Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Nonviral Transfer Technology
  • Published:

Sustained delivery and expression of DNA encapsulated in polymeric nanoparticles

Gene Therapy volume 7, pages 1896–1905 (2000)Cite this article

Abstract

Sustained release polymeric gene delivery systems offer increased resistance to nuclease degradation, increased amounts of plasmid DNA (pDNA) uptake, and the possibility of control in dosing and sustained duration of pDNA administration. Furthermore, such a system lacks the inherent problems associated with viral vectors. Biodegradable and biocompatible poly(DL-lactide-co-glycolide) polymer was used to enacapsulate pDNA (alkaline phosphatase, AP, a reporter gene) in submicron size particles. Gene expression mediated by the nanoparticles (NP) was evaluated in vitro and in vivo in comparison to cationic-liposome delivery. Nano size range (600 nm) pDNA-loaded in poly(DL-lactide-co-glycolide) polymer particles with high encapsulation efficiency (70%) were formulated, exhibiting sustained release of pDNA of over a month. The entrapped plasmid maintained its structural and functional integrity. In vitro transfection by pDNA-NP resulted in significantly higher expression levels in comparison to naked pDNA. Furthermore, AP levels increased when the transfection time was extended, indicating sustained activity of pDNA. However, gene expression was significantly lower in comparison with standard liposomal transfection. Seven days after i.m. injections in rats, naked pDNA and pDNA-NP were found to be significantly more potent (1–2 orders of magnitude) than liposomal pDNA. Plasmid DNA-NP treatment exhibited increased AP expression after 7 and 28 days indicating sustained activity of the NP.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  1. Mahato RI, Smith LC, Rolland A . Pharmaceutical perspectives of nonviral gene therapy Adv Genet 1999 41: 95–156

    Article  CAS  Google Scholar 

  2. Ledley FD . Non-viral gene therapy Curr Opin Biotechnol 1994 5: 626–636

    Article  CAS  Google Scholar 

  3. Ledley FD . Nonviral gene therapy: the promise of genes as pharmaceutical products Hum Gene Ther 1995 6: 1129–1144

    Article  CAS  Google Scholar 

  4. Rolland AP . From genes to gene medicines: recent advances in nonviral gene delivery Crit Rev Ther Drug Carrier Syst 1998 15: 143–198

    Article  CAS  Google Scholar 

  5. Roy K, Mao H-Q, Huang S-K, Leong KW . Oral gene delivery with chitosan-DNA nanoparticles generates immunologic protection in a murine model of peanut allergy Nature Med 1999 5: 387–391

    Article  CAS  Google Scholar 

  6. Chonn A, Cullis PR . Recent advances in liposome technologies and their applications for systemic gene delivery Adv Drug Deliv Rev 1998 30: 73–83

    Article  CAS  Google Scholar 

  7. Bonadio J, Goldstein SA, Levy RJ . Gene therapy for tissue repair and regeneration Adv Drug Deliv Rev 1998 33: 53–69

    Article  CAS  Google Scholar 

  8. Labhasetwar V et al. A DNA controlled-release coating for gene transfer: transfection in skeletal and cardiac muscle J Pharm Sci 1998 87: 1347–1350

    Article  CAS  Google Scholar 

  9. Shea LD, Smiley E, Bonadio J, Mooney DJ . DNA delivery from polymer matrices for tissue engineering Nat Biotechnol 1999 17: 551–554

    Article  CAS  Google Scholar 

  10. Smith T, Zhang Y, Niven R . Toward development of a non-viral gene therapeutic Adv Drug Deliv Rev 1997 26: 135–150

    Article  CAS  Google Scholar 

  11. Mathiowitz E et al. Biologically erodable microspheres as potential oral drug delivery systems Nature 1997 386: 410–414

    Article  CAS  Google Scholar 

  12. Hedley ML, Curley J, Urban R . Microspheres containing plasmid-encoding antigenes elicit cytotoxic T-cell responses Nature Med 1998 4: 365–368

    Article  CAS  Google Scholar 

  13. Leong KW et al. DNA-polycation nanospheres as non-viral gene delivery vehicles J Control Release 1998 53: 183–193

    Article  CAS  Google Scholar 

  14. Labhasetwar V, Bonadio J, Goldstein AS, Levy RJ . Gene transfection using biodegradable nanospheres: results in tissue culture and a rat osteotomy model Colloids and Surfaces B: Biointerfaces 1999 16: 281–290

    Article  CAS  Google Scholar 

  15. Ledley FD . Pharmaceutical approach to somatic gene therapy Pharm Res 1996 13: 1595–1614

    Article  CAS  Google Scholar 

  16. Jong YS et al. Controlled release of plasmid DNA J Control Release 1997 47: 123–134

    Article  CAS  Google Scholar 

  17. Luo D, Woodrow Mumford K, Belcheva N, Saltzman WM . Controlled DNA delivery systems Pharm Res 1999 16: 1300–1308

    Article  CAS  Google Scholar 

  18. Ando S, Putnam D, Pack DW, Langer R . PLGA microspheres containing plamid DNA: preservation of supercoiled DNA via cryopreparation and carbohydrate stabilization J Pharm Sci 1999 88: 126–130

    Article  CAS  Google Scholar 

  19. Anderson JM, Shive MS . Biodegradation and biocompatibility of PLA and PLGA microspheres Adv Drug Deliv Rev 1997 28: 5–24

    Article  CAS  Google Scholar 

  20. Hsu YY, Hao T, Hedley ML . Comparison of process parameters for microencapsulation of plasmid DNA in poly(D,L-lactic-co-glycolic) acid microspheres J Drug Target 1999 7: 313–323

