Nanooncology Engineering nanomaterials for cancer therapy
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Gil Gonçalves Gerard Tobias
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Nanooncology
Engineering Nanomaterials for Cancer
Therapy and Diagnosis
123
Editors
Gil Gonçalves Gerard Tobias
Institut de Ciència de Materials Institut de Ciència de Materials
de Barcelona (ICMAB-CSIC) de Barcelona (ICMAB-CSIC)
Bellaterra Bellaterra
Spain Spain
ISSN 2194-0452 ISSN 2194-0460 (electronic)
Nanomedicine and Nanotoxicology
ISBN 978-3-319-89877-3 ISBN 978-3-319-89878-0 (eBook)
https://doi.org/10.1007/978-3-319-89878-0
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Contents
Gold Nanoparticles for Imaging and Cancer Therapy . . . . . . . . . . . . . . 1
Marc-André Fortin, Teresa Simão and Myriam Laprise-Pelletier
Liposomes-Based Nanoparticles for Cancer Therapy
and Bioimaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Calvin Cheung and Wafa T. Al-Jamal
Quantum Dots for Cancer Therapy and Bioimaging . . . . . . . . . . . . . . . 89
Fu-Gen Wu, Xiaodong Zhang, Xiaokai Chen, Wei Sun, Yan-Wen Bao,
Xian-Wu Hua, Ge Gao and Hao-Ran Jia
Polymeric Nanoparticles for Cancer Therapy and Bioimaging . . . . . . . . 137
Eva Espinosa-Cano, Raquel Palao-Suay, María Rosa Aguilar,
Blanca Vázquez and Julio San Román
Imaging and Treating Cancer with Carbon
Nanotube Technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 173
Nicholas G. Zaibaq, Sakineh E. Moghaddam and Lon J. Wilson
Micellar-Based Nanoparticles for Cancer Therapy
and Bioimaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 211
Fernanda Andrade, Andreia Almeida, Diana Rafael,
Simo Schwartz, Jr and Bruno Sarmento
Magnetic Nanoparticles for Cancer Therapy and Bioimaging . . . . . . . . 239
Ester Polo, Pablo del Pino, Alberto Pardo, Pablo Taboada
and Beatriz Pelaz
Dendrimers-Based Nanoparticles for Cancer Therapy
and Bioimaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 281
Jørn B. Christensen
Porous Silicon Particles for Cancer Therapy and Bioimaging . . . . . . . . 305
Steven J. P. McInnes, Abel Santos and Tushar Kumeria
v
vi Contents
Metal/Metal Oxide Nanoparticles for Cancer Therapy . . . . . . . . . . . . . . 341
M. P. Vinardell and M. Mitjans
Reconfigurable Nucleic Acid Materials for Cancer Therapy . . . . . . . . . 365
Morgan Chandler, Weina Ke, Justin R. Halman, Martin Panigaj
and Kirill A. Afonin
Fullerenes for Cancer Therapy and Bioimaging . . . . . . . . . . . . . . . . . . . 387
Xuejiao J. Gao, Xiaomei Shen, Gengmei Xing and Xingfa Gao
Carbon Nano-onions for Bioimaging and Cancer Therapy
Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 417
Adalberto Camisasca and Silvia Giordani
Gold Nanoparticles for Imaging
and Cancer Therapy
Marc-André Fortin, Teresa Simão and Myriam Laprise-Pelletier
Abstract Gold nanoparticles have properties useful in biomedical imaging and
cancer therapy. This biocompatible metal has been used for centuries in medicine.
