Марквичева Елена Арнольдовна

Личная информация

Под ее руководством защищено 25 дипломных работ и 10 кандидатских диссертаций.

Научные интересы

Ее научные интересы связаны с получением новых биоматериалов для биомедицины (системы с контролируемой доставкой лекарств, нано-капсулирование биоактивных пептидов и белков, микрокапсулирование животных клеток, биодеградируемые матриксы (скаффолды)  для репарации тканей и др..

Основные научные результаты

Автор более 200 публикаций, среди которых более 80 статей, 7 патентов, 7 глав в книгах, вышедших в России и за рубежом.

Членство в научных обществах

Участвует в работе русских научных и зарубежных обществ. Является представителем и главным координатором международного общества Bioencapsulation Research Group в России, представляет Россию (является экспертом и входит в координационный комитет) в международных программах COST (840 и 865).

Избранные публикации

  1. Drozdova M.G., ZaytsevaZotova D.S., Akasov R.A., Golunova A.S., Artyukhov A.A., Udartseva O.O., Andreeva E.R., Lisovyy D.E., Shtilman M.I., Markvicheva E.A. (2017). Macroporous modified poly (vinyl alcohol) hydrogels with charged groups for tissue engineering: Preparation and in vitro evaluation. Mater Sci Eng C Mater Biol Appl 75, 1075–1082 [+]

    Poly(vinyl alcohol) (PVA) hydrogels are widely employed for various biomedical applications, including tissue engineering, due to their biocompatibility, high water solubility, low protein adsorption, and chemical stability. However, non-charged surface of PVA-based hydrogels is not optimal for cell adhesion and spreading. Here, cross-linked macroporous hydrogels based on low molecular weight acrylated PVA (Acr-PVA) was synthesized by modification of the pendant alcohol groups on the PVA with glycidyl methacrylate (GMA). To enhance cell affinity, charged groups were introduced to the hydrogel composition. For this purpose, Acr-PVA was copolymerized with either negatively charged acrylic acid (AA) or positively charged 2-(diethylamino) ethyl methacrylate (DEAEMA) monomers. A surface charge of the obtained hydrogels was found to be in function of the co-monomer type and content. Confocal microscopy observations confirmed that adhesion and spreading of both mouse fibroblasts (L929) and human mesenchymal stem cells (hMSC) on the modified Acr-PVA-AA and Acr-PVA-DEAEMA hydrogels were better than those on the non-modified Acr-PVA hydrogel. The increase of DEAEMA monomer content from 5 to 15mol% resulted in the enhancement of cell viability which was 1.5-fold higher for Acr-PVA-DEAEMA-15 hydrogel than that of the non-modified Acr-PVA hydrogel sample.

  2. Haq S., Samuel V., Haxho F., Akasov R., Leko M., Burov S.V., Markvicheva E., Szewczuk M.R. (2017). Sialylation facilitates self-assembly of 3D multicellular prostaspheres by using cyclo-RGDfK(TPP) peptide. Onco Targets Ther 10, 2427–2447 [+]

    Prostaspheres-based three dimensional (3D) culture models have provided insight into prostate cancer (PCa) biology, highlighting the importance of cell-cell interactions and the extracellular matrix (EMC) in the tumor microenvironment. Although these 3D classical spheroid platforms provide a significant advance over 2D models mimicking in vivo tumors, the limitations involve no control of assembly and structure with only limited spatial or glandular organization. Here, matrix-free prostaspheres from human metastatic prostate carcinoma PC3 and DU145 cell lines and their respective gemcitabine resistant (GemR) variants were generated by using cyclic Arg-Gly-Asp-D-Phe-Lys peptide modified with 4-carboxybutyl-triphenylphosphonium bromide (cyclo-RGDfK(TPP)).

  3. Akasov R., Gileva A., ZaytsevaZotova D., Burov S., Chevalot I., Guedon E., Markvicheva E. (2016). 3D in vitro co-culture models based on normal cells and tumor spheroids formed by cyclic RGD-peptide induced cell self-assembly. Biotechnol. Lett. , [+]


    To design novel 3D in vitro co-culture models based on the RGD-peptide-induced cell self-assembly technique.


    Multicellular spheroids from M-3 murine melanoma cells and L-929 murine fibroblasts were obtained directly from monolayer culture by addition of culture medium containing cyclic RGD-peptide. To reach reproducible architecture of co-culture spheroids, two novel 3D in vitro models with well pronounced core-shell structure from tumor spheroids and single mouse fibroblasts were developed based on this approach. The first was a combination of a RGD-peptide platform with the liquid overlay technique with further co-cultivation for 1-2 days. The second allowed co-culture spheroids to generate within polyelectrolyte microcapsules by cultivation for 2 weeks. M-3 cells (a core) and L-929 fibroblasts (a shell) were easily distinguished by confocal microscopy due to cell staining with DiO and DiI dyes, respectively.


    The 3D co-culture spheroids are proposed as a tool in tumor biology to study cell-cell interactions as well as for testing novel anticancer drugs and drug delivery vehicles.

