Cytotoxicity of Carbon Nanotubes
The industrial applications of carbon nanotubes (CNTs) for creating of new kinds of materials are limited because of their potential toxicity. There are many data concerning CNTs influences on the living body–from quite negative ones to the possibility of CNTs use in medicine. In a given work, the c...
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Інститут металофізики ім. Г.В. Курдюмова НАН України
2012
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| Cite this: | Cytotoxicity of Carbon Nanotubes / Yu. Sementsov, T. Aleksyeyeva, I. Yermolenko, V. Tin’kov, L. Ieleiko, V. Mikhailenko // Наносистеми, наноматеріали, нанотехнології: Зб. наук. пр. — К.: РВВ ІМФ, 2012. — Т. 10, № 1. — С. 177-183. — Бібліогр.: 7 назв. — англ. |
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| author | Sementsov, Yu. Aleksyeyeva, T. Yermolenko, I. Tin’kov, V. Ieleiko, L. Mikhailenko, V. |
| author_facet | Sementsov, Yu. Aleksyeyeva, T. Yermolenko, I. Tin’kov, V. Ieleiko, L. Mikhailenko, V. |
| citation_txt | Cytotoxicity of Carbon Nanotubes / Yu. Sementsov, T. Aleksyeyeva, I. Yermolenko, V. Tin’kov, L. Ieleiko, V. Mikhailenko // Наносистеми, наноматеріали, нанотехнології: Зб. наук. пр. — К.: РВВ ІМФ, 2012. — Т. 10, № 1. — С. 177-183. — Бібліогр.: 7 назв. — англ. |
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| description | The industrial applications of carbon nanotubes (CNTs) for creating of new kinds of materials are limited because of their potential toxicity. There are many data concerning CNTs influences on the living body–from quite negative ones to the possibility of CNTs use in medicine. In a given work, the cell toxicity of CNTs on the example of healthy hepatocytes and Eirlich adenocarcinoma cells (EAC) is studied. As shown, the cells’ contact with CNTs suspension leads to the radicals’ release from cells. This radical release depends on both the time of cells’ contact with suspension and the CNTs concentration. As also shown, the influence of CNTs on EAC is essentially higher than that on healthy hepatocytes.
Промислове використання вуглецевих нанорурок (ВНР) для виготовлення нових композиційних матеріялів обмежене їх потенційною токсичністю. В літературі є дані, які уможливлюють тлумачити вплив ВНР на живий організм від вкрай неґативного до можливого використання їх в медицині. Дану роботу присвячено вивченню токсичности ВНР на прикладі здорових клітин печінки (гепатоцитів) та клітин аденокарциноми Ерліха (ЕАК). Показано, що контакт клітин з суспензією ВНР призводить до викиду ними перекисних радикалів, а рівень викиду цих радикалів залежить від часу контакту і концентрації ВНР. Також було встановлено, що вплив ВНР на ЕАК є більш явним у порівнянні з гепатоцитами.
Промышленное использование углеродных нанотрубок (УНТ) для создания новых композиционных материалов ограничено их потенциальной токсичностью. В литературе представлены данные, которые позволяют рассматривать влияние УНТ на живой организм как крайне негативное, так и позволяющее их использование в медицине. Данная работа посвящена изучению токсичности УНТ на примере здоровых клеток печени (гепатоцитов) и клеток аденокарциномы Эрлиха (ЭАК). Показано, что контакт клеток с суспензией УНТ приводит к выбросу клетками перекисных радикалов, уровень чего зависит от времени контакта и концентрации УНТ. Также показано, что влияние УНТ на ЭАК – более выраженное, чем на гепатоциты.
