Abstract
Purpose. The gastrointestinal tract poses a variety of morphological and physiological barriers to the expression of a target gene. In this work, N-acetylated chitosan is used as a gene delivery carrier for solving this problem.
Methods. Plasmid DNAs carrying the lacZ gene and interluekin-10 (IL-10) gene were mixed with N-acetylated chitosan. The N-acetylated chitosan/plasmid DNA complex was mixed into a food paste to feed mice. The transport and distribution characteristics of the plasmid along the intestinal mucosa were identified by β-galactosidase assay. In addition, the stomach and intestines were subjected to analysis for the production of IL-10.
Results. The efficiency of N-acetylated chitosan-mediated gene delivery to the intestines was observed to be higher than that of chitosan alone. In particular, this result was most significant in the case of the duodenum, where the LacZ gene was expressed most effectively through the use of N-acetylated chitosan. It was also demonstrated that the IL-10 gene was successfully transferred to intestines through this method.
Conclusions. A plasmid DNA was able to be orally delivered to the intestines using N-acetylated chitosan as a carrier. Thus, we have developed a dietary dose system for delivering a DNA vaccine for treating gastrointestinal diseases.
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references
J. M. Wilson. Adenoviruses as gene-delivery vehicles. N. Engl. J. Med. 334:1185–1187 (1996).
M. Lee, J. W. Nah, Y. Kwon, J. J. Koh, K. S. Ko, and S. W. Kim. Water-soluble and low molecular weight chitosan-based plasmid DNA delivery. Pharm. Res. 18:427–431 (2001).
C. W. Pouton and L. W. Seymour. Key issues non-viral gene delivery. Adv. Drug Deliv. Rev. 34:3–19 (1998).
K. A. Janes, P. Calvo, and M. J. Alonso. Polysaccharide colloidal particles as delivery sysytem for macromolecules. Adv. Drug Deliv. Rev. 47:83–97 (2001).
C. M. Lehr, J. A. Bowstra, E. H. Schacht, and H. E. Junginger. In vitro evaluation of mucoadhesive properties of chitosan and some other natural polymers. Int. J. Pharm. 78:43–48 (1992).
R. J. Mumper, J. Wang, J. M. Claspell, and A. P. Rolland. Novel polymeric condensing carriers for gene transfer. Proc. Natl. Symp. Control. Rel. Bioact. Mater. 22:178–179 (1995).
K. Roy, H. Q. Mao, S. K. Huang, and K. W. Leong. Oral gene delivery with chitosan-DNA nanoparticles generates immunologic protection in a murine model of peanut allergy. Nat. Med. 5:387–391 (1999).
S. Hirschfeld and H. R. Clearfield. Pharmacologic therapy for inflammatory bowel disease. Am. Fam. Physician 51:1971–1975 (1995).
Y. Shibata, L. A. Foster, W. J. Metzger, and Q. N. Myrvik. Alveolar macrophage priming by intravenous administration of chitin Particles, polymers of N-acetyl-D-glucosamine, in mice. Infect. Immun. 65:1734–1741 (1997).
M. J. Skeen, M. A. Miller, T. M. Shinnick, and H. K. Ziegler. Regulation of murine macrophage IL-12 production. Activation of macrophages in vivo, restimulation in vitro, and mudulation by other cytokines. J. Immunol. 156:1196–1206 (1996).
S. Aiba. Studies on chitosan: 2. Solution stability and reactivity of partially N-acetylated chitosan derivatives in aqueous media. Int. J. Biol. Macromol. 11:249–252 (1989).
K. Kofuji, T. Ito, Y. Murata, and S. Kawashima. Biodegradation and drug release of chitosan gel beads in subcutaneous air pouches of mice. Bio. Pharm. Bull. 24:205–208 (2001).
C. Yomota, T. Komuro, and T. Kimura. Studies on the degradation of chitosan films by lysozyme and release of loaded chemicals. Yakugaku Zasshi 110:442–448 (1990).
P. J. Hendrikx, A. C. Martens, F. W. Schultz, J. W. Visser, and A. Hagenbeek. Monitoring of leukemia growth in a rat model using a highly sensitive assay for the detection of LacZ marked leukemic cells. Leukemia 9:1954–1960 (1995).
I. Lalani, K. Bhol, and A. R. Ahmed. Inerleukin-10: biology, role in inflammation and autoimmunity. Ann. Allergy Asthama. Immunol. 79:469–483 (1997).
P. M. Mathisen, M. J. Yu, J. M. Johnson, J. A. Drazba, and V. K. Tuohy. Treatment of experimental autoimmune encephalomyelitis with genetically meodified memory T cells. J. Exp. Med. 186:159–164 (1997).
A. K. Singla and M. Chawla. Chitosan: some pharmaceutical and biological aspects—an update. J. Pharm. Pharmacol. 5:1047–1067 (2001).
I. Azuma. Synthetic immunoajuvants: application to non-specific host stimulation and potentiation of vaccine immunogenicity. Vaccine 10:1000–1006 (1992).
K. Nishimura, S. Nishimura, H. Seo, N. Nishi, S. Tokura, and I. Azuma. Macrophage activation with multi-porous beads prepared from partially deacetylated chitin. J. Biomed. Mater. Res. 20:1359–1372 (1986).
K. Nishimura, S. Nishimura, H. Seo, N. Nishi, S. Tokura, and I. Azuma. Effect of multiporous microspheres derived from chitin and partially deacetylated chitin on the activation of mouse peritoneal mactophages. Vaccine 5:136–140 (1987).
M. Ito, A. Ban, and M. Ishihara. Anti-ulcer Effects of Chitin and Chitosanm Healthy Foods, in Rat. Jpn. J. Pharmacol. 82:218–225 (2000).
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Kai, E., Ochiya, T. A Method for Oral DNA Delivery with N-Acetylated Chitosan. Pharm Res 21, 838–843 (2004). https://doi.org/10.1023/B:PHAM.0000026437.32238.6f
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DOI: https://doi.org/10.1023/B:PHAM.0000026437.32238.6f