RESEARCH PAPER

Gold nanoparticles induce DNA damage in the bloodand liver of rats

Eria Cardoso • Eduardo Londero • Gabriela Kozuchovski Ferreira •

Gislaine Tezza Rezin • Elton Torres Zanoni • Frederico de Souza Notoya •

Daniela Dimer Leffa • Adriani Paganini Damiani • Francine Daumann •

Paula Rohr • Luciano da Silva • Vanessa M. Andrade • Marcos Marques da Silva Paula

Received: 5 August 2014 / Accepted: 30 October 2014

� Springer Science+Business Media Dordrecht 2014

Abstract The potential of gold nanoparticles

(GNPs) for use in different biological applications

has led to a strong interest in the study of their possible

deleterious effects in biological systems and how these

effects may be mitigated. This study was undertaken

to investigate the effects of the acute and chronic

administration of GNPs with mean diameters of 10

and 30 nm on deoxyribonucleic acid (DNA) damage

in the blood and liver of adult rats. For the acute

administration, Wistar adult rats received a single

intraperitoneal injection of either GNPs or a saline

solution. For the chronic administration, Wistar adult

rats received a daily single injection of the same GNPs

or saline solution for 28 days. Twenty-four hours after

either the single (acute) or final injection (chronic), the

rats were euthanised by decapitation, and the blood

and liver were isolated for the evaluation of DNA

damage. In this study, we demonstrated that the acute

and chronic administration of GNPs 10 and 30 nm in

size increased the frequency of DNA damage and the

damage index in the blood and liver of adult rats.

These findings suggest that the DNA damage may be

caused by oxidative stress, which occurred regardless

of the type of administration and GNP size.

Keywords Gold nanoparticles � DNA damage �Comet assay � Blood � Liver

Introduction

Gold nanoparticles (GNPs) possess unique properties,

such as biocompatibility, high surface reactivity,

resistance to oxidation, flexibility in functionalisation

and a wide range of delivery targets (Sonavane et al.

2008; Aschner 2009; Zhang et al. 2010). GNPs are

useful for the delivery and controlled release of a

variety of chemical agents, including anticancer drugs,

antibiotics, amino acids, peptides, glucose, antioxi-

dants, nucleic acids and isotopes (Oberdorster et al.

2005). Although GNPs are recognised as being as

E. Cardoso � E. Londero � G. K. Ferreira �E. T. Zanoni � F. de Souza Notoya �L. da Silva � M. M. da Silva Paula (&)

Laboratorio de Sıntese de Complexos Multifuncionais,

PPGCS, Universidade do Extremo Sul Catarinense,

Criciuma, SC 88806-000, Brazil

e-mail: bocaocao@gmail.com

E. Cardoso

Instituto Federal de Educacao, Ciencia e Tecnologia

Catarinense, Campus Sombrio, Sombrio, SC 88960-000,

Brazil

G. T. Rezin

Laboratorio de Fisiopatologia Clınica e Experimental,

PPGCS, Universidade do Sul de Santa Catarina, Tubarao,

SC 88704-9000, Brazil

D. D. Leffa � A. P. Damiani � F. Daumann �P. Rohr � V. M. Andrade

Laboratorio de Biologia Celular e Molecular, PPGCS,

Universidade do Extremo Sul Catarinense, Av.

Universitaria, 1105, Criciuma, SC 88806-000, Brazil

123

J Nanopart Res (2014) 16:2727

DOI 10.1007/s11051-014-2727-1

nontoxic (Merchant 1998; Connor et al. 2005; Shukla

et al. 2005), some reports have suggested that they are

toxic; this has been shown to depend on the physical

dimensions, surface chemistry and shape of the GNPs.

Their potential for use in different biological applica-

tions has led to a strong interest in the study of their

possible deleterious effects in biological systems and

how these effects may be mitigated (Pernodet et al.

2006; Chithrani and Chan 2007; Pan et al. 2007).

