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.
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.
Authors: Manuela Raviña; Eva Cubillo; David Olmeda; Ramón Novoa-Carballal; Eduardo Fernandez-Megia; Ricardo Riguera; Alejandro Sánchez; Amparo Cano; María José Alonso Journal: Pharm Res Date: 2010-09-21 Impact factor: 4.200
Authors: Michael D Buschmann; Abderrazzak Merzouki; Marc Lavertu; Marc Thibault; Myriam Jean; Vincent Darras Journal: Adv Drug Deliv Rev Date: 2013-07-18 Impact factor: 15.470
Authors: Julio C Fernandes; Xingping Qiu; Francoise M Winnik; Mohamed Benderdour; Xiaoling Zhang; Kerong Dai; Qin Shi Journal: Int J Nanomedicine Date: 2012-11-23