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Galactosylated chitosan (GC)-graft-poly(vinyl pyrrolidone) (PVP) as hepatocyte-targeting DNA carrier:in vitro transfection

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Abstract

Galactosylated chitosan-graft-poly(vinyl pyrrolidone) (GCPVP) was synthesized and characterized for hepatocyte-targeting gene carrier. GCPVP itself as well as GCPVP/DNA complex had negligible cytotoxicity regardless of the concentration of GCPVP and the charge ratio, but GCPVP/DNA complex had slightly cytotoxic effect on HepG2 cells only in the case of the higher charge ratio and 20 mM of Ca2+ concentration used. Through the confocal laser scanning microscopy, it is shown that the endocytosis by interaction between galactose ligands of GCPVP and ASGPR of the hepatocytes was the major route of transfection of GCPVP/F-plasmid complexes.

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References

  1. Crrotto, E. and Aros, R., Quantitative determination of chitosan and the percentage of free amino groups.Anal. Biochem., 211, 240–241 (1993).

  2. Crystal, R. G., Transfer of genes to humans: early lessons and obstacles to success.Science, 270, 404–410 (1995).

  3. Dodane, V. and Vilivalam, V.D., Pharmaceutical applications of chitosan.Pharm. Sci. Technol. Today, 1, 246–253 (1998).

  4. Erbacher, P., Zou, S. M., Bettinger, T., Steffan, A.-M., and Remy, J.-S., Chitosan-based vector/DNA complexes for gene delivery: biophysical characteristics and transfection ability.Pharm. Res., 15, 1332–1339 (1998).

  5. Ishii, T., Okahata, Y., and Sato, T., Facile preparation of a fluorescence-labeled plasmid.Chem. Lett., 386–387 (2000).

  6. Jiang, H. L., Park, I. K., Shin, N. R., Yoo, H. S., Akaike, T., and C. S. Cho, Controlled release of vaccine from bordetella bronchiseptica dermonecrotoxinloaded chitosan microspheresin vitro.Arch. Pharm. Res., 27, 346–350 (2004).

  7. Kamada, H., Tsutsumi, Y., Tsunoda, S.I., Kihira, T., Kaneda, Y., Yamamoto, Y., Nakagawa, S., Horisawa, Y., and Mayumi, T., Molecular design of conjugated tumor necrosis factor-: synthesis and characteristics of polyvinyl pyrrolidone modified tumor necrosis factor-α.Biochem. Bioph. Res. Comm., 257, 448–453 (1999).

  8. Kim, S. H., Goto, M., Cho, C. S., and Akaike, T., Specific adhesion of primary hepatocytes to a novel glucose-carrying polymer.Biotechnol. Lett., 22, 1049–1057 (2000).

  9. Kim, S. H., Goto, M., and Akaike, T., Specific binding of glucose-derivatized polymers to the asialoglycoprotein receptor of mouse primary hepatocytes.J. Biol. Chem., 276, 35312–35319 (2001).

  10. Lee, K. Y., Kwon, I. C., Kim, Y. H., Jo, W. H., and S. Y. Jeong, Preparation of chitosan self-aggregates as a gene delivery system.J. Control Release, 51, 213–220 (1998).

  11. Mu, Y., Kamada, H., Kodaira, H., Sato, K., TsuTsumi, Y., Maeda, M., Kawasaki, K., Nomizu, M., Yamada, Y., and Mayumi, T., Bioconjugation of laminin-related peptide YIGSR with polyvinyl pyrrolidone increases its antimetastatic effect due to a longer plasma half-life.Biochem. Biophys. Res. Comm., 264, 763–767 (1999).

  12. Niidome, T. and Huang, L., Gene therapy progress and prospects: nonviral vectors.Gene ther., 9, 1647–1652 (2002).

  13. Park, I. K., Park, Y. H., Shin, B. A., Choi, E. S., Kim, Y. R., Akaike, T., and Cho, C. S., Galactosylated chitosan-graft-dextran as hepatocyte-targeting DNA carrier.J. Control Release, 69, 97–108 (2000).

  14. Park, I. K., Kim, T. H., Park, Y. H., Shin, B. A., Choi, E. S., Chowdhury, E. H., Akaike, T., and Cho, C. S., Galactosylated chitosan-graft-poly(ethylene glycol) as hepatocyte-targeting DNA carrier.J. Control Release, 76, 349–362, (2001).

  15. Park, I. K., Ihm, J. E., Park, Y. H., Choi, Y. J., Kim, S. I., Kim, W. J., Akaike, T., and Cho, C. S., Preparation and characterization of galactosylated chitosan-graft-poly(vinyl pyrrolidone) as hepatocyte-targeting gene carrier.J. Control Release, 86, 349–359 (2003).

  16. Park, E. I., Manzella, S. M., and Baenziger, J. U., Rapid clearance of sialylated glycoproteins by the asialoglycoprotein receptor.J. Biol. Chem., 278, 4597–4602 (2003).

  17. Park, I. K., Yang, J., Jeong, H. J., Bom, H.S., Harada, I., Akaike, T., Kim, S. I., and Cho, C. S., Galactosylated chitosan as a synthetic extracellular matrix for hepatocytes attachment.Biomaterials, 24, 2331–2337 (2003).

  18. Park, I. K., Kim, T. H., Kweon, H. Y., Park, Y. H., Kim, W. J., Akaike, T., and Cho, C. S., Visualization of transfection of galactosylated chitosan-graft-poly(ethylene glycol) /DNA complexes into hepatocytes by confocal laser scanning microscopy.Int. J. Pharm., 257, 103–110 (2003).

  19. Stockert, R. J., The asialoglycoprotein receptor: relationships between structure, function and expression.Physiol. Rev., 75, 591–609 (1995).

  20. Tripathy, S. K., Black, H. B., Goldwasser, E., and Leiden, J. M., Immune responses to transgene-encoded proteins limit the stability of gene expression after injection of replication-defective adenovirus vectors.Nat. Med., 2, 545–550 (1996).

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Correspondence to In-Kyu Park or Hu-Lin Jiang or Seung-Eun Cook or Myung-Haing Cho or Su-ll Kim or Hwan-Jeong Jeong or Toshihiro Akaike or Chong-Su Cho.

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Park, I., Jiang, H., Cook, S. et al. Galactosylated chitosan (GC)-graft-poly(vinyl pyrrolidone) (PVP) as hepatocyte-targeting DNA carrier:in vitro transfection. Arch Pharm Res 27, 1284 (2004). https://doi.org/10.1007/BF02975895

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Key words

  • Galactosylated chitosan
  • Poly(vinyl pyrrolidone)
  • Hepatocyte
  • Gene delivery