Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro
Creation of new nanostructures possessing high antitumor activity is an important problem of modern biotechnology. Aim. To evaluate cytotoxicity of created complex of pristine C₆₀ fullerene with the anthracycline antibiotic doxorubicin (Dox), as well as of free C₆₀ fullerene and Dox, towards differe...
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Інститут молекулярної біології і генетики НАН України
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| Cite this: | Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro / S.V. Prylutska, G.V. Didenko, G.P. Potebnya, K.I. Bogutska, Yu.I. Prylutskyy, U. Ritter, P. Scharff // Вiopolymers and Cell. — 2014. — Т. 30, № 5. — С. 372-376. — Бібліогр.: 25 назв. — англ. |
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nasplib_isofts_kiev_ua-123456789-1545532025-02-09T17:09:29Z Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro Токсичний ефект комплексу фулерен C₆₀–доксорубіцин на нормальні і пухлинні клітини in vitro Токсический эффект комплекса фуллерен C₆₀–доксорубицин на нормальные и опухолевые клетки in vitro Prylutska, S.V. Didenko, G.V. Potebnya, G.P. Bogutska, K.I. Prylutskyy, Yu.I. Ritter, U. Scharff, P. Bioorganic Chemistry Creation of new nanostructures possessing high antitumor activity is an important problem of modern biotechnology. Aim. To evaluate cytotoxicity of created complex of pristine C₆₀ fullerene with the anthracycline antibiotic doxorubicin (Dox), as well as of free C₆₀ fullerene and Dox, towards different cell types – tumor, normal immunocompetent and hepatocytes. Methods. Measurement of size distribution for particles in C₆₀ + Dox mixture was performed by a dynamic light scattering (DLS) technique. Toxic effect of C₆₀+ Dox complex in vitro towards tumor and normal cells was studied using the MTT assay. Results. DLS experiment demonstrated that the main fraction of the particles in C₆₀+ Dox mixture had a diameter in the range of about 132 nm. The toxic effect of C₆₀ + Dox complex towards normal (lymphocytes, macrophages, hepatocytes) and tumor (Ehrlich ascites carcinoma, leukemia L1210, Lewis lung carcinoma) cells was decreased by ~10–16 % and ~7–9 %, accordingly, compared with the same effect of free Dox. Conclusions. The created C₆₀ + Dox composite may be considered as a new pharmacological agent that kills effectively tumor cells in vitro and simultaneously prevents a toxic effect of the free form of Dox on normal cells. Створення нових наноструктур з високою протипухлинною активністю є важливою проблемою сучасної біотехнології. Мета. Оцінити цитотоксичність створеного комплексу фулерену C₆₀ з антибіотиком антрациклінового ряду доксорубіцином (Докс) на різні типи клітин (пухлинні, імунокомпетентнi і гепатоцити) та порівняти одержані результати з токсичною дією вільного фулерену C₆₀ і Докс за умов in vitro. Методи. Розподіл за розміром частинок у C₆₀+ Докс суміші вимірювали методом динамічного розсіювання світла (ДРС). Токсичний ефект комплексу C₆₀+ Докс щодо нормальних і пухлинних клітин вивчали in vitro з використанням МТТ-тесту. Результати. Встановлено, що в суміші C₆₀+ Докс в основному реєструються частинки з діаметром 132 нм. Токсична дія комплексу C₆₀+ Докс щодо нормальних (лімфоцити, макрофаги, гепатоцити) і пухлинних (асцитна карцинома Ерліха, лейкоз L1210, карцинома легені Льюїс) клітин виявилася меншою приблизно на 10–16 і 7–9 % відповідно порівняно із впливом вільного Докс. Висновки. Розроблений комплекс C₆₀ + Докс можна розглядати як новий фармакологічний препарат, що за умов in vitro здатний ефективно знищувати пухлинні клітини і одночасно запобігати побічним токсичним ефектам, притаманним традиційному протипухлинному препарату Докс щодо нормальних клітин. Создание новых наноструктур с высокой противоопухолевой активностью является важной проблемой современной