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IET Nanobiotechnology
Research Article
Comparison effects of titanium dioxide
nanoparticles on immune cells in adaptive
and innate immune system
ISSN 1751-8741
Received on 13th October 2016
Revised 9th February 2017
Accepted on 8th May 2017
E-First on 1st August 2017
doi: 10.1049/iet-nbt.2016.0205
www.ietdl.org
Patinya Sukwong1, Supunsa Kongseng1, Sunisa Chaicherd1, Krongtong Yoovathaworn1, Suchakree
Tubtimkuna2, Dakrong Pissuwan1,3,4
1Toxicology
Program, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
of Chemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
3Center of Excellence on Environmental Health and Toxicology, Mahidol University, Bangkok 10400, Thailand
4Materials Science and Engineering Program, Multidisciplinary Unit, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
E-mail: dakrong.pis@mahidol.ac.th
2Department
Abstract: Titanium dioxide nanoparticles (TiO2-NPs) have been increasingly mixed in food and daily use products. Therefore,
the investigation of cytotoxic effects of TiO2-NPs is required to allay concerns of health effects related to contact with products
containing TiO 2-NPs. In this study, the authors demonstrated how TiO2-NPs impact on two main sub-types of immune cells that
play a major role in adaptive and innate immune system. Human T-lymphocytes (Jurkat cells) and murine macrophages (RAW
264.7 cells) were used in this study. The authors results showed that cell viability of Jurkat and RAW 264.7 cells were
significantly decreased, when cells were treated with TiO2-NPs at 250 and 500 µg/ml. However, the decrease of cell viability of
RAW 264.7 cells was higher than that of Jurkat cells. A similar trend was also found in DNA fragmentation. An induction of
reactive oxygen species was detected in both cells treated with TiO2-NPs at concentrations ≥25 µg/ml. A significant induction of
tumour necrosis factor alpha (TNF-α) was found in Jurkat and RAW 264.7 cells treated with 25 µg/ml TiO2-NPs. In contrast,
there was no significant induction of interleukin-6 (IL-6) in both cells that were treated with different concentrations of TiO2-NPs.
1 Introduction
Titanium dioxide nanoparticles (TiO2-NPs) are one of the metalbased nanoparticles that have been extensively used as white
pigments in various industries [1]. In food, daily use products, and
pharmaceutical applications, TiO2-NPs have been used as food
colour agents, food preservatives, and antimicrobial/antifungal
agents. Furthermore, TiO2-NPs have been used in orthopedic and
dental implants to induce tissue formation [1] and reduce bacterial
infection [2]. This can contribute to high exposure in humans [3].
Thus, a lot of concerns regarding toxic effects of TiO2-NPs have
been raised. There have been a considerable number of reports on
toxicity of TiO2-NPs on various types of cells lines [4–7]. Immune
cells have received substantial interest for investigation on the
adverse effect of nanoparticles [8–13] because they play an
important role in the body defence mechanism. Immune cells can
be activated by pathogens such as bacteria, virus, and fungi [14–
16] and also nanoparticles [17, 18]. When immune cells are
activated by foreign substances, they can release several
inflammatory cytokines such as interferon, tumour necrosis factor
alpha (TNF-α), and various interleukins resulting in inflammation
induction. The chronic inflammation may lead to cancer formation
[19–21]. It is commonly known that the immune system can be
divided into two groups as innate and adaptive immune systems.
Both systems contain different types of immune cells and have
different mechanisms to protect the body from pathogenic
microorganisms and foreign materials. The innate immune system
acts as the first defender of the body defence system without an
antigen-specific mechanism. Examples of immune cells in the
innate immunity system are: macrophage, dendritic cell, monocyte,
mast cell, and granulocyte. Dissimilar to the innate immune
system, the adaptive immune system needs antigen-specific
interaction to activate immune responses. Major immune cells in
the adaptive immune system are lymphocyte cells [13].