  4. Akasov R., Haq S., Haxho F., Samuel V., Burov S.V., Markvicheva E., Neufeld R.J., Szewczuk M.R. (2016). Sialylation transmogrifies human breast and pancreatic cancer cells into 3D multicellular tumor spheroids using cyclic RGD-peptide induced self-assembly. Oncotarget , [+]

    Multicellular tumor spheroids (MTS) have been at the forefront of cancer research, designed to mimic tumor-like developmental patterns in vitro. Tumor growth in vivo is highly influenced by aberrant cell surface-specific sialoglycan structures on glycoproteins. Aberrant sialoglycan patterns that facilitate MTS formation are not well defined. Matrix-free spheroids from breast MCF-7 and pancreatic PANC1 cancer cell lines and their respective tamoxifen (TMX) and gemcitabine (Gem) resistant variants were generated using the RGD platform of cyclic Arg-Gly-Asp-D-Phe-Lys peptide modified with 4-carboxybutyl-triphenylphosphonium bromide (cyclo-RGDfK (TPP)). MCF-7 and MCF-7 TMX cells formed tight spheroids both in the classical agarose- and RGD-based platforms while all PANC1 cells formed loose aggregates. Using lectin histochemistry staining, sialidase assay, neuraminidase (Vibrio cholerae) and oseltamivir phosphate (OP) neuraminidase inhibitor treatments, MCF-7 and PANC1 cells and their drug-resistant variants expressed different sialic acid (SA) content on their cell surfaces. α-2,3- and α-2,6-sialic acid surface residues facilitated spheroid formation under cyclo-RGDfK(TPP)-induced self-assembly. Pretreatment with α-2,3-SA specific Maackia amurensis (MAL-II) lectin, α-2,6-SA specific Sambucus nigra (SNA) lectin, and exogenous α-2,6-SA specific neuraminidase (Vibrio cholerae) dose-dependently reduced spheroid volume. OP enhanced cell aggregation and compaction forming spheroids. PANC1 and MDA-MB231 xenograft tumors from untreated and OP-treated RAGxCγ double mutant mice expressed significantly higher levels of α-2,3-SA over α-2,6-SA. MCF-7 spheroids also expressed a high α-2,3-SA to α-2,6-SA ratio. These results suggest that the relative levels of specific sialoglycan structures on the cell surface correlate with the ability of cancer cells to form avascular multicellular tumor spheroids and in vivo xenograft tumors.

  5. Akasov R., ZaytsevaZotova D., Burov S., Leko M., Dontenwill M., Chiper M., Vandamme T., Markvicheva E. (2016). Formation of multicellular tumor spheroids induced by cyclic RGD-peptides and use for anticancer drug testing in vitro. Int J Pharm 506 (1-2), 148–157 [+]

    Development of novel anticancer formulations is a priority challenge in biomedicine. However, in vitro models based on monolayer cultures (2D) which are currently used for cytotoxicity tests leave much to be desired. More and more attention is focusing on 3D in vitro systems which can better mimic solid tumors. The aim of the study was to develop a novel one-step highly reproducible technique for multicellular tumor spheroid (MTS) formation using synthetic cyclic RGD-peptides, and to demonstrate availability of the spheroids as 3D in vitro model for antitumor drug testing. Cell self-assembly effect induced by addition of both linear and cyclic RGD-peptides directly to monolayer cultures was studied for 12 cell lines of various origins, including tumor cells (e.i. U-87 MG, MCF-7, M-3, HCT-116) and normal cells, in particular L-929, BNL.CL2, HepG2. Cyclo-RGDfK and its modification with triphenylphosphonium cation (TPP), namely cyclo-RGDfK(TPP) in a range of 10-100μM were found to induce spheroid formation. The obtained spheroids were unimodal with mean sizes in a range of 60-120μm depending on cell line and serum content in culture medium. The spheroids were used as 3D in vitro model, in order to evaluate cytotoxicity effects of antitumor drugs (doxorubicin, curcumin, temozolomide). The developed technique could be proposed as a promising tool for in vitro test of novel antitumor drugs.

  6. Attia M.F., Anton N., Akasov R., Chiper M., Markvicheva E., Vandamme T.F. (2016). Biodistribution and Toxicity of X-Ray Iodinated Contrast Agent in Nano-emulsions in Function of Their Size. Pharm. Res. 33 (3), 603–14 [+]

    This study aimed to investigate the impact of the size of X-ray iodinated contrast agent in nano-emulsions, on their toxicity and fate in vivo.

  7. Demina T.S., Akopova T.A., Vladimirov L.V., Zelenetskii A.N., Markvicheva E.A., Grandfils C.h. (2016). Polylactide-based microspheres prepared using solid-state copolymerized chitosan and d,l-lactide. Mater Sci Eng C Mater Biol Appl 59, 333–8 [+]

    Amphiphilic chitosan-g-poly(d,l-lactide) copolymers have been manufactured via solid-state mechanochemical copolymerization and tailored to design polyester-based microspheres for tissue engineering. A single-step solid-state reactive blending (SSRB) using low-temperature co-extrusion has been used to prepare these copolymers. These materials have been valorized to stabilize microspheres processed by an oil/water emulsion evaporation technique. Introduction of the copolymers either in water or in the oil phase of the emulsion allowed to replace a non-degradable emulsifier typically used for microparticle preparation. To enhance cell adhesion, these copolymers were also tailored to bring amino-saccharide positively charged segments to the microbead surface. Size distribution, surface morphology, and total microparticle yield have been studied and optimized as a function of the copolymer composition.