биотехнологии. Цель. Оценить цитотоксичность созданного комплекса фуллерена C₆₀ с антибиотиком антрациклинового ряда доксорубицином (Докс) на разные типы клеток (опухолевые, иммунокомпетентные, гепатоциты) и сравнить полученные результаты с токсическим действием свободного фуллерена C₆₀и Докс в условиях in vitro. Методы. Распределение по размеру частиц в смеси C₆₀ + Докс измеряли методом динамического рассеивания света (ДРС). Токсический эффект комплекса C₆₀ + Докс на нормальные и опухолевые клетки in vitro изучали с использованием МТТ-теста. Результаты. Установлено, что в смеси C₆₀ + Докс регистрируются в основном частицы с диаметром 132 нм. Токсическое действие комплекса C₆₀+ Докс по отношению к нормальным (лимфоциты, макрофаги, гепатоциты) и опухолевым (асцитная карцинома Эрлиха, лейкоз L1210, карцинома легкого Льюис) клеткам оказалось меньшим на ~ 10–16 и 7–9 % соответственно по сравнению с действием свободного Докс. Выводы. Разработанный комплекс C₆₀ + Докс можно рассматривать как новый фармакологический препарат, способный в условиях in vitro эффективно уничтожать опухолевые клетки и одновременно предотвращать побочные токсические эффекты, присущие традиционному противоопухолевому препарату Докс относительно нормальных клеток. 2014 Article Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro / S.V. Prylutska, G.V. Didenko, G.P. Potebnya, K.I. Bogutska, Yu.I. Prylutskyy, U. Ritter, P. Scharff // Вiopolymers and Cell. — 2014. — Т. 30, № 5. — С. 372-376. — Бібліогр.: 25 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.0008B4 https://nasplib.isofts.kiev.ua/handle/123456789/154553 57.085.23 + 576.3: 616-006.04 en Вiopolymers and Cell application/pdf Інститут молекулярної біології і генетики НАН України |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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| topic |
Bioorganic Chemistry Bioorganic Chemistry |
| spellingShingle |
Bioorganic Chemistry Bioorganic Chemistry Prylutska, S.V. Didenko, G.V. Potebnya, G.P. Bogutska, K.I. Prylutskyy, Yu.I. Ritter, U. Scharff, P. Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro Вiopolymers and Cell |
| description |
Creation of new nanostructures possessing high antitumor activity is an important problem of modern biotechnology. Aim. To evaluate cytotoxicity of created complex of pristine C₆₀ fullerene with the anthracycline antibiotic doxorubicin (Dox), as well as of free C₆₀ fullerene and Dox, towards different cell types – tumor, normal immunocompetent and hepatocytes. Methods. Measurement of size distribution for particles in C₆₀ + Dox mixture was performed by a dynamic light scattering (DLS) technique. Toxic effect of C₆₀+ Dox complex in vitro towards tumor and normal cells was studied using the MTT assay. Results. DLS experiment demonstrated that the main fraction of the particles in C₆₀+ Dox mixture had a diameter in the range of about 132 nm. The toxic effect of C₆₀ + Dox complex towards normal (lymphocytes, macrophages, hepatocytes) and tumor (Ehrlich ascites carcinoma, leukemia L1210, Lewis lung carcinoma) cells was decreased by ~10–16 % and ~7–9 %, accordingly, compared with the same effect of free Dox. Conclusions. The created C₆₀ + Dox composite may be considered as a new pharmacological agent that kills effectively tumor cells in vitro and simultaneously prevents a toxic effect of the free form of Dox on normal cells. |
| format |
Article |
| author |
Prylutska, S.V. Didenko, G.V. Potebnya, G.P. Bogutska, K.I. Prylutskyy, Yu.I. Ritter, U. Scharff, P. |
| author_facet |
Prylutska, S.V. Didenko, G.V. Potebnya, G.P. Bogutska, K.I. Prylutskyy, Yu.I. Ritter, U. Scharff, P. |
| author_sort |
Prylutska, S.V. |
| title |
Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro |
| title_short |
Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro |
| title_full |
Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro |
