Publications on the project |
101 Oligoperoxide Based Physically Detectable Nanocomposites for Cell Targeting, Visualization and Treatment |
Authors: | A. Zaichenko, N. Mitina, O. Shevchuk, O. Shapoval, N. Boiko, R. Bilyy, R. Stoika, A. Voloshinovskii and D. Horak | |
Summary: | Novel promising routes for the obtaining luminescent, magnetic, plasmon resonance capable nanocomposites as well as drug delivery systems are proposed and studied. These routes are based on the synthesis and application of original type of functional oligoperoxides, their coordinating complexes of transition including rare earth metal cations for controlled synthesis of block, comb‐like or nanogel oligoelectrolytes as well as for template synthesis of functional polymer and polymer‐mineral nanoparticles. Developed nanoscale delivery systems were tested as medicines for cancer administration in vitro and in vivo. Physically detectable lectin‐conjugated nanocomposites were studied as biomarkers for pathological cell targeting and visualization. | |
Keywords: | oligoperoxide surfactants, oligoelectrolytes, drug and gene delivery systems, physically detectable nanocomposites, bioconjugates, targeting, labeling and visualization | |
Edition: | AIP Conference Proceedings | | | 2010,
178-182,English |
101 Immobilization of doxorubicin on the olygoeleсtrolytic polymeric carrier VEP-GMA-PEG increases its and anticancer activity cellular uptake |
Authors: | Yu. V. Senkiv, A. R. Ryabtseva, P. Heffeter, E. A. Shlyakhtina, N. E. Mitina, W. Berger, O. S. Zaichenko, R. S. Stoika | |
Summary: | Application of special systems for drug delivery into target cells might be useful for overcoming several problems in treatment of dangerous diseases. These are: consequences of nonspecific negative effects of drugs towards healthy cells, low sensitivity of cancer cells to anticancer drugs used in doses that are non-toxic for the organism, resistance of tumor cells to anticancer drugs and of the pathogenic microorganisms to the antibiotics. Here we studied the efficiency of application of novel nanoscale drug delivery system in tumor cell lines, including drug-resistant ones. The polyethylene glycol (PEG)-modified polymeric carrier VEP-GMA used in this study, was synthesized at the Department of Organic Chemistry of Lviv National Polytechnic University. We compared
the effect of free doxorubicin and of this anticancer drug immobilized on the polymeric carrier, towards human tumor cells. It was found that such immobilization of doxorubicin significantly enhanced the cytotoxic action of this drug towards human lung carcinoma A549 cells, human colorectal carcinoma HCT116 cells, human breast carcinoma MCF-7 cells and their doxorubicin-resistant MCF-7/ADR subline. The results of our studies demonstrated that using doxorubicin complex with novel polymeric nanoscale carrier VEP-GMA-PEG permitted reducing the active dose of doxorubicin in cancer cells at least 10 times, comparing with such dose of this anticancer drug used in free form.
Since the antineoplastic effect of carrier-immobilized doxorubicin was maintained, these results suggest a potential reduction of negative side effects of the corresponding chemotherapy.
