Chemosphere 208 (2018) 196e206

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Chemosphere
journal homepage: www.elsevier.com/locate/chemosphere

Effects of coating materials on antibacterial properties of industrial

and sunscreen-derived titanium-dioxide nanoparticles on Escherichia
coli
Soyoung Baek a, Sung Hee Joo a, *, Pat Blackwelder b, c, Michal Toborek d
a
Department of Civil, Architectural, and Environmental Engineering, University of Miami, 1251 Memorial Dr. McArthur Engineering Building, Coral Gables,
FL 33146-0630, USA
b
Department of Marine Geosciences, University of Miami, 4600 Rickenbacker Causeway Miami, FL 33149-1098, USA
c
University of Miami Center for Advanced Microscopy (UMCAM), Department of Chemistry, 1301 Memorial Dr, Coral Gables, FL 33146-0630, USA
d
Department of Biochemistry and Molecular Biology, School of Medicine, University of Miami, 1011 NW 15th Street, Miami, FL, 33136, USA

h i g h l i g h t s

The coating with CMC, PVP, or SiO2 enhanced toxicity of TiO2 NPs on E. coli.

Toxicity of CMC or PVP coating was linked to size reduction of TiO2 NPs.

The presence of silica increased the generation of ROS from TiO2 NPs.

More ROS generation by silica was correlated with TiO2 NP toxicity.

a r t i c l e i n f o a b s t r a c t

Article history: Organic or inorganic stabilizers are often used for coating nanoparticles (NPs) in consumer products.
Received 29 October 2017 However, upon release of stabilized NPs into the environment, uncertainty exists as to the antimicrobial
Received in revised form properties of NPs due to stabilizers and the resultant bioaccumulation in organisms. This study in-
12 May 2018
vestigates antibacterial effects and subsequent mechanisms of TiO2 NPs on Escherichia coli (E. coli) in the
Accepted 27 May 2018
Available online 28 May 2018
presence and absence of stabilizers (CMC, PVP, and SiO2) commonly used in consumer products.
Compared with uncoated TiO2 NPs, the presence of any stabilizers tested in this study increased toxicity
Handling Editor: Tamara S. Galloway of NPs and enhanced growth inhibition in E. coli. While the particle sizes of TiO2 were smaller as the
result of coating with PVP or CMC and appeared to contribute to E. coli cell damage, the generation of
Keywords: reactive oxygen species (ROS) was independent of stabilizer type. In fact, coating with PVP and CMC
Antibacterial effect exerted ROS scavenging properties. In contrast, increased ROS production was observed at higher con-
Carboxymethyl cellulose (CMC) centrations of TiO2 and upon coating with SiO2. This impact of SiO2 can be related to the formation of a
E. coli TieOeSi chemical bond. The results of the present study emphasize the importance of nanoparticle
polyvinylpyrrolidone (PVP)
coating to their anti-bacterial activity and toxicity.
Reactive oxygen species (ROS)
© 2018 Elsevier Ltd. All rights reserved.
TiO2 nanoparticles

1. Introduction causing unknown and unintended environmental effects (Hegde

Growing employment of nanotechnology leads to increasing
releases of nanoparticles (NPs) into the natural environment,
Abbreviations: CMC, Carboxymethyl cellulose; PVP, Polyvinylpyrrolidone; E. coli,
Escherichia coli; IND TiO2 NPs, Industrial TiO2 NPs; SiO2þ IND TiO2 NPs, Industrial
TiO2 NPs in the presence of silica; SUN TiO2 NPs, Sunscreen-derived TiO2 NPs; NPs,
Nanoparticles; TiO2, Titanium dioxide.
* Corresponding author.
E-mail address: s.joo1@miami.edu (S.H. Joo).
et al., 2016; Padmavathy and Vijayaraghavan, 2008; Wang et al.,
2016a). Due to the versatile properties of NPs, including surface
area, particle size, and quantum effects, NPs are widely used in
commercial products (Jeon et al., 2016; Wang et al., 2016b). TiO2
NPs are mainly used in personal-care products, coatings, cleaning
agents, and paints, producing around 10,000 tons per year globally
(Piccinno et al., 2012; Weir et al., 2012). Such wide and increasing
productions also raise concerns regarding potential toxicity to
living organisms (Jeon et al., 2016; Leung et al., 2016). NPs can
negatively affect human health by targeting both the human host

