Colloid and Polymer Science

, Volume 287, Issue 7, pp 759–765 | Cite as

Self-assembling chitosan/poly-γ-glutamic acid nanoparticles for targeted drug delivery

  • Zsolt Keresztessy
  • Magdolna Bodnár
  • Elizabeth Ber
  • István Hajdu
  • Min Zhang
  • John F. Hartmann
  • Tamara Minko
  • János BorbélyEmail author
Original Contribution


For the purpose of targeted drug delivery, composite biodegradable nanoparticles were prepared from chitosan and the poly-γ-glutamic acid via an ionotropic gelation process. These stable self-assembled nanoparticles were characterized by dynamic light scattering, transmission electron microscopy, and atomic force microscopy, which demonstrated that the nanosystem consists of spherical particles with a smooth surface both in aqueous environment and in dried state. Toxicity measurements showed that the composition is nontoxic when tested either on cell cultures or in animal feeding experiments. To evaluate the potential of the nanosystem for intracellular drug delivery, the nanoparticles were fluorescently labeled and folic acid was attached as a cancer cell-specific targeting moiety. The ability of the particles to be internalized was tested using confocal microscopic imaging on cultured A2780/AD ovarian cancer cells, which overexpress folate receptors. The quantitative data obtained by digital processing of the intensity of green color of each pixel in the pictures inside the cell boundaries and total intensity of fluorescence inside the cells showed that “targeted” particles internalized into the cells significantly faster and the total accumulation of these particles was substantially higher in the cancer cells when compared with “nontargeted” particles, which may facilitate effective and specific cytoplasmic delivery of anticancer agents loaded into such nanoparticles.


Chitosan Poly-γ-glutamic acid PGA Nanoparticles Cytotoxicity In vivo toxicity Cellular uptake 



This research was supported by the grant of Regional University Knowledge Center of Debrecen (Genomnanotech Debrecen, contract number RET-06/432/2004) and a grant from the New Jersey Commission on Science and Technology.


