Electroinduced and Adhesive Properties

Acting Head of Department

Assoc. Prof. Natalia Krasteva, PhD

  • E-mail: natalybio21.bas.bg
  • Phone: (359 2) 9792622, 9793242
  • Fax: (359 2) 8723787
  • Address: “Acad. G. Bonchev” Str., Block 21, Room 311
    Sofia 1113, Bulgaria

Staff

Laboratory “Biomimetic Membranes”

  • Prof. Galya Staneva, PhD, g_stanevayahoo.com
  • Senior Assist. Prof. Rusina Hazarosova, PhD, r_hazarosovaabv.bg
  • Assist. Prof. Vesela Yordanova, v.v.yordanova.bulabv.bg
  • Ralitsa Veleva, biologist
  • Prof. George Altunkov, DSc., Associated member, altunkovabv.bg
  • Main research topics

    Research Topics

    Electroporation, electrofusion and electrotransformatiom. Electrotransfer of macromolecules through lipid membranes. Electrical properties of biological membranes. Electrochemotherapy of tumour cells.

    Cell adhesion and reorganization of cytoskeleton after electrotreatment. Influence of low voltage, low frequency electric field on the adhesive properties of nanostructures (tissue engineering).

    Experimental Methods

    Electroporation and electrotransformation methods, patch clamp, fluorescent method for measuring of transmembrane potentials, radioisotope methods, determination of particle electrophoretic mobility, DNA fragmentation, MMT test, FACS analysis.

    Cell Adhesion:

    1. Colorimetric assays for cell adhesion, growth and viability.
    2. Immunofluorescence microscopy
      • Determining the extend of cell spreading and the strength of cell adhesion
      • Visualization of cell cytoskeleton
      • Dynamic of cell receptors upon cell adhesion
    3. Enzyme immunoassay
    4. Western Blotting
    5. Immunoprecipitation
    6. Zymography

    Research projects

    Current projects

    1. Role of substrate elasticity in the ostheogenic differentiation of mesenchyme stem cells
      Bulgarian National Science Fund, № DFNI-B01/18
      Coordinator: Assoc. Prof. Natalia Krasteva
    2. Studying the adsorption of plasma proteins and cell adhesion onto conductive 2D and 3D dimensional polymer composite surfaces, with application to regenerative medicine
      Bulgarian National Science Fund, № DNTS/Germany/01/0003
      Coordinator: Assoc. Prof. Rumiana Tzoneva

    Past projects

    1. Vesicles and Membranes
      French-Bulgarian Laboratory, 1997-2002.
    2. Biophysical Mechanism of the effect of electromagnetic field on biological and model membrane
      EU project-COST 244, 1994-1996.
    3. Mechanism of electrical permeabilisation
      DFG, 436-100, 1993-1998 (2 projects).
    4. Mechanism of electroporative DNA and oligo transport through cell membranes and lipid bilayers
      DFG 436/113, 1999-2001.
    5. Transmembrane transport of anticancer drugs and DNA by electrical pulses
      French-Bulgarian project, (CNRS-BAS), 2003-2005, Toulouse, France.
    6. Electrotransfer of anticancer drugs and DNA
      French-Bulgarian project, (CNRS-BAS), 2006-2007, Toulouse, France.
    7. Electrotransfer of macromolecules
      Bulgarian National Science Fund № K-31, 1991-1994.
    8. Effect of electrical pulses at the interaction and transfer of poly ions through biological membranes
      Bulgarian National Science Fund, № K-411, 1994-1997.
    9. Family of apparatus for electrochemotherapy of malignant tumors: laboratory and clinical studies – prototypes and construction
      Bulgarian National Science Fund, № N 5011, 1998-1999.
    10. Effect of electric field on macromolecules in biological and model membranes
      Bulgarian National Science Fund № K-707, 1997-2002.
    11. Transmembrane transfer of drugs and DNA by electrical pulses
      Bulgarian National Science Fund № K-1303, 2003-2006.
    12. Exploitation and characterization of nanostrctured biomimetic surfaces  as substrates for cultivating of osteoblasts
      Bulgarian National Science Fund № DAAD-15, 2007.
    13. Study of the interaction cell-nanostructured biomaterial (nanofiber nonwoven textile)
      Bulgarian National Science Fund № НТ-4, 2006-2008.
    14. Treatment of cancer cells and solid tumors by suitable choice of electrical pulses
      Bulgarian National Science Fund № D0 02/178
      Coordinator: Prof. Iana Tsoneva
    15. Study of the influence of an applied electric field on the adsorption of adhesive proteins and  endotelisation of nanostructure biomaterials for needs of tissue engineering and regenerative medicine
      Bulgarian National Science Fund and DAAD, № Д0 02/326
      Coordinator: Assoc. Prof. Rumiana Tzoneva