Owing to the chance that TiO2-NPs can be transferred into the
body, it increases greatly the possibility of TiO2-NP and immune
IET Nanobiotechnol., 2017, Vol. 11 Iss. 7, pp. 759-765
© The Institution of Engineering and Technology 2017
cell interactions. Although many researches have shown the effect
of TiO2-NPs in various types of cells; to our knowledge, the
investigation of cytotoxic effects of TiO2-NPs in comparison with
cellular innate and adaptive immune responses is limited.
Therefore, in this study we investigated how immune cells in
innate and adaptive immune systems responded to TiO2-NPs at
different concentrations. We used murine macrophage cells (RAW
264.7) and human Jurkat T cells as a representative cell for innate
and adaptive immune cells, respectively.
2 Materials and methods
2.1 Characterisation and preparation of nanoparticles
TiO2-NPs suspension (20% w/v) was a commercial product from
Hangzhou Wanjing New Material Co., Ltd. Size and shape of
TiO2-NPs were characterised by transmission electron microscope
(TEM; JEM-1230). The suspension of TiO2-NPs (2500 μg/ml) was
centrifuged at 10,000 × g for 15 min. The pellet was dispersed in
200 µl of water and then a small volume of this suspension was
dropped on a copper grid coated with carbon. The sample was
dried and then observed under TEM. The average size of TiO2-NPs
was measured from TEM images.
2.2 Cell culture
Human T-lymphocytes (Jurkat cells) were cultured in RPMI1640
medium supplemented with 10% fetal bovine serum (FBS) and 1%
antibiotic-antimycotic solution. In the case of RAW 264.7 cells,
they were cultured in DMEM medium supplemented with 10%
FBS and 1% antibiotic-antimycotic solution. Cells were incubated
at 37°C in a humidified atmosphere of 5% CO2. All instances for
treating cells with TiO2-NPs were performed in cell culture media
without FBS. TiO2-NPs were also dispersed in cell culture media
without FBS at desired concentrations.
759
2.3 Determination of cell viability
Cells were seeded into a 96-well culture plate at a concentration of
1 × 105 cells per well and incubated at 37°C overnight. After
incubation, the cells were treated with TiO2-NPs at concentrations
of 5, 25, 250, and 500 μg/ml for 24 h. After 24 h treatment, cell
viability was determined using CellTiter-Glo® luminescent
viability assay (Promega, WI, USA). Untreated cells were prepared
as a control cell
2.4 Apoptosis cell death by DNA fragmentation assessment
Jurkat and RAW 264.7 cells were separately seeded into a six-well
plate, incubated overnight at 37°C, and treated with TiO2-NPs at
concentrations of 25, 250, and 500 µg/ml for 24 h. Cells without
treatment of TiO2-NPs were used as control cells. After treatment,
Jurkat cells were re-suspended in 10 mM TE buffer (pH 8.3) and
then lysed with lysis buffer (100 mm Tris-HCl pH 8, 10 mm EDTA
and 1% SDS). In the case of RAW 264.7 cells, cells were detached
before suspending in TE buffer. Following this, lysed cells were
incubated at 50°C for 1 h to remove histone molecules from DNA
and then 20 µg/ml of ribonuclease A was added. The lysed cell
sample was further incubated for 1 h at 37°C. Thereafter, the
proteinase K (0.1 mg/ml) was added into the lysed cell sample and
further incubated at 50°C overnight. To extract the chromosomal
DNA of the lysed cell sample, the mixture of phenol/chloroform/
isoamyl alcohol was added and then precipitated with ethanol.
Finally, DNA samples were separated by 1.75% agarose gel
electrophoresis and then stained with ethidium bromide. The
fragmentation of DNA was visualised under UV light using a
Syngene gel documentation (Syngene, Frederick, MD) [22, 23].
2.5 Measurement reactive oxygen species (ROS) generation
in cells treated with TiO2-NPs
Intracellular ROS generation was observed by using the oxidantsensitive dye 2′, 7′-dichlorofluorescin-diacetate (DCFH-DA).