In the last 20 years, the rapid developments in nanotechnology have revealed several
applications of nanosized gold, which are now being evaluated for clinical proce-
dures. For instance, gold nanoparticles can be used to develop sensors due to their
optical properties; they also make possible the development of new hyperthermia
and drug delivery treatments. However, gold nanoparticles could find more imme-
diate and direct applications in medical physics procedures, such as X-ray imaging
and radiotherapy. First, this chapter provides an overview of the different synthesis
routes for the production of biomedical gold nanoparticles. Then, an overview of the
physical principles of photon–matter interactions, that are fundamental to the con-
cept of X-ray attenuation in biological tissues, is presented. The properties of gold
nanoparticles as contrast agents for X-ray and computed tomography (CT) imaging
are reviewed, along with the principles of the radiosensitization effect useful in medi-
cal physics and oncology. The main mechanisms leading to dose enhancement, to cell
damage and to cell death, are described in the light of the specific interactions taking
place between ionizing photons and high-Z materials such as gold (Au) when these
are distributed in biological tissues such as tumours. Finally, the performance of gold
nanoparticles as CT contrast agents and radiosensitizers in oncology is discussed, in
the perspective of their consideration for clinical applications.
Keywords Gold nanoparticles · Radiotherapy · Brachytherapy · Computed
tomography · Radiosensization · Cancer treatment · Theranostics
M.-A. Fortin (B) · T. Simão · M. Laprise-Pelletier
Centre de Recherche du Centre Hospitalier, Universitaire de Québec-Université Laval (CR CHU
de Québec), axe Médecine Régénératrice, Québec G1L 3L5, QC, Canada
e-mail:
[email protected]M.-A. Fortin · T. Simão · M. Laprise-Pelletier
Department of Mining, Metallurgy and Materials Engineering, Université Laval, Québec, QC
G1V 0A6, Canada
M.-A. Fortin · T. Simão · M. Laprise-Pelletier
Centre de Recherche sur les Matériaux Avancés (CERMA), Université Laval, Québec, QC G1V
0A6, Canada
© Springer International Publishing AG, part of Springer Nature 2018 1
G. Gonçalves and G. Tobias (eds.), Nanooncology, Nanomedicine
and Nanotoxicology, https://doi.org/10.1007/978-3-319-89878-0_1
2 M.-A. Fortin et al.
1 Introduction
Over the last decades, gold nanoparticles (Au NPs) have emerged as a new promising
material for a large variety of medical applications [13, 41, 51, 52, 55, 78, 95, 97,
100, 120, 131, 142, 176, 178, 190, 197, 199, 200]. Au NPs bear several physico-
chemical properties useful for the development of diagnosis tools as well as for
imaging contrast agents. They can also be used as radiosensitizers: these are products
that, when they are injected in biological tissues, have the capacity to increase the
impact of radiotherapy [25, 39, 45, 76, 111, 204, 208]. Finally, nanosized gold is
resistant to oxidation in biological media and overall, it is accepted by the cells at
relatively high concentrations [64, 95, 113, 173].
A very large number of studies and reviews have been written about the properties
of Au NPs used as ‘plasmonic’ materials [57, 186, 200]. Plasmons are oscillations
of free electrons taking place at the surface of conductor nanomaterials (such as
gold). The surface electrons of Au NPs can couple with electromagnetic radiation
of certain wavelengths that are far larger than the particle. Plasmonic nanoparti-
cles exhibit interesting scattering, absorbance and coupling properties, and several
proof-of-concept studies useful to the biomedical field have been made by exploiting
the resonant properties of Au NPs irradiated by visible and near-infrared light. For
instance, new sensor technologies have been developed relying on the principle that
molecules reacting with a specific molecular function present at the surface of Au
NPs, induce slight changes to specific resonance peaks. These can be sensitively
detected with appropriate spectroscopic tools. The resonant properties of Au NPs
under visible and infrared light can also be exploited to increase the temperature of
cells, a phenomenon referred to as ‘hyperthermia’ [53, 182].
Although the plasmonic characteristics of Au NPs appear very useful for in vitro
applications, their potential for in vivo procedures has always been somewhat limited
to skin diseases and to superficial cancer (e.g. skin cancer). In fact, visible and near-
infrared photons diffuse strongly in the biological tissues. Diffusion limits the depth
of penetration of such low-energy photons in biological tissues to a few millimetres
only. Therefore, to be useful for a large variety of applications in vivo, Au NPs must
preferably be irradiated with mid-to-high energy electromagnetic radiation—mainly
photons—that have the capacity to penetrate deep enough in the body.
