Advertisement

AAPS PharmSciTech

, Volume 12, Issue 1, pp 192–200 | Cite as

Development and Evaluation of Chitosan-Coated Liposomes for Oral DNA Vaccine: The Improvement of Peyer’s Patch Targeting Using a Polyplex-Loaded Liposomes

  • Sunee Channarong
  • Wanpen Chaicumpa
  • Nuttanan Sinchaipanid
  • Ampol MitrevejEmail author
Research Article

Abstract

The aim of this study was to develop chitosan-coated and polyplex-loaded liposomes (PLLs) containing DNA vaccine for Peyer’s patch targeting. Plain liposomes carrying plasmid pRc/CMV-HBs were prepared by the reverse-phase evaporation method. Chitosan coating was carried out by incubation of the liposomal suspensions with chitosan solution. Main lipid components of liposomes were phosphatidylcholine/cholesterol. Sodium deoxycholate and dicetyl phosphate were used as negative charge inducers. The zeta potentials of plain liposomes were strongly affected by the pH of the medium. Coating with chitosan variably increased the surface charges of the liposomes. To increase the zeta potential and stability of the liposome, chitosan was also used as a DNA condensing agent to form a polyplex. The PLLs were coated with chitosan solution. In vivo study of PLLs was carried out in comparison with chitosan-coated liposomes using plasmid encoding green fluorescence protein as a reporter. A single dose of plasmid equal to 100 μg was intragastrically inoculated into BALB/c mice. The expression of green fluorescence protein (GFP) was detected after 24 h using a confocal laser scanning microscope. The signal of GFP was obtained from positively charged chitosan-coated liposomes but found only at the upper part of duodenum. With chitosan-coated PLL540, the signal of GFP was found throughout the intestine. Chitosan-coated PLL demonstrated a higher potential to deliver the DNA to the distal intestine than the chitosan-coated liposomes due to the increase in permanent positive surface charges and the decreased enzymatic degradation.

KEY WORDS

chitosan DNA vaccine liposome oral immunization polyplex-loaded liposome 

Notes

ACKNOWLEDGMENTS

The authors would like to acknowledge the Thailand Research Fund for financial support through the Royal Golden Jubilee Ph.D. Program (grant no. PHD/0222/2543). The authors wish to express their sincere thanks to the Molecular Immunology, Faculty of Allied Health Sciences, Thammasat University for partial support of the facilities.