Jurkat and RAW 264.7 cells were separately seeded into a culture
plate at a density of 1 × 105 cells per well and allowed to adhere
overnight. After incubation, the cell culture media was removed
and cells were treated with TiO2-NPs at concentrations of 25 and
250 µg/ml for 3 h. Following treatment, cells were further stained
with 10 µm DCFH-DA in a dark condition at 37°C for 30 min.
Thereafter, cells were washed and observed under fluorescence
microscope to detect formation of fluorescent compound 2′, 7′DCF occurring from ROS production. Untreated cells were
prepared as a control.
To measure ROS levels, both cells were separately seeded in
each well of a 96-well culture plate at a density of 1 × 105 cells per
well and allowed to grow overnight. The cell culture media was
removed after incubation. Then, cells were treated with TiO2-NPs
at concentrations of 5, 25, 250, and 500 µg/ml for 1 and 3 h. Cells
were then stained with DCFH-DA in dark conditions as mentioned
previously. The stained cells were washed with PBS and were
lysed by lysis buffer (PBS containing 10% Tween20 for Jurkat
cells or PBS containing 20% Tween20 for RAW 264.7 cells). The
presence of ROS generation was detected from a fluorescent signal
of DCF substance. This fluorescent signal was detected at 492 nm
excitation and 595 nm emissions using a SpectraMax M3
Microplate Reader with SoftMax® Pro software.
2.6 Inflammatory cytokine release of cells treated with TiO2NPs
Jurkat and RAW 264.7 cells (1 × 105 cells per well) were cultured
and treated with TiO2-NPs. Jurkat cells were treated with 5, 25, and
250 µg/ml TiO2-NPs for 24 h. In the case of RAW 264.7 cells,
TiO2-NPs at concentrations of 25, 250, and 500 µg/ml were
applied to cells. Following incubation, cell supernatants were
collected and stored at −80°C for analysing the amounts of IL-6
and TNF-α using ELISA kits (eBioscience, San Diego, CA).
Concanavalin A (Con A; 1 µg/ml) and lipopolysaccharide (LPS; 5 760
µg/ml) were used as a positive control for Jurkat and RAW 264.7
cells, respectively. ELISA analysis was performed by following the
instruction from the manufacturer. The intensity of colour
development was measured using a microplate reader (Molecular
devices, SpectraMax® M series, CA, USA) at a wavelength of 450 nm.
2.7 Detection of Ti content using inductively coupled
plasma–mass spectroscopy (ICP-MS)
Jurkat and RAW cells at a density of 1 × 105 cells/well were treated
with TiO2-NPs at concentrations of 5 and 250 µg/ml in cell culture
media without FBS for 24 h. Following this, cells were prepared
for Ti content detection by slightly modifying the protocol
published in Thurn et al. [24] and Hsiao et al. [25]. First, cells were
centrifuged. In the case of RAW264.7 cells, cells were trypsinised
with 0.5% trypsin in EDTA before centrifugation. Next, cell pellets
of both cells were washed by centrifugation three times with PBS
and finally washed with 3% nitric acid (HNO3). After washing,
cells were digested by incubating at 65°C in 65% HNO3 overnight.
Thereafter, digested cells were diluted by adding Miil-Q water to
have a final volume at 50 ml. A titanium standard within a range of
0–20 ppb was also prepared. Then, the contents of Ti in all
prepared samples were analysed using ICP-MS (ELAN® DRC-e,
PerkinElmer SCIEX) and 49Ti was selected as an isotope.
2.8 Statistical analysis
All data were expressed as mean ± standard error. The one-way
analysis of variance and Turkey's multiple comparison test at p < 0.05 in GraphPad Prism®, Version 5.0 was used for this analysis.
