AAPS PharmSciTech

, 10:900 | Cite as

Amyloid-Beta Associated with Chitosan Nano-Carrier has Favorable Immunogenicity and Permeates the BBB

  • Zhang Songjiang
  • Wu LixiangEmail author
Brief/Technical Note


Subfragments of amyloid-beta (Aβ) appear to protect neurons from Alzheimer’s disease (AD). The permeability of the blood–brain barrier (BBB) has limited in vivo research. The aim of this study is to explore permeation of the BBB by chitosan nanoparticles loaded with Aβ and to evaluate immunogenicity of these particles. Chitosan microspheres were prepared by mechanical stirring emulsification methods combined with chemical crosslinking. Morphological characteristics of the nanoparticles were examined using high-resolution transmission electron microscopy. The peptide association efficiency was determined by high-performance liquid chromatography. Fluorescently labeled chitosan nanoparticle-intramembranous fragments of Aβ (NP-IF-A) were administered systemically to mice in order to evaluate brain translocation by fluorescence microscopy. The immunogenicity of the nano-vaccine was determined by enzyme-linked immunosorbent assay (ELISA). All nanoparticles analyzed were well-separated, roughly spherical structures with uniform particle size distribution in the range of 15.23 ± 10.97 nm. The peptide association efficiency was 78.4%. The brain uptake efficiency of nano-antigen was 80.6%; uptake efficiency of antigen alone was only 20.6%. ELISA showed that the nano-vaccine had favorable immunogenicity. A chitosan nano-carrier for Aβ allowed permeation of the BBB and was non-immunogenic. These findings indicate that this novel targeted nano-vaccine delivery system can be used as a carrier for Aβ. This system will further research of peptide vaccines for AD.

Key words

amyloid-beta blood–brain barrier immunogenicity nanoparticle 



This work was supported by the Construct Program of the Key Discipline in Hunan Province, Natural Science Foundation of Hunan Province, China (No.08JJ307), and National High Tech 863 Grant (Project No. 2007AA021901).


