Basic Research
Copyright ©The Author(s) 2004. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jan 1, 2004; 10(1): 112-116
Published online Jan 1, 2004. doi: 10.3748/wjg.v10.i1.112
Transfection of mEpo gene to intestinal epithelium in vivo mediated by oral delivery of chitosan-DNA nanoparticles
Jing Chen, Wu-Li Yang, Ge Li, Ji Qian, Jing-Lun Xue, Shou-Kuan Fu, Da-Ru Lu
Jing Chen, Ge Li, Ji Qian, Jing-Lun Xue, Da-Ru Lu, State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life sciences, Fudan University, Shanghai 200433, China
Wu-Li Yang, Shou-Kuan Fu, The Key Laboratory of Molecular Engineering of Polymers under Ministry of Education, Department of Macromolecular Science, Fudan University, Shanghai, 200433, China
Author contributions: All authors contributed equally to the work.
Supported by the State High Technology Development Program 863 (2001AA217181), the National Natural Science Foundation of China (No.50233030), Foundation of Doctor Degree Thesis from Ministry of Education (199925), Encourage Project of Teaching and Research of University Excellent Youth Teacher, and the Youth Foundation of Fudan University
Correspondence to: Dr. Da-Ru Lu, State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China. drlu@fudan.edu.cn Telephone: +86-21-65642424 Fax: +86-21-65642799
Received: May 12, 2003
Revised: July 20, 2003
Accepted: July 19, 2003
Published online: January 1, 2004
Abstract

AIM: To prepare the chitosan-pmEpo nanoparticles and to study their ability for transcellular and paracellular transport across intestinal epithelia by oral administration.

METHODS: ICR mice were fed with recombinant plasmid AAV-tetO-CMV-mEpo (containing mEpo gene) or pCMVβ (containing LacZ gene), whether it was wrapped by chitosan or no. Its size and shape were observed by transmission electron microscopy. Agarose gel electrophoresis was used to assess the efficiency of encapsulation and stability against nuclease digestion. Before and after oral treatmant, blood samples were collected by retro-orbital puncture, and hematocrits were used to show the physiological effect of mEpo.

RESULTS: Chitosan was able to successfully wrap the plasmid and to protect it from DNase degradation. Transmission electron microscopy showed that freshly prepared particles were approximately 70-150 nm in size and fairly spherical. Three days after fed the chitosan-pCMVβ complex was fed, the mice were killed and most of the stomach and 30% of the small intestine were stained. Hematocrit was not modified in naive and ‘naked’ mEpo-fed mice, a rapid increase of hematocrit was observed during the first 4 days of treatment in chitosan-mEpo-fed animals, reaching 60.9 ± 1.2% (P < 0.01), and sustained for a week. The second feed (6 days after the first feed) was still able to promote a second hematocrit increase in chitosan-mEpo-fed animals, reaching 65.9 ± 1.4% (P < 0.01), while the second hematocrit increase did not appear in the ‘naked’ mEpo-second-fed mice.

CONCLUSION: Oral chitosan-DNA nanoparticles can efficiently deliver genes to enterocytes, and may be used as a useful tool for gene transfer.

Keywords: $[Keywords]