Sonavane et al. (2008) demonstrated the presence

of a wide distribution of gold particles inside the living

system. Additionally, Sadauskas et al. (2007) studied

the biodistribution of colloidal GNPs administered

intravenously and intraperitoneally to adult female

mice. In all of the mice exposed to the GNPs,

accumulations of nanoparticles were traced to Kupffer

cells, i.e. the macrophages of the liver. No accumu-

lation was observed in cells other than macrophages in

any of the organs examined in this study. However, it

is essential to understand the interactions of GNPs

with vital organs, such as the blood and liver (de

Lamirande and Gagnon 1993; Sadauskas et al. 2007;

Sadauskas et al. 2009).

Studies have strongly focused on the potential of

nanomaterials to cause oxidative stress (de Lamirande

and Gagnon 1993; Butterfield and Stadtman 1997;

Porter and Janicke 1999; Karihtala et al. 2009; Ng

et al. 2010), and an excessive production of reactive

oxygen species (ROS) can lead to reactions with

macromolecules, such as deoxyribonucleic acid

(DNA), lipids and proteins. It has been proposed that

DNA damage contributes to cellular dysfunction.

Cellular DNA is a sensitive target of damage follow-

ing oxidative stress. This damage can include chem-

ical and structural modifications to purine and

pyrimidine bases and 20-deoxyribose and the forma-

tion of single- and double-strand breaks. Strand breaks

within DNA can occur either directly, due to damage

from free radical exposure, or indirectly, due to the

cleavage of the DNA backbone during base excision

repair (BER) (Liu and Martin 2001; Martin and Liu

2002). Continued oxidative damage to DNA can alter

signalling cascades and gene expression and cause

replication errors and genomic instability (Karihtala

et al. 2009). The comet assay, which is also called

single-cell gel electrophoresis, is considered to be the

most sensitive quantitative method for measuring

early damage to the genomic DNA of eukaryotic cells

or disaggregated tissues (Ng et al. 2010; Collins 2014).

Because the widespread use of nanoparticles (NPs)

in the future will probably have an enormous impact

on human health, it is essential to understand their

effects on the blood and liver. This study was

undertaken to investigate the effects of the acute and

chronic administration of GNPs with mean diameters

of 10 and 30 nm on DNA damage in the blood and

livers of adult rats using the alkaline comet assay.

Materials and methods

Synthesis and characterisation of gold

nanoparticles

GNPs of both 10 and 30 nm were synthesised as

described by Turkevich et al., with minor modifica-

tions (CooperaStevenson 1951). Tetrachloroauric acid

(HAuCl4) was acquired from Sigma-Aldrich (St.

Louis, MO, USA), and sodium citrate (Na3C6H5O7.2-

H2O), which is a reducing agent and stabiliser, was

acquired from Nuclear (Diadema, SP, Brazil). GNP

size was controlled using distinct sodium citrate

concentrations. Initially, 100 ml of 0.50 mM tetra-

chloroauric acid was transferred to a round-bottomed

flask and heated to 90 �C with mechanical stirring at

700 rpm. The sodium citrate solution was then added,

and the system was stirred at 90 �C for another

20 min. Sodium citrate solutions at concentrations of

136.0 and 8.5 mM were used to obtain GNPs-10 and

GNPs-30, respectively. The 10 and 30 nm GNPs

possessed pH values of 6.5 and 2.8, respectively. The

pH values of both were adjusted to physiologic pH

with a buffer solution. Next, the GNP solutions were

centrifuged (13,000 rpm for 15 min), washed twice

with ultrapure water and dispersed in saline solution.

The electronic spectra were obtained using a Shima-

dzu instrument model UV-1800 (Shimadzu Corp.,

Kyoto, Japan), which showed SPR (surface plasmon

resonance) bands with kmax values of 517 and 547 nm

for GNPs-10 and GNPs-30, respectively. X-ray dif-

fraction measurements were performed to determine

the mean diameters of GNPs-10 and GNPs-30, which

were calculated using Scherer’s equation from the

signal at 2h = 38� (major relative intensity) (Hirn

et al. 2011). These values were confirmed by the

transmission electron microscopy (TEM) analysis

images obtained using a JEOL Titan 80–300 kV.