    International collaboration

    • Prof. E. Peycheva, M. Georgieva, Ph. D., National Centre of Oncology, Sofia.
    • Prof. M. P. Rols, CNRS, Toulouse, France.
    • Prof. E. Neumann, Chemical Faculty, University of Bielefeld, Germany.
    • Prof. M. Berger, German Cancer Research Center, Heidelberg, Germany.
    • Prof. G. Altankov, Institute for Bioengineering of  Catalonia, Spain.
    • Prof. T. Groth, Martin-Luter-Univirsitetat, Halle-Wintenberg, Germany.
    • Dr Barbara Seifert, Helmholz-Zentrum Geesthacht, Centre for Material and Coastel, Teltow, Germany

    Selected publications

    1. Tsoneva, I., Iordanov, I., Berger, A., Tomov, T., Nikolova, B., Mudrov N., Berger, M. (2010) Electrodelivery of drugs into cancer cells in the presence into cancer cells in the presence of poloxamer 188, Journal of Biomedicine and Biotechnology, (electronic): 1110-7251.
    2. Tzoneva R, Weckwerth C, Seifert B, Behl M, Heuchel M, Tsoneva I, Lendlein A. (2010) In vitro evaluation of elastic multiblock co-polymers as a scaffold material for reconstruction off blood vessels, J Biomater Sci Polym Ed., 2010, Nov 11.
    3. Tzoneva R., Seifert B., Albrecht W., Richau K., Groth T., Lendlein A. (2008) Hemocompatibility of poly(ether imide) membranes functionalized with carboxylic groups, J Mater Sci Mater Med, 19, 10, 3203-3210.
    4. Tomov, T., Nikolova, B., (2010) Electrically induced colloid-osmotic lysis of erythrocytes in the presenceof macromolecular polymers, Compt. Rend. Acad. Bulg. Sci., v.63, N5, 709-712.
    5. Pehlivanova V., Tsoneva I., Tzoneva R. (2011) Influence of electroporation on cell adhesion, growth and viability of cancer cells and fibroblasts, Compt. Rend. Acad. Bulg. Sci., 4, 64.
    6. Krasteva V., Pehlivanova V., Seifert B, Lützow K., Tsoneva I., Richau K., Lendlein A., Tzoneva R. (2011) Influence of AC electric fields on the adsorption of plasma proteins onto nanofiber biomaterials, Compt. Rend. Acad. Bulg. Sci., 4, 64,.
    7. Peycheva Е, Daskalov I, Tsoneva I (2007) Electrochemotherapy of Mycosis fyngoides by interferon-alfa, Bioelectrichemistry 70, 283-286.
    8. Tsoneva I, Nikolova B, Georgieva M, Guenova M, Tomov T, Rols M-P, Berger MR (2005) Induction of apoptosis by electrotransfer of positively charged proteins as Cytochrom C and Histone H1 into cells. Biochem. Biophys. Acta 1721, 55-64.
    9. Berger M, Tsoneva I, Konstantinov S, Eibl H (2003) Induction of apoptosis by EPC3 is associated with changes in signal molecule expression and location. Ann NY Acad. Sci. 1010, 307-311.
    10. Dimitrov V, Stoimenova M, Tsoneva I (2002) Electrically induced concentration fluctuations in E. coli suspensions. Colloid and Surfaces, A Physicochem and Eng. Aspects 209, 201-205.
    11. Hristova N, Angelova M, Tsoneva I (2002) An experimental approach for direct observation of the interaction of polyanions with spingosine-containing giant vesicles. Bioelectrochem. 58, 65-73.
    12. Nikolova B, Dimitrov V, Tsoneva I (2000) Mechanism of DNA transfer to E. coli by weak alternating electric. J. Balkan Ecol. 3, 48-56.
    13. Tomov T, Tsoneva I (2000) Are the stainless steel electrodes inert? Bioelectrochem. 51, 207-209.
    14. Angelova M, Tsoneva I. (1999) Interaction of DNA with giant liposomes. Chem. Phys. Lipids. 101, 123-137.
    15. Angelova M, Hristova N, Tsoneva I. (1999) DNA -induced endocytosis upon local microinjection to giant unilamellar cationic vesicles. Eur. Biophys. J. 28, 142-150.
    16. Phylipova E, Suss M, Tomov T, Tsoneva I (1997) Effect of external electric fields on the activity of membrane bound enzymes at transmembrane potential equal to zero. Compt. Rend. Bulg. Acad. Sci. 50, 105-108.
    17. Hristova N, Tsoneva I, Neumann E (1997) Sphingosine-mediated electroporative DNA transfer through lipid bilayer. FEBS Lett. 415, 81- 86.
    18. Neumann E, Kakorin S, Tsoneva I, Nikolova B, Tomov T (1996) Calcium mediated DNA adsorption to yeast cells and kinetics of cell transformation by electroporation. Biophys. J. 71, 868- 877.
