
after intratracheal instillation. Toxicological Sci 77:126
134.
Lam CW, James J, McCluskey R, Sivaram A, Hunter R. 2006. A
review of carbon nanotube toxicity and assessment of potential
occupational and environmental health risks. Crit Rev Toxicol
36(3):189
217.
Lead JR, Muirhead D, Gibson CT. 2005. Characterization of
freshwater natural aquatic colloids by atomic force microscopy
(AFM). Environ Sci Technol 39:6930
6936.
Lee S-H. 2005. Inactivation of bacterial endospores by photo-
catalytic nanocomposites. Colloid Surface 40:93
98.
Li N, Sioutas C, Cho A, Schmiz D, Misra C, Sempf J, Wang M,
Oberley T, Froines JR, Nel A. 2003. Ultrafine particulate
Toxicology of nanoparticles: A historical perspective 23
pollutants induce oxidative stress and mitochondrial damage.
Environmental Health Perspectives 111(4):455
460.
Lisunova MO, Lebovka NI, Melezhyk OV, Boiko YP. 2006.
Stability of the aqueous suspensions of nanotubes in the
presence of nonionic surfactant. J Colloid Interface Sci
299:740
746.
Lovern SB, Klaper RD. 2006. Daphia magna mortality when
exposed to titanium dioxide and fullerene (C60) nanoparticles.
Environ Toxicol Chem 25:1132
1137.
Lundborg M, Eklund A, Lind B, Camner P. 1985. Dissolution of
metals by human and rabbit alveolar macrophages. Brit J Ind
Med 42:642
645.
Malsch I, Oud M. 2004. Outcome of the Open Consultation on
the European Strategy for Nanotechnology. December, 2004;
www.nanoforum.org
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 A67:87

107.
McCarthy JF, Yurek GJ, Elliott JF, Amdur MO. 1982. Generation
and characterization of submicron aerosols of zinc oxide. Am
Ind Hyg Assoc J 43:880
886.
McCarthy JF, Zachara JM. 1989. Subsurface transport of con-
taminants: mobile colloids in the subsurface environment may
alter the transport of contaminants. Environ Sci Technol
23:496
502.
McConnell EE, Axten C, Hesterberg TW, Chevalier J, Miller
WC, Everitt J, Oberdöster G, Chase GR, Thevenaz P, Kotin P.
1999. Studies on the inhalation toxicology of two Fiberglasses
and Amosite Asbestos in the Syrian Golden Hamster. Part II.
Results of chronic exposure 1. Inhal Toxicol 11(9):785
835.
Mehta D, Bhattacharya J, Matthay MA, Malik AB. 2004.
Integrated control of lung fluid balance. Am J Physiol Lung
Cell Mol Physiol 287:L1081
1090.
Meyer M, Schreck R, Baeuerle PA. 1993. H2O2 and antioxidants
have opposite effects on activation of NF-kappa B and AP-1 in
intact cells: AP-1 as secondary antioxidant-responsive factor.
EMBO J 12(5):2005
2015.
Mills A, Hunte SLJ. 1997. An overview of semiconductor
photocatalysis. J Photochem Photobiol A: Chem 108:1
35.
Mills NL, Amin N, Robinson SD, Anand A, Davies J, Patel D, de
la Fuente JM, Cassee FR, Boon NA, MacNee W, Millar AM,
Donaldson K, Newby DE. 2006. Do inhaled carbon nanopar-
ticles translocate directly into the circulation in humans? Am J
Respir Crit Care Med 173:426
431.
Mossman B, Landesman J. 1983. Importance of oxygen free
radicals in asbestos-induced injury to airway epithelial cells.
CHEST 83:50S
51.
Mossman B, Marsh J, Shatos M. 1986. Alteration of superoxide
dismutase activity in tracheal epithelialcells by asbestos and
inhibition of cytotoxicity by antioxidants. Lab Invest 54:204