It was shown that the uptake by tumor cells of the carrier- immobilized doxorubicin was significantly enhanced comparing with such uptake of free doxorubicin. Our data demonstrated that neither macropinocytosis, nor endocytosis can be responsible
for the uptake of doxorubicin that is immobilized on the nanosized polymeric carrier. Our future experiments are focused on the improvement of characteristics of this carrier by means of its specific functionalization aimed at reaching its addressed action towards tumor cells in vitro and in vivo. | |
Keywords: | nanosized polymeric drug carriers, doxorubicin, human tumor cells, cell resistance to drugs | |
Edition: | Studia Biologica | | | 2012,
5-16,Ukrainian |
101 Structural and colloidal-chemical characteristics of nanosized drug delivery systems based on pegylatedcomb-like carriers |
Authors: | A. Riabtseva, N. Mitina, N. Boiko, S. Garasevich, I. Yanchuk, R. Stoika, O. Slobodyanyuk, A. Zaichenko | |
Summary: | Novel comb-like polymeric and oligomeric
drug carriers combining backbone – copolymers of 5-tertbutylperoxy-5-methyl-1-hexene-3-yne
(VEP) and glycidyl
methacrylate (GMA) – and side PEG chains of various
lengths were synthesized. Nanosized delivery systems
containing conjugates of water soluble anticancer drug
Doxorubicin were developed. The structures of
copolymers and their conjugates with drugs were
confirmed by IR-spectroscopy. Structural and colloidalchemical
properties of water drug delivery systems were
studied using photoluminescent (PL), UV-spectroscopy
techniques, surface tension measurements and dynamic
light scattering. The scheme of the immobilization of
water soluble doxorubicin on developed PEGylated
polymeric carriers was assumed. | |
Keywords: | polymeric carriers, drug delivery systems, anticancer preparations | |
Edition: | Chemistry & Chemical Technology | | | 2012,
291–295,English |
101 Polyplex formation by novel surface active comb-like polyamfolytes and plasmid DNA |
Authors: | N. S. Finiuk, T. Y. Vitak, N. Y. Mitina, Y. Z. Filyak, O. S. Zaichenko,
R. S. Stoika | |
Summary: | Formation of the interpolyelectrolytic complexes (polyplexes) of plasmid DNA and novel surface active comb-like polyampholytic carriers was studied. To do that, the method of determi -
ning DNA retardation during its electrophoresis in the agarose gel was used. Optimal conditions for the formation of such polylexes were defined: PC concentration 0.1–0.003 %, рН 7.4, 20 min,
24 °С. It was found that polyampholyte possessing quaternized amino-containing side chains is capable to form the most stable polyplexes with plasmid DNA. The association of DNA with
polyampholytic carriers and its release from such complex do not cause changes in DNA structure. Therefor, the polyampholytic carriers under study protected DNA from its nuclease
cleavage. Thus, novel surface active comb-like polyampholytes are perspective carriers for delivering DNA to the recipient cells. | |
Keywords: | plasmid DNA, polyampholytic carriers, stability of polyplexes, DNA electrophoresis | |
Edition: | Biotechologia | | | 2012,
,Ukrainian |
101 A novel method for genetic transformation of yeast cells using oligoelectrolyte polymeric nanoscale carriers |
Authors: | Ye. Filyak, N. Finiuk, N. Mitina, O. Bilyk, V. Titorenko, O. Hrydzhuk, A. Zaichenko, R. Stoika | |
Summary: | The genetic transformation of target cells is a key tool in modern biological research, as well as in many gene therapy and biotechnology applications. Here we describe a new method for delivery of DNA into several industrially important species of yeast, including Saccharomyces cerevisiae. Our method is based on the use of a novel nanoscale oligoelectrolyte polymer possessing a comb-like structure as a carrier molecule. Direct comparisons to standard transformation methods clearly show that our approach: (i) yields two times more transformants of Hansenula polymorpha NCYC 495 compared to electroporation approaches and 15 times more transformants compared to lithium acetate protocols, as well as (ii) 5 times more Pichia pastoris GS115 transformants compared to electroporation and 79 times more transformants compared to lithium acetate. Taken together, these results clearly indicate genetic transformation of yeasts using oligoelectrolyte polymer carriers is a highly effective means of gene delivery. | |
Keywords: | genetic transformation; yeast; gene delivery; nanoscale system; comb-like oligoelectrolyte polymer | |
Edition: | BioTechniques | | | 2013,
35–43,English |
101 Biophysical study of novel oligoelectrolyte-based nonviral gene delivery systems for mammalian cells |
Authors: | S. Z. Ficen, Z. Guler, N. Mitina, N. Finіuk, R. Stoika, A. Zaichenko, S. E. Ceylan | |
Summary: | Background
Gene therapy is an important treatment for genetic and acquired diseases. The success of gene therapy is largely dependent on the development of suitable vectors for gene transfer. Vectors are needed to overcome cellular barriers and to achieve efficient DNA delivery with low cytotoxicity. In the present study, we synthesized and characterized a novel comb-like oligoelectrolyte nanocarrier, BG-2, as a nonviral gene delivery vector.