In fact, biomedical imaging and cancer treatment by radiotherapy are two areas of
medicine where the injection of Au NPs in vivo could be recommended for clinical
diagnoses and therapies [45, 70, 71, 76]. High-energy photons (from 1 keV and
above) are already exploited in medicine. This is precisely the range of energies
useful to medical physics. Irradiated with photons in the ionizing energy range, Au
NPs can be used as effective contrast agents for X-ray computed tomography (CT),
as well as for ‘boosting’ the radiation dose delivered in radiotherapy treatments [25,
84, 131].
Therefore, in vivo applications of Au NPs based on their interactions with higher
energy photons, have emerged in modern medicine and in particular for the treatment
of cancer. Au NPs can be used to enhance the differences of density between biologi-
Gold Nanoparticles for Imaging and Cancer Therapy 3
cal tissues (e.g. using X-ray and CT imaging). They can also be used in radiosensitiz-
ing procedures to induce more damages to cells irradiated with radiotherapy beams or
with internal radioactive sources. In fact, Au NPs are a very dense material; they can
attenuate high-energy photons in the biological tissues even if they are distributed at
relatively moderate concentrations. Also, photons interacting with high-Z materials
such as gold generate many by-products such as low-energy photons and electrons.
These ‘secondary products’ have the capacity to increase the therapeutic impact of
the primary photons beams, as well as the radioactive sources used in radiotherapy.
In medical physics, Au NPs are often referred to as ‘radiosensitizers’, i.e. products
that have the capacity to enhance radiotherapy treatments.
This chapter begins with an introduction to the different synthesis routes leading
to the production of Au NPs, as well as to their surface functionalization. Then, an
overview about the physical mechanisms by which photons (~10–500 keV) interact
with the atoms present in biological tissues is given. Photons of energy higher than
10 keV can penetrate deep enough in the biological tissues (e.g. sub-visible light
wavelength range), and therefore can be exploited either for imaging or therapeutic
applications. Finally, the applications of Au NPs as contrast agents for CT (X-ray
imaging), and as radiosensitizers in radiotherapy (medical physics in oncology), are
presented in the context of their ongoing evaluation towards clinical applications.
2 Synthesis of Gold Nanoparticles for Biomedical
Applications
Colour change is the most evident signature of colloidal gold. Depending on the
size of the nanoparticles, gold presents either an intense red colour for small to
medium-sized particles, purple for larger particles and blue for aggregates [52, 78].
Because the properties and the applications of Au NPs so closely depend on their
size and shape, the colour of aqueous suspensions of Au NPs provides very efficient
indications to the chemist at every step of the nanoparticle preparation.
The first experiments for the synthesis of gold sols under controlled conditions
were reported by Michael Faraday in the 1850s [58, 152]. The scientist accidentally
generated a ruby red solution while mounting pieces of gold leaves onto microscope
slides. He referred to this solution as ‘activated gold’. Then, he used phosphorus
to reduce a solution of gold chloride. Already interested in the properties of light
and matter, Faraday further investigated the optical properties of these colloidal gold
solutions. For a long time, the composition of ‘ruby’ or ‘activated’ gold was unclear,
but the colour-particle size relationship was already acknowledged. The studies on Au
NPs did not make significant advances until the end of the twentieth century, with the
exponential development of advanced analytical technologies, such as atomic force
microscopy and electron microscopy. Due to their relatively easy synthesis routes,
high stability and high density per particle, colloidal gold has also been used in the
4 M.-A. Fortin et al.
development of physical separation techniques such as ultracentrifugation, as well
as electron microscopy dyes.
Gold nanoparticles produced for biomedical applications are usually generated
from precursor solutions containing chloroauric acid (H[AuCl4 ]). After H[AuCl4 ]
is dissolved in water, the solution is rapidly stirred while a reducing agent is added.