  1. 1.
    Selkoe DJ. Physiological production of the beta-amyloid protein and the mechanism of Alzheimer’s disease. Trends Neurosci. 1993;16:403–9. doi: 10.1016/0166-2236(93)90008-A.PubMedCrossRefGoogle Scholar
  2. 2.
    Lopez OL, Rabin BS, Huff FJ, Rezek D, Reinmuth OM. Serum autoantibodies in patients with Alzheimer’s disease and vascular dementia and in nondemented control subjects. Stroke. 1992;23:1078–83.PubMedGoogle Scholar
  3. 3.
    Morgan D, Diamond DM, Gottschall PE, Ugen KE, Dickey C, Hardy J, et al. A beta peptide vaccination prevents memory loss in an animal model of Alzheimer’s disease. Nature. 2000;408:982–5. doi: 10.1038/35050116.PubMedCrossRefGoogle Scholar
  4. 4.
    Soto C. Plaque busters: strategies to inhibit amyloid formation in Alzheimer’s disease. Mol Med Today. 1999;5:343–50.PubMedCrossRefGoogle Scholar
  5. 5.
    Fradinger EA, Monien BH, Urbanc B, Lomakin A, Tan M, Li H, et al. C-terminal peptides coassemble into Abeta42 oligomers and protect neurons against Abeta42-induced neurotoxicity. Proc Natl Acad Sci U S A. 2008;105:14175–80. doi: 10.1073/pnas.0807163105.PubMedCrossRefGoogle Scholar
  6. 6.
    Bernkop-Schnurch A, Krajicek ME. Mucoadhesive polymers as platforms for peroral peptide delivery and absorption: synthesis and evaluation of different chitosan–EDTA conjugates. J. Control. Release. 1998;50:215–23. doi: 10.1016/S0168-3659(97)00136-3.PubMedCrossRefGoogle Scholar
  7. 7.
    Borges O, Borchard G, de Sousa A, Junginger HE, Cordeiro-da-Silva A. Induction of lymphocytes activated marker CD69 following exposure to chitosan and alginate biopolymers. Int. J. Pharm. 2007;337:254–64. doi: 10.1016/j.ijpharm.2007.01.021.PubMedCrossRefGoogle Scholar
  8. 8.
    Jameela SR, Kumary TV, Lal AV, Jayakrishnan A. Progesterone-loaded chitosan microspheres: a long acting biodegradable controlled delivery system. J Control Release. 1998;52:17–24. doi: 10.1016/S0168-3659(97)00187-9.PubMedCrossRefGoogle Scholar
  9. 9.
    Aktaş Y, Yemisci M, Andrieux K, Gürsoy RN, Alonso MJ, Fernandez-Megia E, et al. Development and brain delivery of chitosan-PEG nanoparticles functionalized with the monoclonal antibody OX26. Bioconjug Chem. 2005;16:1503–11.PubMedCrossRefGoogle Scholar
  10. 10.
    Felt O, Buri P, Gurny R. Chitosan: a unique polysaccharide for drug delivery. Drug Dev. Ind. Phar. 1998;24:979–93.CrossRefGoogle Scholar
  11. 11.
    Illum L. Chitosan and its use as a pharmaceutical excipient. Pharm. Res. 1998;15:1326–31.PubMedCrossRefGoogle Scholar
  12. 12.
    Singla AK, Chawla M. Chitosan: some pharmaceutical and biological aspects—an update. J. Pharm. Pharmacol. 2001;53:1047–67.PubMedCrossRefGoogle Scholar
  13. 13.
    Kumbar SG, Kulkarni AR, Minabhavi MA. Crosslinked chitosan microspheres for encapsulation of diclofenac sodium: effect of crosslinking agent. J. Microencapsul. 2002;19:173–80.PubMedCrossRefGoogle Scholar
  14. 14.
    Jameela SR, Kumary TV, Lal AV, Jayakrishnan A. Progesterone-loaded chitosan microspheres: a long acting biodegradable controlled delivery system. J. Control Rel. 1998;52:17–24. doi: 10.1016/S0168-3659(97)00187-9.CrossRefGoogle Scholar
  15. 15.
    Huang RG, Schwartz JB, Ofner CM III. Microencapsulation of chlorpheniramine maleate-resin particles with crosslinked chitosan for sustained release. Pharm. Dev. Technol. 1999;4:107–15.PubMedGoogle Scholar
  16. 16.
    Pavanetto F, Perugini P, Conti B, Modena T, Genta I. Evaluation of process parameters involved in chitosan microspheres preparation by the o/w/o multiple emulsion method. J. Microencaps. 1996;13:679–88.CrossRefGoogle Scholar
  17. 17.
    Prabha S, Zhou WZ, Panyam J, Labhasetwar V. Size-dependency of nanoparticle-mediated gene transfection: studies with fractionated nanoparticles. Int J Pharm. 2002;244:105–15. doi: 10.1016/S0378-5173(02)00315-0.PubMedCrossRefGoogle Scholar
  18. 18.
    Schenk D, Barbour R, Dunn W, Gordon G, Grajeda H, Guido T, et al. Immunization with amyloid-beta attenuates Alzheimer-disease-like pathology in the PDAPP mouse. Nature. 1999;400:173–77. doi: 10.1038/22124.PubMedCrossRefGoogle Scholar
  19. 19.
    Janus C, Pearson J, McLaurin J, Mathews PM, Jiang Y, Schmidt SD, et al. A beta peptide immunization reduces behavioural impairment and plaques in a model of Alzheimer’s disease. Nature. 2000;408:979–82. doi: 10.1038/35050110.PubMedCrossRefGoogle Scholar
  20. 20.
    Kim MS, Sung MJ, Seo SB, Yoo SJ, Lim WK, Kim HM. Water-soluble chitosan inhibits the production of pro-inflammatory cytokine in human astrocytoma cells activated by amyloid beta peptide and interleukin-1beta. Neuroscience Letters. 2002;321:105–9. doi: 10.1016/S0304-3940(02)00066-6.PubMedCrossRefGoogle Scholar
  21. 21.
    Mori T, Okumura M, Matsuura M, Ueno K, Tokura S, Okamoto Y, et al. Effects of chitin and its derivatives on the proliferation and cytokine production of fibroblasts in vitro. Biomaterials. 1997;18:947–51. doi: 10.1016/S0142-9612(97)00017-3.PubMedCrossRefGoogle Scholar
  22. 22.
    Pae HO, Seo WG, Kim NY, Oh GS, Kim GE, Kim YH, et al. Induction of granulocytic differentiation in acute promyelocytic leukemia cells (HL-60) by water-soluble chitosan oligomer. Leuk. Res. 2001;25:339–46. doi: 10.1016/S0145-2126(00)00138-7.PubMedCrossRefGoogle Scholar
  23. 23.
    Nishimura K, Nishimura S, Seo H, Nishi N, Tokura S, Azuma I. Effect of multiporous microspheres derived from chitin and partially deacetylated chitin on the activation of mouse peritoneal macrophages. Vaccine. 1987;5:136–40.PubMedCrossRefGoogle Scholar
  24. 24.
    Shibata Y, Foster LA, Metzger WJ, Myrvik QN. Alveolar macrophage priming by intravenous administration of chitin particles, polymers of N-acetyl-d-glucosamine, in mice. Infect. Immunol. 1997;65:1734–41.Google Scholar
  25. 25.
    Ballabh P, Braun A, Nedergaard M. The blood–brain barrier: An overview, structure, regulation and clinical implications. Neurobiol. Dis. 2004;16:1–13. doi: 10.1016/j.nbd.2003.12.016.PubMedCrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2009

Authors and Affiliations

  1. 1.Department of Physiology, Xiangya Medical CollegeCentral South UniversityChangshaChina
  2. 2.Department of PhysiologyHenan College of Traditional Chinese MedicineZhengzhouChina

Personalised recommendations