212.
Mossman BT, Marsh JP, Sesko A, Hill S, Shatos MA, Doherty J,
Petruska J, Adler KB, Hemenway D, Mickey R, Vacek P, Kagan
E. 1990. Inhibition of lung injury, inflammation, and interstitial
pulmonary fibrosis by polyethylene glycol-conjugated catalase
in a rapid inhalation model of asbestosis. Am Rev Respiratory
Dis 141:1266
1271.
Müller RH, Keck CM. 2004. Drug delivery to the brain

realization by novel drug carriers. J Nanosci Nanotechnol
4:471
483.
Muller J, Huaux F, Moreau N, Misson P, Heilier J-F, Delos M,
Arras M, Fonseca A, Nagy JB, Lison D. 2005. Respiratory
toxicity of multi-all carbon nanotubes. Toxicol Appl Pharmacol
207:221
231.
24 G. Oberdörster et al.
National Nanotechnology Initiative (NNI). 2006. NNI: Environ-
mental, Health and Safety Research Needs for Engineered
Nanoscale Materials. NNI White paper issued by the Office of
the President of the United States, September, 2006.
Nel A, Xia T, Mädler L, Li N. 2006. Toxic potential of materials
at the nano level. Science 311(5761):622
627.
Nemmar A, Hoet PHM, Vanquickenborne B, et al. 2002. Passage
of inhaled particles into the blood circulation in humans.
Circulation 105:411
414.
Nemmar A, Hoylaerts MF, Hoet PHM, Nemery B. 2004.
Possible mechanism of the cardiovascular effects of inhaled
particles: Systemic translocation and prothrombotic effects.
Toxicol Lett 149:243
253.
Oberdörster E. 2004. Manufactured nanomaterials (Fullerenes,
C60) induce oxidative stress in the brain of juvenile largemouth
bass. Environ Health Perspect 112:1058
1062.
Oberdörster G, Hochrainer D, Ma RH. 1979. Zinc oxide aerosols:
Generation, lung clearance and effects of lung clearance.
Gesellschaft für Aerosolforschung 7:132
137.
Oberdörster G, Ferin J, Finkelstein J, Wade P, Corson N. 1990.
Increased pulmonary toxicity of ultrafine particles? II. Lung
lavage studies. J Aerosol Sci 21:384
387.
Oberdörster G, Ferin J, Gelein R, Soderholm SC, Finkelstein J.
1992. Role of the alveolar macrophage in lung injury: Studies
with ultrafine particles. Environ Health Perspect 97:193
197.
Oberdörster G, Ferin J. 1992. Metal compounds used in new
technologies: Metal oxides of ultrafine particles have increased
pulmonary toxicity. In: Merian E, Haerdi W, editors. Metal
compounds in environment & life. Vol. 4. Science & Technol-
ogy Letters. Northwood, UK: Science Reviews. pp 443
450.
Oberdörster G, Gelein R, Ferin J, Weiss B. 1995. Association of
particulate air pollution and acute mortality: Involvement of
ultrafine particles? Inhalation Toxicol 7:111
124.
Oberdörster G, Sharp Z, Atudorei V, Elder A, Gelein R, Kreyling
W, Cox C. 2004. Translocation of inhaled ultrafine particles to
the brain. Inhalation Toxicol 16(6/7):437
445.
Oberdörster G, Oberdörster E, Oberdörster J. 2005. Invited
review: Nanotoxicology: An emerging discipline evolving from
studies of ultrafine particles. Environ Health Perspect
113:823
839.
Oberdörster E, Zhu S, Blickley TM, McClellan-Green P, Haasch
ML. 2006. Ecotoxicology of carbon-based engineered nano-
particles: Effects of fullerene (C60) on aquatic organisms.