Methods
A novel surface-active oligoelectrolyte of comb-like structure was synthesized via controlled radical copolymerization using oligoperoxide Cu+2 coordinating complex as a multisite initiator of graft copolymerization. The critical micellar concentration was determined by Nile Red fluorescence. Complex formation of DNA with BG-2 was determined by YOYO-1 fluorescence. The physicochemical properties of DNA in complex with BG-2 have been investigated by electrophoresis, dynamic light scattering and fluorescence spectroscopy. The BG-2/DNA complex was demonstrated by scanning electron microscopy. Interactions between BG-2/DNA complex and model membranes were also studied. The sensitivity of the DNA molecule, complexed with BG-2, against deoxyribonuclease I and serum nucleases was assessed by agarose gel electrophoresis. BG-2 efficiency in the transfection of HeLa cells was determined by measuring luciferase gene expression using a luminometer and cytotoxicity was also evaluated.
Results
BG-2 oligoelectrolyte was successful in overcoming cellular barriers as a result of forming stable and small sized complexes with DNA, interacting with model membranes in a desirable manner and protecting DNA from nuclease. The transfection efficiency was quite high and cytotoxicity was low.
Conclusions
BG-2 appears to be a promising nonviral vector with low cytotoxicity and efficient transfection properties. | |
Keywords: | gene delivery;nanobiotechnology;transfection;vector-non viral;vector-polymeric | |
Edition: | Journal of Gene Medicine | | | 2013,
193–204,English |
101 Enhanced cytotoxicity of anticancer drug delivered by novel nanoscale polymeric carrier |
Authors: | R. Stoika, N. Boiko, Y. Senkiv, Y. Shlyakhtina, R. Panchuk, N. Finiuk, Y. Filyak, R. Bilyy, Y. Kit, N. Skorohyd, O. Klyuchivska, A. Zaichenko, N. Mitina, A. Ryabceva | |
Summary: | We compared in vitro action of highly toxic anticancer drug doxorubicin under its delivery to the mammalian tumor cells in free form and after encapsulation in novel bio-functionalized nanoscale polymeric carrier. Such encapsulation was found to enhance significantly drug uptake by the targeted cells, as well as its cytotoxic action. 10 times higher cytotoxicity of the carrier-immobilized doxorubicin comparing to its free form was demonstrated by direct cell counting, and 5 times higher cytotoxicity of encapsulated doxorubicin was shown by FACS analysis. The polymeric carrier itself did not possess significant toxicity in vitro or in vivo (laboratory mice). The carrier protected against negative side effects of doxorubicin in mice with experimental NK/Ly lymphoma. The life duration of tumor-bearing animals treated with doxorubicin-carrier complex was significantly longer than life duration in animals treated with free doxorubicin. Besides, the effective treatment dose of the carrier-delivered doxorubicin in tumor-bearing mice was 10 times lower than such dose of free doxorubicin. Thus, novel nanoscale polymers possess high potential as drug carrier. | |
Keywords: | | |
Edition: | Journal of Physics: Conference Series | | | 2013,
012038,English |
101 Action of free and polymer carrier encapsulated doxorubicin towards HCT 116 cells of human colorectal carcinoma |
Authors: | Yu.V. Senkiv, P. Heffeter, A.O. Riabtseva, N.M. Boiko, O.S. Zaichenko, N.Ye. Mitina, W. Berger, R.S. Stoika | |
Summary: | Development of novel nanoscale functionalized
carriers is nowadays one of the most urgent
problems in cancer treatment.