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177
PACS numbers: 61.48.De, 81.05.ub,82.70.Kj,87.17.-d,87.19.X-,87.64.-t, 87.85.jj
Cytotoxicity of Carbon Nanotubes
Yu. Sementsov, T. Aleksyeyeva*, I. Yermolenko*, V. Tin’kov*,
L. Ieleiko**, and V. Mikhailenko**
O. O. Chuiko Institute of Surface Chemistry,
N.A.S. of Ukraine,
17 General Naumov Str.,
03164 Kyyiv, Ukraine
*G. V. Kurdyumov Institute for Metal Physics,
N.A.S. of Ukraine,
36 Academician Vernadsky Blvd.,
UA-03680 Kyyiv-142, Ukraine
**R. E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology,
N.A.S. of Ukraine,
45 Vasylkivska Str.,
03133 Kyyiv, Ukraine
The industrial applications of carbon nanotubes (CNTs) for creating of new
kinds of materials are limited because of their potential toxicity. There are
many data concerning CNTs influences on the living body–from quite nega-
tive ones to the possibility of CNTs use in medicine. In a given work, the cell
toxicity of CNTs on the example of healthy hepatocytes and Eirlich adenocar-
cinoma cells (EAC) is studied. As shown, the cells’ contact with CNTs suspen-
sion leads to the radicals’ release from cells. This radical release depends on
both the time of cells’ contact with suspension and the CNTs concentration.
As also shown, the influence of CNTs on EAC is essentially higher than that
on healthy hepatocytes.
Промислове використання вуглецевих нанорурок (ВНР) для виготовлен-
ня нових композиційних матеріялів обмежене їх потенційною токсичніс-
тю. В літературі є дані, які уможливлюють тлумачити вплив ВНР на жи-
вий організм від вкрай неґативного до можливого використання їх в ме-
дицині. Дану роботу присвячено вивченню токсичности ВНР на прикладі
здорових клітин печінки (гепатоцитів) та клітин аденокарциноми Ерліха
(ЕАК). Показано, що контакт клітин з суспензією ВНР призводить до ви-
киду ними перекисних радикалів, а рівень викиду цих радикалів зале-
жить від часу контакту і концентрації ВНР. Також було встановлено, що
вплив ВНР на ЕАК є більш явним у порівнянні з гепатоцитами.
Наносистеми, наноматеріали, нанотехнології
Nanosystems, Nanomaterials, Nanotechnologies
2012, т. 10, № 1, сс. 177—183
© 2012 ІМФ (Інститут металофізики
ім. Г. В. Курдюмова НАН України)
Надруковано в Україні.
Фотокопіювання дозволено
тільки відповідно до ліцензії
178 CYTOTOXICITY OF CARBON NANOTUBES
Промышленное использование углеродных нанотрубок (УНТ) для созда-
ния новых композиционных материалов ограничено их потенциальной
токсичностью. В литературе представлены данные, которые позволяют
рассматривать влияние УНТ на живой организм как крайне негативное,
так и позволяющее их использование в медицине. Данная работа посвя-
щена изучению токсичности УНТ на примере здоровых клеток печени
(гепатоцитов) и клеток аденокарциномы Эрлиха (ЭАК). Показано, что
контакт клеток с суспензией УНТ приводит к выбросу клетками перекис-
ных радикалов, уровень чего зависит от времени контакта и концентра-
ции УНТ. Также показано, что влияние УНТ на ЭАК – более выражен-
ное, чем на гепатоциты.
Key words: carbon nanotubes, cells, cytotoxic effect.
(Received September 5, 2011)
1. INTRODUCTION
Carbon nanotubes (CNTs) are extensively used in different fields of
science and industry last decade. Such a multidisciplinary application
of CNTs raises the question of their safety for human health. There is
no accurate data of CNTs influence on living body. A lot of information
in literature varies from their toxicity to possible use in pharmacolo-
gy. Thus, this study is carried out to evaluate the CNTs effect on cells
of different nature.
2. MATERIALS AND METHODS
Multiwall CNTs (Fig. 1) were synthesized by CVD method on equip-
Fig. 1. Multiwall CNTs (TEM data).
Yu. SEMENTSOV, T. ALEKSYEYEVA, I. YERMOLENKO et al. 179
ment with manufacturing capacity 1.0—1.5 kg per day [1]. The compo-
site of oxides like Al2O3—MoO3—Fe2O3 was used as catalytic agent. Pro-
pylene was the source of carbon. CNTs were obtained as agglomerates
of entangled tubes with 20—500 μm in size. Additives were eliminated
by CNTs treatment with hydrofluoric acid. Ultrasound drying of CNTs
resuspended in saline solution was carried out using UZDN-2 disperga-
tor. Size distribution function of particle agglomerates was deter-
mined on laser correlation spectrometer ‘ZetaSizer-3’ with type 7032 s
multicomputing correlator (Malvern Instrument, Great Britain).