| title_fullStr |
Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro |
| title_full_unstemmed |
Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro |
| title_sort |
toxic effect of c₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
2014 |
| topic_facet |
Bioorganic Chemistry |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/154553 |
| citation_txt |
Toxic effect of C₆₀ fullerene-doxorubicin complex towards tumor and normal cells in vitro / S.V. Prylutska, G.V. Didenko, G.P. Potebnya, K.I. Bogutska, Yu.I. Prylutskyy, U. Ritter, P. Scharff // Вiopolymers and Cell. — 2014. — Т. 30, № 5. — С. 372-376. — Бібліогр.: 25 назв. — англ. |
| series |
Вiopolymers and Cell |
| work_keys_str_mv |
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| fulltext |
BIOORGANIC CHEMISTRY
UDC 57.085.23 + 576.3: 616-006.04
Toxic effect of C60 fullerene-doxorubicin complex
towards tumor and normal cells in vitro
S. V. Prylutska1, G. V. Didenko2, G. P. Potebnya2, K. I. Bogutska1, Yu. I. Prylutskyy1,
U. Ritter3, P. Scharff3
1Educational and Scientific Center "Institute of Biology",
Taras Shevchenko National University of Kyiv
64/13, Volodymyrska Str., Kyiv, Ukraine, 01601
2R. E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine
45,Vasylkivska Str., Kyiv, Ukraine, 03022
3Institute of Chemistry and Biotechnology,
Technical University of Ilmenau
25, Weimarer Str., Ilmenau, Germany, 98693
psvit@bigmir.net
Creation of new nanostructures possessing high antitumor activity is an important problem of modern biotech-
nology. Aim. To evaluate cytotoxicity of created complex of pristine C
60
fullerene with the anthracycline antibio-
tic doxorubicin (Dox), as well as of free C
60
fullerene and Dox, towards different cell types – tumor, normal im-
munocompetent and hepatocytes. Methods. Measurement of size distribution for particles in C
60
+ Dox mixture
was performed by a dynamic light scattering (DLS) technique. Toxic effect of C
60
+ Dox complex in vitro towards
tumor and normal cells was studied using the MTT assay. Results. DLS experiment demonstrated that the main
fraction of the particles in C
60
+ Dox mixture had a diameter in the range of about 132 nm. The toxic effect of C
60
+
Dox complex towards normal (lymphocytes, macrophages, hepatocytes) and tumor (Ehrlich ascites carcinoma,
leukemia L1210, Lewis lung carcinoma) cells was decreased by ~10–16 % and ~7–9 %, accordingly, compared
with the same effect of free Dox. Conclusions. The created C
60
+ Dox composite may be considered as a new phar-
macological agent that kills effectively tumor cells in vitro and simultaneously prevents a toxic effect of the
free form of Dox on normal cells.
Keywords: C60 fullerene-doxorubicin complex, normal and tumor cells, cytotoxicity, MTT assay, dynamic light
scattering.
Introduction. The important problem in nanobiotech-
nology is to solve a complex task at the intersection of
chemistry, physics, materials science, biology and me-
dicine, which involves the targeted design, synthesis
and study of functional properties of nanomaterials
(with at least one of the dimensions in the size range up
to 100 nm), which are characterized by low toxicity and
high specific bioactivity, for their application in the
treatment of common diseases. The proposed unique na-
nobiotechnology is expected in the nearest future to sol-
ve the problem of early diagnosis of various pathologi-
es with the identification of their localization and selec-
tive delivery of drugs to the target organs. C60 fullerene
has an eminent position among the available promising
and effective biomedical compounds [1]. Due to their
nanoscale dimension, combination of strength with low
weight [2], strong antioxidant properties [3, 4], acces-
sibility for cellular uptake [5–7], the pristine C60 fulle-
renes are considered as pharmaceutically valuable com-
pounds of a new class [8–10].
