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The Development and Mechanism Studies of Cationic Chitosan-Modified Biodegradable PLGA Nanoparticles for Efficient siRNA Drug Delivery

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ABSTRACT

Purpose

In order to improve siRNA delivery for possible clinical applications, we developed biodegradable chitosan-modified poly(D,L-lactide-co-glycolide) (CHT-PLGA) nanoparticles with positive surface charge, high siRNA loading, high transfection efficiency and low toxicity.

Methods

CHT-PLGA nanoparticles were prepared, and siRNA was loaded by emulsion evaporation method with poly(vinyl alcohol) (PVA) as emulsifier. siRNA loading efficiency, particle size, and Zeta potential of nanoparticles were measured. Gel retardation and protection assays were conducted to determine the loading and binding of siRNA in the formulation. Cell transfection was performed to study in vitro siRNA silencing efficiency. XTT assay was used to evaluate the cytotoxicity.

Results

It was found that the nanoparticle diameter and positive Zeta potential increase as the chitosan coating concentration increases. CHT-PLGA nanoparticles showed excellent siRNA binding ability and effective protection of oligos from RNase degradation. siRNA-loaded nanoparticles were successfully delivered into the HEK 293 T cell line, and the silencing of green fluorescence protein (GFP) expression was observed using fluorescent microscopy and flow cytometry. In addition, the cytotoxicity assay revealed that CHT-PLGA nanoparticles had relatively low cytotoxicity.

Conclusion

This study suggests that biodegradable cationic CHT-PLGA nanoparticles possess great potential for efficient and safer siRNA delivery in future clinical applications.

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ACKNOWLEDGEMENTS

The authors thank Ms. Myongcha Shin for her technical assistance in conducting FACS study at Columbia University. We would also like to thank Dr. Robert Chapman in the College of Pharmacy at Midwestern University for taking time to review the manuscript and giving us invaluable advice.

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Authors and Affiliations

  1. Division of Pharmaceutical Sciences, Arnold & Marie Schwartz College of Pharmacy, Long Island University, 75 DeKalb Avenue, Brooklyn, New York, 11201-5497, USA

    Xudong Yuan, Bruhal A. Shah, Naimesh K. Kotadia & Zhiqian Wu

  2. Nanotarget, 10054 Mesa Ridge Court, San Diego, California, 92121, USA

    Jian Li

  3. Department of Microbiology, College of Physicians and Surgeons, Columbia University, 701 West 168th Street, New York City, New York, 10032, USA

    Hua Gu

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  1. Xudong Yuan
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  2. Bruhal A. Shah
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  5. Hua Gu
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Correspondence to Xudong Yuan.

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Yuan, X., Shah, B.A., Kotadia, N.K. et al. The Development and Mechanism Studies of Cationic Chitosan-Modified Biodegradable PLGA Nanoparticles for Efficient siRNA Drug Delivery. Pharm Res 27, 1285–1295 (2010). https://doi.org/10.1007/s11095-010-0103-0

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  • DOI: https://doi.org/10.1007/s11095-010-0103-0

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