Abstract
Abstract
Purpose
Cationic polymers have many advantages as vectors for mediated cellular entry and delivery of siRNA. However, toxicity related to their cationic charge has compromised clinical use. It is hypothesized that the siRNA-vector complex composition and properties can be controlled to optimize therapeutic performance. Here we investigate siRNA complexes with branched polyethylenimine (bPEI) versus generation 4 polyamidoamine dendrimers (PAMAM) on interactions with immobilized lipid membranes, and cellular uptake and toxicity.
Methods
A model siRNA was complexed with either PAMAM or bPEI, and their size and zeta-potential characterized. Interaction of the complexes and parent polymers with lipid bilayers was investigated using atomic force microscopy and correlated with the uptake and toxicity in HeLa cells.
Results
PAMAM and its siRNA complexes formed circular shaped micron-sized holes in lipid bilayers, while bPEI formed nanoscale holes. Flow cytometry and fluorescence microscopy demonstrated PAMAM-siRNA complexes to have a higher cellular uptake than bPEI-siRNA complexes. bPEI-siRNA complexes did not impact on viability, however PAMAM-siRNA complexes demonstrated increasing cell toxicity as N/P ratio increased. PAMAM-siRNA complexes accumulated around the cell nucleus, while PEI-siRNA complexes were located closer to the cell wall.
Conclusion
Complexation of PAMAM dendrimer or bPEI with siRNA modified physicochemical properties of the parent polymer, however it did not impact on the mechanism of interaction with model lipid bilayers or how the polymer/siRNA complex interacted and was internalized by HeLa cells. Interaction of siRNA polymer complexes with cells is related to the action of the parent polymer.
Graphical abstract
Funder
none
University of South Australia
Publisher
Springer Science and Business Media LLC
Subject
Pharmacology (medical),Organic Chemistry,Pharmaceutical Science,Pharmacology,Molecular Medicine,Biotechnology
Cited by
3 articles.
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