  8. Akasov R., Borodina T., Zaytseva E., Sumina A., Bukreeva T.V., Burov S., Markvicheva E. (2015). Ultrasonically Assisted Polysaccharide Microcontainers for Delivery of Lipophilic Antitumor Drugs: Preparation and in vitro Evaluation. ACS Appl Mater Interfaces 7 (30), 16581–9 [+]

    High toxicity, poor selectivity, severe side effects are major drawbacks of anticancer drugs. Various drug delivery systems could be proposed to overcome these limitations. The aim of the study was to fabricate polysaccharide microcontainers (MC) loaded with thymoquinone (TQ) by one-step ultrasonication technique and to study their cellular uptake and cytotoxicity in vitro. Two MC fractions with a mean size of 500 nm (MC-0.5) and 2 µM (MC-2) were prepared and characterized. Uptake of the MC by mouse melanoma M-3 cells was evaluated in both 2D (monolayer culture) and 3D (multicellular tumor spheroids) models by confocal microscopy, flow cytometry and fluorimetry. The higher cytotoxicity of the TQ-MC-0.5 sample than that of the TQ-MC-2 fraction was in a good correlation with higher MC-0.5 accumulation in the cells. The MC-0.5 beads were more promising than the MC-2 particles because of a higher cellular uptake in both 2D and 3D models, an enhanced antitumor effect and a lower non-specific toxicity.

  9. Privalova A., Markvicheva E., Sevrin C.h., Drozdova M., Kottgen C., Gilbert B., Ortiz M., Grandfils C.h. (2015). Biodegradable polyester-based microcarriers with modified surface tailored for tissue engineering. J Biomed Mater Res A 103 (3), 939–48 [+]

    Microcarriers have been proposed in tissue engineering, namely for bone, cartilage, skin, vascular, and central nervous system. Although polyester-based microcarriers have been already used for this purpose, their surface properties should be improved to provide better cell growth. The goal of this study was to prepare microbeads based on poly(D,L-lactide) acid, poly(L-lactide) acid, and to study cell behavior (adhesion, spreading, growth, and proliferation) in function of microbead topography and surface chemistry. To improve L-929 fibroblasts adhesion, microbead surface has been modified with three polycations: chitosan, poly(2-dimethylamino ethylmethacrylate) (PDMAEMA), or chitosan-g-oligolactide copolymer (chit-g-OLA). Although modification of the microbead surface with chitosan and PDMAEMA was performed through physical adsorption on the previously prepared microbeads, chit-g-OLA copolymer was introduced directly during microbead processing. This simple approach (1) bypass the use of an emulsifier (polyvinyl alcohol, PVA); (2) avoid surface "contamination" with PVA molecules limiting a control of the surface characteristics. In vitro study of the growth of mouse fibroblasts on the microbeads showed that both surface topography and chemistry affected cell attachment, spreading, and proliferation. Cultivation of L-929 fibroblasts for 7 days resulted in the formation of a 3D cell-scaffold network.

  10. Privalova A.M., Uglanova S.V., Kuznetsova N.R., Klyachko N.L., Golovin Yu.I., Korenkov V.V., Vodovozova E.L., Markvicheva E.A. (2015). Microencapsulated Multicellular Tumor Spheroids as a Tool to Test Novel Anticancer Nanosized Drug Delivery Systems In Vitro. J. Nanosci. Nanotechnol. 15 (7), 4806–4814 [+]

    In the study, MCF-7 human breast adenocarcinoma cells were used to study cytotoxicity of novel anticancer nanosized formulations, such as docetaxel-loaded nanoemulsion and liposomal formulation of a lipophilic methotrexate (MTX) prodrug. In Vitro study of cytotoxicity was carried out in 2 models, namely using 3D In Vitro model based on multicellular tumor spheroids (MTS) and 2D monolayer culture. MTS were generated by tumor cell cultivation within alginate-oligochitosanmicro-capsules. In the case of the monolayer culture, cell viability was found to be 25, 18 and 12% for the samples containing nanoemulsion at concentrations 20, 300 and 1000 nM of docetaxel, respectively, after 48 hs incubation. For MTS these values were higher, namely 33, 23 and 18%, respectively. Cytotoxicity of liposomal MTX prodrug-based formulation with final concentration of 1, 2, 10, 50, 100 and 1000 nM in both models was also studied. MTX liposomal formulation demonstrated lower cytotoxicity on MTS compared to intact MTX. Moreover, MTS were also more resistant to both liposomal formulation and intact MTX than the monolayer culture. Thus, at 1000 nM MTX in the liposomal form, cell viability in MTS was 1.4-fold higher than that in the monolayer culture. MTS could be proposed as a promising tool to test novel anticancer nanosized formulations In Vitro.