Carbon 44(6):1112
1120.
Panté N, Kann M. 2002. Nuclear pore complex is able to
transport macromolecules with diameters of 39 nm. Molec
Biol Cell 13:425
434.
Peters A, von Klot S, Heier M, Trentinaglia I, Hörmann A,
Wichmann HE, Löwel H. 2004. Exposure to traffic and the
onset of myocardial infarction. N Eng J Med 351:1721
1730.
Quinlan TR, Marsh JP, Janssen YW, Borm PA, Mossman BT.
1994. Oxygen radicals and asbestos-mediated disease. Environ
Health Perspect 102:107
110.
Roco MC. 2003a. The future of the National Nanotechnology
Initiative. (http://www.nano.gov/html/res/slides.pdf).
Roco MC. 2003b. Converging science and technology at the
nanoscale: Opportunities for education and training. Nat
Biotechnol 21:1247
1249.
Rosenkranz P, Fernandes TF, Ford AT, Chaudhry Q, Stone V.
Uptake and fate of fluorescent nanoparticles in aquatic
invertebrates. SETAC abstract, The Hague May 2006.
Rouse JG, Yang J, Ryman-Rasmusen JP, Barrow AR, Monteior-
Riviere NA. 2007. Effects of mechanical flexion on the
penetration of fullerene amino acid-derivatized peptide nano-
particles through skin. NanoLetters 7(1):155
160.
Royal Society and Royal Academy of Engineers. 2004. Na-
noscience and nanotechnologies: opportunities and uncertain-
ties. 2004. London, The Royal Society.
Rückerl R, Ibald-Mulli A, Koenig W, Schneider A, Woelke G,
Cyrys J, Heinrich J, Marder V, Frampton M, Wichmann HE,
Peters A. 2006. Air pollution and Markers of inflammation and
coagulation in patients with coronary heart disease. Am J
Respir Crit Care Med 173:432
441.
Ruska H, von Borries B, Ruska E. 1940. Die Bedeutung der
Übermikroskopie für die Virusforschung. Arch Ges Virusforsch
1:155
169.
Ryman-Rasmussen JP, Riviere JE, Monteiro-Riviere NA. 2006.
Penetration of intact skin by quantum dots with diverse
physiochemical properties. Tox Sci 91:159
165.
Scientific Committee on Emerging and Newly Identified Health
Risks (SCENIHR). 2005. http://files.nanobio-raise.org/Down-
loads/scenihr.pdf accessed 22/01/2007.
Sato Y, Yokoyama A, Shibata K, Akimoto Y, Ogino S, Nodasaka
Y, Kohgo T, Tamura K, Akasaka T, Uo M, Motomiya K,
Jeyadevan B, Ishiguro M, Hatakeyama R, Watari F, Tohji K.
2005. Influence of length on cytotoxicity of multi-walled
carbon nanotubes against human acute monocytic leukemia
cell line THP-1 in vitro and subcutaneous tissue of rats in vivo.
Mol Biosyst 1(2):176
182.
Sayes CM, Fortner JD, Guo W, Lyon D, Boyd AM, Ausman KD,
Tao YJ, Sithaaraman B, Wilson LJ, Hughes JB, West JL, Colvin
VL. 2004. The differential cytotoxicity of water-soluble full-
erenes. Nano Lett 4(10):1881
1887.
Sayes CM, Liang F, Hudson JL, Mendez J, Guo W, Beach JM,
Moore VC, Doyle CD, West JL, Billups WE, Ausman KD.
2006. Functionalization density dependence of single-walled
carbon nanotubes cytotoxicity in vitro. Toxicol Lett 161(2):
135
142.
Schiller CF, Gebhart J, Heydeer J, Rudolf G, Stahlhofen W. 1986.
Factors influencing total deposition of ultrafine aerosol parti-
cles in the human respiratory tract. J Aerosol Sci 17(3):328