REFERENCES

  1. 1.
    Montgomery DL, Ulmer JE, Donnelly JJ, Liu MA. DNA vaccines. Pharmacol Ther. 1997;74:195–205.PubMedCrossRefGoogle Scholar
  2. 2.
    Dertzbaugh MT. Genetically engineered vaccines: an overview. Plasmid. 1998;39:100–13.PubMedCrossRefGoogle Scholar
  3. 3.
    Nugent J, Po AL, Scott EM. Design and delivery of non-parenteral vaccines. J Clin Pharm Ther. 1998;23:257–85.PubMedGoogle Scholar
  4. 4.
    Kirman JR, Seder RA. DNA vaccination: the answer to stable, protective T-cell memory? Curr Opin Immunol. 2003;15:471–6.PubMedCrossRefGoogle Scholar
  5. 5.
    Takahashi I, Nochi T, Yuki Y, Kiyono H. New horizon of mucosal immunity and vaccines. Curr Opin Immunol. 2009;21:352–8.PubMedCrossRefGoogle Scholar
  6. 6.
    Yeh P, Ellens H, Smith PL. Physiological considerations in the design of particulate dosage forms for oral vaccine delivery. Adv Drug Del Rev. 1998;34:123–33.CrossRefGoogle Scholar
  7. 7.
    Jain S, Singh P, Mishra V, Vyas SP. Mannosylated niosomes as adjuvant-carrier system for oral genetic immunization against hepatitis B. Immunol Lett. 2005;101:41–9.PubMedCrossRefGoogle Scholar
  8. 8.
    Conacher M, Alexander J, Brewer JM. Oral immunization with peptide and protein antigens by formulation in lipid particles incorporating bile salts (bilosomes). Vaccine. 2001;19:2965–74.PubMedCrossRefGoogle Scholar
  9. 9.
    Guliyeva Ü, Öner F, Özsoy S, Haziroglu R. Chitosan microparticles containing plasmid DNA as potential oral gene delivery system. Eur J Pharm Biopharm. 2006;62:17–25.PubMedCrossRefGoogle Scholar
  10. 10.
    Mann JFS, Scales HE, Sharkir E, Alexander J, Carter KC, Mullen AB, et al. Oral delivery of tetanus toxoid using particles significant systemic and mucosal immunity. Methods. 2006;38:90–5.PubMedCrossRefGoogle Scholar
  11. 11.
    Mann JFS, Shakir E, Carter KC, Mullen AB, Alexander J, Ferro VA. Lipid vesicle size of an oral influenza vaccine delivery vehicle influences the Th1/Th2 bias in the immune response and protection against infection. Vaccine. 2009;27:3643–9.PubMedCrossRefGoogle Scholar
  12. 12.
    Khatri K, Goyal AK, Gupta PN, Mishra N, Mehta A, Vyas SP. Surface modified liposomes for nasal delivery of DNA vaccine. Vaccine. 2008;26:2225–33.PubMedCrossRefGoogle Scholar
  13. 13.
    Page DT, Cudmore S. Innovations in oral gene delivery: challenge and potentials. DDT. 2001;6(2):92–101.PubMedGoogle Scholar
  14. 14.
    Filion MC, Phillips NC. Major limitations in the use of cationic liposomes for DNA delivery. Int J Pharm. 1998;162:159–70.CrossRefGoogle Scholar
  15. 15.
    Dokka S, Toledo D, Shi X, Castranova V, Rojanasakul Y. Oxygen radical-mediated pulmonary toxicity induced by some cationic liposomes. Pharm Res. 2000;17:521–5.PubMedCrossRefGoogle Scholar
  16. 16.
    Bivas-Benita M, Laloup M, Versteyhe S, Dewit J, de Braekeleer J, Jongert E, et al. Generation of Toxoplasma gondii GRA1 protein and DNA vaccine loaded chitosan particles: preparation, characterization, and preliminary in vivo studies. Int J Pharm. 2003;266:17–27.PubMedCrossRefGoogle Scholar
  17. 17.
    Howard KA, Li XW, Somavarapu S, et al. Formulation of a microparticle carrier for oral polyplex-based DNA vaccines. Biochim Biophy Acta. 2004;1674:149–57.Google Scholar
  18. 18.
    Almofti MR, Harashima H, Shinohara Y, Almofti A, Baba Y, Kiwada H. Cationic liposome-mediated gene delivery: biophysical study and mechanism of internalization. Arch Biochem Biophys. 2003;410(2):246–53.PubMedCrossRefGoogle Scholar
  19. 19.
    Jayakumar R, Chennazhi KP, Muzzarelli RAA, Tamura H, Nair SV, Selvamurugan N. Chitosan conjugated DNA nanoparticles in gene therapy. Carbohydr Polym. 2010;79:1–8.CrossRefGoogle Scholar
  20. 20.