    Article  CAS  Google Scholar 

  21. Capan Y et al. Influence of formulation parameters on the characteristics of poly(D,L-lactide-co-glycolide) microspheres containing poly(L-lysine) complexed plasmid DNA J Control Release 1999 60: 279–286

    Article  CAS  Google Scholar 

  22. Wang D, Robinson DR, Kwon GS, Samuel J . Encapsulation of plasmid DNA in biodegradable poly(D,L-lactic-coglycolic acid) microspheres as a novel approach for immunogen delivery J Control Release 1999 57: 9–18

    Article  CAS  Google Scholar 

  23. Cherng JY et al. Effect of DNA topology on the tranfection efficiency of poly((2-dimethylamino)ethyl methacrylate)-plasmid complexes J Control Release 1999 60: 343–353

    Article  CAS  Google Scholar 

  24. Adami CA et al. Stability of peptide-condensed plasmid DNA formulations J Pharm Sci 1998 87: 678–683

    Article  CAS  Google Scholar 

  25. Capan Y et al. Preparation and characterization of poly(D,L-lactide-co-glycolide) microspheres for controlled release of poly(L-lysine) complexed plasmid DNA Pharm Res 1999 16: 509–513

    Article  CAS  Google Scholar 

  26. Wolfert MA, Seymour LW . Atomic force microscopic analysis of the influence of the molecular weight of poly(L)lysine on the size of polyelectrolyte complexes formed with DNA Gene Therapy 1996 3: 269–273

    CAS  Google Scholar 

  27. Weintraub H, Cheng PF, Conrad K . Expression of transfected DNA depends on DNA topology Cell 1986 46: 115–122

    Article  CAS  Google Scholar 

  28. Langer R . New methods of drug delivery Science 1990 249: 1527–1533

    Article  CAS  Google Scholar 

  29. Cohen S et al. Controlled delivery systems for proteins based on poly (lactic/glycolic) microsphers Pharm Res 1991 8: 713–720

    Article  CAS  Google Scholar 

  30. Crystal RG . Transfer of genes to humans: early lessons and obstacles to success Science 1995 270: 404–410

    Article  CAS  Google Scholar 

  31. Colin WP, Leonard WS . Key issues in non-viral gene delivery Adv Drug Deliv Rev 1998 34: 3–19

    Article  Google Scholar 

  32. Ertl HCJ, Verma P, He Z, Xiang ZQ . Plasmid vectors as anti-viral vaccines Ann NY Acad Sci 1995 772: 77–87

    Article  CAS  Google Scholar 

  33. Cohen H, Golomb G, Levy R . Gene delivery by polymer-chencapsulated DNA Proc 26th Intern Symp Control Rel Bioact Mater 1999 6458–6459

  34. Yang YW, Yang JC . Characterization of calcium phosphate as a gene carrier (II): zeta potential and DNA transfection Drug Del 1996 3: 181–186

    Article  CAS  Google Scholar 

  35. Slomkowski S, Winnik MA, Furlong P, Reynolds WF . Synthesis of low-polydispersity poly(tetramethylene oxide) using benzil and pyrene derivatives as initiators Macromolecules 1989 22: 503–509

    Article  CAS  Google Scholar 

  36. Hedin U, Thyberg J . Receptor-mediated endocytosis of immunoglobulin-coated colloidal gold particles in cultured mouse peritoneal macrophages. Chloroquine and monensin inhibit transfer of the ligand from endocytic vesicles to lysosomes Eur J Cell Biol 1985 39: 130–135

    CAS  PubMed  Google Scholar 

  37. Kato T, Okada S, Yutaka T, Yabuuchi H . The effects of sucrose loading on lysosomal hydrolases Mol Cell Biochem 1984 60: 83–98

    Article  CAS  Google Scholar 

  38. Levy MY, Barron LG, Meyer KB, Szoka FC . Characterization of plasmid DNA transfer into mouse skeletal muscle: evaluation of uptake mechanism, expression and secretion of gene products into blood Gene Therapy 1996 3: 201–211

    CAS  Google Scholar 

Download references

Acknowledgements

The authors gratefully acknowledge technical assistance from M Tarshish and E Rahamim, and thank A Honigman and M Levy for helpful discussions. This work was supported in part by a grant from the DKFZ, Germany and the Israeli Ministry of Science (01680). G Golomb is affiliated with the David R Bloom Center of Pharmacy at the School of Pharmacy, The Hebrew University of Jerusalem. RJ Levy's contributions were supported by grants from the National Institute of Health (HL41663), and The William J Rashkind Endowment of the Children's Hospital of Philadelphia.

Author information

Authors and Affiliations

  1. Department of Pharmaceutics, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel

    H Cohen, J Gao, I Fishbein, V Kousaev & G Golomb

  2. Division of Pediatric Cardiology, Children's Hospital of Philadelphia, Philadelphia, PA, USA

    R J Levy

  3. Center of Molecular and Macromolecular Studies, Polish Academy of Sciences, Lodz, Poland

    S Sosnowski & S Slomkowski

Authors
  1. H Cohen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R J Levy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. I Fishbein
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. V Kousaev
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S Sosnowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. S Slomkowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. G Golomb
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cohen, H., Levy, R., Gao, J. et al. Sustained delivery and expression of DNA encapsulated in polymeric nanoparticles. Gene Ther 7, 1896–1905 (2000). https://doi.org/10.1038/sj.gt.3301318

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301318

Keywords

This article is cited by

Search

Advanced search

Quick links