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Chitosan-Thiamine Pyrophosphate as a Novel Carrier for siRNA Delivery

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Abstract

Purpose

A novel siRNA carrier was formulated between chitosan (CS) and thiamine pyrophosphate (TPP). Their ability to deliver siRNA were evaluated in stable and constitutive EGFP-expressing HepG2 cells.

Methods

CS-TPP was prepared by dissolving CS in TPP solution at a CS:TPP molar ratio of 1.5:1. Complexes of CS-TPP/siRNA were formed at varying weight ratios and characterized using gel electrophoresis. Their morphologies and particle sizes were evaluated, and the transfection efficiency and cytotoxicity of CS-TPP/siRNA complexes were examined in stable and constitutive EGFP-expressing HepG2 cells.

Results

Gel electrophoresis results indicated that binding of CS-TPP and siRNA depended on the molecular weight (MW) and weight ratio of CS, and the particle sizes of CS-TPP/siRNA complexes were in nano-size. The CS-TPP-mediated siRNA silencing of the endogenous EGFP gene occurred maximally with 70–73% efficiency. The CS-TPP/siRNA complex with the lowest MW of CS (20 kDa) at a weight ratio of 80 showed the strongest inhibition of gene expression, which was higher than Lipofectamine 2000™. Over 90% the average cell viabilities of the complexes were observed by MTT assay.

Conclusions

This study suggests that CS-TPP is straightforward to prepare, safe and exhibits significantly improved siRNA delivery potential in vitro.

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Acknowledgements

The authors are grateful for financial support by Genetic Engineering and Biotechnology (BIOTEC), Thailand (grant number: BT-B-01-MG-16–4812), the Commission of Higher Education (Thailand), the Thailand Research Funds and National Research Council of Thailand.

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Authors and Affiliations

  1. Faculty of Pharmacy, Silpakorn University, Nakhonpathom, 73000, Thailand

    Theerasak Rojanarata, Praneet Opanasopit, Sunee Techaarpornkul & Tanasait Ngawhirunpat

  2. National Nanotechnology Center, Thailand Science Park, Pathumthani, Thailand

    Uracha Ruktanonchai

Authors
  1. Theerasak Rojanarata
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  2. Praneet Opanasopit
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  3. Sunee Techaarpornkul
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  4. Tanasait Ngawhirunpat
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Correspondence to Praneet Opanasopit.

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Rojanarata, T., Opanasopit, P., Techaarpornkul, S. et al. Chitosan-Thiamine Pyrophosphate as a Novel Carrier for siRNA Delivery. Pharm Res 25, 2807–2814 (2008). https://doi.org/10.1007/s11095-008-9648-6

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  • DOI: https://doi.org/10.1007/s11095-008-9648-6

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