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Development and Evaluation of Chitosan-Coated Liposomes for Oral DNA Vaccine: The Improvement of Peyer’s Patch Targeting Using a Polyplex-Loaded Liposomes

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Abstract

The aim of this study was to develop chitosan-coated and polyplex-loaded liposomes (PLLs) containing DNA vaccine for Peyer’s patch targeting. Plain liposomes carrying plasmid pRc/CMV-HBs were prepared by the reverse-phase evaporation method. Chitosan coating was carried out by incubation of the liposomal suspensions with chitosan solution. Main lipid components of liposomes were phosphatidylcholine/cholesterol. Sodium deoxycholate and dicetyl phosphate were used as negative charge inducers. The zeta potentials of plain liposomes were strongly affected by the pH of the medium. Coating with chitosan variably increased the surface charges of the liposomes. To increase the zeta potential and stability of the liposome, chitosan was also used as a DNA condensing agent to form a polyplex. The PLLs were coated with chitosan solution. In vivo study of PLLs was carried out in comparison with chitosan-coated liposomes using plasmid encoding green fluorescence protein as a reporter. A single dose of plasmid equal to 100 μg was intragastrically inoculated into BALB/c mice. The expression of green fluorescence protein (GFP) was detected after 24 h using a confocal laser scanning microscope. The signal of GFP was obtained from positively charged chitosan-coated liposomes but found only at the upper part of duodenum. With chitosan-coated PLL540, the signal of GFP was found throughout the intestine. Chitosan-coated PLL demonstrated a higher potential to deliver the DNA to the distal intestine than the chitosan-coated liposomes due to the increase in permanent positive surface charges and the decreased enzymatic degradation.

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ACKNOWLEDGMENTS

The authors would like to acknowledge the Thailand Research Fund for financial support through the Royal Golden Jubilee Ph.D. Program (grant no. PHD/0222/2543). The authors wish to express their sincere thanks to the Molecular Immunology, Faculty of Allied Health Sciences, Thammasat University for partial support of the facilities.

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Authors and Affiliations

  1. Department of Industrial Pharmacy, Faculty of Pharmacy, Mahidol University, Bangkok, Thailand

    Sunee Channarong, Nuttanan Sinchaipanid & Ampol Mitrevej

  2. Faculty of Medicine Siriraj Hospital Mahidol University, Bangkok, Thailand

    Wanpen Chaicumpa

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  1. Sunee Channarong
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  2. Wanpen Chaicumpa
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  3. Nuttanan Sinchaipanid
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  4. Ampol Mitrevej
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Correspondence to Ampol Mitrevej.

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Channarong, S., Chaicumpa, W., Sinchaipanid, N. et al. Development and Evaluation of Chitosan-Coated Liposomes for Oral DNA Vaccine: The Improvement of Peyer’s Patch Targeting Using a Polyplex-Loaded Liposomes. AAPS PharmSciTech 12, 192–200 (2011). https://doi.org/10.1208/s12249-010-9559-9

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  • DOI: https://doi.org/10.1208/s12249-010-9559-9

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