    Yamamoto H, Kuno Y, Sugimoto S, Takeuchi H, Kawashima Y. Surface-modified PLGA nanosphere with chitosan improved pulmonary delivery of calcitonin by mucoadhesion and opening of the intercellular tight junctions. J Control Release. 2005;102:373–81.PubMedCrossRefGoogle Scholar
  21. 21.
    Prego C, Garcí M, Torres D, Alonso MJ. Transmucosal macromolecular drug delivery. J Control Release. 2005;101:151–62.PubMedCrossRefGoogle Scholar
  22. 22.
    Van der Merwe SM, Verhoef JC, Verheijden JHM, Kotzé AF, Junginger HE. Trimethylated chitosan as polymeric absorption enhancer for improved peroral delivery of peptide drugs. Eur J Pharm Biopharm. 2004;58:225–35.PubMedCrossRefGoogle Scholar
  23. 23.
    Takeuchi H, Matsui Y, Sugihara H, Yamamoto H, Kawashima Y. Effectiveness of submicron-sized, chitosan-coated liposomes in oral administration of peptide drugs. Int J Pharm. 2005;303:160–70.PubMedCrossRefGoogle Scholar
  24. 24.
    Alpar HO, Somavarapua S, Atuahb KN, Bramwell VW. Biodegradable mucoadhesive particulates for nasal and pulmonary antigen and DNA delivery. Adv Drug Del Rev. 2005;57:411–43.CrossRefGoogle Scholar
  25. 25.
    Davis HL, Michel ML, Whalen RG. DNA-based immunization for hepatitis B induces continuous secretion of antigen and high levels of circulating antibody. Hum Mol Genetics. 1993;2:1847–51.CrossRefGoogle Scholar
  26. 26.
    Mao HQ, Roy K, Troung-Le VL, et al. Chitosan-DNA nanoparticles as gene carrier: synthesis, characterization and transfection efficiency. J Control Release. 2001;70:399–421.PubMedCrossRefGoogle Scholar
  27. 27.
    Cócera M, López O, Coderch L, Parra JL, de la Maza A. Permeability investigations of phospholipid liposomes by adding cholesterol. Colloid Surf A. 2003;221:9–17.CrossRefGoogle Scholar
  28. 28.
    Moghimi SM, Patel HM. Tissue specific opsonins for phagocytic cells and their different affinity for cholesterol-rich liposomes. FEBS Lett. 1988;233(1):143–7.PubMedCrossRefGoogle Scholar
  29. 29.
    Guo J, Ping Q, Jiang G, Huang L, Tong Y. Chitosan-coated liposomes: characterization and interaction with leuprolide. Int J Pharm. 2003;260:167–73.PubMedCrossRefGoogle Scholar
  30. 30.
    Mumper RJ, Wang J, Claspell JM, Rolland AP. Novel polymeric condensing carriers for gene delivery. Proc Int Symp Control Rel Bioact Mater. 1996;22:178–9.Google Scholar
  31. 31.
    Kiang T, Wen J, Lim HW, Leong KW. The effect of the degree of chitosan deacetylation on the efficacy of gene transfection. Biomaterials. 2004;25:5293–301.PubMedCrossRefGoogle Scholar
  32. 32.
    Liu W, Sun S, Cao Z, Zhang X, Yao K, Lu WW, et al. An investigation on the physicochemical properties of chitosan/DNA polyelectrolyte complexes. Biomaterials. 2005;26(15):2705–11.PubMedCrossRefGoogle Scholar
  33. 33.
    Takeuchi K, Ishihara K, Kawaura C, Noji M, Furuno T, Nakanishi M. Effect of zeta potential of cationic liposomes containing cationic cholesterol derivatives on gene transfection. FEBS Lett. 1996;397:207–9.PubMedCrossRefGoogle Scholar
  34. 34.
    Bowman K, Sarkar R, Raut S, Leong KW. Gene transfer to hemophilia A mice via oral delivery of FVIII-chitosan nanoparticles. J Control Release. 2008;132:252–9.PubMedCrossRefGoogle Scholar
  35. 35.
    Hejazi R, Amiji M. Chitosan-based gastrointestinal delivery systems. J Control Release. 2003;89:151–65.PubMedCrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2010

Authors and Affiliations

  • Sunee Channarong
    • 1
  • Wanpen Chaicumpa
    • 2
  • Nuttanan Sinchaipanid
    • 1
  • Ampol Mitrevej
    • 1
    Email author
  1. 1.Department of Industrial Pharmacy, Faculty of PharmacyMahidol UniversityBangkokThailand
  2. 2.Faculty of Medicine Siriraj Hospital Mahidol UniversityBangkokThailand

Personalised recommendations