However, along with significant prospects of use of
these substances for the prevention and treatment of di-
seases, there are also some problems and cautions. Thus,
some results of biological studies of C60 fullerene aque-
ous dispersions indicate their possible toxic effects on
the human organism [11]. On the other hand, the pristi-
372
ISSN 0233–7657. Biopolymers and Cell. 2014. Vol. 30. N 5. P. 372–376 doi: http://dx.doi.org/10.7124/bc.0008B4
� Institute of Molecular Biology and Genetics, NAS of Ukraine, 2014
ne C60 fullerenes were demonstrated not to possess any
toxicity at low concentrations, or at least they showed
no acute toxic effect in the in vitro systems and in vivo
[12, 13]. The toxicity of C60 molecules was identified to
depend strongly on their surface modification, synthe-
sis and treatment conditions, concentration of nanopar-
ticles in solvent medium and, consequently, a size of
the formed aggregates (clusters) [11]. It is assumed that
the main mechanisms of cytotoxic action of the C60
fullerene derivatives are the evoked lipid peroxidation
and consecutive progress of oxidative stress and related
effects, including DNA damage and necrosis [11, 14].
Doxorubicin (Dox) is an anthracycline antibiotic
that is one of the most common therapeutic agents in
cancer chemotherapy [15]. Dox binds non-covalently to
DNA, blocks the synthesis of nucleic acids, demonst-
rates high anti-mitotic activity and pronounced mutage-
nic effects, but also exerts toxic effects towards normal
tissues and cells [16]. The free radicals which are formed
during Dox chemotherapy, inactivate enzymes of antioxi-
dant protection and invoke immediate oxidative damage
of cells with high oxidative metabolism and activity of
the mitochondrial respiratory chain, particularly cardiac
myocytes and hepatocytes.
One of the ways in protection against Dox-induced
chemical insults of normal tissues is a combined use of
cytostatics together with antioxidants of different nature
[17]. One can assume that immobilization of Dox on C60
fullerene [18] will prevent its toxic action on normal cells
and enhance its uptake by the target cells that is impor-
tant for the biomedical application of C60 fullerene-drug
conjugates [19, 20].
The aim of this study was to evaluate the in vitro to-
xicity of the C60 fullerene with doxorubicin (C60 + Dox)
complex towards different cell types (tumor, immuno-
competent, hepatocytes) and compare it with the effect
of C60 fullerene and free Dox under in vitro conditions.
Materials and methods. Material preparation and
characterization. A highly stable reproducible C60 fulle-
rene aqueous colloid solution (C60FAS) was prepared
according to protocol [21, 22]. In our experiments the
C60FAS sample with 0.15 mg/ml concentrations of C60
fullerene was used. The resulting probe microscopy ima-
ges clearly indicate the presence in water of]individual
C60 fullerenes and their aggregates with a typical size up
to 100 nm [21–23].
Dox («Doxorubicin-TEVA», «Pharmachemie B.
V.», Netherlands, 10 mg of lyophilized powder), dissol-
ved in saline (0.9 % NaCl), with an initial concentration
0.15 mg/ml was used in experiments.
Dox was immobilized on the C60 fullerene accor-
ding to the following protocol: C60FAS (0.15 mg/ml)
and Dox (0.15 mg/ml) were mixed in 1:2 volume ratio,
the resulting mixture was treated in the ultrasonic dis-
perser for 20 min, and afterwards left for 12 h magnetic
stirring at room temperature. The absorption spectra of
native Dox and C60 + Dox mixture were measured in the
wavelength range � = 400–600 nm at room tempera-
ture. The pronounced hypochromic effect observed in
the experiment indicates the formation of a stable comp-
lex between Dox and C60 fullerene [18].
Measurement of the size distribution of particles in
C60 + Dox mixture was performed by a dynamic light
scattering (DLS) at T = 298 K on a Zetasizer Nano-
ZS90 (UK). DLS instrument equipped with a He-Ne la-
ser (max 5 mW) operating at the wavelength of 633 nm
was used.
Cell culture experiments in vitro. Immunocompetent
cells and hepatocytes were isolated from intact mice
(males of Balb/c line 2.0–2.5 months of age, weight
20–25 g), who were kept at 25 ± 1 °C on a standard diet
of vivarium of the R. E. Kavetsky Institute of Expe-
rimental Pathology, Oncology and Radiobiology, NAS
of Ukraine (R. E. Kavetsky IEPOR, Kyiv). All experi-
ments were conducted in accordance with the standards
of the European Convention for the Protection of Verte-
brate Animals under supervision of bioethical commit-
tee of the abovementioned institution.