The aim of our study was to compare the
antineoplastic effect of free doxorubicin and its
complex with a nanoscale polymeric carrier towards
HTC116 colorectal carcinoma cells. It was
established that application of the complex of
poly(5-tret-butylperoxy)-5-methyl-1-hexene-3-inco-glycydyl
metacrylat)-graft-polyethyleneglycol
(poly(VEP-GMA-PEG)-graft-PEG), where
VEP – 5-tret-butylperoxy)-5-methyl-1-hexene-
3-in; GMA – glycydyl metacrylat; graft-PEG –
graft-polyethyleneglycol accordingly, functionalized
with phosphatidylcholine for doxorubicin
delivery increased 10 times the efficiency of cytotoxic
action of this drug, as compared wich such
efficiency in case of the action of free doxorubicin.
The encapsulated form of doxorubicin caused more intensive cleavage of the reparation enzyme
PARP and longer delay in G2/M cell cycle arrest,
compared to such effects of free doxorubicin.
The developed carrier itself is non-toxic to the
used mammalian cells and does not cause impairment
in their cell cycle. A deletion in both alleles
of p53 gene did not affect the antineoplastic action
of doxorubicin that was immobilized on the
nanoscale carrier. Thus, p53-dependent signaling
pathways are not involved in the cytotoxic action
of doxorubicin-carrier complex. It is suggested
that novel nanoscale polymeric carrier poly(VEPGMA-PEG)-graft-PEG
functionalized with phosphatidylcholine
could be a promising carrier for
targeted delivery of anticancer drugs. | |
Keywords: | human colorectal carcinoma cells, doxorubicin, nanoscale polymeric drug carrier, cell cycle, PARP, apoptosis | |
Edition: | Ukrainian Biochemical Journal | | | 2013,
33–44,Ukrainian |
101 Action of doxorubicin delivered to tumor cells in vitro and in vivo by novel nanoscale oligoelectrolytic carrier |
Authors: | N. M. Boiko, Yu. V. Senkiv, Ye. A. Shlyakhtina, O. Y. Kluchivska,
N. P. Skorohid, N. E. Mitina, T. V. Skorohoda, M. M. Moskvin, O. S. Zaichenko, R. S. Stoika | |
Summary: | The main tasks of modern biopharmaceutics
are focused at the development of new nanoscale
carriers with low toxicity, given size, regulated
response to local and remote effects, and capability
of visualization of drug action and diagnostic
results. A novel oligomeric carrier VAMANG-MP
for delivery of the anticancer drug
doxorubicin to tumor cells of different lines have
been used. The synthesized oligomeric carrier
was additionally functionalized by phosphatitylcholine.
It was demonstrated that such delivery
of doxorubicin to the target cells permits 10
times decreasing of its acting cytotoxic dose
comparing with such dose of free doxorubicin,
with preserving similar level of the antineoplastic
effect. This dose-specific effect was demonstrated
both in vitro towards various mammalian
tumor cells, and in vivo towards mice with experimental
NK/Ly lymphoma was shown. Action of
immobilized doxorubicin was followed by more
intense formation of vesicles on a surface of the
target cells in vitro, and by their inter-nucleosomal
DNA fragmentation as compared to action of
free doxorubicin, as well as by appearance of
higher amount of dead cells in ascytic fluid of
the treated NK/Ly lymphoma mice. Thus, the
synthesized nanoscale olygoelectrolytic carrier
is a perspective system for delivery of anticancer
drugs to target cells. | |
Keywords: | doxorubicin delivery, tumor cells, olygoelectrolytes, nanoscale particles | |
Edition: | Biotechnologia Acta | | | 2013,
53–62,Ukrainian |
101 Evaluation of cytotoxic and mutagenic action of novel surface active comb-like polyampholytes that are used for delivery of nucleic acids to target cells |
Authors: | N. S. Finiuk, Y. Z. Filyak, N. M. Boiko, N. Y. Mitina, O. S. Zaichenko, R. S. Stoika | |
Summary: | Rapid development of new nanomaterials and nanotechnologies is accompanied
by significant achievements in various fields of medicine, industry and precise technology,
as well as by undesirable effects on human health and environment. Our studies
were focused on determination of toxicity in vitro and evaluation of the mutagenic activity
of novel polyampholyte carriers of BG-2 type. It was shown that they did not exert
cytotoxic and could be used in the in vitro experiments at a concentration less than
0.01%. The carriers were not capable of triggering gene mutations in the Ames test. At
the absence of metabolic activation, all studied carriers lacked genotoxic effects. No
mutagenic effect was observed for novel carriers after adding of the microsomal fraction
of rat liver for both strains of Salmonella typhimurium, namely TA98 and TA100. This