To prevent the particles from aggregating, a stabilizing agent is usually added for
coating the nanoparticle surface. A comprehensive review of the different classes of
Au NPs synthesis was written by Daniel et al. [48]. A selection of the most significant
ones is reviewed here.
2.1 Direct Reduction: The Turkevitch Method
The chemical reduction of AuCl4 by citric acid in a hot aqueous solution was inves-
tigated in 1951 [185]. In this synthesis, citrate acts as the reducing agent, then as
the capping agent that stabilizes the Au NPs through electrostatic interactions. The
Turkevitch method allows the growth of NPs with many different shapes and diam-
eters usually ranging from 10 to 50 nm (Fig. 1). A significant increase in the particle
diameters is observed at lower citrate concentration, leading to the generally accepted
conclusion that the total particle surface area is determined by the number of citrate
ions available to cover it [65].
More recently, an alternative approach showed that a high concentration of Cl−
ions also leads to larger sizes for Au NPs reduced by citric acid, since it decreases
their surface charge and thereby promotes coarser particle size and even aggregation
[205]. A similar result was obtained by increasing the pH of the NP solution just
after initiating the reduction step. An increase of the pH limits the nucleation process
and decreases the final number of grown Au NPs [167]. Although simple, the direct
chemical reduction synthesis route usually leads to the production of NPs of relatively
large sizes, showing anisotropy and a certain degree of size polydispersity.
2.2 Seed-Mediated Growth for Smaller and Narrower
Particle Size Distributions
The constantly increasing demand for Au NPs in photonics, biology and medicine,
has led to the development of new synthesis routes for achieving better control over
size distributions. One of the most promising approaches in this goal is the seed-
mediated growth technique. In this concept, small nanoparticle seeds of a typical size
smaller than 15 nm, act as catalytic nucleation centres for the growth of larger NPs
[86]. The nucleation of very small NP seeds is usually made through the reduction
of gold ions by sodium borohydride (NaBH4 ), a relatively toxic reducing agent that
is tolerated as an initiator of Au NPs seeds. However, it is not indicated in the final
Gold Nanoparticles for Imaging and Cancer Therapy 5
synthesis steps of biomedical Au NPs. Shape uniformity in the seed suspension is
of paramount importance to achieve both shape and size uniformity of the final NPs
[141].
Fig. 1 Gold nanoparticles synthesized by the Turkevitch method in citrate: a, b as-synthesized; c,
d grafted with polyethylene glycol PEG 1000 mw; and e, f grafted with polyethylene glycol PEG
5000 mw. The longer the chains, the larger the spacing between the particles. Source The authors,
following procedures adapted from [185] including the use of PEG-thiol as a surfactant
6 M.-A. Fortin et al.
Several articles have been published on the topic of seed-mediated growth of
Au NPs. One of the most comprehensive ones is a three-step process that was used
to produce uniform Au NPs with a diameter up to 300 nm (Fig. 2) [209]. The main
limitation of this method is that an important population of smaller NPs often remains
in solution additionally to the grown seeds, requiring further purification to achieve
very narrow particle size distributions. Perrault et al. overcame this problem by using
hydroquinone as a reducing agent [147]. Hydroquinone reduces selectively the gold
ions that are located in the immediate vicinity of Au0 seed nanoclusters, leaving
isolated gold ions unreduced. This chemical route allows a selective growth of the
seeds while avoiding further nucleation of Au clusters in the reaction fluid.