The Au concentrations were measured by atomic

2727 Page 2 of 8 J Nanopart Res (2014) 16:2727

123

absorption spectroscopy (AAS) (Varian model AA

240Z; Varian Medical Systems, Inc., Palo Alto, CA,

USA). The value obtained for both original solutions

was 70 mg l-1. The zeta potentials of the solutions

were also measured (Zeta Potential Analyser, Zeta

PALS Brookhaven Instruments, Holtsville, NY, USA)

at 25 �C, and the values at -30 and -35 mV for

GNPs-10 and GNPs-30, respectively, were obtained.

After all of the characterisation tests, we observed that

the GNPs were compliant with previous reports by

Cardoso et al. (2014).

Animals

Adult (60 days old) male Wistar rats (250–300 g) were

obtained from the central animal house of the Univer-

sidade do Extremo Sul Catarinense. A total of 36

animals were used and were divided into six animals per

group. The animals had free access to food and water

and were maintained on a 12-h light–dark cycle (lights

on 7:00 am) at 23 ± 1 �C. All experimental procedures

were carried out in accordance with the National

Institutes of Health Guide for the Care and Use of

Laboratory Animals and the Brazilian Society for

Neuroscience and Behavior (SBNeC) recommenda-

tions for animal care with the approval of the UNESC

ethics Committee (protocol number 03/2012). More-

over, all efforts were made to minimise animal suffering

and reduce the number of experimental animals.

Acute GNP administration

Animals received a single intraperitoneal administration

of saline solution, GNPs-10 or GNPs-30, 1 ml kg-1 at

concentrations of 70 mg l-1 and 70 lg kg-1. Twenty-

four hours after administration, the animals were killed

by decapitation, and the blood and liver were isolated for

analysis using the Comet assay.

Chronic GNP administration

Animals received, a once daily for 28 days intraperi-

toneal administration of saline solution (0,9 %), or

GNPs-10 (1 ml�kg-1) or GNPs-30 (1 ml�kg-1) at

concentrations of 70 mg l-1 or 70 lg kg-1. Twenty-

four hours after the final administration, the animals

were killed by decapitation, and the blood and liver

were isolated for analysis of index and frequency of

damage using the Comet assay.

Comet assay

The comet assay was carried out under alkaline

conditions as described by Singh et al. (1988), with

some modifications suggested by Tice et al. (2000).

Cells from different tissues were obtained according to

the modifications suggested by Tice and colleagues.

The blood and livers were placed in cold phosphate-

buffered saline (PBS) and finely minced through a

syringe plungerto obtain cell suspensions. The cells

that were isolated from the tissues (20 ll aliquots)

were embedded in low-melting-point agarose (0.75 %

w/v, 95 ll or 80 ll), and the mixture was added to a

microscope slide pre-coated with normal-melting-

point agarose (1.5 % w/v) and covered with a

coverslip (two slides per donor). Each slide was

briefly placed on ice for the agarose to solidify, and the

coverslip was carefully removed. Each slide was also

immersed in freshly prepared lysis solution (2.5 M

NaCl, 100 mM EDTA, 10 mM Tris; pH 10.0–10.5).

The slides were then incubated in freshly prepared

alkaline buffer solution (300 mM NaOH, 1 mM

EDTA; pH [ 13) for 20 min for DNA unwinding,

and electrophoresis was performed using the same

solution. Electrophoresis was carried out for 15 min at

300 mA and 25 V (0.7 V/cm). All of these steps were

performed under dim, indirect light.

After electrophoresis, the slides were neutralised in

400 mM Tris (pH 7.5). They were stained with silver

nitrate, as described previously by VVillela et al.