Obtained autocorrelation function (ACF) was calculated using soft-
ware PCS-Size mode v 1.61. System with concentration 0.2 wt.% of
CNTs in saline solution shows two sizes of particles: 0.01—0.10 μ and
1.0—5.0 μ. Parameters of multiwall CNTs were determined by TEM, x-
ray fluorescence analysis, Raman scattering spectrometry, differen-
tial thermal analysis, differential thermogravimetric analysis, АFM,
and x-ray photoelectron spectroscopy methods. Ash values of purified
CNTs were < 1.0 wt.%, specific surface measured by argon was 220
m2/g, the 5 wt.% losses were determined at 605°С during heating rate
10°С per minute. Amorphous carbon was not present in studied CNTs.
Their diameter was 10—20 nm, number of layers 5—10. In addition to
main x-ray reflex 002, 100, 101, 110, 112 reflexes were observed that
indicated of 3D regulating of graphite-like lattice. In this case, inter-
planar spacing d002 is in the range 0.3436—0.3453 nm. Equal intensity
of G and D modes were observed in Raman scattering spectra. Accord-
ing to x-ray photoelectron spectroscopy data, oxygen was determined
on CNTs surface and characterized according to type its active centres
and oxidized ones (electrochemically or after heat treatment on air) as
0.6, 1.1, 2.3 at.%, respectively. This is considerably lower as com-
pared to carbon fibres [2].
The concentration of oxygen containing centres determined from
1sC-electrons band (Eb) is presented in Table [3, 4]. The concentration
TABLE. Relative concentrations of oxygen containing centres on MWNT and
their classification by energy of 1sO-electrons.
MWNT
samples
Relative concentration, %
Еb = 286.1—
286.3 eV;
phenol,
alcohol (C—OH)
Еb = 287.3—
287.6 eV;
carbonyl,
quinone (C=O)
Еb = 288.4—
288.9 eV;
carboxyl,
ether (C—OOH)
Еb = 290.4—
290.8 eV;
carbonate and/or
absorbed СО, СО2
Initial 49.1 17.2 17.2 16.5
Anode
oxidation
53.8 19.8 13.6 12.8
Heat
treatment
51.1 23.4 15.3 10.2
180 CYTOTOXICITY OF CARBON NANOTUBES
of oxygen-containing centres was determined by the binding energy of
the 1sC-electrons.
3. IN VIVO EXPERIMENT
Experiments were carried out on white inbred male mice with body
weight 20—22 g. Cells of Ehrlich ascetic carcinoma (EAC) (106
cells/mice) were injected intraperitoneally (i.p.) in the volume of 0.4
ml of saline solution. Culture of Ehrlich ascetic carcinoma was ob-
tained in cell lines bank IEPOR. All experiments on mice tumour bear-
ing were carried out during 7 days after the EAC transplantation. The
CNTs suspension in saline solution was i.p. administered in concentra-
tion 0.75 and 1.5 mg per mouse for 24 h.
Fluorescent probe 2′,7′-dichlorofluorescein diacetate (DCFDA, λex =
= 488 nm, λem = 525 nm) (absorption at 485 nm, emission at 528 nm)
was used for measurement of reactive oxygen (ROS) and nitrogen
(RNS) species in the EAC cells and hepatocytes.
The obtained EAC cells and hepatocytes were three times washed and
precipitated by centrifugation for 5 min at 1 500 r. per minute. Evalu-
ation of ROS and RNS was carried out on analyser Synergy HT Multi-
Detection Reader, in plate for 90 min. at 37°С). Reaction mixture is as
follows: 250 μl of PBS (phosphate buffer saline), 25 μl cell suspension
with concentration of 2⋅106/ml, 25 μl 6 mM DCFDA. Measurement was
carried out every 10 minutes during 90 min.