2.3 Syntheses Taking Place in Organic Media
Nanoparticle synthesis routes performed in aqueous conditions are easy to
implement, and as a major advantage, they avoid the introduction of potentially
toxic solvents into the chain of NP production. Unfortunately, the size distribution
Fig. 2 TEM images of a 15 ± 2 nm seeds, b 31 ± 3 nm, c 69 ± 3 nm, d 121 ± 10 nm, and e
151 ± 8 nm and SEM image of f 294 ± 17 nm gold nanoparticles. Scale bars are 200 nm for parts
a–c and 500 nm for parts d–f. Reprinted with permission from [209]. Copyright 2011 American
Chemical Society
Gold Nanoparticles for Imaging and Cancer Therapy 7
of Au NPs synthesized in aqueous media is relatively difficult to keep to narrow
and small values. It is generally well established and acknowledged in the field of
nanotechnology, that syntheses performed in organic media often result in much
smaller and narrower particles size distributions. The Brust–Schiffrin method is one
of the first reported to synthesize Au NPs in organic media, and it is until now one
of the most widely performed to synthesize small particles of narrow distributions
(Fig. 3) [22]. Typically, it consists of using tetraoctylammonium bromide (TOAB)
to transfer the gold salts into an organic phase (e.g. toluene, ethanol and ethylene
glycol). Then, NPs are nucleated through a reduction step through thiol ligands and
NaBH4 . This approach produces very small and stable NPs ranging from 1.5 to 5 nm
that can be easily functionalized with other ligands. Recent developments related to
the Brust–Schiffrin method include mostly fundamental studies of mechanisms such
as charge transfer, nucleation and growth during the synthesis process [54, 146].
Another class of synthesis procedures taking place in an organic solvent is the
so-called polyol process. It allows the synthesis of monodisperse Au NPs, or alterna-
tively to particles of various well-controlled geometries [60]. In this procedure, gold
salts are dispersed in high boiling point alcohols (e.g. ethylene glycol), which also act
as a reducing and as a capping agent. By using a polymeric stabilizer, highly symmet-
rical polyhedron-shaped Au NPs with a narrow size distribution can be obtained in a
wide variety of sizes. Xia et al. have used a polyol process with polyvinylpyrrolidone
(PVP) as the stabilizer to synthesize very uniform silver nanocubes with 50–115 nm
of edge length, and used them as sacrificial templates in a solution of HAuCl4 for
the production of gold nanoboxes with perfectly smooth surfaces [179]. Song et al.
dissolved HAuCl4 and PVP in 1,5-pentanediol, instead of ethylene glycol for its
higher boiling point, and added small concentrations of AgNO3 [165]. Depend-
ing on the concentration of silver ions, octahedral (ratio Ag/Au 1/600) or cubic
(Ag/Au 1/200) shape nanostructures were obtained, among others. This chemical
Fig. 3 Gold nanoparticles synthesized by the Brust–Schiffrin method. Among the different col-
loidal synthesis techniques, this one usually leads to very small particle size distributions. Source
The authors, following procedures described in [22]
8 M.-A. Fortin et al.
behaviour was attributed to the selective deposition of silver species on the seed
surface during the reaction. Overall, the polyol process is a convenient and low-cost
technology for the large-scale production of highly symmetrical Au NPs of homo-
geneous sizes. However, the complete elimination of the solvent residues, as well as
the presence of potentially toxic chemicals, necessitates the introduction of tedious
filtration, dialysis or chromatography procedures that are not necessarily easy to
upscale.
2.4 Au NPs Purification Prior to Coatings
and Functionalization
The purification and functionalization steps to remove potentially toxic reagents
and to cover the NPs surface with biocompatible ligands are very important in the
development of stable as well as functional Au NP formulations for biomedical
applications. The purification is usually performed by dialysis, chromatography or
centrifugation. Those methods are generally sufficient to remove the majority of the
contaminants. As mentioned above, the majority of Au NPs used for biomedical
applications are synthesized either by the Turkevitch [56] method, by the seed-
mediated growth approach [150] or by the Brust–Schiffrin [42] technique. In the
first case, citrate molecules cover the surface of the Au NP. In the second one,
hexadecyltrimethylammonium bromide (CTAB) is usual, whereas alkanethiols are
employed in the Brust–Schiffrin method. For biomedical applications, the Au NPs
must be dispersed in aqueous solutions, and they must be free of even low traces
of contaminants (e.g. organic solvents, reduction agents and excess of surfactants).