https://doi.org/10.1016/j.chemosphere.2018.05.167
0045-6535/© 2018 Elsevier Ltd. All rights reserved.
 S. Baek et al. / Chemosphere 208 (2018) 196e206
and its microbiomes.
Escherichia coli (E. coli) is often found as single cell bacteria in the
intestines of animals and humans and in an aquatic environment. It
is the most commonly used prokaryotic organism used in research
and an excellent host for producing various proteins. Research has
revealed that the toxicity of TiO2 NPs to E. coli is increased by small
particle sizes and large surface areas (Smijs and Pavel, 2011). In
addition, the degree of toxicity in E. coli was found to be influenced
by ionic strength and electrolytes, which influence the aggregation
of TiO2 (Lewicka et al., 2011). Given that it is widely used in labo-
ratory to study the toxicity of NPs and celleNP interaction (Lin et al.,
2014; Baek et al., 2017), E. coli was chosen as the model bacteria
over other bacteria.
Research has revealed increasing toxicity of TiO2 NPs to E. coli at
small particle sizes and large surface areas (Smijs and Pavel, 2011).
In addition, the degree of toxicity in E. coli was influenced by ionic
strength and electrolytes that affect the aggregation of TiO2
(Lewicka et al., 2011). As with other types of NPs, TiO2 NPs are often
coated with stabilizers to prevent aggregation (Labille et al., 2010;
Lewicka et al., 2011; Smijs and Pavel, 2011; Jeon et al., 2016). Thus,
TiO2 may be present in a bare or coated form of NPs in the aquatic
environment (Dalai et al., 2012). Engineered polymers or organic
and inorganic substances may serve as coating material and act as
stabilizers for dispersal of colloidal NPs. Stabilizers have been found
to modify the physicochemical properties of NPs, thereby affecting
particle stability and mobility by electrosteric repulsion (Jafry et al.,
2011; Dalai et al., 2012; Wang and Fan, 2014), and resulting in
altered interaction with organisms (Batley et al., 2012).
According to the DLVO theory, nanoparticle aggregation is
controlled by Van der Waals attractive and electrostatic double-
layer forces (Hotze et al., 2010). Thus, stabilizers influence the
properties of NPs by adsorption of coating materials through
enhancing NPs’ surface charge or presenting steric or electrosteric
repulsion (Huynh and Chen, 2011). However, the impact of poly-
meric organic and inorganic matter coatings on TiO2 NPs toxicity
and biological effects has not been fully understood (Li, 2011).
Although interactions of TiO2 NPs with bacterial cells have been
examined, the majority of these studies have been limited to
toxicity impact. Few reports have evaluated the effect of coating
materials on NP-cell interactions, which is critical in environmental
health and safety. In the present study, we investigated the inter-
action between TiO2 NPs and bacterial cells by the model coating
materials to evaluate the antibacterial impact on E. coli, with further
implications to the toxicity of NPs in the environment.
Several hypotheses are drawn in our study. First, there is an
impact of polymeric organic- and inorganic-matter coatings on
TiO2 NPs’ toxicity and biological effects on E. coli. In addition,
providing insight into this mechanism is a goal since the mecha-
nism is not fully understood (Li, 2011). Although interactions of
TiO2 NPs with bacterial cells have been examined, the majority of
those studies has been limited to toxicity impact of non-coated NPs.
Second, the interaction between TiO2 NPs and bacterial cells is
influenced by the model coating materials. This approach enables
us to evaluate the antibacterial impact of these coated NPs on E. coli,
with further implications to the toxicity of NPs in the environment.
Additionally, TiO2 NPs coated with carboxymethyl cellulose
(CMC) and polyvinylpyrrolidone (PVP) (both neutrally charged
polymers), as well as inorganic coating (SiO2), exhibit bacterial
growth inhibition, ROS generation, and changes in morphology as a
function of size, exposure time and concentration. The organic
coating materials chosen in our study (CMC and PVP) are
commonly used in food-additive and personal-care products (Dalai
et al., 2012; Wang and Fan, 2014).
SiO2 was examined since it is another coating material widely
employed to improve the photocatalytic activity of TiO2 NPs in
197
various commercial products (Nur, 2006; Jafry et al., 2011). Studies
were conducted on industrial SiO2, including SiO2-coated TiO2 NPs
(Jafry et al., 2011; Siddiquey et al., 2008). Since TiO2 NPs are
commonly used in consumer products, comparing the toxicity of
SiO2-coated sunscreen-derived TiO2 with industrial SiO2-coated
TiO2 could exhibit the extent of toxicity upon exposure to
sunscreen-derived TiO2. The SiO2eTiO2 NPs extracted from the
sunscreen and industrial SiO2eTiO2 NPs were then investigated to
determine physicochemical characteristics of the NPs and the de-
gree of nanotoxicity on bacteria.
2. Materials and methods
2.1. Nanoparticle preparation
Industrial and sunscreen-derived TiO2 NPs were used in all ex-
periments. Industrial TiO2 NPs (TiO2, >99.7% purity, <25 nm size,
45e55 m2 g1 surface area, anatase), CMC (average MW: ~90,000),
PVP (average MW: ~10,000), and silica NPs (SiO2: >99.8% purity,
<14 nm size, 175e225 m2 g1) were purchased from Sigma-Aldrich
(St. Louis, MO). Commercially available sunscreen products con-
taining TiO2 NPs were purchased at a local drugstore in Miami, FL.
The CMC- and PVP-coated TiO2 NPs were prepared by dissolving
0.2% (w/v) of CMC or PVP in deionized (DI) water using a magnetic
stirrer for 12 h. After complete dissolution, TiO2 NPs were added to
each solution and dispersed under sonication for 3 min.
To identify the effect of silica present in sunscreen on antibac-
terial properties of TiO2 NPs, the TiO2 NPs were extracted from the
sunscreen product using a similar method as described in the
literature (Nischwitz and Goenaga-Infante, 2012). Briefly, extrac-
tions of NPs were performed by a series of ethanol and DI water
washes, followed by washing with hexane and subsequent soni-
cation and centrifugation. After discarding supernatants, suspen-
sions were washed in ethanol and DI water five times, and the
pellet dried in an oven at 100  C for 12 h. Industrial-grade SiO2 and
TiO2 NPs were mixed in DI water and sonicated for 3 min. Prior to
experimentation, all nanoparticle suspensions were subjected to
dispersion using a sonication probe for 1 min.
2.2. Characterization of TiO2 NPs
Nanoparticle surface morphology, physicochemical properties,
elemental analysis, and interactions with bacterial cells were
examined. Nanoparticle suspensions were analyzed using Energy
Dispersive Spectroscopy (EDS), Field Emission Scanning Electron
Microscopy (SEM, Philips XL-30 FEG), Transmission Electron Mi-
croscopy (JEOL 1400 TEM), Malvern Zetasizer Nano ZS90 analyzer,
and Fourier-Transform Infrared Spectroscopy (FTIR). For SEM and
TEM control cells and nanoparticle-treated E. coli cultures were
suspended in 2% glutaraldehyde in a phosphate-buffered saline
(PBS) buffer overnight. The pellet was then washed three times
with the PBS buffer. A series of graded ethanol at 20, 50, 70, 95, and
100% was used for dehydration in three changes of 5 min each. For
SEM the samples were then dried in three changes of hexame-
thyldisilazane (HMDS) on glass coverslips. After outgas sing over-
night, the samples were placed in a Cressington Sputter Coater and
coated with palladium (Pd). They were then imaged in the Philips
XL-30 Field Emission SEM equipped with an Oxford/Link EDS sys-
tem. To quantify average particle sizes and particle-cell in-
teractions, TEM (Philips CM 10, Eindhoven, Netherlands) analyses