Lymphocytes and hepatocytes were obtained by cent-
rifugation (1,500 rpm, 40 min) of cell suspensions of
spleen and liver, respectively, in Ficoll-Hypaque density
gradient (� = 1.077) [24]. Peritoneal macrophages were
obtained from the abdominal cavity of animals by the
treatment with 89 % RPMI 1640 medium supplemented
with 10 % fetal calf serum and 1 % heparin (5 U/ml) and
subsequent centrifugation (1,000 rpm, 10 min).
For comparative evaluation of the cytotoxic effect
of C60 + Dox complex and free C60 fullerene and Dox,
the cells of Ehrlich ascites carcinoma, L1210 leukemia
and Lewis lung carcinoma were used. The cells were ob-
tained from the bank of R. E. Kavetsky IEPOR. 10 µl of
C60 + Dox mixture or C60FAS, or Dox was added to 100
373
TOXIC EFFECT OF C60 FULLERENE-DOXORUBICIN COMPLEX TOWARDS TUMOR AND NORMAL CELLS IN VITRO
µl of cell suspension in the amount of ~3 �105 cells and
incubated for 18 h. The toxic effect of studied drugs was
evaluated using the MTT assay [25], based on the
ability of the mitochondrial respiratory chain dehydro-
genases to convert 3-(4,5-dimethylthiazol-2-yl)-2,5-di-
phenyl-2H-tetrazol bromide (MTT) to formazan, which
crystallizes in cells. Cell suspension containing the stu-
died compound was incubated in the presence of MTT
for 3.5 h at the temperature of 37 °C. Formazan precipi-
tate was dissolved in a concentrated solution of DMSO.
Extinction measurement was performed on a digital
spectrophotometer («�Quant, BioTEK», USA) at the
wavelength of 540 nm. Cytotoxic activity of the studi-
ed compound was calculated using the formula: (1 –
�/�0) �100 %, where �0 and � are the extinctions of cont-
rol and test sample, respectively. The MTT-test was re-
peated triple for different experiments.
MTT dye was used for visualization of viable tu-
mor cells.
Statistics. Statistical analysis of the experimental
data was performed using a Student t-test (the level of
significance was p � 0.05).
Results and discussion. A typical result of DLS ex-
periment is presented in Fig. 1 and shows the distribu-
tion of the number of light scattering particles accor-
ding to their hydrodynamic diameters in the studied sys-
tem. As one can see, the main fraction of the particles
had diameters in the range of 132 nm for C60 + Dox mix-
ture and 33 nm for C60 fullerene dissolved in saline (for
comparison). Thus, one can assume that C60 + Dox mix-
ture contains individual C60 + Dox complexes as well as
their clusters. Previously, based on a detailed analysis
of quantum-chemical calculations, we have shown [18]
that three Dox molecules may simultaneously bind with
one C60 fullerene without sterical overlapping; a diame-
ter of such complex is 1.38 nm.
Table present the data on survival of different cell
types (in % of control) under the action of studied com-
pounds.
The results obtained from the evaluation of cytotoxic
activity of C60 fullerene (0.15 mg/ml) towards normal
cells (Table) have shown that C60 fullerene does not have
any toxic effect towards macrophages and hepatocytes.
On the contrary, it stimulates lymphocytes growth by ~
15 %, which is probably caused by enhancing metabo-
lism leading to an increase in their proliferative activity.
The toxic effect of the Dox sample (0.15 mg/ml) is
high for all types of normal cells (~ 48–66 %; Table).
The toxic effect of the C60 + Dox (0.05 + 0.1 mg/ml)
complex is kept in respect of all types of normal cells
(~ 38– 50 %; Table).
However, in this case the number of dead cells was
lower by ~ 10–16 % compared with the same effect of
free Dox that is a very important applied result. We be-
lieve that reducing the toxic effect of this complex on
normal cells compared with the effect of free Dox might
be associated with the antioxidant activity of the pristi-
ne C60 fullerene [3, 4].