3 Results and discussion
3.1 Characterisation of TiO2-NPs, cellular uptake, and their
effect on cell viability
From the TEM image of TiO2-NPs (Fig. 1), it shows that the
morphology of TiO2-NPs is spherical in shape. The average size of
TiO2-NPs used in this study was around 26.0 ± 0.9 nm. In
comparison with untreated cells, the cell viabilities of Jurkat cells
(Fig. 2a) treated with TiO2-NPs at concentrations of 250 and 500 µg/ml for 24 h were significantly reduced to 74.4 ± 4.5% and 61.5 ± 3.1%, respectively. A slight decrease of cell viability was found
in Jurkat cells treated with 5 µg/ml TiO2-NPs (96.4 ± 7.3%) and 25 µg/ml TiO2-NPs (94.2 ± 6.1%). The cell viability of RAW 264.7
cells was decreased to 84.2 ± 2.4% after treating with 25 µg/ml
TiO2-NPs. A significant decrease of RAW 264.7 cell viability
(Fig. 2b) was found in cells treated with 250 µg/ml TiO2-NPs
(60.2 ± 2.7%) and 500 µg/ml TiO2-NPs (52.5 ± 5.9%). Our results
here demonstrate that RAW 264.7 cells treated with TiO2-NPs at
high concentrations were more susceptible than Jurkat cells. It is
well known that macrophages (also called innate effector cells) are
phagocytic cells and can phagocytose non-targeted foreign
materials through toll-like receptors (TLRs) [12]. Therefore, the
cellular uptake of TiO2-NPs by RAW 264.7 cells should be higher
than Jurkat cells. With ICP-MS measurement, the content of Ti
detected in RAW 264.7 cells treated with 5 and 250 µg/ml TiO2NPs was higher than that of Jurkat cells treated with TiO2-NPs at
the same concentrations (Fig. 3). The amounts of Ti detected in
RAW 264.7 cells treated with 5 and 250 µg/ml were ∼40.3 and
181.2 ppb respectively. In contrast, very low Ti amounts in Jurkat
cells treated with 5 µg/ml (∼0.2 ppb) and 250 µg/ml (∼0.7 ppb)
were detected. These results provide evidence that RAW 264.7
macrophage cells had a higher cellular uptake ability of TiO2-NPs
than Jurkat cells.
A high uptake of TiO2-NPs could lead to cytolysis explaining
these cases of experiments with a high number of dead cells. It was
reported that the induction of cytolysis could occur when
macrophages interacted with TiO2-NPs [26]. Sohaebuddin et al.
[26] found that RAW 264.7 cells had a greater number of dead
IET Nanobiotechnol., 2017, Vol. 11 Iss. 7, pp. 759-765
© The Institution of Engineering and Technology 2017
Fig. 1 TEM image of TiO2-NPs
cells than other cell types after treatment with high concentrations
of TiO2-NPs. Therefore, in this study the lower number of dead
cells detected in Jurkat cells treated with the same concentration of
TiO2-NPs applied in RAW 264.7 cells could be from the low
uptake of TiO2-NPs into cells.
3.2 Effect of TiO2-NPs on intracellular ROS generation
In normal metabolic processes ROS are generally produced, but
excessive production of ROS can cause oxidative stress and lead to
cell death. Data obtained in previous studies [3, 10, 27] have
provided much evidence that TiO2-NPs could generate ROS in
cells. Here we also investigated the generation of intracellular ROS
mediated by TiO2-NPs in both types of immune cells. As can be
seen in Figs. 4a and b, strong fluorescent signals of ROS
production were detected in Jurkat and RAW 264.7 cells, when
cells were treated with TiO2-NPs at concentrations of 25 and 250 µg/ml. Next, we further examined our hypothesis by measuring the
amount of intracellular ROS generation. A significant increase of
ROS generation was found in Jurkat cells treated with TiO2-NPs at
concentrations of 25 µg/ml (148.9 ± 7.8%), 250 µg/ml (200.9 ± 6.5%), and 500 µg/ml (214.4 ± 10.0%) (Fig. 4c). The induction of
ROS in Jurkat cells could occur through the interaction of TiO2NPs with the cell membrane. Consistent with previous
investigations by others, ROS could be generated in lymphocyte
cells treated with TiO2-NPs [28]. The induction of ROS production
in RAW 264.7 cells was increased when concentrations of TiO2NPs were increased from 5 to 25, 250, and 500 µg/ml (Fig. 4d).