were carried out. Cells were prepared as in the above described
protocol for SEM. However, after the last 100% ethanol wash cells
were embedded in Spurr Resin. The blocks were then sectioned in a
Porter Blum Ultramicrotome, placed on copper grids, and imaged in
a Philips CM-10 TEM (Eindhoven, Netherlands) at the Miller School
198 S. Baek et al. / Chemosphere 208 (2018) 196e206
of Medicine TEM Core at the University of Miami.
Digitally acquired micrographs were analyzed using Image J
software developed at the National Institutes of Health, USA to
calculate average NP diameter, size distribution, and standard de-
viation. A Malvern Zetasizer Nano ZS90 (Malvern Instruments,
Westborough, MA) was employed to measure the hydrodynamic
diameter of particles in suspension using dynamic light scattering
(DLS) at 25  C, and the zeta potential of the NPs in DI water.
For FTIR measurements, nanoparticle suspensions were made in
DI water and dried using a fume hood overnight at room temper-
ature. Using a Frontier series FTIR spectrometer by PerkinElmer
(Frontier, Perkin Elmer, Norwalk, CT), 50e100 mg of the powdered
TiO2 sample was applied to the clean crystal surface so that the
crystal was completely covered with sample before the force gauge
was applied and the spectrum taken. The crystal was thoroughly
cleaned between samples to ensure no cross contamination
occurred. Before each measurement, a blank measurement was
taken. Each sample was scanned four times, and the summation of
its spectra was analyzed.
2.3. E. coli DH5-Alpha cultivation and cell-viability analysis
E. coli cultured for characterizing the nanoparticle-cell in-
teractions was grown as follows. A pUC19 vector was transformed
into Max Efficiency DH5a™ Competent Cells (Invitrogen, CA). The
transformed cells were cultivated on Luria Bertani (LB) agar plates
containing ampicillin. The E. coli colonies were incubated in bac-
terial culture tubes with LB at 37  C for 12e14 h in an incubator
(MaxQ 4000, Thermo Scientific, NY). The growth inhibition of E. coli
was assessed by exposing cultivated cells to different concentra-
tions of TiO2 NP suspensions (10, 50, 250 mg L1) for 6 h. A bacterial
cell culture free of TiO2 NPs was used as a control.
The growth inhibition of E. coli was measured by optical den-
sities of UVevis absorption at 600 nm (OD600). E. coli were inocu-
lated in LB and incubated for 12e16 h at 37  C in an incubator with
shaking at 225 rpm. The cultivated E. coli cells (with an OD600 of
0.45) were exposed to different concentrations (0, 10, 50, or
250 mg L1) of TiO2 nanoparticle suspension in LB for 6 h at 37  C in
an incubator at 225 rpm. Then, the aliquots of 200 mL were added
into a 96-well plate, and OD600 was measured using a microplate
reader (Spectra MAX 190, Molecular Devices, CA) with an interval
of 2 h. The growth inhibition of E. coli was calculated as percentage
of survived cells compared to control cells after 6 h of exposure.
Samples were plated in triplicate, and all experiments were per-
formed under sterile conditions.
2.4. Quantification of ROS generation
To measure the generation of ROS from TiO2 NPs, a fluorogenic
dye 20 ,70 -dichlorofluorescin diacetate (DCFDA) was used following
the manufacturer's instructions (Abcam Inc, Toronto, ON, Canada).
After diffusion into the cell, DCF-DA is deacetylated by cellular es-
terases to a non-fluorescent compound that is oxidized by ROS into
20 , 70 - dichlorofluorescein (DCF). The method allows to assess hy-
droxyl, peroxyl and other ROS activity within cells. Briefly, culti-
vated cells exposed to different coating materials and TiO2 NP
concentrations were seeded in the 96-well plate (black color, clear
bottom). The treated cells were washed with PBS, followed by an
addition of the DCF-DA dye (10 mM) to each well and incubation for
30 min at 37  C. DCF fluorescence was detected by fluorescence
spectroscopy with excitation and emission spectra of 485 nm and
535 nm, respectively.
2.5. Statistical analysis
All the experiments were performed in a triplicate. The statis-
tical student's t-test with Graph Pad Prism 3.0 analysis (Graph Pad
Software Inc., San Diego, CA) was used for the statistical analysis. All
the experimental values were compared to their corresponding
control values and showed statistical significance at p < 0.05.
3. Results and discussion
3.1. Characterization of TiO2 NPs with and without coatings
The morphology and size of industrial and sunscreen-derived
TiO2 NPs were analyzed using SEM and TEM in the presence and
absence of coating materials (CMC, PVP, and SiO2). As shown in
Fig. 1, spheres which were, in some cases, aggregated were
observed in all samples. However, sunscreen-derived (SUN) TiO2
NPs exhibited the largest particle size (up to 284 nm), (Table 1).
Industrial TiO2 NPs (IND) in the absence of CMC or PVP exhibited
significant aggregation (Fig. 1), larger particle sizes, and low zeta
potential values (Table 1) as compared to coated TiO2 NPs.
In contrast, the presence of CMC or PVP significantly enhanced
the stability of IND TiO2 NPs as determined by the zeta potential
values measured at pH 7 (Table 1), with substantial shape changes
characterized by spherical particles and relatively smaller TiO2 NP
sizes (Fig. 1A and Table 1). Such substantial shape transformation by
stabilizers has been found in other studies (Othman et al., 2012;
Tejamaya et al., 2012). TiO2 NPs have also been found to exhibit a
fully dispersed and stabilized form in natural water containing
organic materials (Planchon et al., 2013). In this study we found the
particle size distributions measured by ImageJ revealed average
sizes of 29.4 nm (IND TiO2 NPs), 16.3 nm (CMC þ TiO2 NPs), and
9.8 nm (PVP þ TiO2 NPs) (Table 1).
In colloidal suspension, the hydrodynamic diameters were on
the order of IND TiO2 NPs (625.7 nm) > SiO2 mixed with IND TiO2
NPs (594.5 nm) > IND TiO2 NPs mixed with CMC (547.1 nm) > SUN
TiO2 NPs (335.7 nm) > IND TiO2 NPs mixed with PVP (206.6 nm).
However, the difference in hydrodynamic diameters of CMC- and
PVP-mixed TiO2 NPs was observed. This suggests an adsorption
effect related to the molecular weight of the stabilizers. For
example, CMC that has a greater molecular weight and longer
chains can adsorb more on the NPs’ surfaces in comparison to PVP,
thereby exhibiting stronger binding with the NPs. This can result in
larger particle sizes. Short chain dispersants may be more effective
in stabilizing NPs in suspension; this has been demonstrated in
other studies (Liufu et al., 2005; Othman et al., 2012).
TiO2 NPs in the absence of coating materials were aggregated
(Fig. 1B). Interestingly, silica-coated IND TiO2 NPs clearly shows the
distribution of silica around the NPs. These bonded clusters appear
to significantly aggregate in comparison with those in the presence
of either CMC or PVP. In addition, TiO2 and SiO2 NPs appear to be
interconnected to form individual spheres. A similar observation
was made in the study in which silica NPs did not coat TiO2 NPs
(Jaroenworaluck et al., 2006).
Increasing the amount of silica in coatings on TiO2 NPs does not
appear to be correlated to change in physical characteristics, e.g.,
forming individual amorphous spheres (Jaroenworaluck et al.,
2006). However, the study by Zhang et al. (2009) showed that
increasing the number of silica NPs can result in a notable disper-
sion effect of the NPs. In the current study, elemental analysis (EDS)
composition of IND TiO2 NPs, IND TiO2 NPs mixed with silica, and
SUN TiO2 NPs were performed. The results confirmed Ti purity in
IND TiO2 NPs as well as 60% Ti and 40% Si from SUN and IND TiO2
NPs mixed with silica NPs (Fig. 1C).
 S. Baek et al. / Chemosphere 208 (2018) 196e206 199