The results obtained on the basis of evaluating the
cytotoxic activity of C60 fullerene (0.15 mg/ml) towards
tumor cells (Table) have shown that C60 fullerene dis-
plays the maximum toxic effect on cells of Ehrlich asci-
te carcinoma (~26 %), does not have this effect on leu-
kemia L1210 cells at all, and conversely, stimulates the
growth of Lewis lung carcinoma cells by ~17 %.
The toxic effect of Dox sample (0.15 mg/ml) is high
for all types of tumor cells (~ 69–93 %; Table).
The toxic effect of C60 + Dox (0.05 + 0.1 mg/ml) com-
plex is kept in respect of all types of tumor cells (~ 62–
84 %; Table). However, in this case the toxic effect of
C60 + Dox complex compared with the same effect of
free Dox is weaker, namely: the number of dead tumor
cells was lower by ~ 7–9 %.
As an example, the microscopic images shown in Fig.
2 illustrate the result of toxic action of studied compounds
towards Ehrlich ascite carcinoma cells after 18 h of in-
cubation.
The cancer cells are known to develop resistance to
traditional chemotherapy drugs, but it is less likely that
374
PRYLUTSKA S. V. ET AL.
N
um
b
er
Size, nm
0
4
8
12
16
20
33 nm 132 nm
1
2
0 100 200 300 400 500 600
Fig. 1. Light scattering distribution of the number of particles accor-
ding to their hydrodynamic diameters for C
60
fullerene dissolved in
saline (1) and C
60
+ Dox mixture (2)
they can develop resistance when multiple drugs, for exa-
mple, C60 fullerene and Dox, are delivered simultaneous-
ly as a C60 + Dox complex. It is important that one of the
potential drugs, for example, C60 fullerene can be a car-
rier of Dox to the nucleus. Finally, C60 fullerene can act
as a strong antioxidant effectively reducing the toxic si-
de effects of anticancer drug Dox towards normal cells.
Acknowledgements. S.V. Prylutska is grateful to
DAAD for support.
Òîêñè÷íèé åôåêò êîìïëåêñó ôóëåðåí Ñ60–äîêñîðóá³öèí íà
íîðìàëüí³ ³ ïóõëèíí³ êë³òèíè in vitro
Ñ. Â. Ïðèëóöüêà, Ã. Â. ijäåíêî, Ã. Ï. Ïîòåáíÿ, Ê. ². Áîãóöüêà,
Þ. ². Ïðèëóöüêèé, Ó. гòòåð, Ï. Øàðô
Ðåçþìå
Ñòâîðåííÿ íîâèõ íàíîñòðóêòóð ç âèñîêîþ ïðîòèïóõëèííîþ àê-
òèâí³ñòþ º âàæëèâîþ ïðîáëåìîþ ñó÷àñíî¿ á³îòåõíîëî㳿. Ìåòà.
Îö³íèòè öèòîòîêñè÷í³ñòü ñòâîðåíîãî êîìïëåêñó ôóëåðåíó C60 ç
àíòèá³îòèêîì àíòðàöèêë³íîâîãî ðÿäó äîêñîðóá³öèíîì (Äîêñ) íà
ð³çí³ òèïè êë³òèí (ïóõëèíí³, ³ìóíîêîìïåòåíòíi ³ ãåïàòîöèòè) òà
ïîð³âíÿòè îäåðæàí³ ðåçóëüòàòè ç òîêñè÷íîþ 䳺þ â³ëüíîãî ôóëå-
ðåíó C60 ³ Äîêñ çà óìîâ in vitro. Ìåòîäè. Ðîçïîä³ë çà ðîçì³ðîì ÷à-
ñòèíîê ó C60 + Äîêñ ñóì³ø³ âèì³ðþâàëè ìåòîäîì äèíàì³÷íîãî
ðîçñ³þâàííÿ ñâ³òëà (ÄÐÑ). Òîêñè÷íèé åôåêò êîìïëåêñó C60 + Äîêñ
ùîäî íîðìàëüíèõ ³ ïóõëèííèõ êë³òèí âèâ÷àëè in vitro ç âèêîðèñòàí-
íÿì ÌÒÒ-òåñòó. Ðåçóëüòàòè. Âñòàíîâëåíî, ùî â ñóì³ø³ C60 +
Äîêñ â îñíîâíîìó ðåºñòðóþòüñÿ ÷àñòèíêè ç ä³àìåòðîì 132 íì.