    19. Spassova M, Petrov AG, Tsoneva I, Petkova JI, Neumann E (1994) Dip patch clamp currents suggest electrodifusive transport of the polyelectrolyte DNA through lipid bilayers. Biophys. Chem. 52, 267-274.
    20. Ganeva V, Tsoneva I (1993) Effect of n-alcohols on the electrotransformation and permeability of Saccharomyces cerevisiae. Appl. Microbiol. Biothechnol. 38, 792- 798.
    21. Tsoneva I, Tomov T, Panova I, Doncheva J (1992) Electrofusion in the presence of divalent cations. Bioelectochem. Bioenerg. 27, 37-40.
    22. Tomov T, Tsoneva I (1992) Influence of electrical pulse on the transmembrane potential in yeast. Bioelectrochem. Bioenerg. 27, 33 -36.
    23. Tsoneva I, Panova I, Tomov T, Strahilov D (1990) Effective production of hybridomas secreting monoclonal antibodies against Hc antigen of Salmonella by electrofusion. Bioelectrochem. Bioenerg. 24, 41-49.
    24. Tzoneva R, Faucheux N, Groth Т (2007) Wettability of substrata controls cell-substrate and cell-cell adhesions”, Biochim Biophys Acta 1770(11), 1538-1547.
    25. Faucheux N, Tzoneva R, Nagel MD, Groth T (2006) The dependence of fibrillar adhesions in human fibroblasts on substratum chemistry”, Biomaterials, 2006, 27, 2, 234-245.
    26. Vladkova TG, Keranov IL, Dineff PD, Youroukov SY, Avramova IA, Krasteva N, Altankov GP (2005) Plasma based Ar+ beam assisted poly(dimethylsiloxane) surface modification Nuclear Instruments and Methods in Physics Research, Section B: Beam Interactions with Materials and Atoms 236 (1-4), pp. 552-562.
    27. Krasteva N, Seifert B, Hopp M, Malsch G, Albrecht W, Altankov G, Groth Th (2005) Membranes for biohybrid liver support: The behaviour of C3A hepatoblastoma cells is dependent on the composition of acrylonitrile copolymers Journal of Biomaterials Science, Polymer Edition 16 (1), pp. 1-22.
    28. Krasteva N, Seifert B, Albrecht W, Weigel T, Schossig M, Altankov G, Groth Th (2004) Influence of polymer membrane porosity on C3A hepatoblastoma cell adhesive interaction and function. Biomaterials 25 (13), pp. 2467-2476.
    29. Tzoneva R, Groth T, Altankov G, Paul D (2002) Remodelling of fibrinogen by endothelial cells in dependence on fibronectin matrix assembly. Effect of substratum wettability, Journal of Materials Science: Materials in Medicine 13, 1235-1244.
    30. Groth T, Seifert B, Malsch G, Albrecht W, Paul D, Kostadinova A, Krasteva N, Altankov G (2002) Interaction of human skin fibroblasts with moderate wettable polyacrylonitrile-copolymer membranes. Journal of Biomedical Materials Research 61 (2), pp. 290-300.
    31. Krasteva N, Harms U, Albrecht W, Seifert B, Hopp M, Altankov G, Groth T (2002) Membranes for biohybrid liver support systems – Investigations on hepatocyte attachment, morphology and growth. Biomaterials 23 (12), pp. 2467-2478.
    32. Vladkova T, Krasteva N, Kostadinova A, Altankov G (1999) Preparation of PEG-coated surfaces and a study for their interaction with living cells. Journal of Biomaterials Science, Polymer Edition 10 (6), pp. 609-620.
    33. Groth T, Altankov G, Kostadinova A, Krasteva N, Albrecht W, Paul D (1999) Altered vitronectin receptor (alpha-v integrin) function in fibroblasts adhering on hydrophobic glass. Journal of Biomedical Materials Research 44 (3), pp.341-351.
    34. Nikolova, B., Tsoneva, I., Peycheva, E. (2011) Treatment of Melanoma by electroporation of bacillus Calmette-Guerin. Biotechnol. & Biotechnol. Eq., 2011, 2, 25, 3, 2522-24.

    Resources

    Video “Electrochemical therapy: An easy, highly efficient and safe method for treating skin tumours”

    The video has been created in the frames of the project Ref. No. DO 02/178 “Treatment of cancer cells and solid tumors by suitable choice of electrical pulses” with the National Science Fund of Bulgaria, with coordinator Prof. Iana Tsoneva, DSc.
    For download (in Bulgarian):

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