(2006), and then fixed for 10 min in a solution

containing 15 % trichloroacetic acid w/v, 5 % zinc

sulphate w/v and 5 % glycerol v/v, rinsed three times

in distilled water and dried for 2 h at 37 �C. The dry

slides were re-hydrated for 5 min in distilled water and

then stained (in 5 % sodium carbonate w/v, 0.1 %

ammonium nitrate w/v, 0.1 % silver nitrate w/v,

0.25 % tungstosilicic acid and 0.15 % formaldehyde

w/v, which was freshly prepared in the dark) and

continuously shaken for 35 min. The stained slides

were then rinsed twice with distilled water, submerged

in the stop solution (1 % acetic acid), rinsed again, and

immediately coded for analysis with an optic micro-

scope. Images of 100 randomly selected cells were

individually analysed. To calculate the damage index

(DI), the cells were visually allocated into five classes

according to tail size (0 = no tails and 4 = maxi-

mum-length tails), which resulted in a single DNA

damage score for each sample, and consequently, for

J Nanopart Res (2014) 16:2727 Page 3 of 8 2727

123

each group studied. Thus, the DI of a group could

range from 0 (completely undamaged = 100

cells 9 0) to 400 (maximum damage = 100

cells 9 4). The damage frequency (DF in percentage)

was calculated for each sample based on the numbers

of cells with versus without tail. International guide-

lines and recommendations for the Comet assay

consider the visual scoring of comets to be a well-

validated evaluation method. This assay has been

demonstrated to have a high correlation value of

1using computer-based image analyses (Collins et al.

1997). Negative and positive controls were used for

each electrophoresis run to ensure the reliability of the

procedure. All slides were coded for a blind analysis.

Statistical analysis

For all analyses of the DNA damage parameters, all

data were expressed as means ± standard deviations.

Statistical analyses of the damage index and damage

frequency, as measured by the Comet assay, were

carried out using one-way analysis of variance

followed by the Tukey test. The analyses were

performed using GraphPad prism version 5.00 for

Windows software.

Results

The results correspondent of characterisation tests of

the GNPs have been published in our previous study

by Cardoso et al. (2014).

Figure 1 shows the comet assay parameters for the

acute treatment with 10 and 30 nm GNP. Animals

treated with 10 nm GNP presented higher DI and DF

values in both evaluated tissues when compared with

the control group (p \ 0.01, ANOVA–Tukey). Treat-

ment with 30 nm GNP showed elevated DI and DF

values in the blood (p \ 0.01, ANOVA–Tukey) and

elevated DI (p \ 0.01, ANOVA–Tukey) and DF

(p \ 0.05, ANOVA–Tukey) values in the liver when

compared with the saline treatment.

The comet assay results for the chronic treatment

with 10 and 30 nm GNP are presented in Fig. 2. The

chronic 10 and 30 nm GNP treatments induced higher

levels of DNA damage, for both DF and DI, in the liver

and blood (p \ 0.01, ANOVA–Tukey) when com-

pared to the control group.

Discussion

The UV–vis spectroscopy and X-ray diffraction

results (showed in Cardoso et al. 2014) agree with

prior studies that showed that the electronic spectra

and, consequently, the colours of colloidal GNP

dispersions depend on the particle geometry and size

(Philip 2008). Colour results from the interactions of

nanoparticle surface electrons with incident light

(Philip 2008; Alkilany and Murphy 2010). The

characteristic SPR band widens and shifts to red and

the XRD peaks narrow as the particle size increases

(Chen et al. 2005; Link and El-Sayed 1999). The

Scherrer equation, based on the most intense peak

(2h = 388), was used to calculate the mean particle

diameters for GNP-10 and GNP-30 dispersions.

Moreover, the XRD spectra of the dried GNPs were

compatible with the standard spectrum of metallic

gold (JCPDS number: 4-0784) (Yan et al. 2005; Singh

et al. 2009; Aromal and Philip 2012).

Morais et al. (2012) evaluated the biodistribution of

GNPs and demonstrated that they are rapidly distrib-

uted, while the liver is the preferential organ of GNP

accumulation. The altered accumulation of GNPs in

various organs and tissues has been related to the size

and surface charge of the NPs that mediate dynamic

protein binding and exchange (Hirn et al. 2011). The

adsorption of plasma proteins onto the surfaces of

NPs, which is known as opsonisation, occurs instantly

when the particles enter the bloodstream (Owens and

Peppas 2006). Considering this and the fact that

oxidative stress can provoke DNA damage, our goal in

this work was to evaluate DNA damage in the blood

and liver of rats after the acute and chronic adminis-

tration of 10 and 30 nm GNPs.