State of the cell surface was examined by SEM. The isolated cells
were fixed with 10% formalin, then dehydrated in battery alcohols,
coated with gold, and studied with JSM-6490LV (JEOL Japan).
4. RESULTS
Exposure to CNTs in concentration of 0.75 and 1.5 mg per mouse was
accompanied by the increase of ROS and RNS production in hepato-
cytes by 102% and 56% (Fig. 2).
Increasing of free radicals level, which produced by normal cells in
24 hours after contacting with CNTs, could be explained by cells’ reac-
tion to exogenous particles. Moreover, findings confirmed hypothesis
that the CNTs influence on cells physiology.
In EAC cells, lower dose of CNTs caused slight increase of ROS level
(by 20%). Increasing the dose of CNTs (1.5 mg per mouse) led to 39%
rise of ROS production (Fig. 3).
Explanation of such disproportion could be found in cell nature. In
addition, as at it will be shown below, the surviving of EAC cells after
exposure with CNTs is rather low.
After exposure to CNTs in concentration 1.5 mg per mouse, the un-
Yu. SEMENTSOV, T. ALEKSYEYEVA, I. YERMOLENKO et al. 181
damaged EAC cells were not detected in visual field (Fig. 4), unlike to
hepatocytes (Fig. 5).
Our research has shown that it is hard to tell whether CNTs pene-
trate in cell membranes. It was speculated in the works [5—7] that
CNTs penetrate through the cell membranes and thus affect on their
physiological cycles. This hypothesis is quite controversial, because
the probability of membrane damaging from the CNTs satisfies their
aspect number (≅ 1000). In addition, it was shown that CNTs in water
suspensions are presented as agglomerates in the form of entangled
tubes (Fig. 1) with sizes ranging from 200 to 300 nm. Attraction of
CNTs to definite receptors on the cell surface may be more probable be-
cause they have different reactive groups (see Table). Complete block-
age of receptors or covalent binding to the active centres on the mem-
brane may explain this CNTs effect on cells viability. This ability of
CNTs to interfere in the physiology of cells opens up new prospects for
Fig. 2. Level of free radicals production in hepatocytes after injection of the
CNTs suspension.
Fig. 3. Level of free radicals production in EAC cells after injection of the
CNTs suspension.
182 CYTOTOXICITY OF CARBON NANOTUBES
their using for targeted drug delivery.
5. CONCLUSION
This study shows that direct contact of CNTs with cells leads to in-
creasing of free radicals production and to injury of cells membrane,
particularly in the case of EAC cells. In any case, CNTs influence on
Fig. 5. Hepatocytes after exposure with CNTs at concentration of 1.5 mg per
mouse (SEM data).
Fig. 4. EAC cells after exposure with CNTs with concentration of 0.75 mg per
mouse (SEM data).
Yu. SEMENTSOV, T. ALEKSYEYEVA, I. YERMOLENKO et al. 183
cells physiology in the manner depending on their nature, and it is nec-
essary to provide further investigations.
REFERENCES
1. Yu. I. Sementsov, O. V. Melezhek, G. P. Prikhod’ko et al., Physical Chemistry
of Nanomaterials and Supramolecular Structures (Eds. A. P. Shpak and P. P.
Gorbyk) (Kyiv: Naukova Dumka: 2007), vol. 2, p. 116.
2. Z. R. Yue, W. Jiang, L. Wang et al., Carbon, 37: 1785 (1999).
3. H. P. Boehm, Carbon, 40: 145 (2002).
4. H. P. Boehm, Carbon, 32: 759 (1994).
5. I. Vakarelski and K. Higashitan, Langmuir, 23 (22), 10893 (2007).
6. L. Monticelli, E. Salonen, P. C. Ke, and I. Vattulainen, Soft Matter, 5, No. 22:
4433 (2009).