  11. Demina T., ZaytsevaZotova D., Yablokov M., Gilman A., Akopova T., Markvicheva E., Zelenetskii A. (2012). DC discharge plasma modification of chitosan/gelatin/PLLA films: Surface properties, chemical structure and cell affinity. Surf. Coating Tech. , [+]

    The work was aimed to study an effect of direct current discharge on chemical structure, surface properties and cell response to the plasma modified composite films. The film samples were prepared by solvent casting from colloidal solution of the ternary blend of chitosan, gelatin and poly(L,L-lactide), PLLA, obtained by solid-state reactive blending (SSRB), in CH2Cl2 and treated with air plasma at pressure of 10–20 Pa and a discharge current of 50 mA for 60 sec. The model film samples casted from the initial components were treated and studied as well. Contact angle of wettability measurements of the films showed that plasma modification led to increase of hydrophilicity and surface energy. Contact angle changes after plasma treatment of chitosan/gelatin/PLLA (CGP) film were more similar to those for the poly(L,L-lactide) film. X-ray photoelectron spectroscopy (XPS) data confirmed that a surface layer of the blend films was enriched with a polyester component. However, study of mouse fibroblasts (L929) attachment and growth on the films showed that the CGP film provided an enhanced cell growth compared to this on the poly(L,L-lactide) film. Plasma modification of the polymer films resulted in a substantial increase in fibroblasts viability on the plasma treated poly(L,L-lactide) films and to a rather strong decrease of cell growth on the plasma treated CGP and chitosan ones. Thus, plasma surface modification could be proposed as a good tool to control cell response on the material surface.

  12. Марквичева Е.А., Дроздова М.Г., Акасов Р.А., ЗайцеваЗотова Д.С. (2011). Биосовместимые материалы в тканевой инженерии, В кн: Клеточные технологии для регенеративной медицины / под ред.: Г.П.Пинаева, М.С.Богдановой, А.М.Кольцовой. – СПБ.: Изд-во Политехн.ун-та. , 103–126 ID:735
  13. Балабашин Д., ЗайцеваЗотова Д., Топорова В., Панина А., Марквичева Е., Свирщевская Е., Алиев Т. (2011). Способы увеличения продукции рекомбинантных антител в клеточных линиях CHO DG44. Современные проблемы науки и образования  (5), [+]

    The cell line CHO DG44 producing recombinant antibodies(Abs) to human tumor necrosis factor-alpha has been obtained. The influence of cell inoculation density and cultivation protocols on the level of Ab biosynthesis has been studied. The highest Ab yields have been observed at the inoculation density 3×106 cells/ml. The alternative method to cells-in-suspension cultivation has been proposed, which is the cell cultivation in calcium alginate hydrogel microgranules or alginate chitosan semipermeable microcapsules. It has been shown that the Ab production level by CHO DG44 cells entrapped into polymer microcapsules exceeds that of the cells-in-suspension cultivation regime.

  14. ZaytsevaZotova D., Balysheva V., Tsoy A., Drozdova M., Akopova T., Vladimirov L., Chevalot I., Marc A., Goergen J.L., Markvicheva E. (2011). Biocompatible Smart Microcapsules Based on Chitosan‐Poly (vinyl alcohol) Copolymers for Cultivation of Animal Cells. Advanced Biomaterials , [+]

    In this study, two novel chitosan-graft-poly(vinyl alcohol) copolymers are synthesized and used as water-soluble at physiological conditions polycations for preparation of smart microcapsules. The microcapsules provide growth and proliferation of eight mammalian cell lines, including hybridoma and tumor cells, at long-term cell cultivation in vitro. The microcapsules are stable in cell culture medium but can be dissolved by changing pH value of the medium (up to 8.0–8.2), thus making possible a simple release of the entrapped cells. Monoclonal antibody production by encapsulated hybridoma cells is demonstrated. Cultivation of tumor cells within the microcapsules allows the formation of 3D multicellular spheroids, which can be proposed as an in vitro model for anticancer drug screening.

  15. ZaytsevaZotova D.S., Udartseva O.O., Andreeva E.R., Bartkowiak A., Bezdetnaya L.N., Guillemin F., Goergen J.L., Markvicheva E.A. (2011). Polyelectrolyte microcapsules with entrapped multicellular tumor spheroids as a novel tool to study the effects of photodynamic therapy. Journal of biomedical materials research. Part B, Applied biomaterials , [+]

    In the current study, semi-permeable alginate-oligochitosan microcapsules for multicellular tumor spheroids (MTS) generation were elaborated and tested, to estimate a response of the microencapsulated MTS (MMTS) to photodynamic therapy (PDT). The microcapsules (mean diameter 600 μm) with entrapped human breast adenocarcinoma MCF-7 cells were obtained using an electrostatic bead generator, and MMTS were generated by in vitro long-term cell cultivation. The formed MMTS were incubated in Chlorin e6 photosensitizer solution and then irradiated using 650-nm laser light. The cell viability was measured by MTT-assay in 24 h after irradiation, and histological analysis was performed. The proposed MTS-based model was found to be more resistant to the PDT than the two-dimensional monolayer cell culture model. Thus, MMTS could be considered as a promising three-dimesional in vitro model to estimate the doses of drugs or parameters for PDT in vitro before carrying out preclinical tests. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2011.