indicates a safety of novel polyampholyte carriers of BG-2 type as a tool for delivery of nucleic acids into target cells. | |
Keywords: | polyampholytic carriers, cytotoxic impact, cultures of mammalian
cells, Ames test | |
Edition: | Studia Biologica | | | 2013,
15-26,Ukrainian |
101 Enhanced Anticancer Activity and Circumvention of Resistance Mechanisms by Novel Polymeric/Phospholipidic Nanocarriers of Doxorubicin |
Authors: | Y. Senkiv, A. Riabtseva, P. Heffeter, N. Boiko, C. R. Kowol, U. Jungwith, Y. Shlyakhtina, S. G. Garasevych, N. Mitina,
W. Berger, A. Zaichenko, R. Stoika | |
Summary: | Severe toxic side effects and drug resistance are the major limitations of doxorubicin (Dox), one of the most potent anticancer agents in clinical use. Nanocarrier preparations offer the opportunity to overcome these drawbacks, which is reflected in the clinical approval of two liposomal Dox preparations. Additionally, there are many attempts to enhance the activity of Dox against multi-drug resistant (MDR) cancer cells. However, most of these strategies resulted in the increased uptake of Dox in resistant cells, only, while it remained unchanged in chemo-sensitive cells. Here, we present a new polymeric-phospholipidic hybrid delivery system which distinctly enhanced the accumulation and activity of Dox in all tested cancer cell lines including several MDR cell models. Notably, the resistance levels against Dox were reduced from about 6-fold to about 2-fold. Moreover, the new nanocarriers were shown to rapidly (within 10 min) and effectively transport Dox into resistant as well as sensitive cancer cells. Consequently, treatment with the new Dox-containing nanocarriers resulted in effective cell cycle arrest in G2/M phase and ROS-induced cell death induction. Finally, the new nanocarriers were tested against NK/Ly lymphoma and L1210 leukemia cells in vivo. In both cell models, the nanoformulation of Dox resulted in 100% cured animals already at low concentrations (0.1 mg/kg), while free Dox solely extended survival time. This indicates that the incorporation of phospholipids into PEGylated polymeric nanocarriers is a promising strategy to enhance efficacy and reduce toxicity of Dox treatment against both sensitive and resistant cancer models in vitro and in vivo. | |
Keywords: | cancer therapy; doxorubicin; drug delivery; drug resistance; nanocarrier | |
Edition: | Journal of Biomedical Nanotechnology | | | 2014,
1369-1381,English |
101 Nanoformulation Improves Activity of the (pre)Clinical Anticancer Ruthenium Complex KP1019 |
Authors: | P. Heffeter, A Riabtseva, Y. Senkiv, C. R. Kowol, W. Körner, U. Jungwith, N. Mitina, B. K. Keppler, T. Konstantinova,
I. Yanchuk, R. Stoika, A. Zaichenko, W. Berger | |
Summary: | Ruthenium anticancer drugs belong to the most promising non-platinum anticancer metal compounds in clinical evaluation. However, although the clinical results are promising regarding both activity and very low adverse effects, the clinical application is currently hampered by the limited solubility and stability of the drug in aqueous solution. Here, we present a new nanoparticle formulation based on polymer-based micelles loaded with the anticancer lead ruthenium compound KP1019. Nanoprepared KP1019 was characterised by enhanced stability in aqueous solutions. Moreover, the nanoparticle formulation facilitated cellular accumulation of KP1019 (determined by ICP-MS measurements) resulting in significantly lowered IC50 values. With regard to the mode of action, increased cell cycle arrest in G2/M phase (PI-staining), DNA damage (Comet assay) as well as enhanced levels of apoptotic cell death (caspase 7 and PARP cleavage) were found in HCT116 cells treated with the new nanoformulation of KP1019. Summarizing, we present for the first time evidence that nanoformulation is a feasible strategy for improving the stability as well as activity of experimental anticancer ruthenium compounds. | |
Keywords: | anticancer cancer therapy; cell death induction; drug delivery; nanocarrier; ruthenium | |
Edition: | Journal of Biomedical Nanotechnology | | | 2014,
877-884,English |
101 Synthesis of novel pegylated carriers for nanoscale drug delivery systems |
Authors: | A. Riabtseva , N. Mitina , N. Boiko , Z. Nadashkevych , R. Stoika , O. Zaichenko | |
Summary: | The results of purposeful synthesis of new oligomeric PEGylated carriers and their properties studied
by chemical and physical-chemical methods are presented in the article. It is shown the opportunity to obtain
nanoscale water systems of conjugates of novel oligomeric carriers with anticancer substances.