They must be stable in physiological media, which are rich in diverse ions, proteins
and many other molecules. Thus, in order to preserve the colloidal stability and assure
biocompatibility, the Au NPs obtained by the previously described methods must be
functionalized with biocompatible molecules.
2.5 Ligand-Free Au NP Suspensions
The conventional Au NP synthesis approaches do have several limitations in terms
of toxicity risks induced by surfactants and chemical residues. For instance, sodium
borohydride (NaBH4 ) is a harsh reagent that must be entirely removed from the
solutions [146]. Then, the gold ions used in most Au NP synthesis techniques usually
come from chloroauric acid, a potent acid which must be entirely cleared from the
biomedical solutions [5]. In addition, ligand exchange procedures involve several
manipulation steps that lead to material loss, as well as to agglomeration. Several
alternatives to the more conventional chemical synthesis approaches have emerged
recently to synthesize purer Au NPs, generally free from ligands or reducing agents,
Gold Nanoparticles for Imaging and Cancer Therapy 9
and therefore ready to be efficiently functionalized. These synthesis techniques could
represent a major advantage in the quest for compounds that limit the toxicity risks
related to the presence of residues in Au NPs compounds.
2.5.1 Pulsed Laser Ablation in Liquid (PLAL)
The synthesis technique that possibly enables the production of the smallest and the
purest Au NPs solutions, is pulsed laser ablation in liquid (PLAL). This synthesis
method was introduced by Cotton and Henglein in 1993 [63, 138]. It is performed
by immersing a gold metal target in a fluid, followed by irradiation of the target
surface for a certain time with a pulsed laser [153]. The target absorbs the laser pulse
energy, resulting in heating and photoionization of the irradiated area. Solid, liquid
and vaporized materials are emitted in the form of a plasma plume, which expands
outwards in a confining liquid [8, 9, 81]. Then, the plasma plume starts to cool down
and a cavitation bubble is formed [8]. At this moment, the supersaturation point
is achieved due to the concentration of metal ions, to the high pressure and to the
temperature attained because of liquid confinement. This causes NPs to nucleate:
metal atoms condense and coalesce in the form of NP nuclei. The newly formed
NPs start to diffuse into the expanding cavitation bubble, where further growth,
coalescence or aggregation happen. At last, when the cavitation bubble collapses,
all the NPs are released into the surrounding liquid [8]. A schematic representation
of PLAL for the synthesis of Au NPs is shown in Fig. 4. The ultra-small Au NPs
colloids synthesized by this technique can be stabilized by PEG grafting to enhance
their stability in biomedical media [169].
Fig. 4 Pulsed laser ablation in liquid for the nucleation of Au NPs in ultrapure water (a), produces
Au colloids of chemically pure surfaces that are stabilized by electrostatic forces (b). To increase
their colloidal stability in biological fluids, the Au NPs are stabilized by polyethylene glycol.
Additionally, the Au NP@PEG was functionalized with DMSA-DTPA-Mn2+ to be applied as a
contrast agent for MRI (c). PLAL synthesis followed by PEG and DMSA-DTPA-Mn2+ grafting
enables the production of very small particle size distributions (d). Reproduced from [169] with
permission from the Royal Society of Chemistry
10 M.-A. Fortin et al.
The laser wavelength, the laser fluence, the repetition rate, the ablation time and
the composition of the liquid solution can be tuned to achieve a specific concentration
of NPs and size distribution [8, 153]. The most suitable wavelengths to synthesize
Au NPs are in the NIR region (e.g. 1064 nm) [184]. Subsequently, post-irradiation
with wavelengths coinciding with plasmon absorption or interband transition of Au
NPs can be used to tune the size and to reduce polydispersity [124].
PLAL allows the production of Au NPs in water or organic solvents [10, 180].