  1. 1.
    McNeil SE (2005) Nanotechnology for the biologists. J Leukocyte Biol 78:585–594CrossRefGoogle Scholar
  2. 2.
    Majoros IJ, Thomas T, Baker JR Jr (2006) Molecular engineering in nanotechnology: engineered drug delivery. In: Reith M, Schommers W (eds) Handbook of theoretical and computational nanotechnology, vol 6. American Scientific, Stevenson Ranch, CA, pp 673–717Google Scholar
  3. 3.
    Vicent MJ, Duncan R (2006) Polymer conjugates: nanosized medicines for treating cancer. Trends Biotech 24:39–47CrossRefGoogle Scholar
  4. 4.
    Duncan R (2003) The dawning era of polymer therapeutics. Nat Rev Drug Discover 2:347–360CrossRefGoogle Scholar
  5. 5.
    Nobs L, Buchegger F, Gurny R, Allémann E (2006) Biodegradable nanoparticles for direct and two-step tumor immunotargeting. Bioconj Chem 17:139–145CrossRefGoogle Scholar
  6. 6.
    Torchilin VP (2006) Multifunctional nanocarriers. Adv Drug Del Rev 58:1532–1555CrossRefGoogle Scholar
  7. 7.
    Ulbrich K, Subr V (2004) Polymeric anticancer drugs with pH-controlled activation. Adv Drug Deliv Rev 56:123–1050CrossRefGoogle Scholar
  8. 8.
    Sawant RM, Hurley JP, Salmaso S, Kale A, Tolcheva E, Levchenko TS, Torchilin VP (2006) “SMART” drug delivery systems: double-targeted pH-responsive pharmaceutical nanocarriers. Bioconj Chem 17:943–949CrossRefGoogle Scholar
  9. 9.
    Grodzinsky P, Silver M, Molnar LM (2006) Nanotechnology for cancer diagnostics: promises and challenges. Expert Rev Mol Diagn 6:307–318CrossRefGoogle Scholar
  10. 10.
    Khan MK, Nigavekar SS, Minc LD, Kariapper MST, Nair BM, Lesniak WG, Balogh LP (2005) In vivo biodistribution of dendrimers and dendrimer nanocomposites—implications for cancer imaging and therapy. Technol Cancer Res Threat 4:603–613Google Scholar
  11. 11.
    Hajdu I, Bodnar M, Filipcsei G, Hartmann JF, Daroczi L, Zrinyi M, Borbely J (2008) Nanoparticles prepared by self-assembly of chitosan and poly-γ-glutamic acid. Coll Polym Sci 286:343–350CrossRefGoogle Scholar
  12. 12.
    Berger J, Reist M, Mayer JM, Felt O, Gurny R (2004) Structure and interactions in chitosan hydrogels formed by complexation or aggregation for biomedical applications. Eur J Pharm Biopharm 57:35–52CrossRefGoogle Scholar
  13. 13.
    Hsieh C-Y, Tsai S-P, Wang D-M, Chang Y-N, Hsieh H-J (2005) Preparation of γ-PGA/chitosan composite tissue engineering matrices. Biomaterials 26:5617–5623CrossRefGoogle Scholar
  14. 14.
    Lin Y-H, Chung CK, Chen C-T, Liang H-F, Chen S-C, Sung H-W (2005) Preparation of nanoparticles composed of chitosan/poly-g-glutamic acid and evaluation of their permeability through caco-2 cells. Biomacromolecules 6:1104–1112CrossRefGoogle Scholar
  15. 15.
    Lin Y-H, Mi F-L, Chen C-T, Chang W-C, Peng S-F, Liang H-F, Sung H-W (2007) Preparation and characterization of nanoparticles shelled with chitosan for oral insulin delivery. Biomacromolecules 8:146–152CrossRefGoogle Scholar
  16. 16.
    Lin WC, Yu DG, Yang MC (2006) Blood compatibility of novel poly(γ-glutamic acid)/polyvinyl alcohol hydrogels. Colloid Surface B 47:43–49CrossRefGoogle Scholar
  17. 17.
    Krecz A, Pocsi I, Borbely J (2001) Preparation and chemical modification of poly-gamma-L-glutamic acid. Folia Microbiol 46:183–186CrossRefGoogle Scholar
  18. 18.
    Borbely M, Nagasaki Y, Borbely J, Fan K, Bhogle A, Sevoian M (1994) Biosynthesis and chemical modification of poly(gamma-glutamic acid). Polym Bull 32:127–132CrossRefGoogle Scholar
  19. 19.
    Bodnár M, Hartmann JF, Borbély J (2005) Preparation and characterization of chitosan-based nanoparticles. Biomacromolecules 6:2521–2527CrossRefGoogle Scholar
  20. 20.
    Bodnár M, Hartmann JF, Borbély J (2006) Synthesis and study of cross-linked chitosan-N-poly(ethylene glycol) nanoparticles. Biomacromolecules 7:3030–3036CrossRefGoogle Scholar
  21. 21.
    Minko T, Kopeckova P, Pozharov V, Kopecek J (1998) HPMA copolymer bound adriamycin overcomes MDR1 gene encoded resistance in a human ovarian carcinoma cell line. J Controlled Release 54:223–233CrossRefGoogle Scholar
  22. 22.
    Minko T, Kopeckova P, Kopecek J (1999) Chronic exposure to HPMA copolymer-bound adriamycin does not induce multidrug resistance in a human ovarian carcinoma cell line. J Controlled Release 59:133–148CrossRefGoogle Scholar
  23. 23.
    Pakunlu RI, Cook TJ, Minko T (2003) Simultaneous modulation of multidrug resistance and antiapoptotic cellular defense by MDR1 and BCL-2 targeted antisense oligonucleotides enhances the anticancer efficacy of doxorubicin. Pharm Res 20:351–359CrossRefGoogle Scholar
  24. 24.
    Dharap SS, Chandna P, Wang Y, Khandare JJ, Qiu B, Stein S, Minko T (2006) Molecular targeting of BCL2 and BCLXL proteins by synthetic BH3 peptide enhances the efficacy of chemotherapy. J Pharmacol Exp Ther 316:992–998CrossRefGoogle Scholar
  25. 25.
    Khandare JJ, Chandna P, Wang Y, Pozharov VP, Minko T (2006) Novel polymeric prodrug with multivalent components for cancer therapy. J Pharmacol Exp Ther 317:929–937CrossRefGoogle Scholar
  26. 26.
    Khandare JJ, Jayant S, Singh S, Chandna P, Wang Y, Vorsa N, Minko T (2006) Dendrimer versus linear conjugate: influence of polymeric architecture on the delivery and anticancer effect of Paclitaxel. Bioconjug Chem 17:1464–1472CrossRefGoogle Scholar
  27. 27.
    Dharap SS, Wang Y, Chandna P, Khandare JJ, Qiu B, Gunaseelan S, Sinko PJ, Stein S, Farmanfarmaian A, Minko T (2005) Tumor-specific targeting of an anticancer drug delivery system by LHRH peptide. Proc Natl Acad Sci USA 102:12962–12967CrossRefGoogle Scholar
  28. 28.
    Pakunlu RI, Wang Y, Saad M, Khandare JJ, Starovoytov V, Minko T (2006) In vitro and in vivo intracellular liposomal delivery of antisense oligonucleotides and anticancer drug. J Controlled Release 114:153–162CrossRefGoogle Scholar
  29. 29.
    Hong S, Leroueil PR, Majoros IJ, Orr BG, Baker JR Jr, Banasak Holl MM (2007) The binding avidity of a nanoparticle-based multivalent targeted drug delivery platform. Chem Biol 14:P107–P115CrossRefGoogle Scholar
  30. 30.
    Kang HS, Park SH, Lee YG, Son TI (2007) Polyelectrolyte complex hydrogel composed of chitosan and poly(γ-glutamic acid) for biological application: preparation, physical properties, and cytocompatibility. J Appl Polym Sci 103(1):386–394CrossRefGoogle Scholar
  31. 31.
    Bhumkar DR, Pokharkar VB (2006) Studies on effect of pH on cross-linking of chitosan with sodium tripolyphosphate: a technical note. AAPS PharmSciTech 7(2):138–143CrossRefGoogle Scholar
  32. 32.
    Novak L, Banyai I, Fleischer-Radu JE, Borbely J (2007) Direct amidation of poly(γ-glutamic acid) with benzylamine in dimethyl sulfoxide. Biomacromolecules 8:1624–1632CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Zsolt Keresztessy
    • 1
  • Magdolna Bodnár
    • 1
  • Elizabeth Ber
    • 2
  • István Hajdu
    • 1
  • Min Zhang
    • 2
  • John F. Hartmann
    • 3
  • Tamara Minko
    • 2
  • János Borbély
    • 1
    • 4
    Email author
  1. 1.Department of Colloid and Environmental ChemistryUniversity of DebrecenDebrecenHungary
  2. 2.Department of Pharmaceutics, RutgersThe State University of New JerseyPiscatawayUSA
  3. 3.ElizaNor Polymer LLCPrinceton JunctionUSA
  4. 4.BBS Nanotechnology Ltd.Debrecen 16Hungary

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