Keywords: polymeric carriers, oligoperoxides, PEGylation, drug delivery systems. | |
Keywords: | polymeric carriers, oligoperoxides, PEGylation, drug delivery systems. | |
Edition: | | | | 2011,
19-27,Ukrainian |
101 PEGylated oligomeric carriers and nanoscale delivery systems of antimicrobial substances |
Authors: | A. Riabtseva , Y. Ostapchuk, N. Mitina , M. Moskvin,N. Boyko , R. Stoika , O. Zaichenko | |
Summary: | The results of purposeful synthesis of new oligomeric PEGylated carriers and their properties studied by chemical and physical-chemical methods are presented in the article. It is shown the opportunity to obtain nanoscale water systems of conjugates of novel oligomeric carriers with antimicrobial substances. | |
Keywords: | polymeric carriers, oligoperoxides, PEGylation, drug delivery systems | |
Edition: | | | | 2011,
367-373,Ukrainian |
101 Nanoscale delivery systems of antimicrobial and anticancer drugs based on new PEGylated oligomeric carriers |
Authors: | A.А. Riabtseva , N.E. Mitina , N.N. Boyko , R.S. Stoika , A.S. Zaichenko | |
Summary: | | |
Keywords: | | |
Edition: | | | | 2011,
121-122,Russian |
101
The events in the framework of the project |
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101 Executant:Institute of Cell Biology, Department of Biochemistry, Physiology and Molecular Biology, Section of Chemical and Biological Sciences 4.1. Biocompatible nanostructured materials: interaction with biological systems, safety use, means and methods controlled delivery Purpose:To develop physical and chemical bases of encapsulating of physiologically active substances, including fluorescent, in polymer nanocomposites with reactive functional shell by mini-emulsion polymerization involving surface active monomers and initiators and multi investigate mechanisms of action at the cellular and molecular levels. Expected results:Release of new product: material Stage 1:Obtaining new nanoscale polymer composites using unsaturated surfactants based oligoperoxides for multi aqueous polymerization; encapsulation in situ fluorescein in synthesized nanocomposites with providing protection against leakage of fluorophores from NC. Stage 2:The use of obtained nanocomposites for the delivery of drugs in the tumor cells of animals and humans in conditions in vitro; comparison of the molecular mechanisms of action of these drugs in conditions of delivery in the free state and in combination with novel NC. Stage 3:The research of mechanisms of internalization of obtained nanocomposites in malignant cells, influence on cell cycle and intensity of induction of apoptosis signaling mechanisms in malignant mammalian cells using Western blot analysis, fluorescence microscopy, flow cytometry. Stage 4:The integration of nucleic acids (plasmid DNA and siRNA) with created nanocomposites thanks to the particular structure of these nanocomposites; these bio-functionalized nanocomposites will be used for genetic modification of different types of target cells (bacteria, yeasts, animal cells and human). Stage 5:The use of obtained nanocomposites for the delivery of drugs (including doxorubicin) in tumor cells of experimental animals; comparison of the molecular mechanisms of action of these drugs in conditions of delivery in the free state and in combination with novel nanocomposites.
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