Òîêñè÷íà ä³ÿ êîìïëåêñó C60 + Äîêñ ùîäî íîðìàëüíèõ (ë³ìôîöèòè,
ìàêðîôàãè, ãåïàòîöèòè) ³ ïóõëèííèõ (àñöèòíà êàðöèíîìà Åðë³õà,
ëåéêîç L1210, êàðöèíîìà ëåãåí³ Ëüþ¿ñ) êë³òèí âèÿâèëàñÿ ìåíøîþ
ïðèáëèçíî íà 10–16 ³ 7–9 % â³äïîâ³äíî ïîð³âíÿíî ³ç âïëèâîì â³ëüíî-
ãî Äîêñ. Âèñíîâêè. Ðîçðîáëåíèé êîìïëåêñ C60 + Äîêñ ìîæíà ðîç-
ãëÿäàòè ÿê íîâèé ôàðìàêîëîã³÷íèé ïðåïàðàò, ùî çà óìîâ in vitro
çäàòíèé åôåêòèâíî çíèùóâàòè ïóõëèíí³ êë³òèíè ³ îäíî÷àñíî çà-
ïîá³ãàòè ïîá³÷íèì òîêñè÷íèì åôåêòàì, ïðèòàìàííèì òðàäèö³é-
íîìó ïðîòèïóõëèííîìó ïðåïàðàòó Äîêñ ùîäî íîðìàëüíèõ êë³òèí.
Êëþ÷îâ³ ñëîâà: êîìïëåêñ ôóëåðåí C
60
–äîêñîðóá³öèí, íîðìàëüí³ ³
ïóõëèíí³ êë³òèíè, öèòîòîêñè÷í³ñòü, ÌÒÒ-òåñò, äèíàì³÷íå ðîçñ³-
ÿííÿ ñâ³òëà.
Òîêñè÷åñêèé ýôôåêò êîìïëåêñà ôóëëåðåí C60–äîêñîðóáèöèí
íà íîðìàëüíûå è îïóõîëåâûå êëåòêè in vitro
Ñ. Â. Ïðèëóöêàÿ, Ã. Â. Äèäåíêî, Ã. Ï. Ïîòåáíÿ, Å. È. Áîãóöêàÿ,
Þ. È. Ïðèëóöêèé, Ó. Ðèòòåð, Ï. Øàðô
Ðåçþìå
Ñîçäàíèå íîâûõ íàíîñòðóêòóð ñ âûñîêîé ïðîòèâîîïóõîëåâîé àê-
òèâíîñòüþ ÿâëÿåòñÿ âàæíîé ïðîáëåìîé ñîâðåìåííîé áèîòåõíî-
375
TOXIC EFFECT OF Ñ60 FULLERENE-DOXORUBICIN COMPLEX TOWARDS TUMOR AND NORMAL CELLS IN VITRO
Cells Ñ60 fullerene (0.15 mg/ml) Dox (0.15 mg/ml) Ñ60 + Dox complex (0.05 + 0.1 mg/ml)
Normal cells
Lymphocytes 114.67 ± 1.37* 43.20 ± 4.61* 56.22 ± 2.02*
Macrophages – 51.52 ± 3.92* 62.38 ± 3.72*
Hepatocytes – 34.43 ± 3.22* 49.53 ± 4.97*
Tumor cells
Ehrlich ascite carcinoma 74.19 ± 2.39* 21.39 ± 2.39* 29.19 ± 2.99*
L1210 Leukemia – 7.47 ± 0.56* 16.44 ± 0.44*
Lewis lung carcinoma 117.47 ± 1.36* 31.09 ± 3.13* 38.42 ± 3.47*
N o t e. The results are statistically significant compared to the corresponding control (without any compound); *ð � 0.05.