7. J. Bentz, Biophysical Journal, 78, No. 2: 886 (2000).
|
| id | nasplib_isofts_kiev_ua-123456789-75221 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1816-5230 |
| language | English |
| last_indexed | 2025-12-07T17:33:36Z |
| publishDate | 2012 |
| publisher | Інститут металофізики ім. Г.В. Курдюмова НАН України |
| record_format | dspace |
| spelling | Sementsov, Yu. Aleksyeyeva, T. Yermolenko, I. Tin’kov, V. Ieleiko, L. Mikhailenko, V. 2015-01-27T17:45:02Z 2015-01-27T17:45:02Z 2012 Cytotoxicity of Carbon Nanotubes / Yu. Sementsov, T. Aleksyeyeva, I. Yermolenko, V. Tin’kov, L. Ieleiko, V. Mikhailenko // Наносистеми, наноматеріали, нанотехнології: Зб. наук. пр. — К.: РВВ ІМФ, 2012. — Т. 10, № 1. — С. 177-183. — Бібліогр.: 7 назв. — англ. 1816-5230 PACS numbers: 61.48.De, 81.05.ub, 82.70.Kj, 87.17.-d, 87.19.X-, 87.64.-t, 87.85.jj https://nasplib.isofts.kiev.ua/handle/123456789/75221 The industrial applications of carbon nanotubes (CNTs) for creating of new kinds of materials are limited because of their potential toxicity. There are many data concerning CNTs influences on the living body–from quite negative ones to the possibility of CNTs use in medicine. In a given work, the cell toxicity of CNTs on the example of healthy hepatocytes and Eirlich adenocarcinoma cells (EAC) is studied. As shown, the cells’ contact with CNTs suspension leads to the radicals’ release from cells. This radical release depends on both the time of cells’ contact with suspension and the CNTs concentration. As also shown, the influence of CNTs on EAC is essentially higher than that on healthy hepatocytes. Промислове використання вуглецевих нанорурок (ВНР) для виготовлення нових композиційних матеріялів обмежене їх потенційною токсичністю. В літературі є дані, які уможливлюють тлумачити вплив ВНР на живий організм від вкрай неґативного до можливого використання їх в медицині. Дану роботу присвячено вивченню токсичности ВНР на прикладі здорових клітин печінки (гепатоцитів) та клітин аденокарциноми Ерліха (ЕАК). Показано, що контакт клітин з суспензією ВНР призводить до викиду ними перекисних радикалів, а рівень викиду цих радикалів залежить від часу контакту і концентрації ВНР. Також було встановлено, що вплив ВНР на ЕАК є більш явним у порівнянні з гепатоцитами. Промышленное использование углеродных нанотрубок (УНТ) для создания новых композиционных материалов ограничено их потенциальной токсичностью. В литературе представлены данные, которые позволяют рассматривать влияние УНТ на живой организм как крайне негативное, так и позволяющее их использование в медицине. Данная работа посвящена изучению токсичности УНТ на примере здоровых клеток печени (гепатоцитов) и клеток аденокарциномы Эрлиха (ЭАК). Показано, что контакт клеток с суспензией УНТ приводит к выбросу клетками перекисных радикалов, уровень чего зависит от времени контакта и концентрации УНТ. Также показано, что влияние УНТ на ЭАК – более выраженное, чем на гепатоциты. en Інститут металофізики ім. Г.В. Курдюмова НАН України Наносистеми, наноматеріали, нанотехнології Cytotoxicity of Carbon Nanotubes Article published earlier |
| spellingShingle | Cytotoxicity of Carbon Nanotubes Sementsov, Yu. Aleksyeyeva, T. Yermolenko, I. Tin’kov, V. Ieleiko, L. Mikhailenko, V. |
| title | Cytotoxicity of Carbon Nanotubes |
| title_full | Cytotoxicity of Carbon Nanotubes |
| title_fullStr | Cytotoxicity of Carbon Nanotubes |
| title_full_unstemmed | Cytotoxicity of Carbon Nanotubes |
| title_short | Cytotoxicity of Carbon Nanotubes |
| title_sort | cytotoxicity of carbon nanotubes |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/75221 |
| work_keys_str_mv | AT sementsovyu cytotoxicityofcarbonnanotubes AT aleksyeyevat cytotoxicityofcarbonnanotubes AT yermolenkoi cytotoxicityofcarbonnanotubes AT tinkovv cytotoxicityofcarbonnanotubes AT ieleikol cytotoxicityofcarbonnanotubes AT mikhailenkov cytotoxicityofcarbonnanotubes |