  16. Borodina T., Grigoriev D., Markvicheva E., Mohwald H., Shchukin D. (2011). Vitamin E Microspheres Embedded Within a Biocompatible Film for Planar Delivery. Advanced Engineering Materials 13 (3), B123–B130 [+]

    We demonstrate a new one-batch approach to the fabrication of a biocompatible Ca-alginate film with embedded vitamin E-loaded microspheres that could be used for planar dermal drug delivery. Stable vitamin E microspheres, coated with gum acacia, are produced by ultrasonic treatment of a two-phase liquid system. The Fourier transform infrared spectroscopy indicates an interaction between biopolymer functional groups induced by ultrasonication. Confocal laser fluorescence microscopy, scanning electron microscopy (SEM) and atomic force microscopy (AFM) demonstrate a homogeneous microsphere distribution within the Ca-alginate polymer film. The kinetics of in vitro vitamin E release found for the polymer film with entrapped microspheres was much more sustained (100% in 96 h) compared to the polymer film with vitamin E embedded in the free state (100% in 5 h). The novelty of the proposed research involves the ultrasonic fabrication of loaded microspheres and formation of biodegradable coating directly doped with microspheres.

  17. Tsoy A., ZaytsevaZotova D., Edelweiss E., Bartkowiak A., Goergen J.L., Vodovozova E., Markvicheva E. (2010). Microencapsulated multicellular tumor spheroids as a novel in vitro model for drug screening. Biochemistry (Moscow) Supplement Series B: Biomedical Chemistry 4 (3), 243–250 [+]

    В работе использован метод микрокапсулирования  клеток аденокарциномы молочной железы человека MCF-7 в биосовместимые альгинат-хитозановые микрокапсулы с целью генерирования на основе этих клеток  мультиклеточных опухолевых сфероидов (МОС)  и их дальнейшего исследования в качестве модели in vitro для тестирования противораковых лекарственных средств. На МОС, полученных на основе клеточной линии MCF-7 (аденокарцинома молочной железы человека), было исследовано цитотоксическое действие метотрексата. В зависимости от времени культивирования клеток в микрокапсулах были  получены МОС со средним размером 150, 200 и 300 мкм. После инкубирования МОС с метотрексатом различных концентраций (1, 2, 10, 50 и 100 нМ) в течение 48 часов оценивали количество жизнеспособных клеток. Показано, что МОС гораздо устойчивее к метотрексату, чем монослойная культура. Так, при концентрации метотрексата 100 нМ в МОС размером 300 мкм доля жизнеспособных клеток в 2,5 раза превышала количество живых клеток в монослойной культуре. Таким образом, было показано, что микрокапсулированные  МОС могут более адекватно отражать состояние клеток  в малых солидных опухолях, чем монослойная культура,  и могут в дальнейшем быть предложены в качестве новой модели in vitro для тестирования противораковых лекарств.

  18. Sukhanova T.V., Prudchenko I.A., Efremov E.S., Uglanova S.V., Filatova L.Y.u., Markvicheva E.A., Klyachko N.L. (2010). Biomolecules in colloid nanocontainers for drug delivery: Entrapment and properties of the delta sleep-inducing peptide. Moscow University Chemistry Bulletin 65 (3), 175–179 [+]

    Nanoemulsions of the water in oil type (w/o) composed of nontoxic components, including low concentrations of AOT (5%) and soy-bean lecithin Lipoid S100 (10%) in eucalyptus oil and limonene, were developed and characterized. These nanoemulsions can be used for the incorporation of biomolecules. As shown, AOT based systems possessed the highest solubilization capacity (12% of an aqueous solution); photon-correlation spectroscopy revealed that the size of nanoemulsions of 5% AOT in eucalyptus oil increased linearly with the growth of the water content. The ability of hydrophilic delta sleep-inducing peptide (DSIP), which is a regulatory neuropeptide, to exist in nanoemulsions of the w/o type in the absence and in the presence of additional biopolymers was demonstrated. DSIP entrapment in a nanoemulsion results in its substan tial stabilization (80–90% after 2 months of incubation at 22°C versus 28% in an aqueous solution). The kinetics of peptide release was studied in in vitro model experiments; a substantial slowing-down of peptide release in nanoemulsions in comparison to aqueous solutions was revealed. DSIP-containing nanoemulsion systems, which can be used in medicine and cosmetology, can serve as a basis for the development of novel stable pharmaceutical dosage forms of prolonged action peptide preparations.

  19. Бовин Н.В., Марквичева Е.А., Селина О.Е. (2009). Сорбент для удаления антител из цельной крови и способ его получения. Патент RU 2360707. , ID:400
  20. Markvicheva E.A., Antonov E.N., Popova A.V., Bogorodsky S.E., Likhareva V.V., Feldman B.M., Strukova S.M., Popov V.K., Rumsh L.D. (2009). BIODEGRADABLE POLYMER MICROPARTICLES WITH ENTRAPED HERBAL EXTRACTS: PREPARATION WITH SUPERCRITICAL CARBON DIOXIDE AND USE FOR TISSUE REPAIR. Biomedical Chemistry 55 (4), 479–488 [+]

    Biodegradable microparticles based on poly-D,L-lactide with entrapped mixture of herbal water-soluble extracts of Plantago major and Calendula officinalis were prepared. For preparation of these microparticles the previously developed method based on the usage of supercritical carbon dioxide (SC-CO2) was proposed. Microparticles were obtained by two techniques: 1) by preparing porous polymer monolith containing entrapped mixture of herbal extracts, which was then reduced to fine microparticles (ca. 0.1 mm) by dry ice grinding (called here as “monolithisation technique”) and 2) by spraying of this polymer/extracts mixture through a jet (spray technique). In vitro release kinetic profile of herbal extract mixture was found to depend on the microparticle preparation technique, on the microparticle structure as well as on the initial ratio polymer/extracts (w/w). The microparticles were used for gastric ulcer treatment in a rat model. The extracts released from microparticles were found to accelerate tissue repair.