However, organic solvents are prone to pyrolysis during laser ablation and the
degraded molecules can adsorb on the surface of the NPs, which may raise bio-
compatibility issues. In pure water, PLAL-synthesized Au NPs show a size range of
10–40 nm; they are electrostatically stabilized by negative charges that result from
the formation of Au–O− species [8, 153, 180]. To improve the stability and to control
the Au NPs size in water, it is possible to use low salt concentration. The anions of
the salts adsorb on the surface of the Au NPs and increase the negative charge den-
sity, thus enhancing the repulsive forces between the NPs. This effect prevents both
the growth of the NPs by avoiding nuclei coalescence and improves the colloidal
stability prior to further grafting of the nanoparticles with biocompatible molecules
(e.g. to create steric hindrance) [130, 153].
In a biomedical context, the main advantage of PLAL over conventional Au NP
synthesis colloidal chemistry routes is the production of NPs directly in water with-
out the use of reducing or stabilizing chemicals (e.g. NaBH4 ) [153]. As a result,
the surface of the NPs is free of ligands and it is thereby readily available for the
conjugation with other molecules [153]. Moreover, many reducing chemicals used
in colloidal Au NP synthesis routes can be toxic, and this hazard is totally eliminated
in laser ablation. Thus, the ligand-free surface allows to avoid extensive purification
processes and aggregation associated with ligand exchange procedures [194]. The
main limitations of PLAL for Au NP synthesis are the necessity to use very expensive
lasers as well as low concentrations of solutions synthesized, which require several
concentration steps in order to reach biomedical applicability.
2.5.2 Atmospheric Plasma Electrochemistry
Atmospheric plasma synthesis, or plasma electrochemistry, allows the rapid nucle-
ation and growth of Au NPs directly in water, without the need of any chemical
reducing agent. Compared to pulsed laser ablation, it has the advantage of producing
more concentrated solutions of Au NPs while requiring less expensive equipment.
Plasma is a charged gas containing free electrons, positive and negative ions, neutral
species in the ground or excited state, and photons. This mixture of charged species
gives plasmas unique physico-chemical properties. The most common and easily
attained method for forming plasma is to apply an electrical voltage between two
electrodes, solid or liquid, separated by a gas. With aqueous solution, plasma elec-
trochemistry must be operated near atmospheric pressure and at low temperature to
avoid solvent evaporation.
Gold Nanoparticles for Imaging and Cancer Therapy 11
A great variety of chemical reactions take place at the plasma–liquid interface. In
the presence of nitrogen as the main plasma component or as a contaminant from the
presence of air, the formation and dissolution of nitric acid can cause a significant
decrease in pH [108]. The electrolysis of water 2H2 O + 2e− → 2OH− + H2 at the
plasma cathode and the formation of chlorine gas (2Cl− → Cl2 + 2e− ) at the anode
were also reported [158]. The formation of reactive ions H+ , H− , O− , OH− , radicals
H· , O· and OH· and H2 O2 is also a very important factor in the cascade of chemistry
events taking place in the nucleation and growth of nanoparticles [66, 125].
The exact mechanism for the nucleation of Au NPs by plasma–liquid electro-
chemistry is still not entirely understood. The electrons generated in the plasma are
accelerated to the surface of the liquid by the electric field between the electrodes and
pass into solution in the form of solvated electrons [1, 125]. Several reactive oxygen
and nitrogen species generated by the plasma treatment act as metal ion reducers.
These ‘fugitive’ or metastable reducing agents make possible the nucleation of Au
NPs while avoiding the use of external toxic chemicals (e.g. NaBH4 ) for the synthe-
sis of water-dispersed Au NPs. Gold ions in solution can reach the plasma–liquid
interface by the combined effects of the electric field, the convection forces and
the concentration gradient [2]. When an ion reaches the interface, it is reduced into
atomic gold directly by the electrons: [AuCl4 ]− + 3e− → Au0 + 4Cl− . Once reduced,
the gold atoms collide and form clusters, which diffuse away from the plasma–liquid
interface. These clusters act as nucleation sites of NPs [2]. Size control is usually
performed by adjusting only the concentrations of metal salt and surfactant in solu-
tion. Increasing the current can also lead to the production of smaller NPs since it
allows a higher nucleation rate [79].