The investigation of ROS induction after treating cells with
different concentrations of TiO2-NPs for 1 h was also done. We
found that the ROS levels in Jurkat and RAW 264.7 cells treated
with TiO2-NPs (25–500 µg/ml TiO2-NPs) for 1 h were higher than
a prolonged treatment (3 h). The increase of ROS at 1 h-treatment
in both cells was in a dose-dependent manner (Figs. 4e and f). The
high levels of ROS generation were detected in RAW 264.7 cells
treated with TiO2-NPs (5–500 µg/ml) for 1 h. As mentioned
previously, macrophage cells are phagocytic cells. Therefore,
Fig. 3 Ti amounts detected in Jurlat and RAW 264.7 cells treated with 5
and 250 µg/ml TiO2-NPs using ICP-MS. Single asterisk symbol indicates
the significant increase of Ti contents compared with untreated cells (p < 0.05; n ≥ 3)
phagocytosis of TiO2-NPs by RAW 264.7 cells could increase
more interaction between cells and TiO2-NPs resulting in enhanced
production of ROS at a higher level than that of Jurkat cells after 1 h-treatment with TiO2-NPs. It was also reported that free radicals
from TiO2-NPs could be generated through Fenton-type reactions
with macromolecules of cells [29]. A high amount of superoxide
ions produced from the reaction between TiO2-NPs and NADPH
oxidase, which is the enzyme in the cell membrane, and cytosolic
components, was reported [30]. These reactions could later lead to
oxidative stress induction in cells and activate the innate immune
system resulting in a high-death rate in RAW 264.7 cells treated
with high concentrations of TiO2-NPs. At a prolonged treatment (3 h), The ROS generation of RAW 264.7 cells was lower than that of
Jurkat cells. The reason for this is unclear and we shall not go into
the details at this point; however, there is some possibility such as
size change of TiO2-NPs occurring from an aggregation with an
increased incubation time that could impact on ROS production
mechanism of phagocytic Raw 264.7 cells. Overall, we found that
the generation of ROS related to the decrease of cell viability of
Jurkat and RAW 264.7 cells.
3.3 DNA fragmentation
It is widely considered that ROS can induce oxidative DNA
damage [31–33]. Therefore, we further investigated the
fragmentation of DNA in Jurkat and RAW 264.7 cells treated with
TiO2-NPs at concentrations of 25, 250, and 500 µg/ml (Fig. 5). Our
results clearly indicate that DNA fragments were detected in Jurkat
and RAW 264.7 cells treated with 250 and 500 µg/ml TiO2-NPs.
The signal of ROS generation in both cells treated with 250 and
500 µg/ml TiO2-NPs was also detected. This implies that the
generation of ROS was involved in DNA fragmentation. With an
innate immune system, it has been recently reported that TLRs,
which are presented on phagocytic cells, play an important role in
innate immunity activation. These receptors are transmembrane
proteins that can recognise foreign materials. It was reported that
TLRs were involved in cellular uptake and cellular response to
Fig. 2 Effect of TiO2-NPs on cell viabilities of Jurkat
(a) and RAW 264.7, (b) cells after treating with 5, 25, 250, and 500 µg/ml TiO2-NPs for 24 h. Single asterisk symbol indicates the significant decrease of cell viability compared
with untreated cells (p < 0.05; n ≥ 7)
IET Nanobiotechnol., 2017, Vol. 11 Iss. 7, pp. 759-765
© The Institution of Engineering and Technology 2017
761
Fig. 4 Effect of TiO2-NPs on intracellular ROS generation in Jurkat
(a, c) and RAW 264.7, (b, d) cells after treating with different concentrations of TiO2-NPs for 3 h (n ≥ 8). The ROS generations in Jurkat, (e) and RAW 264.7 cells, (f) treated with
TiO2-NPs for 1 h (n ≥ 4). Single asterisk symbol indicates the significant increase of ROS compared with untreated cells (p < 0.05)