The survival of normal and tumor cells (in %) under the action of various compounds (ÌÒÒ-test)
a b c d
Fig. 2. Toxic effect of Ñ
60
fullerene (b), Dox (c) and Ñ
60
+ Dox complex (d) towards cells of Ehrlich ascite carcinoma after 18 h treatment compared
with the control (à) ( 160)
376
PRYLUTSKA S. V. ET AL.
ëîãèè. Öåëü. Îöåíèòü öèòîòîêñè÷íîñòü ñîçäàííîãî êîìïëåêñà
ôóëëåðåíà Ñ60 ñ àíòèáèîòèêîì àíòðàöèêëèíîâîãî ðÿäà äîêñîðó-
áèöèíîì (Äîêñ) íà ðàçíûå òèïû êëåòîê (îïóõîëåâûå, èììóíîêîì-
ïåòåíòíûå, ãåïàòîöèòû) è ñðàâíèòü ïîëó÷åííûå ðåçóëüòàòû ñ
òîêñè÷åñêèì äåéñòâèåì ñâîáîäíîãî ôóëëåðåíà C60 è Äîêñ â óñëî-
âèÿõ in vitro. Ìåòîäû. Ðàñïðåäåëåíèå ïî ðàçìåðó ÷àñòèö â ñìåñè
Ñ60 + Äîêñ èçìåðÿëè ìåòîäîì äèíàìè÷åñêîãî ðàññåèâàíèÿ ñâåòà
(ÄÐÑ). Òîêñè÷åñêèé ýôôåêò êîìïëåêñà Ñ60 + Äîêñ íà íîðìàëüíûå
è îïóõîëåâûå êëåòêè in vitro èçó÷àëè ñ èñïîëüçîâàíèåì ÌÒÒ-òåñ-
òà. Ðåçóëüòàòû. Óñòàíîâëåíî, ÷òî â ñìåñè Ñ60 + Äîêñ ðåãèñòðè-
ðóþòñÿ â îñíîâíîì ÷àñòèöû ñ äèàìåòðîì 132 íì. Òîêñè÷åñêîå
äåéñòâèå êîìïëåêñà Ñ60 + Äîêñ ïî îòíîøåíèþ ê íîðìàëüíûì
(ëèìôîöèòû, ìàêðîôàãè, ãåïàòîöèòû) è îïóõîëåâûì (àñöèòíàÿ
êàðöèíîìà Ýðëèõà, ëåéêîç L1210, êàðöèíîìà ëåãêîãî Ëüþèñ)
êëåòêàì îêàçàëîñü ìåíüøèì íà ~ 10–16 è 7–9 % ñîîòâåòñòâåííî
ïî ñðàâíåíèþ ñ äåéñòâèåì ñâîáîäíîãî Äîêñ. Âûâîäû. Ðàçðàáîòàí-
íûé êîìïëåêñ Ñ60 + Äîêñ ìîæíî ðàññìàòðèâàòü êàê íîâûé ôàð-
ìàêîëîãè÷åñêèé ïðåïàðàò, ñïîñîáíûé â óñëîâèÿõ in vitro ýôôåê-
òèâíî óíè÷òîæàòü îïóõîëåâûå êëåòêè è îäíîâðåìåííî ïðåäîò-
âðàùàòü ïîáî÷íûå òîêñè÷åñêèå ýôôåêòû, ïðèñóùèå òðàäèöèîí-
íîìó ïðîòèâîîïóõîëåâîìó ïðåïàðàòó Äîêñ îòíîñèòåëüíî íîð-
ìàëüíûõ êëåòîê.
Êëþ÷åâûå ñëîâà: êîìïëåêñ ôóëëåðåí C
60
–äîêñîðóáèöèí, íîðìàëü-
íûå è îïóõîëåâûå êëåòêè, öèòîòîêñè÷íîñòü, ÌÒÒ-òåñò, äèíàìè-
÷åñêîå ðàññåÿíèå ñâåòà.
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Received 25.01.14
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