In the last 10 years, new ‘cold plasma’ reactors operating at atmospheric pressure
brought the possibility to synthesize NPs from metal salts directly in water. The
first type of atmospheric plasma reactor that was used to nucleate Au NPs was the
microplasma [104, 160], and the most commonly used geometry of microplasma
is the microhollow cathode [20, 104, 155]. It consists of a gas stream flowing in a
cylindrical hollow cathode placed a few millimetres above a liquid solution contain-
ing metal ions. A DC voltage, typically several thousand V/cm, is applied between
the cathode and the solution. The plasma discharge generated at the output of the
microhollow cathode thus extends to the surface of the liquid, where the reduction
of the metal ions and the nucleation of NPs take place.
Richmonds and Sankaran demonstrated the possibility to synthesize gold and
silver NPs from the dissolution of a metal sheet in a slightly acidic aqueous solution
[155]. Using fructose as a stabilizing agent, the reduction by argon plasma has formed
10 nm diameter NPs for both metals. This device shows the conventional concept
of an electrochemical cell, where the metal film acts as a sacrificial anode, and
the microplasma as a cathode [160]. This concept was adapted several times for
the synthesis of gold and silver NPs of different sizes and morphologies, with and
without stabilizer [127, 188, 189]. Unfortunately, the microplasma technologies have
a rather limited surface coverage, and other types of plasma must be considered to
allow the development of technologies enabling the synthesis of large volumes of
solutions.
12 M.-A. Fortin et al.
A promising alternative to the microplasma is the dielectric barrier discharge
(DBD). Through the DBD technology, plasma–liquid interface area of several cm2
can be generated [16]. This technology was recently employed to synthesize radioac-
tive Au NPs for applications in oncology [19]. The main limitation of Au NPs syn-
thesized by plasma electrochemistry remains the relatively high polydispersity of
particles, requiring further separation steps. Further work aimed at understanding
the exact electrochemical mechanisms behind the reduction of Au ions should help
reaching a better control over size.
2.6 Surface Treatment of Au NPs
The size, shape and surface properties of the Au NPs are determinant factors for their
successful biomedical applications [4, 35, 40, 55, 59, 95, 134]. To be used in vivo, the
particles should be dispersible in water; they should form a stable colloid in biolog-
ical media, which are aqueous solutions with significant ionic strengths and rich in
proteins; they should not induce cytotoxicity; they should not adsorb too many pro-
teins on their surfaces to avoid uptake by the mononuclear phagocytic system (MPS)
cells; and they should be easily conjugated with specific ligands for targeting studies
[35, 59, 172]. Therefore, the Au NPs produced by the previous techniques should
undergo ligand exchange processes for their surface to be coated with different types
of biocompatible, antifouling and/or biologically active molecules [119, 159].
The surface of Au NPs can be functionalized using molecules containing thi-
ols, amines, carboxylic acids and phosphines [140]. The most resistant coatings are
formed by making use of ligands that have the highest affinity to Au. Therefore, thiol
modification is usually preferred, as the sulphur atoms form a coordinate covalent
bond with the metal surface [119]. The surface of Au NPs stabilized with citrate
or CTAB can be easily functionalized by exchanging these molecules with thiol
containing ligands [159]. The initial stabilizing agent is quickly replaced during the
adsorption of sulphur atoms, followed by a slower reorganization and packing of
the incoming molecules [52, 119]. The surface of Au NPs stabilized by alkanethiols
can be modified by thiol–thiol exchange [159]. This process requires higher molar
excess of the incoming ligands [119]. In addition, this method can also be used to
produce mixed organic monolayers containing different types of ligands at specific
ratios [159]. Finally, polyethylene glycol (PEG) is a commonly used antibiofouling
ligand for Au NPs [52]. When this highly hydrophobic polymer is attached at the
surface of Au NPs, the long chains prevent the aggregation of the nanoparticles by
steric repulsion. PEG also prevents the adhesion of proteins at the surface of the NPs.