DNA damage is defined as any modification to

DNA that changes its coding properties or the normal

functioning of transcription or replication (Rao 1993;

Morais et al. 2012). Environmental and endogenous

agents, ROS, reactive nitrogen species and other

factors, such as temperature, errors in DNA replication

and repair, and methylation, can lead to damaged

DNA in cells (Tice et al. 2000; Collins 2014). It has

been estimated that metabolism-generated ROS can

cause approximately 10,000 lesions per day in the

genome of a human non-neuronal cell and that purine

base turnover in DNA, resulting from hydrolytic

depurination and its subsequent repair, occurs in

2727 Page 4 of 8 J Nanopart Res (2014) 16:2727

123

approximately 2,000–10,000 bases per day (Rao

1993).

Ours results demonstrated that the acute and

chronic administration of 10 and 30 nm GNPs caused

DNA damage in the blood and liver of the rats, which

was determined using the alkaline comet assay that

measures DNA strand breaks in single cells. The

in vitro study by Kang et al. (2009) showed that DNA

damage did not occur in L5178Y cells containing

GNPs that were 60 nm or smaller GNPs, as

Fig. 1 Effects of the acute

administration of 10 and

30 nm GNPs on the damage

index and damage frequency

(mean ± SD) in the liver

(a–b) and peripheral blood

(c–d) of rats. *Data

significantly differ in

relation to the control group

(p \ 0.05, ANOVA–

Tukey). **Data significantly

differ in relation to the

control group (p \ 0.01,

ANOVA–Tukey)

Fig. 2 Effects of the

chronic administration of 10

and 30 nm GNPs on the

damage index and damage

frequency (mean ± SD) in

the liver (a–b) and

peripheral blood (c–d) of

rats. **Data significantly

differ in relation to the

control group (p \ 0.01,

ANOVA–Tukey)

J Nanopart Res (2014) 16:2727 Page 5 of 8 2727

123

determined by the Comet assay; damage only occurred

with 100 nm GNPs. However, other studies have

reported DNA damage in association with 20 nm (Li

et al. 2008) and 8 nm (Auffan et al. 2009) GNPs,

although no damage was reported in a study using

10 nm GNPs (Singh et al. 2010).

Hillyer and Albrecht (2001) showed that after

administration, GNPs appeared in various tissues in

mice and that the amount of absorption and distribu-

tion in the body inversely correlated with the size of

the particles and surface properties. However, in our

study was demonstrated that both sizes of GNP

analysed (10 and 30 nm) caused DNA damage.

Additionally, studies have demonstrated binding

affinities between GNP and thiol and amine groups

that promote combinations with biomolecules (Katz

and Willner 2004; Ojea-Jimenez and Puntes 2009) and

the free radical formation due to GNP (Siddiqi et al.

2012). Previous works show that the very large surface

areas of ultra-small particles can result in the direct

formation of ROS that can cause cellular damage by

attacking DNA, proteins and membranes and affect

the cytoplasm, mitochondrial function and the nucleus

(Brown et al. 2001; Lewinski et al. 2008). Therefore,

oxidative stress is a possible mechanism for induced

toxicity by acute and chronic administration of GNP of

10 and 30 nm on DNA.

Additionally, GNPs can cause the generation of

caspase-3, interferon-c and 8-hydroxydeoxyguanosine

(8OHdG), which may lead to inflammation and DNA

damage/cell death, which was also demonstrated in the

recent study by Cardoso et al. (2014). Thus, oxidative

stress has been emphasized as a likely mechanism of NP

toxicity (Li et al. 2008), and it may be hypothesised from

the present results that DNA damage results from the

excessive production of free radicals, generating oxida-

tive damage to cells (Halliwell and Gutteridge 2001). In

addition, studies involving experimental animals have

shown that augmented ROS, and particularly superox-

ide (•O2–), can interact with DNA, resulting in oxidative

damage and DNA fragmentation-mediated cellular

injury (Gill and Wilcox 2006; Touyz 2004; Zhong and

Xu 2008; Madsen-Bouterse et al. 2010). Chueh et al.