  21. Selina O.E., Markvicheva E.A., Belov S.Y.u., Vlasova N.N., Balysheva V.I., Churin A.I., Bartkoviak A., Sukhorukov G.B. (2009). BIODEGRADABLE MICROCAPSULES WITH ENTRAPPED DNA FOR DEVELOPMENT OF NEW DNA VACCINES. Russ. J. Bioorgan. Chem. 35 (1), 103–110 [+]

    In this study an universal method for preparation of biodegradable microcapsules for antigen entrapment was proposed and optimized. The multilayer microcapsules were prepared by layer-by-layer adsorption of various polyelectrolytes (such as alginate, poly-L-lysine, κ-carrageenan, chitosan and dextran derivatives). High entrapment efficiency of protein and plasmid DNA (non less than 90%) was shown. To carry out in vivo tests, a set of microcapsules with entrapped pTKShi plasmid encoding the E2 polypeptide of classical swine fever was prepared. It was shown that an injection of these microcapsules into mice induced an immune response. The highest antibody titers of mouse blood sera were got after immunization by microcapsules based on modified dextran/carrageenan and modified chitosan/carrageenan. The proposed method for antigen entrapment in biodegradable microcapsules could be used for development of encapsulated vaccines of a new generation (DNA-vaccines).

  22. Selina O.E., Chinarev A.A., Obukhova P.S., Bartkowiak A., Bovin N.V., Markvicheva E.A. (2009). Alginate-chitosan microspheres for the specific sorption of antibodies. Russ. J. Bioorgan. Chem. 34 (4), 468–474 [+]

    Potentially hemocompatible alginate-chitiosan microparticles and microcapsules coated with a semipermeable membrane with incorporated glycoconjugates were synthesized. The membrane acts as a barrier, which keeps the incorporated glycoconjugate from going outside but permits antibodies to penetrate inside and specifically bind to antigens, high-molecular polysaccharide conjugates. The supports obtained are highly competitive in sorption capacity with Sepharose modified by the same oligosaccharides.

  23. Балышева В.И., Марквичева Е.А., Власова Н.Н., Сухоруков Г.Б., Селина О.Е. (2008). Способ доставки ДНК в макроорганизм для разработки вакцин и соматической генной терапии. Патент RU 2336090C2. , ID:380
  24. Антонов Е.Н., Богородский С.Е., Фельдман Б.М., Марквичева Е.А., Румш Л.Д., Попов В.К. (2008). Получение биодеградируемх микрочастиц с биоактивными компонентами с помощью сверхкритического диоксида углерода. Сверхкритические Флюиды : Теория и Практика 3 (1), 34–42 [+]

    В работе представлены результаты разработки процесса сверхкритического флюидного (СКФ) синтеза биорезорбируемых полимерных микрочастиц, наполненных смесью биоактивных экстрактов подорожника и календулы. Процесс основан на распылении пластифицированной сверхкритическим диоксидом углерода порошковой смеси аморфного алифатического полиэфира (D,L-полилактид) и мелкодисперсного водорастворимого растительного экстракта в ресивер низкого давления через сопло определенного профиля и диаметра. Исследована кинетика выхода растительного экстракта из микрочастиц, полученных при различных давлениях сверхкритического СО2. Показано, что скорость выхода ЭПК в физиологический раствор зависит как от величины рабочего давления диоксида углерода, так и от исходного соотношения полимер/экстракт.

  25. Borodina T.N., Rumsh L.D., Kunizhev S.M., Sukhorukov G.B., Vorozhtsov G.N., Feldman B.M., Rusanova A.V., Vasileva T.V., Strukova S.M., Markvicheva E.A. (2008). Entrapment of herbal extracts into biodegradable microcapsules. Biochemistry (Moscow) Supplemental Series B: Biomedical Chemistry 2 (2), 176–182 [+]

    The microcapsules with entrapped herbal water-soluble extracts of plantain Plantago major and calendula Calendula officinalis L. (PCE) were prepared by layer-by-layer (LbL)-adsorption of carrageenan and  oligochitosan onto CaCO3 microparticles with their subsequent dissolving after the treatment of EDTA. Entrapment of PCE was performed by using adsorption and co-precipitation techniques. The co-precipitation provided better entrapment of PCE into the carbonate matrix compared to adsorption. In vitro release kinetics (AGJ) was studied using artificial gastric juice. Using the model of acetate ulcer in rats it has been demonstrated that PCE released from the microcapsules accelerates gastric tissue repair.

  26. Антонов Е.Н., Баграташвили В.Н., Бочкова С.А., Попов В.К., Попова А.В., Марквичева Е.А., Бородина Т.Н., Румш Л.Д., Фельдман Б.М. (2008). Влияние селективного лазерного спекания на активность Трипсина, инкапсулированного в полилактид. Альманах клинической медицины 17 (2), 30–32 [+]

    Включение биологически активного вещества, трипсина в частицы полилактида осуществлялось путем обработки смеси порошков в сверхкритическом СО2. Полученный композит затем спекался методом поверхностно селективного лазерного спекания. Проведено исследование зависимости активности трипсина от параметров лазерного спекания. Показано, что при энергиях лазерного излучения, используемых для формирования твердых трехмерных структур на базе биорезорбируемых полимеров, трипсин сохраняет более 80% своей первоначальной активности. Таким образом, метод поверхностно селективного лазерного спекания может быть эффективно использован для обеспечения управляемого пролонгированного выхода биологически активных веществ в тканевой инженерии.