Fig. 1. SEM (A) and TEM (B) images of 5 types of TiO2 NPs: (a) IND TiO2 (Industrial TiO2 NPs), (b) CMC þ IND TiO2 NPs (Industrial TiO2 NPs mixed with CMC), (c) PVP þ IND TiO2 NPs
(Industrial TiO2 NPs mixed with PVP), (d) SiO2þIND TiO2 NPs (Industrial TiO2 NPs mixed with silica NPs), and (e) SUN TiO2 NPs (Sunscreen-derived TiO2 NPs); TiO2 suspension
concentration: 50 mg L1. EDS images of TiO2 NPs (C); (a) IND TiO2, (b) CMC þ IND TiO2 NPs, (c) PVP þ IND TiO2 NPs, (d) SiO2þIND TiO2 NPs, (e) SUN TiO2 NPs.

3.2. The impact of coating materials on antibacterial properties of
TiO2 NPs in E. coli
We propose that the differences in morphology, size, zeta po-
tential, and stability of TiO2 NPs in the presence of coating materials
(CMC, PVP, and SiO2) may affect its antibacterial activity. Therefore,
the effect of these commonly used coating materials in consumer
products on the antibacterial properties of TiO2 NPs was investi-
gated. E. coli was exposed to TiO2 NPs for 6 h and growth inhibition
was recorded as a function of time and concentration (Fig. 2A).
Overall, bacterial growth was inhibited over time, and cell viability
decreased significantly at the highest TiO2 NP concentration
(250 mg L1).
Above 50 mg L1 of TiO2 coated with PVP, a slightly higher
antibacterial effect was evident compared to IND TiO2 NPs and CMC-
coated TiO2 NPs (Fig. 2A). At 250 mg L1 of TiO2 NP suspension, the
coating materials (CMC and PVP) induced higher toxicity to E. coli
compared to non-coated TiO2 NPs. The greatest antibacterial effect
was found in the order of PVP þ TiO2 NPs > CMC þ TiO2 NPs > IND
TiO2 NPs. This order of toxicity appeared to correlate with particle
sizes (IND TiO2 NPs > CMC þ TiO2 NPs > PVP þ TiO2 NPs). Thus,
stabilizers such as CMC and PVP appear to increase the toxicity of
TiO2 NPs because they play a role as dispersants that prevent
nanoparticle aggregation (Liufu et al., 2005; Othman et al., 2012).
The addition of dispersant resulted in the production of more
dispersed, more stabilized aqueous suspension, and thus smaller
nanoparticle sizes. Such particle sizes indicated a variance among
different coating materials and their properties. For instance, PVP-
200 S. Baek et al. / Chemosphere 208 (2018) 196e206
Fig. 1. (continued).
mixed TiO2 NPs exhibited a smaller particle size compared to that of
CMC-mixed TiO2 NPs because CMC has a heavier weight and longer
chains in comparison to PVP, which can adsorb more on NPs. Small
NPs have more reactivity and a higher probability to be internalized
by uptake of NPs than large ones. Similar correlations between
smaller particle sizes and higher toxicity were also observed in other
studies in which such an effect was linked to increased generation of
ROS (Pagnout et al., 2012; Lin et al., 2014) or the degree of nano-
particle aggregation (Chowdhury et al., 2012).
The antibacterial effect of TiO2 on E. coli in the presence of silica
was examined, and similar trends as those observed in Fig. 2A were
found (Fig. 2B). Specially, increasing toxicity over longer time
exposure and significant growth inhibition at the highest TiO2
concentration (250 mg L1) were found. A significant antibacterial
effect (the lowest cell viability) from either SiO2 mixed TiO2 NPs
(SiO2þTiO2) or SUN TiO2 NPs at 250 mg L1 was observed (Fig. 2B)
with the order of SiO2 coated TiO2 NPs > SUN TiO2 NPs > IND TiO2
NPs. These results confirm that anti-bacterial impact is influenced
by the type of surface coating and particle sizes. Indeed, the cata-
lytic activity of TiO2 was demonstrated to be particle size depen-
dent, and the addition of silica NPs exhibited a scattering effect,
(Van Grieken et al., 2002). In addition, TiO2 NPs coated with silica
exhibited higher toxicity than uncoated TiO2 NPs due to an
enhanced surface area, resulting in increased adsorption and
reactivity (Jafry et al., 2011).
The slightly different toxicity effects between silica-coated IND
TiO2 NPs and SUN TiO2 NPs could be caused, at least in part, by their
physicochemical characteristics. For instance, the significantly
larger particle size observed in SUN TiO2 NPs (Fig. 1B) compared to
the particle size in SiO2-coated IND TiO2 NPs (Table 1) may have
resulted in a lower antibacterial effect. In addition, the higher
growth inhibition by SUN TiO2 NPs compared to IND TiO2 NPs could
be caused by the chemical composition effect in the presence of
silica (e.g., complexation or adsorption of Si on TiO2) rather than
physicochemical properties of TiO2 NPs.
3.3. Effects of coated and uncoated TiO2 NPs on ROS generation in
E.coli
The release of ROS (hydroxyl, superoxide radicals, hydrogen
peroxide, and singlet oxygen) occurs when TiO2 absorbs UV light as
a semiconductor, generating electrons and positively charged holes
(hþ Þ (Carlotti et al., 2009; Gupta and Tripathi, 2011). TiO2 NPs are
known to induce oxidative stress and DNA damage in E. coli cells
(Kumar et al., 2011; Dasari et al., 2013); however, the impact of
coating materials on TiO2 NP-induced antibacterial effect on E. coli
is not fully understood. Therefore, ROS generation was assessed
following the exposure of E. coli to TiO2 NPs in the presence and
absence of the coating materials. Overall, ROS generation appeared
to increase as the function of TiO2 nanoparticle concentration and
exposure time. However, ROS generation was similar among the
three types of TiO2 NPs studied in the present paper. These results
are in contrast to the toxicity effects that were most significant in
PVP-coated IND TiO2 NPs, followed by CMC-coated IND TiO2 NPs,
and IND TiO2 NPs, suggesting that an increase in ROS may not be
the primary mechanism responsible for toxicity of TiO2 NPs.
The lack of differences in oxidative stress induced by the studied
TiO2 NPs suggests that CMC and PVP coating materials may react
with ROS generated from TiO2 NPs, scavenging ROS as per the
following chemical reaction (Joo and Zhao, 2008):
 S. Baek et al. / Chemosphere 208 (2018) 196e206 201

Fig. 2. Growth curves of E. coli at different concentrations (control as well as 10, 50, and 250 mg L1) and cell-growth inhibition rate (%) of (A) TiO2 NPs with and without organic
coating materials: IND TiO2 NPs, CMC þ IND TiO2 NPs, and PVP þ IND TiO2 NPs, (B) TiO2 NPs with and without inorganic coating materials; IND TiO2 NPs, SiO2þIND TiO2 NPs, and
SUN TiO2 NPs derived from sunscreen. The asterisk indicates a significant difference relative to the control (*, p < 0.05) based on the Student's t-test. The error bars represent the
standard deviation (n ¼ 3).

Table 1
Zeta potential, hydrodynamic diameter (nm), and TEM particle size (nm) of TiO2 NPs.