(2014) showed that genes involved in BER and

homologous recombination pathways were significantly

enhanced by GNPs, indicating that intracellular GNPs

may cause base damage. Considering these data collec-

tively, we presume that DNA damage is likely to be

related to blood GNP levels, which may explain the

effects of GNPs on the DNA in the blood and liver of rats

after the acute and chronic administration of GNPs of

different diameters. Although our previous study has

demonstrated differences related to chronic and acute

treatments in cerebral cortex (Cardoso et al. 2014), the

present study not shows difference between these

administrations in the blood and liver. So, we suggest

that GNPs have easy access to blood and liver, since that

not present neither impediment, as the blood brain

barrier.

Conclusions

In conclusion, the present study demonstrated that the

acute and chronic administration of 10 and 30 nm

GNPs caused DNA damage in the blood and liver of

the treated rats. These findings suggest that the DNA

damage may be caused by oxidative stress, which

occurs regardless of the type of administration and the

size of the GNP.

Acknowledgments This work was supported by grants from

Conselho Nacional de Pesquisa e Desenvolvimento (CNPq),

CAPES and the Universidade do Extremo Sul Catarinense

(UNESC). The authors would also like to thank CNPq and

UNESC for the fellowship support.

References

Alkilany AM, Murphy CJ (2010) Toxicity and cellular uptake of

gold nanoparticles: what we have learned so far? J Nano-

particle 12:2313–2333

Aromal SA, Philip D (2012) Green synthesis of gold nanopar-

ticles using Trigonella foenum-graecum and its size-

dependent catalytic activity. Spectrochimistry Acta 97:1–5

Aschner M (2009) Chapter 8 - nanoparticles: transport across

the olfactory epithelium and application to the assessment

of brain function in health and disease. Prog Brain Res

180:141–152

Auffan M, Rose J, Orsiere T, De Meo M, Thill A, Zeyons O,

Proux O, Masion A, Chaurand P, Spalla O (2009) CeO2

nanoparticles induce DNA damage towards human dermal

fibroblasts in vitro. Nanotoxicology 3:161–171

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

Butterfield DA, Stadtman ER (1997) Protein oxidation pro-

cesses in aging brain. Adv Cell Aging Gerontol 2:161–191

Cardoso E, Rezin GT, Zanoni ET, Notoya FS, Leffa DD,

Damiani AP, Daumann F, Rodriguez JCO, Benavides R,

2727 Page 6 of 8 J Nanopart Res (2014) 16:2727

123

Silva L (2014) Acute and chronic administration of gold

nanoparticles cause dna damage in the cerebral cortex of

adult rats. Mutat Res 766–767:25–30

Chen Y, Gu X, Nie CG, Jiang ZY, Xie ZX, Lin CJ (2005) Shape

controlled growth of gold nanoparticles by a solution

synthesis. Chem Commun 33:4181–4183

Chithrani BD, Chan WC (2007) Elucidating the mechanism of

cellular uptake and removal of protein-coated gold nano-

particles of different sizes and shapes. Nano Lett

7:1542–1550

Chueh PJ, Liang RY, Lee YH, Zeng ZM, Chuang SM (2014)

Differential cytotoxic effects of gold nanoparticles in dif-

ferent mammalian cell lines. J Hazard Mater 264:303–312

Collins AR (2014) Measuring oxidative damage to DNA and its

repair with the comet assay. Biochim Biophys Acta

1840:794–800

Collins A, Dusinska M, Franklin M, Somorovska M, Petrovska

H, Duthie S, Fillion L, Panayiotidis M, Raslova K,

Vaughan N (1997) Comet assay in human biomonitoring

studies: reliability, validation, and applications. Environ

Mol Mutagen 30:139–146

Connor EE, Mwamuka J, Gole A, Murphy CJ, Wyatt MD (2005)

Gold nanoparticles are taken up by human cells but do not

cause acute cytotoxicity. Small 1:325–327

CooperaStevenson P (1951) A study of the nucleation and

growth processes in the synthesis of colloidal gold. Discuss

Faraday Soc 11:55–75

de Lamirande E, Gagnon C (1993) Human sperm hyperactiva-

tion in whole semen and its association with low super-

oxide scavenging capacity in seminal plasma. Fertil Steril

59:1291–1295

Gill PS, Wilcox CS (2006) NADPH oxidases in the kidney.