  27. Stashevskaya K., Markvicheva E., Strukova S., Prudchenko I., Zubov V., Grandfils C.h. (2007). Thrombin receptor agonist peptide etrapped in poly(d,l,-lactide-co-glycolide) microcapsules:preparation and characterization. J Microencapsulation 24 (2), 129–142 [+]

    Thrombin receptor agonist peptide (TRAP-6) could advantageously replace thrombin in terms of accelerating wound healing being less expensive and more stable. To promote TRAP-6 pharmacological action as a tissue reconstruction stimulator this study investigated its entrapment within poly(D,L)-lactide-co-glycolide (PLGA) microparticles. Due to its low molecular weight and water solubility, TRAP-6 microencapsulated form is expected to be more useful. This paper reports TRAP-6 microencapsulation by a double (w/o/w) emulsion-vaporation technique. TRAP-6 release kinetics were evaluated by both chemical (HPLC) and biological assays in vitro. The results revealed a high level of TRAP-6 sensitivity to physico-chemical events during the microencapsulation. The surface morphology difference between control microparticles (without TRAP-6) and microparticles with entrapped TRAP-6 during in vitro degradation highlighted a particular role of TRAP-6. The results can allow one to optimize the microencapsulation procedure and to encounter a new promising approach to development of biodegradable polymer drug delivery systems for wound healing.

  28. Borodina T., Markvicheva E., Kunizhev S., Moewald H., Sukhorukov G., Kreft O. (2007). Controlled release of DNA from self-degrading microcapsules. Macromol Rap Communications 28, 1894–1899 [+]

    Self-disintegrating microcapsules were prepared by encapsulating a highly active mix of proteases (Pronase®) into biodegradable polyelectrolyte shells. Pronase was captured by micron-sized calcium carbonate particles that were subsequently embedded into onion-like shells of poly(L-arginine) and poly(L-aspartic acid). EDTA treatment was used to extract the calcium carbonate constituents from the resulting core-shell particles. As a consequence, Pronase was released into the capsule interior and started to digest the surrounding polyelectrolyte shell. Lifetimes of such self-disintegrating capsules could be successfully adjusted to seconds, hours or days by varying the amount of encapsulated Pronase. The enzyme-mediated, sustained release of encapsulated DNA is presented as a prospective application in drug delivery.

  29. Балышева В.И., Власова Н.Н., Селина О.Е., Белов С.Ю., Марквичева Е.А., Сухоруков Г.Б. (2007). Микрокапсулирование ДНК как способ доставки ДНК-вакцин. Российский ветеринарный журнал, Мелкие домашние и дикие животные 1, 25–26 [+]

    The report is devoted to the increase of efficacy of a biological material (i.e., DNA) targeted delivery into the macroorganism. The nucleic acid delivery system as developed by the authors comprises DNA incorporation into biologically compatible and biologically degradable microspheres formed by polymers.

  30. Rusanova A., Makarova A., Gorbacheva L., Vasileva T., Markvicheva E., Grandfils C.h., Bespalova Z., Umarova B., Strukova S. (2007). Thrombin receptor agonist peptide as a novel antiulcerogenic factor, Chapter 11. In : Novel Aspects of Biotechnology and Medicine (Eds A.Egorov, E. Zaikov), Nova Science Publishers, Inc.: New-York , 93–101 ID:382
  31. Markvicheva E., Stashevskaya K., Strukova S., Prudchenko I., Rusanova A., Makarova A., Vasilieva T., Bespalova J., Grandfils C.h. (2006). Biodegradable microparticles loaded with thrombin receptor agonist peptide for gastric ulcer treatment in rats. J. Drug Del. Sci. Tech 16 (4), 321–325 [+]



    The aim of the current paper was to elaborate an immobilization method of thrombin receptor agonist peptide (TRAP-6) in biodegradable biocompatible poly(d,l)-lactide-co-glycolide (PLGA) microparticles and to demonstrate the effect of the entrapped peptide for tissue repair, namely for a gastric ulcer treatment in rats. TRAP-6 was entrapped in polymer using w/o/w double emulsion-evaporation technique. The morphology of empty and TRAP-6 loaded microparticles was evaluated by light and scanning electron microscopy (SEM). In vitro release kinetics profile of TRAP-6 from microparticles was studied by HPLC. To investigate gastric mucosal protection effect in vivo, TRAP-6-loaded microparticles were administered in a rat stomach after a previous mucosal injury (a gastric ulcer). Microparticles with entrapped TRAP-6 were found to reduce both an inflammation and proliferation phases of wound healing, and thus accelerated tissue repair in rats.