332.
Schreck R, Albermann K, Baeuerle PA. 1992. Nuclear factor
kappa B: an oxidative stress-responsive transcription factor of
eukaryotic cells (a review). Free Radic Res Commun
17(4):221
237.
Seaton A, MacNee W, Donaldson K, Godden D. 1995. Particu-
late air pollution and acute health effects. Lancet 345(Jan.
21):176
178.
Selikoff IJ, Lee DHK. 1978. Asbestos and disease. New York:
Academic Press.
Semmler M, Seitz J, Erbe F, Mayer P, Heyder J, Oberdörster G,
Kreyling W. 2004. Long-term clearance kineic of inhaled
ultrafine insoluble iridium particles from the rat lung, including
transient translocation into secondary organs. Inhalation Tox-
icol 16(6/7):453
459.
Shen Z, Parker VD, Aust AE. 1995. Mediated, thin-layer cell,
coulometric determination of redox-active iron on the surface
of asbestos fibers. Anal Chem 67(2):307
311.
Shvedova AA, Kisin ER, Mercer R, Murray AR, Johnson VJ,
Potapovich AI, Tyurina YY, Gorelik O, Arepalli S, Schwegler-
Berry D, Hubbs AF, Antonini J, Evans DE, Ku BK, Ramsey D,
Maynard A, Kagan VE, Castranova V, Baron P. 2005. Unusual
inflammatory and fibrogenic pulmonary responses to single-
walled carbon nanotubes in mice. Am J Physiol Lung Cell Mol
Physiol 289(5):L698
708.
Silva VM, Corson N, Elder A, Oberdörster G. 2005. The rat ear
vein model for investigating in vivo thrombogenicity of ultrafine
particles (UFP). Toxicological Sci 85:983
989.
Smith AE, Helenius A. 2004. How viruses enter animal cells.
Science 304:237
242.

Sondi I, Salopek-Sondi B. 2004. Silver nanoparticles as anti-
microbial agent: A case study on E. coli as a model for Gram-
negative bacteria. J Colloid Interface Sci 275:177
182.
Spadaro JA, Berger TJ, Barranco SD, Chapin SE, Becker RO.
1974. Antibacterial effects of silver electrodes with weak-direct
current. Microb Agents Chemother 6:637
642.
Stanley WM. 1935. Isoltion of a crystalline protein assessing the
properties of tobacco mosaic virus. Science 81(2113):644

645.
Stone V, Shaw J, Brown DM, MacNee W, Faux SP, Donaldson K.
1998. The role of oxidative stress in the prolonged inhibitory
effect of ultrafine carbon black on epithelial cell function.
Toxicol In Vitro 12:649
659.
Sunada K, Kikuchi Y, Hashimoto K, Fujishima A. 1998.
Bactericidal and detoxification effects of TiO2 thin film
photocatalysts. Environ Sci Technol 32:726
728.
Swift DL, Montassier N, Hopke PK, Karpen-Hayes K, Cheng Y-
S, Su YF, Yeh HC, Strong JC. 1992. Inspiratory deposition of
ultrafine particles in human nasal replicate cast. J Aerosol Sci
23:65
72.
Swift DL, Montassier N, Hopke PK, Karpen-Hayes K, Cheng Y-
S, Su YF, Yeh HC, Strong JC. 1994. Inspiratory deposition of
ultrafine particles in human nasal replicate cast. J Aerosol Sci
23:65
72.
Teague SV, Raabe OG. 1980. Generation of fume aerosols of zinc
oxide. Am Ind Hyg Assoc J 41:680
683.
Timbrell V. 1982. Deposition and retention of fibres in the human
lung. Ann Occup Hyg 26:347
369.
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.
Tratnjek PG, Johnson RL. 2006. Nanotechnologies for environ-
mental cleanup. Nanotoday 1:44
48.
Tremblay JF. 2003. Fullerenes by the ton. Chemical & Engineer-
ing News 81:13
14.
Tsoli M, Kuhn H, Brandau W, Esche H, Schmid G. 2005.
Cellular uptake and toxicity of Au55 clusters. Small 1(8