Antioxid Redox Signal 8:1597–1607

Halliwell B, Gutteridge JMC (2001) Oxidative stress: adapta-

tion, damage, repair and death. In: Halliwell B, Gutteridge

JMC (eds) Free radicals in biology and medicine. Oxford

University Press, Oxford, pp 246–350

Hillyer JF, Albrecht RM (2001) Gastrointestinal persorption and

tissue distribution of differently sized colloidal gold

nanoparticles. J Pharm Sci 90:1927–1936

Hirn S, Semmler-Behnke M, Schleh C, Wenk A, Lipka J,

Schaffler M, Takenaka S, Moller W, Schmid G, Simon U,

Kreyling WG (2011) Particle size-dependent and surface

charge-dependent biodistribution of gold nanoparticles

after intravenous administration. Eur J Pharm Biopharm

77:407–416

Kang SJ, Yum YN, Kim JH, Song H, Jeong J, Lim YT, Chung

BH, Park SN (2009) Induction of DNA damage in L5178Y

cells treated with gold nanoparticle. Biomol Ther 17:6

Karihtala P, Soini Y, Vaskivuo L, Bloigu R, Puistola U (2009)

DNA adduct 8-hydroxydeoxyguanosine, a novel putative

marker of prognostic significance in ovarian carcinoma. Int

J Gynecol Cancer 19:1047–1051

Katz E, Willner I (2004) Integrated nanoparticle-biomolecule

hybrid systems: synthesis, properties, and applications.

Angew Chem Int 43:6042–6108

Lewinski N, Colvin V, Drezek R (2008) Cytotoxicity of nano-

particles. Small 4:26–49

Li JJ, Zou L, Hartono D, Ong CN, Bay BH, Lanry Yung LY

(2008) Gold nanoparticles induce oxidative damage in lung

fibroblasts in vitro. Adv Mater 20:138–142

Link S, El-Sayed MA (1999) Size and temperature dependence

of the plasmon absorption of colloidal gold nanoparticles.

J Phys Chem B103:4212–4217

Liu Z, Martin LJ (2001) Motor neurons rapidly accumulate

DNA single-strand breaks after in vitro exposure to nitric

oxide and peroxynitrite and in vivo axotomy. J Comp

Neurol 432:35–60

Madsen-Bouterse SA, Zhong Q, Mohammad G, Ho YS,

Kowluru RA (2010) Oxidative damage of mitochondrial

DNA in diabetes and its protection by manganese super-

oxide dismutase. Free Radic Res 44:313–321

Martin LJ, Liu Z (2002) DNA damage profiling in motor neu-

rons: a single-cell analysis by comet assay. Neurochem Res

27:1093–1104

Merchant B (1998) Gold, the noble metal and the paradoxes of

its toxicology. Biologicals 26:49–59

Morais T, Soares ME, Duarte JA, Soares L, Maia S, Gomes P,

Pereira E, Fraga S, Carmo H, Bastos Mde L (2012) Effect

of surface coating on the biodistribution profile of gold

nanoparticles in the rat. Eur J Pharm Biopharm 80:185–193

Ng CT, Li JJ, Bay BH, Yung LY (2010) Current studies into the

genotoxic effects of nanomaterials. J Nucleic Acids

21:2010

Oberdorster G, Maynard A, Donaldson K, Castranova V, Fitz-

patrick J, Ausman K, Carter J, Karn B, Kreyling W, Lai D,

Olin S, Monteiro-Riviere N, Warheit D, Yang H (2005)

Principles for characterizing the potential human health

effects from exposure to nanomaterials: elements of a

screening strategy. Part Fibre Toxicol 2:8

Ojea-Jimenez I, Puntes V (2009) Instability of cationic gold

nanoparticle bioconjugates: the role of citrate ions. J Am

Chem Soc 131:13320–13327

Owens DE, Peppas NA (2006) Opsonization, biodistribution,

and pharmacokinetics of polymeric nanoparticles. Int J

Pharm 307:93–102

Pan Y, Neuss S, Leifert A, Fischler M, Wen F, Simon U, Schmid

G, Brandau W, Jahnen-Dechent W (2007) Size-dependent

cytotoxicity of gold nanoparticles. Small 3:1941–1949

Pernodet N, Fang X, Sun Y, Bakhtina A, Ramakrishnan A,

Sokolov J, Ulman A, Rafailovich M (2006) Adverse effects

of citrate/gold nanoparticles on human dermal fibroblasts.