  32. Marc A., Markvicheva E., Jourdain C., Bezdetnaya L., Merlin J.L., Guillemin F., Zubov V., Goergen J.L. (2005). Spheroids formation by encapsulation of cancer cells to mimic small size tumors, In : Animal Cell Technology meets Genomics. Godia F. and Fussenger M. (Eds.), Springer, Netherlands , 261–263 ID:398
  33. Markvicheva E., Lozinsky V., Plieva F., Kochetkov K., Rumsh L., Zubov V., Kumar R., Parmar V., Belokon Y.u. (2005). Gel-immobilized enzymes as promising biocatalysts for enantioselective hydrolysis in water/organic media. Pure and Applied Chemistry 77 (1), 227–236 [+]

    Chemo-enzymatic methods constitute a promising approach to obtain various biologically active compounds, including enantiomerically pure substances. Entrapment in gels is one of the most convenient methods to stabilize enzymes for their application in water/organic media. Proteases and lipases are widely used for enantioselective transformations of various organic compounds in water-poor media. In this study, chymotrypsin was entrapped into a composite poly(N-vinyl caprolactam)-calcium alginate (PVCL-CaAlg) and covalently attached to poly(vinyl alcohol) (PVA) cryogel beads. Lipase was immobilized by covalently attaching to aldehyde-bearing PVA cryogel beads. The activities of the entrapped biocatalysts were studied. Both entrapped α-chymotrypsin and lipase retained high activity in acetonitrile/water medium (water content 0.5-20 %) and displayed high storage stability for several months. The high operational stability of immobilized α-chymotrypsin and lipase in a cyclic process (up to 912 h in total) was also demonstrated. Gel-immobilized enzymes were successfully used to obtain optically pure L-phenylalanine (ee 98.6 and 83 % in the case of α-chymotrypsin and lipase, respectively) by enantioselective hydrolysis of Schiff's base of amino acid ethyl ester in an acetonitrile/water system.

  34. Markvicheva E., Dugina T., Grandfils C.h., Lange M., Stashevskaya K., Vasilieva T., Rumsh L., Strukova S. (2004). of bioencapsulated proteinases and peptides for wound healing, Charter 12, In: Advanced Biomaterials for Medical Applications, Series II Mathematics, Physics and Chemistry. Thomas D.W. (Ed.), Kluwer Academic Publishers: Dordrecht-Boston-London , 165–176 ID:397
  35. Markvicheva E., Grandfils C.h. (2004). Microcarriers for animal cell culture, In: Fundamentals of Cell Immobilisation Biotechnology, Series Focus on Biotechnology. Willaert R., Nedovich V. (Eds) Kluwer Academic Publishers: Dordrecht-Boston-London 8A, 441–461 ID:396
  36. Markvicheva E., Bezdetnaya L., Bartkowiak A., Marc A., Goergen J.L., Guillemin F., Poncelet D. (2003). Encapsulated multicellular tumor spheroids as a novel in vitro model to study small size tumors. Hemijska industrija 57 (12), 585–588 [+]

    Presently multicellular tumor spheroids (MTS) are being widely used in various aspects of tumor biology, including studies in biology and photodynamic therapy. The cellular organization of spheroids allows the recreation of in vivo small tumors much better than all common two-dimensional in vitro models. The cell encapsulation method could be proposed as a novel technique to quickly and easily prepare a large number of spheroids with narrow size distribution within a desirable diameter range. Moreover, the proposed technique for spheroid generation using encapsulated growing tumor cells could provide entirely new avenues to develop a novel spheroid co-culture model (for instance, the in vitro co-cultvation of tumor cells and monocytes, or epithelial cells, or fibroblasts etc). The current research was aimed at developing a simple and reliable method to encapsulate tumor cells and to cultivate them in vitro. In order to generate spheroids, MCF-7 cells were encapsulated and cultivated in 200 ml T-flasks in a 5% CO2 atmosphere at 37°C for 4-5 weeks. The cell proliferation was easily observed using a light microscope. The cells grew in aggregates increasing in size with time. The cell growth resulted in the formation of large cell clusters (spheroids) which filled the whole microcapsule volume in 4-5 weeks.

  37. Vilchez C., Garbayo I., Markvicheva E., Galvan F., Leon R. (2001). Studies on the suitability of alginate-entrapped Chlamydomonas reinhardtii cells for sustaining nitrate consumption process. Bioresource Technology 78, 55–61 [+]

    Some aspects of the suitability of alginate beads entrapping Chlamydomonas reinhardtii cells for nitrate consumption from nitrate-containing waters were studied and discussed. Among 14 different metal cations tested as gel bead stabilizing agents, only calcium and barium formed beads showing nitrate-consuming activity. Pure calcium alginate cell entrapment resulted in the most suitable method for active cell immobilization compared to alginate-composite-gel beads based on poly-vinylcaprolactam (PVCL) and poly-vinylpyrrolidone (PVP). To perform a continuous nitrate consumption process, calcium alginate-entrapped cells were first grown in a 2.5 l airlift-loop reactor. A cell loading of about 150 μg Chl. g−1 gel was achieved. Afterwards, five days nitrate consumption processes were performed and three different dilution rates were applied: (i) D<μ; (ii) D=μ; (iii) D>μ, where μ is the specific growth rate (h−1). The maximum consumption rates calculated for each dilution rate were: (i) 3.8, (ii) 6.4 and (iii) 7.2 mg nitrate mg−1 Chl. h−1. For low dilution rates (D<μ) some nitrite (<300 μM) was excreted into the culture medium. However, this concentration of nitrite was not high enough to inhibit nitrate consumption.