TiO2-NPs [34]. A high Ti content detected in phagocytic RAW
264.7 cells treated with TiO2-NPs illustrated that TLRs on
phagocytic RAW 264.7cells could promote the cellular uptake of
TiO2-NPs into cells. The involvement of TLRs in DNA damage
enhanced by TiO2-NPs was also reported [35]. Therefore, the ROS
generation that leads to oxidative stress-induced DNA cleavages in
RAW 264.7 cells could occur from this molecular mechanism. In
contrast, the cellular uptake of TiO2-NPs Jurkat cells was very low
compared with RAW 264.7 cells (Fig. 3). According to the study
investigated by Eom and Choi [36], the mechanism of TiO2-NPs
induced DNA damage in Jurkat cells might occur through p38
MAPK pathway after the generation of ROS, which lead to
breaking of the DNA strand. This available evidence and our
results suggest that the DNA damage of Jurkat and RAW cells
treated with 250 and 500 µg/ml TiO2-NPs could occur from a
different mechanism. Furthermore, as we can see from all results, a
significant reduction of cell viability and an induction of
internucleosomal DNA fragmentation (180–200 bp) were detected
in Jurkat and RAW 264.7 cells treated with 250 and 500 µg/ml
TiO2-NPs. This implies that the cell viability and DNA
fragmentation are correlated.
762
3.4 TiO2-NPs induced pro-inflammation cytokine release
The investigation of how TiO2-NPs affect cytokine induction in
Jurkat and RAW 264.7 cells is also important. It is generally
known that cytokines play a major role as a mediator/regulator in
the immune system. We therefore measured pro-inflammatory
cytokines released by Jurkat and RAW 264.7 cells. We focused on
TNF-α and IL-6 cytokines. The high level of TNF-α could cause
cell death [37], septic shock, and autoimmune disease [38]. The
problem of aging and chronic morbidity can be modulated by a
high expression of IL-6 [39]. We investigated the release of both
cytokines by Jurkat cells treated with TiO2-NPs at concentrations
of 5, 25, and 250 μg/ml. High concentrations of TiO2-NPs (25,
250, and 500 μg/ml) were applied to RAW 264.7 cells. The
outcomes of cell viability, DNA fragmentation, and ROS results
indicate that high concentrations of TiO2-NPs might induce
deleterious effects in RAW 264.7 cells. The results from our study
showed that the release of TNF-α from Jurkat and RAW 264.7 cells
was significantly enhanced to a 13.8- and 2.5-fold increase
(compared with untreated cells) after treating with 25 μg/ml TiO2NPs for 24 h (Fig. 6). At 250 μg/ml of TiO2-NPs, the significant
induction of TNF-α was also detected in Jurkat cells but the TNF-α
level dropped from a 13.8 to a 8.9 fold increase. A reduction of
IET Nanobiotechnol., 2017, Vol. 11 Iss. 7, pp. 759-765
© The Institution of Engineering and Technology 2017
Fig. 5 The effect of TiO2-NPs at concentrations of 25, 250, and 500 µg/ml on DNA fragmentation induction in Jurkat
(a) and RAW 264.7, (b) cells
Fig. 6 Effect of TiO2-NPs on the release of cytokines in Jurkat
(a) and RAW 264.7, (b) cells. Cells were treated with different concentrations of TiO2-NPs for 24 h. Concanavalin A and lipopolysaccharide were used as a positive control
treatment for Jurkat and RAW 264.7 cells respectively. *Significant difference of cell events compared with control (untreated) cells (p < 0.05, n ≥ 3)
TNF-α levels was found in RAW 264.7 cells treated with 250 and
500 μg/ml TiO2-NPs. This might be due to the increase of dead
cells after treating RAW 264.7 cells at the same concentrations
(Fig. 2). However, it is worth noting that TiO2-NPs can adsorb
cytokines [40, 41]. Specifically in our study, we investigated the
effect of TiO2-NPs on both immune cells in serum free conditions.