NPs Zeta potential (mV) Hydrodynamic diameter (nm) TEM particle size (nm)

IND TiO2 14.23 625.7 29.4

CMC þ IND TiO2 61.5 547.1 16.3
PVP þ IND TiO2 43.6 206.6 9.8
SiO2þIND TiO2 15 594.5 29.6
SUN TiO2 14.8 335.7 284.5
202 S. Baek et al. / Chemosphere 208 (2018) 196e206

OH þ CMC / H2O þ CMC

(1)
The pseudo-first-order rate constant of this reaction was re-
ported as 1  109 M1 s1 (Wach et al., 2004). Interactions of TiO2
NPs with stabilizers have been shown to inhibit ROS generation
(Tuchina and Tuchin, 2010). Other studies proposed the generation
of ROS from NPs, and described the interaction of NPs with cellular
components (Hegde et al., 2016; Wang et al., 2016a, 2016b). For
example, it was reported that polymer coatings blocked or elimi-
nated the photocatalytic activity of TiO2 NPs, thereby minimizing
free radical formation on TiO2 surfaces (Lee et al., 2007b).
Similar observations were made in the reports in which the
correlation between growth inhibition of treated E. coli and ROS
generation was inconsistent (Kumar et al., 2011; Dalai et al., 2012).
Instead of the ROS effect, particle size may have attributed to the
toxicity. Kumar et al. (2011) confirmed using electron microscopy
that a significant uptake of TiO2 NPs occurred on E. coli through the
adhesion of aggregated NPs on the surface of bacteria and the
subsequent changes in morphology (Kumar et al., 2011). In addi-
tion, NPs may penetrate bacterial cell membranes and inhibit their
metabolic enzymes (Pal et al., 2007).
In contrast to lack of influence of uncoated or polymer-coated
IND TiO2 NPs on oxidative stress (Fig. 3A), a statistically signifi-
cant generation of ROS was observed in E. coli cells treated with
SiO2-coated IND TiO2 NPs at 250 mg L1 (Fig. 3B). The explanation of
these results can be related to an increase in the surface area and
the band structure through the quantum-size effect, leading to
enhanced photocatalytic activity of TiO2 (Kim et al., 2005). The
quantum-size effect, i.e. the energy gap between the valence and
conduction bands is enlarged as the particle size of TiO2 decreases
(Zhang et al., 2009). In support of our observations, enhanced band-
gap energy of TiO2 on the silica surface has been described in other
studies (Kim et al., 2005; Park and Kang, 2005; Zhang et al., 2009).
The lower ROS generation from SUN TiO2 NPs compared to that
from SiO2-coated IND TiO2 NPs could be due to the larger particle
size or different chemical compositions (Xiong et al., 2011). As
indicated in other studies and demonstrated in our present work,
the correlation between ROS generation and antibacterial activity is
influenced by physicochemical properties and chemical composi-
tion of the coating materials (Park et al., 2013; Leung et al., 2016).
Indeed, it was reported that NPs can induce ROS-independent
antibacterial impact due to their interactions with cell mem-
branes (Leung et al., 2014).
3.4. SEM and TEM analyses of E.coli cells exposed to TiO2 NPs
To gain further insight into TiO2 antibacterial effects on E.coli
related to the coating materials, SEM and TEM images of E. coli after
exposure to TiO2 NPs at different concentrations in the presence
and absence of the coating materials were examined. At the high
nanoparticle concentration (250 mg L1), more aggregated forms
appeared, and more NP particles were attached to cells, whereas at
the low concentration (50 mg L1), fewer aggregations were seen
(Fig. 4). Attachment of NPs to cell membranes and indications of

Fig. 3. ROS generation assessment of five types of TiO2 NPs (50 and 250 mg L1) by fluorescence intensity ratio (%) after exposure to E. coli over 6 h in the presence of (A) organic
coating materials and (B) inorganic coating materials. The asterisk indicates a significant difference relative to the control (*, p < 0.05) based on the Student's t-test. The error bars
represent the standard deviation (n ¼ 3).

Fig. 4. SEM images of E. coli after exposure to different TiO2 NPs (in the absence and in the presence of organic and inorganic coating materials) in different concentrations:
Industrial TiO2 NPs, CMC þ IND TiO2 NPs, PVP þ IND TiO2 NPs, SiO2þIND TiO2 NPs, and SUN TiO2 NPs derived from sunscreen at 50 and 250 mg L1 concentrations.
 S. Baek et al. / Chemosphere 208 (2018) 196e206 203

damage were particularly notable at the high concentration of TiO2
NPs. Such marked cell disruption seen at high concentrations of NPs
have also been found in other studies (Kim et al., 2005; Hegde et al.,
2016; Leung et al., 2016).
Fig. 5 illustrates the ultrastructural relationships in fixed and
sectioned bacterial cells after treatment with 250 mg L1 IND TiO2
NPs coated with CMC, PVP, or SiO2, and SUN TiO2 NPs. While IND
TiO2 nanoparticle–exposed E.coli cells showed significant adsorp-
tion of NPs, several nanoparticle aggregated forms surrounded the
cells (Fig. 5A) In addition, some NPs penetrated into the bacterial
cells and were associated with cell damage (Fig. 5B).
CMC-coated IND TiO2 NPs showed aggregates in the cells similar
to those induced by IND TiO2 NP. These images include adsorption
of NPs on the cells, intracellular interactions between NPs and the
cells, cell damage, and penetration of the NPs into the cells (Fig. 5C
and D). Some of the cells appeared dead with clear intracellular
space, while some of the apparently living cells remained sur-
rounded by NPs (Fig. 5C). Significant numbers of dead cells
observed in both CMC- and PVP-coated IND TiO2 NPs could be
associated with smaller particle sizes that enabled them to cause
more toxicity. Dead cells appeared to show leakage of cell contents
(Fig. 5D). Importantly, images in Fig. 5 correspond to the previously
described antibacterial effects of NPs (Fig. 2). These effects include
lower toxicity from IND TiO2 NPs compared to that from CMC- or
PVP-coated IND TiO2 NPs. Similarly, PVP-coated IND TiO2 NPs
adhered to cell surfaces. Cells surrounded by NPs with transparent
intracellular space appeared to be dead, while other cells not in
direct contact with the NPs remained intact (Fig. 5E and F).
Direct physical deformation such as membrane deformation and
cell wall damage were observed in PVP-coated IND TiO2 NPs with
black colors showing the adhesion of the NPs onto the cell surfaces
(Fig. 5). While intracellular interactions were not apparent in PVP-