9):841
844.
Vicent MJ, Duncan R. 2006. Polymer conjugates: Nanosized
medicines for treating cancer. Trends Biotechnol 24:39
47.
Vogelson CT. 2001. Advances in drug delivery systems. Mod
Drug Discov 4:49
50.
Wagner JC, Skidmore JW, Hill RJ, Griffiths DM. 1985. Erionite
exposure and mesotheliomas in rats. Br J Cancer 51(5):727

730.
Wang IC, Tai LA, Lee DD, Kanakamma PP, Shen CK, Luh TY,
Cheng CH, Hwang KC. 1999. C(60) and water-soluble
View publication stats
Toxicology of nanoparticles: A historical perspective 25
fullerene derivatives as antioxidants against radical-initiated
lipid peroxidation. J Med Chem 42(22):4614
4620.
Wang ZL. 2004. Functional oxide nanobelts: Materials, proper-
ties and potential applications in nanosystems and biotechnol-
ogy 4261. Annu Rev Phys Chem 55:159
196.
Warheit DB, Chang LY, Hill LH, Hook GR, Crapo JD, Brody AR.
1984. Pulmonary macrophage accumulation and asbestos-
induced lesions at sites of fiber deposition. Am Rev Respiratory
Dis 129:301
310.
Warheit DB, Driscoll KE, Oberdoerster G, Walker C, Kuschner
M, Hesterberg TW. 1995. Contemporary issues in fiber
toxicology. Fundamental & Applied Toxicology 25(2):171

183.
Warheit DB, Laurence BR, Reed KL, Roach DH, Reynolds
GAM, Webb TR. 2004. Comparative pulmonary toxicity
assessment of single-wall carbon nanotubes in rats. Toxicolo-
gical Sci 77:117
125.
Warheit DB, Webb TR, Sayes CM, Colvin VL, Reed KL. 2006.
Pulmonary instillation studies with nanoscale TiO2 rods and
dots in rats: Toxicity is not dependent upon particle size and
surface area. Toxicol Sci 91(1):227
236.
Whitby KT, Clark WE, Marple VA, Sverdrup GM, Sem GJ,
Willeke K, Liu BYH, Pui DYH. 1975. Characterization of
California aerosols Size distribution of freeway aerosol. Atmo-
spheric Environ 9:463
482.
Wilson MR, Donaldson K, Stone V. 2001. Oxidative interactions
between ultrafine particles and metals. Am J Resp Crit Care
Med 163(5):A495 p.
Xiao L, Takada H, Maeda K, Haramoto M, Miwa N. 2005.
Antioxidant effects of water-soluble fullerene derivatives
against ultraviolet ray or peroxylipid through their action of
scavenging the reactive oxygen species in human skin kerati-
nocytes. Biomed Pharmacother 59(7):351
358.
Yang L, Watts DJ. 2005. Particle surface characteristics may play
an important role in phytotoxicity of alumina nanoparticles.
Toxicol Lett 158:122
132.
Zhang W-S. 2003. Nanoscale iron particles for environmental
remediation: An overview. J Nanopart Res 5:323
332.
Zhang Q, Kusaka Y, Sato K, Nakakuki K, Kohyama N,
Donaldson K. 1998. Differences in the extent of inflammation
caused by intratracheal exposure to three ultrafine metals: Role
of free radicals. J Toxicol Environ Health 53(6):423
438.
Zhu S, Oberdörster E, Haasch ML. 2006. Toxicity of an
engineered nanoparticle (fullerene, C(60)) in two aquatic
species, Daphnia and fathead minnow. Mar Environ Res
Zhuang J, Qi J, Jin Y. 2005. Retention and transport of
amphiphilic colloids under unsaturated flow conditions: Effect
of particle size and surface property. Environ Sci Technol
39:7853
7859.