Therefore, the binding of TNF-α molecules and TiO2-NPs might
reduce the TNF-α level particularly in Jurkat cells treated with 250 μg/ml of TiO2-NPs. As expected, a high induction of TNF-α was
detected in cells treated with concanavalin A (for Jurkat cells) and
lipopolysaccharide (for RAW 264.7 cell) that was used as a
positive control for this study. The IL-6 levels of Jurkat and RAW
264.7 cells treated with TiO2-NPs at all concentrations used in this
study were not significantly different from untreated cells. These
IET Nanobiotechnol., 2017, Vol. 11 Iss. 7, pp. 759-765
© The Institution of Engineering and Technology 2017
results suggest that TiO2-NPs at concentrations of 5–250 and 25–
500 μg/ml had no impact on IL-6 induction in Jurkat and RAW
264.7 cells respectively. Similar to the TNF-α, an induction of IL-6
levels was found in cells treated with concanavalin A and
lipopolysaccharide.
The induction of TNF-α level in RAW 264.7 cells treated with
TiO2-NPs (25 μg/ml) could occur from the interaction between
RAW 264.7 cells and TiO2-NPs and then stimulate cells to secrete
TNF-α through the TLR signalling pathway in the innate immune
system. The uptake of TiO2-NPs by phagocytic RAW 264.7 cells
could induce innate defence resulting in releasing of TNF-α. In the
adaptive immune response, the key factor to activate T
lymphocytes or other adaptive immune cells is a ligand-specific
receptor. This binding is needed in the adaptive immune system to
generate immunological memory. Due to the small size and surface
763
Fig. 7 Schematic represents the possible mechanism of TiO2-NPs to induce cellular responses of innate immune cell (RAW 264.7 cell) and adaptive immune
cell (Jurkat cell)
property of TiO2-NPs, they could react with Jurkat cells and
significantly stimulate TNF-α release. The mechanism is still
unclear. However, we proposed two possibilities for TNF-α
induction in Jurkat cells by TiO2-NPs: (i) TiO 2-NPs might have the
specific structure that can react with receptors on Jurkat cells and
(ii) other proteins or biological molecules released by Jurkat cells
might bind to TiO2-NPs and then react with cells to induce cellular
activities. Together with these two possibilities; it may impact on
stimulation of the adaptive immune responses. An overview of the
impact of TiO2-NPs on Jurkat and RAW 264.7 cells is presented in
Fig. 7.
4 Conclusions
[3]
[4]
[5]
[6]
[7]
[8]
In summary, we have demonstrated that TiO2-NPs at
concentrations of 25–500 μg/ml can induce adverse effects in
innate and adaptive immune cells. The concentration of TiO2-NPs
has a strong impact on cell viability and DNA damage. Our results
indicate that TiO2-NPs impacted on cellular activities of both
innate immune cells (RAW 264.7) and adaptive immune cells
(Jurkat). RAW 264.7 cells were more sensitive to TiO2-NPs than
Jurkat cells. This might be due to the fact that RAW 264.7 cells are
in the innate immune system, which is the first line of defence
against foreign materials or pathogens. Nevertheless, TiO2-NPs
could impact on innate and adaptive immune cells. Therefore, there
is need for concern the concentration of TiO2-NPs that are applied
to or are in contacted with human.
[9]
5 Acknowledgments
[16]
This work was supported by Center of Excellence on
Environmental Health and Toxicology (EHT); grants from Faculty
of Science; Toxicology and Materials Science and Engineering
Graduate Program Faculty of Science, Mahidol University. The
authors also thank the funding support from the Office of the
Higher Education Commission (OHEC and Talent Management
Project, Mahidol University. The authors thank Mr Preecha
Sowanthip for a technical support. The authors also acknowledge
Prof Dr Pawinee Piyachaturawat and Assist. Prof Dr Toemsak
Srikhirin for kindly providing materials used in this work.
[17]
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