Fig. 5. E. coli ultrastructure (TEM) after exposure to three different TiO2 NPs (A) in the absence and in the presence of organic coating materials: IND TiO2 NPs, CMC þ IND TiO2 NPs,
and PVP þ IND TiO2 NPs, (B) in the absence and in the presence of inorganic coating materials: IND TiO2 NPs, SiO2þIND TiO2 NPs, SUN TiO2 NPs derived from sunscreen at
250 mg L1 of NP concentration.
204 S. Baek et al. / Chemosphere 208 (2018) 196e206
coated TiO2 NPs, more severe membrane damage occurred
compared to those caused by CMC-coated IND TiO2 NPs, or un-
coated IND TiO2 NPs. Adsorption of NPs on cells may inhibit bac-
terial movement, causing cellular metabolic disturbance and cell
death. Similar observations of the disruption of bacterial mem-
branes were also found in other studies (Dalai et al., 2012; Lin et al.,
2014; Leung et al., 2016; Wang et al., 2016a). Coating materials may
facilitate more interactions on the bacteria surfaces, and ROS
generated from the NPs may result in disruption of the bacteria cell
membranes (Lin et al., 2014). Studies reveal that interactions be-
tween cells and NPs vary depending on the NP composition, coating
materials, and model organisms used (Xiong et al., 2011; Batley
et al., 2012).
In contrast, images of the ultrastructure of sectioned bacteria
cells treated with 250 mg L1 of SUN TiO2 NPs and SiO2-coated IND
TiO2 NPs did not exhibit cell attachment (Fig. 5GeJ). Although they
appeared to form aggregates, SiO2-coated IND TiO2 NPs were not
attached to the bacteria membranes (Fig. 5 GeH). Similarly, larger
aggregates of NPs around the cells were observed from SUN TiO2
NPs, but these NPs were not interacting with the cells (Fig. 5 IeJ).
Such large SUN TiO2 NP aggregates are consistent with their rela-
tively large particle size and TEM particle-size distribution analysis.
Notably, while CMC- or PVP-coated IND TiO2 NPs induced cell
distortion, internalization, and membrane damage, the presence of
silica (either on IND or SUN TiO2 NPs) prevented penetration of NPs
into the bacterial membrane.
3.5. FTIR analyses of the coated TiO2 NPs
FTIR spectroscopy was employed to gain insight into chemical
interactions of the coating materials with NPs. The FTIR patterns of
TiO2 NPs with organic coating materials (CMC and PVP; Fig. 6A) and
silica (Fig. 6B) were examined. As shown in Fig. 6A, IND, CMC-IND
TiO2 NPs, and PVP-IND TiO2 NPs had similar patterns without
notable peaks. The broad band at approximately 650e800 cm1
was attributed to the absorption peak of TieO, confirming the
presence of TiO2 in all the samples (Mirabedini et al., 2011; Guo
et al., 2013; Zhang et al., 2014). The absorption peak (COO) at
1631 cm1 suggests possible surface reaction between carboxyl
groups of CMC and hydroxyl groups of TiO2 from CMC-coated IND
Fig. 6. FTIR spectra of TiO2 NPs (A) in the absence and in the presence of organic coating materials (CMC and PVP): IND TiO2 NPs, CMC-coated IND TiO2 NPs, and PVP-coated IND
TiO2 NPs; (B) in the absence and in the presence of inorganic coating materials: IND TiO2 NPs, SiO2-coated IND TiO2 NPs, and SUN TiO2 NPs.
TiO2 NPs (Liang et al., 2004; Liu et al., 2016). The carbonyl band (C]
O) of PVP from PVP-coated IND TiO2 NPs was observed at
1656 cm1 (Cai et al., 2015; Wu et al., 2017). In addition, Fig. 6B
shows absorption peaks of SieOeSi at 1081 cm1 from SiO2-coated
IND TiO2 NPs and at 1016 cm1 from SUN TiO2 NPs due to the
presence of silica in solution (Feng et al., 2015; Wang and Chen,
2015).
Another absorption peak of TieOeSi at 924 cm1 occurred in
SUN TiO2 NPs. These observations are consistent with other studies
that have described the formation of TieOeSi bonds, which are
related to the photocatalytic activity between 910 and 960 cm1
(Lee et al., 2007a; Rasalingam et al., 2014). SiO2-coated IND TiO2
NPs could modify the band structure that reflects the band gap
energy of TiO2eSiO2 through the chemical bond (TieOeSi) at the
interface of a silica-coated layer or the surface of TiO2 NPs (Kim
et al., 2005; Park and Kang, 2005; Zhang et al., 2009). As shown
in Fig. 5B, TiO2 NPs were evenly distributed in the presence of SiO2
NPs. This suggests that silica is a catalyst that increases hydro-
phobic properties of NPs, thus enhancing the reactivity of TiO2 by
stabilizing its particles (Rasalingam et al., 2014).
4. Conclusion
The addition of coating materials such as CMC, PVP and silica
were found to prevent significant TiO2 aggregation through
dispersion. Such stabilizers changed physicochemical properties
(particle sizes and zeta potential) of TiO2 NPs. Although CMC- or
PVP-coated TiO2 NPs produced stable TiO2 suspension, with a
cluster size smaller than that of uncoated TiO2 NPs, there was no
difference in ROS generation between uncoated and coated TiO2
NPs, indicating possible scavenging of ROS by CMC and PVP. On the
other hand, coated TiO2 induced significant cell death through in-
teractions with bacteria membranes and/or penetration into the
cells. This finding is consistent with cell culture results showing
significant growth inhibition of E. coli at the highest concentration
of IND TiO2 NPs coated with CMC or PVP.
Significant antibacterial growth inhibition was observed in
silica-coated IND TiO2 NPs and SUN TiO2 NPs but not in uncoated
IND TiO2 NPs at the highest concentration of TiO2 NPs. However, in
contrast to CMC- and PVP-coated IND, substantial ROS generation
 S. Baek et al. / Chemosphere 208 (2018) 196e206
was observed in silica-coated IND TiO2 NPs, notably at the highest
concentration of IND TiO2 NPs. This effect may result from the
impact of increased reactivity of TiO2 NPs due to stabilization by
silica.
Our findings have implications to the development of nano-
products that use coating materials, because 1) the types of coating
materials affect the stability and the reactivity of TiO2 NPs, 2)
smaller and more stable particle sizes exhibit higher toxicity, and 3)
silica-coated TiO2 NPs have a more significant antibacterial effect
than that of CMC- or PVP-coated TiO2 NPs. Future research needs to
evaluate the accumulation of NPs in microorganisms using