Small 2:766–773

Philip D (2008) Synthesis and spectroscopic characterization of

gold nanoparticles. Spectrochim Acta 71:80–85

Porter AG, Janicke RU (1999) Emerging roles of caspase-3 in

apoptosis. Cell Death Differ 6:99–104

Rao KS (1993) Genomic damage and its repair in young and

aging brain. Mol Neurobiol 7:23–48

Sadauskas E, Wallin H, Stoltenberg M, Vogel U, Doering P,

Larsen A, Danscher G (2007) Kupffer cells are central in

the removal of nanoparticles from the organism. Part Fibre

Toxicol 4:10

Sadauskas E, Danscher G, Stoltenberg M, Vogel U, Larsen A,

Wallin H (2009) Protracted elimination of gold nanopar-

ticles from mouse liver. Nanomedicine 5:162–169

Shukla R, Bansal V, Chaudhary M, Basu A, Bhonde RR, Sastry

M (2005) Biocompatibility of gold nanoparticles and their

endocytotic fate inside the cellular compartment: a

microscopic overview. Langmuir 21:10644–10654

Siddiqi NJ, Abdelhalim MA, El-Ansary AK, Alhomida AS, Ong

WY (2012) Identification of potential biomarkers of gold

J Nanopart Res (2014) 16:2727 Page 7 of 8 2727

123

nanoparticle toxicity in rat brains. J Neuroinflammation

9:123

Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple

technique for quantitation of low levels of DNA damage in

individual cells. Exp Cell Res 175:184–191

Singh P, Kumari K, Katyal A, Kalra R, Chandra R (2009)

Synthesis and characterization of silver and gold nano-

particles in ionic liquid. Spectrochim Acta A73:218–220

Singh S, D’Britto V, Prabhune A, Ramana A, Dhawan A, Prasad

B (2010) Cytotoxic and genotoxic assessment of glyco-

lipid-reduced and-capped gold and silver nanoparticles.

New J Chem 34:294–301

Sonavane G, Tomoda K, Makino K (2008) Biodistribution of

colloidal gold nanoparticles after intravenous administra-

tion: effect of particle size. Colloids Surf B Biointerfaces

66:274–280

Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A,

Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF

(2000) Single cell gel/comet assay: guidelines for in vitro

and in vivo genetic toxicology testing. Environ Mol

Mutagen 35:206–221

Touyz RM (2004) Reactive oxygen species and angiotensin II

signaling in vascular cells–implications in cardiovascular

disease. Braz J Med Biol Res 37:1263–1273

Villela IV, de Oliveira IM, da Silva J, Henriques JA (2006) DNA

damage and repair in haemolymph cells of golden mussel

(Limnoperna fortunei) exposed to environmental contam-

inants. Mutat Res 605:78–86

Yan WF, Petkov V, Mahurin SM, Overbury SH, Dai S (2005)

Powder XRD analysis and catalysis characterization of

ultra-small gold nanoparticles deposited on titania-modi-

fied SBA-15. Catalisys Commun 6:404–408

Zhang XD, Wu HY, Wu D, Wang YY, Chang JH, Zhai ZB,

Meng AM, Liu PX, Zhang LA, Fan FY (2010) Toxicologic

effects of gold nanoparticles in vivo by different adminis-

tration routes. Int J Nanomedicine 5:771–781

Zhong N, Xu J (2008) Synergistic activation of the human

MnSOD promoter by DJ-1 and PGC-1alpha: regulation by

SUMOylation and oxidation. Hum Mol Genet

17:3357–3367

2727 Page 8 of 8 J Nanopart Res (2014) 16:2727

123