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Pharmaceutical Research

, Volume 24, Issue 1, pp 157–167 | Cite as

Thiolated Chitosan/DNA Nanocomplexes Exhibit Enhanced and Sustained Gene Delivery

  • Dongwon Lee
  • Weidong Zhang
  • Shawna A. Shirley
  • Xiaoyuan Kong
  • Gary R. Hellermann
  • Richard F. Lockey
  • Shyam S. MohapatraEmail author
Research Paper

Abstract

Purpose

Thiolated chitosan appears to possess enhanced mucoadhesiveness and cell penetration properties, however, its potential in gene-drug delivery remains unknown. Herein, we report on a highly effective gene delivery system utilizing a 33-kDa thiol-modified chitosan derivative.

Methods

Thiolated chitosan was prepared by the reaction with thioglycolic acid. Nanocomplexes of unmodified chitosan or thiolated chitosan with plasmid DNA encoding green fluorescenct protein (GFP) were characterized for their size, zeta potential, their ability to bind and protect plasmid DNA from degradation. The transfection efficiency of thiolated chitosan and sustained gene expression were evaluated in various cell lines in vitro and in Balb/c mice in vivo.

Results

Thiolated chitosan–DNA nanocomplexes ranged in size from 75 to 120 nm in diameter and from +2.3 to 19.7 mV in zeta potential, depending on the weight ratio of chitosan to DNA. Thiolated chitosan, CSH360, exhibited effective physical stability and protection against DNase I digestion at a weight ratio ≥ 2.5:1. CSH360/DNA nanocomplexes induced significantly (P < 0.01) higher GFP expression in HEK293, MDCK and Hep-2 cell lines than unmodified chitosan. Nanocomplexes of disulphide-crosslinked CSH360/DNA showed a sustained DNA release and continuous expression in cultured cells lasting up to 60 h post transfection. Also, intranasal administration of crosslinked CSH360/DNA nanocomplexes to mice yielded gene expression that lasted for at least 14 days.

Conclusions

Thiolated chitosans condense pDNA to form nanocomplexes, which exhibit a significantly higher gene transfer potential and sustained gene expression upon crosslinking, indicating their great potential for gene therapy and tissue engineering.

Key words

chitosan gene transfer thiolation crosslinking nanocomplexes 

Notes

Acknowledgment

This work was supported by NIH (5RO1HL71101-01A2) awarded to SSM and Culverhouse Endowment to the Division of Allergy and Immunology.

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Dongwon Lee
    • 1
  • Weidong Zhang
    • 1
  • Shawna A. Shirley
    • 1
  • Xiaoyuan Kong
    • 1
  • Gary R. Hellermann
    • 1
  • Richard F. Lockey
    • 1
  • Shyam S. Mohapatra
    • 1
    Email author
  1. 1.Division of Allergy and Immunology and Joy McCann Culverhouse Airway Disease Center, Department of Internal MedicineUniversity of South Florida College of Medicine and James A. Haley Veteran’s HospitalTampaUSA

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