different types of coated TiO2 nanoproducts under well-controlled
environmental conditions, such as variability in exposure media,
UV radiation, temperature, and co-existing contaminants that may
contribute to synergistic microorganism growth inhibition.
References
Baek, S., Joo, S.H., Kumar, N., Toborek, M., 2017. Antibacterial effect and toxicity
pathways of industrial and sunscreen ZnO nanoparticles on Escherichia coli.
Journal of Environmental Chemical Engineering 5, 3024e3032.
Batley, G.E., Kirby, Jason K., McLaughlin, Michael J., 2012. Fate and risks of nano-
materials in aquatic and terrestrial environments. Accounts Chem. Res. 46,
854e862.
Cai, Y., Liu, M., Song, X., Zhang, J., Wei, Q., Zhang, L., 2015. A form-stable phase
change material made with a cellulose acetate nanofibrous mat from bicom-
ponent electrospinning and incorporated capricemyristicestearic acid ternary
eutectic mixture for thermal energy storage/retrieval. RSC Adv. 5,
84245e84251.
Carlotti, M.E., Ugazio, E., Sapino, S., Fenoglio, I., Greco, G., Fubini, B., 2009. Role of
particle coating in controlling skin damage photoinduced by titania nano-
particles. Free Radic. Res. 43, 312e322.
Chowdhury, I., Cwiertny, D.M., Walker, S.L., 2012. Combined factors influencing the
aggregation and deposition of nano-TiO2 in the presence of humic acid and
bacteria. Environ. Sci. Technol. 46, 6968e6976.
Dalai, S., Pakrashi, S., Kumar, R.S.S., Chandrasekaran, N., Mukherjee, A., 2012.
A comparative cytotoxicity study of TiO2 nanoparticles under light and dark
conditions at low exposure concentrations. Toxicol. Res. 1, 116.
Dasari, T.P., Pathakoti, K., Hwang, H.-M., 2013. Determination of the mechanism of
photoinduced toxicity of selected metal oxide nanoparticles (ZnO, CuO, Co3O4
and TiO2) to E. coli bacteria. J. Environ. Sci. 25, 882e888.
Feng, X., Zhang, S., Wu, H., Lou, X., 2015. A novel folic acid-conjugated TiO(2)-SiO(2)
photosensitizer for cancer targeting in photodynamic therapy. Colloids and
surfaces. B. Biointerfaces 125, 197e205.
Guo, H., Wang, W., Liu, L., He, Y., Li, C., Wang, Y., 2013. Shape-controlled synthesis of
Ag@TiO2 cage-bell hybrid structure with enhanced photocatalytic activity and
superior lithium storage. Green Chem. 15.
Gupta, S.M., Tripathi, M., 2011. A review of TiO2 nanoparticles. Chin. Sci. Bull. 56,
1639e1657.
Hegde, K., Brar, S.K., Verma, M., Surampalli, R.Y., 2016. Current understandings of
toxicity, risks and regulations of engineered nanoparticles with respect to
environmental microorganisms. Nanotechnol. Environ. Eng. 1.
Hotze, E.M., Phenrat, T., Lowry, G.V., 2010. Nanoparticle aggregation: challenges to
understanding transport and reactivity in the environment. J. Environ. Qual. 39,
1909.
Huynh, K.A., Chen, K.L., 2011. Aggregation kinetics of citrate and poly-
vinylpyrrolidone coated silver nanoparticles in monovalent and divalent elec-
trolyte solutions. Environ. Sci. Technol. 45, 5564e5571.
Jafry, H.R., Liga, M.V., Li, Q., Barron, A.R., 2011. Simple route to enhanced photo-
catalytic activity of p25 titanium dioxide nanoparticles by silica addition. En-
viron. Sci. Technol. 45, 1563e1568.
Jaroenworaluck, A., Sunsaneeyametha, W., Kosachan, N., Stevens, R., 2006. Char-
acteristics of silica-coated TiO2 and its UV absorption for sunscreen cosmetic
applications. Surf. Interface Anal. 38, 473e477.
Jeon, S.-k., Kim, E.-j., Lee, J., Lee, S., 2016. Potential risks of TiO2 and ZnO nano-
particles released from sunscreens into outdoor swimming pools. J. Hazard
Mater. 317, 312e318.
Joo, S.H., Zhao, D., 2008. Destruction of lindane and atrazine using stabilized iron
nanoparticles under aerobic and anaerobic conditions: effects of catalyst and
stabilizer. Chemosphere 70, 418e425.
Kim, H.-J., Shul, Y.-G., Han, H., 2005. Photocatalytic properties of silica-supported
TiO2. Top. Catal. 35, 287e293.
Kumar, A., Pandey, A.K., Singh, S.S., Shanker, R., Dhawan, A., 2011. Engineered ZnO
and TiO2 nanoparticles induce oxidative stress and DNA damage leading to
reduced viability of Escherichia coli. Free Radical Biol. Med. 51, 1872e1881.
Labille, J., Feng, J., Botta, C., Borschneck, D., Sammut, M., Cabie, M., Auffan, M.,
Rose, J., Bottero, J.Y., 2010. Aging of TiO2 nanocomposites used in sunscreen.
Dispersion and fate of the degradation products in aqueous environment. En-
viron. Pollut. 158, 3482e3489.
205
Lee, J.-W., Kong, S., Kim, W.-S., Kim, J., 2007a. Preparation and characterization of
SiO2/TiO2 core-shell particles with controlled shell thickness. Mater. Chem.
Phys. 106, 39e44.
Lee, W.A., Pernodet, N., Li, B., Lin, C.H., Hatchwell, E., Rafailovich, M.H., 2007b.
Multicomponent polymer coating to block photocatalytic activity of TiO2
nanoparticles. Chem. Commun. 4815e4817.
Leung, Y.H., Ng, A.M., Xu, X., Shen, Z., Gethings, L.A., Wong, M.T., Chan, C.M.,
Guo, M.Y., Ng, Y.H., Djurisic, A.B., Lee, P.K., Chan, W.K., Yu, L.H., Phillips, D.L.,
Ma, A.P., Leung, F.C., 2014. Mechanisms of antibacterial activity of MgO: non-
ROS mediated toxicity of MgO nanoparticles towards Escherichia coli. Small
10, 1171e1183.
Leung, Y.H., Xu, X., Ma, A.P., Liu, F., Ng, A.M., Shen, Z., Gethings, L.A., Guo, M.Y.,
Djurisic, A.B., Lee, P.K., Lee, H.K., Chan, W.K., Leung, F.C., 2016. Toxicity of ZnO
and TiO2 to Escherichia coli cells. Sci. Rep. 6, 35243.
Lewicka, Z.A., Benedetto, A.F., Benoit, D.N., Yu, W.W., Fortner, J.D., Colvin, V.L., 2011.
The structure, composition, and dimensions of TiO2 and ZnO nanomaterials in
commercial sunscreens. J. Nanoparticle Res. 13, 3607e3617.
Li, Z., 2011. Mechanistic Insight into the Effect of Polymer and NOM Coatings on
Adhesion and Interactions between Nanoparticles and Bacteria.
Liang, H.F., Hong, M.H., Ho, R.M., Chung, C.K., Lin, Y.H., Chen, C.H., Sung, H.W., 2004.
Novel method using a temperature-sensitive polymer (methylcellulose) to
thermally gel aqueous alginate as a pH-sensitive hydrogel. Biomacromolecules
5, 1917e1925.
Lin, X., Li, J., Ma, S., Liu, G., Yang, K., Tong, M., Lin, D., 2014. Toxicity of TiO 2
nanoparticles to Escherichia coli: effects of particle size, crystal phase and water
