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Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae
Published on: March 25, 2015
Evaluation of antisense oligonucleotide loaded chitosan nanoparticles; characterization and antisense effect
S Ozbaş-Turan1, J Akbuğa, B Enneli
1Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Marmara University, Istanbul, Turkey.
Die Pharmazie
|January 26, 2010
Summary
Chitosan nanoparticles effectively delivered antisense oligonucleotides (AsODN) to inhibit beta-galactosidase (beta-gal) gene expression in HeLa cells. Phosphorothioate (PS) AsODN nanoparticles showed superior encapsulation and gene silencing compared to phosphodiester (PO) formulations.
Area of Science:
- Biotechnology
- Nanomedicine
- Gene Therapy
Background:
- Antisense oligonucleotides (AsODN) offer therapeutic potential but require effective delivery systems.
- Chitosan nanoparticles (NPs) are promising carriers due to their biocompatibility and mucoadhesive properties.
- Optimizing formulation parameters is crucial for enhancing AsODN loading, stability, and in vitro efficacy.
Purpose of the Study:
- To investigate the impact of formulation variables on the physicochemical and antisense properties of AsODN-loaded chitosan NPs.
- To compare the preparation methods for phosphodiester (PO) and phosphorothioate (PS) AsODN-loaded NPs.
- To evaluate the gene silencing efficacy of these NPs targeting the beta-galactosidase (beta-gal) gene in HeLa cells.
Main Methods:
- Chitosan nanoparticles loaded with AsODN targeting the beta-galactosidase gene were prepared using varying chitosan molecular weights and concentrations, tripolyphosphate (TPP) concentrations, and alginate inclusion.
- Preparation methods for both PO-AsODN and PS-AsODN NPs were compared.
- In vitro transfection studies were conducted in HeLa cells, with beta-galactosidase activity assayed spectrophotometrically.
Main Results:
- AsODN-NPs exhibited positive surface charges and sizes ranging from 221.4-525.7 nm, influenced by formulation parameters.
- Encapsulation efficiency was higher for PS-AsODN (91-98%) compared to PO-AsODN (78-94%).
- PS-NPs demonstrated significantly higher beta-gal inhibition (up to 90.71% with formulation PT-2) than PO-NPs, with PS-adsorbed NPs achieving an 88% reduction.
Conclusions:
- Chitosan nanoparticles are effective carriers for AsODN delivery, with formulation parameters significantly influencing their properties.
- PS-AsODN loaded NPs demonstrate superior encapsulation efficiency and gene silencing activity compared to PO-AsODN NPs.
- These findings support the potential of chitosan-based NPs as a viable in vivo delivery system for AsODN therapeutics.