chemistry. PLoS One 9, e110247.
Liu, X., Lin, B., Zhang, Z., Lei, H., Li, Y., 2016. Copper(ii) carboxymethylcellulose
(CMC-CuII) as an efficient catalyst for aldehydeealkyneeamine coupling under
solvent-free conditions. RSC Adv. 6, 94399e94407.
Liufu, S., Xiao, H., Li, Y., 2005. Adsorption of poly(acrylic acid) onto the surface of
titanium dioxide and the colloidal stability of aqueous suspension. J. Colloid
Interface Sci. 281, 155e163.
Mirabedini, A., Mirabedini, S.M., Babalou, A.A., Pazokifard, S., 2011. Synthesis,
characterization and enhanced photocatalytic activity of TiO2/SiO2 nano-
composite in an aqueous solution and acrylic-based coatings. Prog. Org. Coating
72, 453e460.
Nischwitz, V., Goenaga-Infante, H., 2012. Improved sample preparation and quality
control for the characterisation of titanium dioxide nanoparticles in sunscreens
using flow field flow fractionation on-line with inductively coupled plasma
mass spectrometry. J. Anal. At. Spectrom. 27, 1084.
Nur, H., 2006. Modification of titanium surface species of titania by attachment of
silica nanoparticles. Mater. Sci. Eng., B 133, 49e54.
Othman, S.H., Abdul Rashid, S., Mohd Ghazi, T.I., Abdullah, N., 2012. Dispersion and
stabilization of photocatalytic TiO2 nanoparticles in aqueous suspension for
coatings applications. J. Nanomater. 2012, 1e10.
Padmavathy, N., Vijayaraghavan, R., 2008. Enhanced bioactivity of ZnO nano-
particlesdan antimicrobial study. Sci. Technol. Adv. Mater. 9, 035004.
Pagnout, C., Jomini, S., Dadhwal, M., Caillet, C., Thomas, F., Bauda, P., 2012. Role of
electrostatic interactions in the toxicity of titanium dioxide nanoparticles to-
ward Escherichia coli. Colloids and surfaces. B. Biointerfaces 92, 315e321.
Pal, S., Tak, Y.K., Song, J.M., 2007. Does the antibacterial activity of silver nano-
particles depend on the shape of the nanoparticle? A study of the Gram-
negative bacterium Escherichia coli. Appl. Environ. Microbiol. 73, 1712e1720.
Park, O.K., Kang, Y.S., 2005. Preparation and characterization of silica-coated TiO2
nanoparticle. Colloid. Surface. Physicochem. Eng. Aspect. 257e258, 261e265.
Park, Y.H., Bae, H.C., Jang, Y., Jeong, S.H., Lee, H.N., Ryu, W.-I., Yoo, M.G., Kim, Y.R.,
Kim, M.K., Lee, J.K., Jeong, J., Son, S.W., 2013. Effect of the size and surface charge
of silica nanoparticles on cutaneous toxicity. Mole. Cell. Toxicol. 9, 67e74.
Piccinno, F., Gottschalk, F., Seeger, S., Nowack, B., 2012. Industrial production
quantities and uses of ten engineered nanomaterials in Europe and the world.
J. Nanoparticle Res. 14, 1109.
Planchon, M., Ferrari, R., Guyot, F., Gelabert, A., Menguy, N., Chaneac, C., Thill, A.,
Benedetti, M.F., Spalla, O., 2013. Interaction between Escherichia coli and TiO2
nanoparticles in natural and artificial waters. Colloids and surfaces. B. Bio-
interfaces 102, 158e164.
Rasalingam, S., Peng, R., Koodali, R.T., 2014. Removal of hazardous pollutants from
wastewaters: applications of TiO2-SiO2Mixed oxide materials. J. Nanomater.
2014, 1e42.
Siddiquey, I.A., Furusawa, T., Sato, M., Honda, K., Suzuki, N., 2008. Control of the
photocatalytic activity of TiO2 nanoparticles by silica coating with poly-
diethoxysiloxane. Dyes Pigments 76, 754e759.
Smijs, T.G., Pavel, S., 2011. Titanium dioxide and zinc oxide nanoparticles in sun-
screens: focus on their safety and effectiveness. Nanotechnol. Sci. Appl. 4,
95e112.
Tejamaya, M., Romer, I., Merrifield, R.C., Lead, J.R., 2012. Stability of citrate, PVP, and
PEG coated silver nanoparticles in ecotoxicology media. Environ. Sci. Technol.
46, 7011e7017.
Tuchina, E.S., Tuchin, V.V., 2010. TiO2 nanoparticle enhanced photodynamic inhi-
bition of pathogens. Laser Phys. Lett. 7, 607e612.
Van Grieken, R., Aguado, J., Lopez-Munoz, M.J., Marugan, J., 2002. Synthesis of size-
controlled silica-supported TiO2 photocatalysts. J. Photochem. Photobiol., A 148,
315e322.
Wach, R.A., Kudoh, H., Zhai, M., Nagasawa, N., Muroya, Y., Yoshii, F., Katsumura, Y.,
2004. Rate constants of carboxymethylcellulose with hydrated electron, hy-
droxyl radical and the decay of CMC macroradicals. A pulse radiolysis study.
206 S. Baek et al. / Chemosphere 208 (2018) 196e206
Polymer 45, 8165e8171.
Wang, D., Lin, Z., Wang, T., Yao, Z., Qin, M., Zheng, S., Lu, W., 2016a. Where does the
toxicity of metal oxide nanoparticles come from: the nanoparticles, the ions, or
a combination of both? J. Hazard Mater. 308, 328e334.
Wang, J., Fan, Y., 2014. Lung injury induced by TiO2 nanoparticles depends on their
structural features: size, shape, crystal phases, and surface coating. Int. J. Mol.
Sci. 15, 22258e22278.
Wang, X., Chen, H., 2015. A new approach to preparation of TiO2@void@SiO2 rattle
type core shell structure nanoparticles via titanyl oxalate complex. Colloid.
Surface. Physicochem. Eng. Aspect. 485, 25e33.
Wang, Y., Zhu, X., Lao, Y., Lv, X., Tao, Y., Huang, B., Wang, J., Zhou, J., Cai, Z., 2016b.
TiO2 nanoparticles in the marine environment: physical effects responsible for
the toxicity on algae Phaeodactylum tricornutum. Sci. Total Environ. 565,
818e826.
Weir, A., Westerhoff, P., Fabricius, L., Hristovski, K., von Goetz, N., 2012. Titanium
dioxide nanoparticles in food and personal care products. Environ. Sci. Technol.
46, 2242e2250.
Wu, Y., Liao, L.-D., Pan, H.-C., He, L., Lin, C.-T., Tan, M.C., 2017. Fabrication and
interfacial characteristics of surface modified Ag nanoparticle based conductive
composites. RSC Adv. 7, 29702e29712.
Xiong, D., Fang, T., Yu, L., Sima, X., Zhu, W., 2011. Effects of nano-scale TiO2, ZnO and
their bulk counterparts on zebrafish: acute toxicity, oxidative stress and
oxidative damage. Sci. Total Environ. 409, 1444e1452.
Zhang, M., Shi, L., Yuan, S., Zhao, Y., Fang, J., 2009. Synthesis and photocatalytic
properties of highly stable and neutral TiO2/SiO2 hydrosol. J. Colloid Interface
Sci. 330, 113e118.
Zhang, W., Shi, J., Wang, X., Jiang, Z., Song, X., Ai, Q., 2014. Conferring an adhesion
layer with mineralization-inducing capabilities for preparing
organiceinorganic hybrid microcapsules. J. Mater. Chem. B 2.