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Cationic lipid-coated PEI/DNA polyplexes with improved efficiency and reduced cytotoxicity for gene delivery into mesenchymal stem cells.

ABSTRACT: BACKGROUND: Effective gene transfection without serum deprivation is a prerequisite for successful stem cell-based gene therapy. Polyethylenimine (PEI) is an efficient nonviral gene vector, but its application has been hindered by serum sensitivity and severe cytotoxicity. METHODS: To solve this problem, a new family of lipopolyplexes was developed by coating PEI/DNA polyplexes with three serum-resistant cationic lipids, namely, lysinylated, histidylated, and arginylated cholesterol. The physical properties, transfection efficiency, cellular uptake, subcellular distribution, and cytotoxicity of the lipopolyplexes was investigated. RESULTS: The outer coat composed of lysinylated or histidylated cholesterol remarkably improved the transfection efficiency of the polyplex with a low PEI/DNA ratio of 2 in the presence of serum. The resulting lysinylated and histidylated cholesterol lipopolyplexes were even more efficient than the best performing polyplex with a high PEI/DNA ratio of 10. Results from cellular uptake and subcellular distribution studies suggest that their higher transfection efficiency may result from accelerated DNA nuclear localization. The superiority of the lipopolyplexes over the best performing polyplex was also confirmed by delivering the therapeutic gene, hVEGF(165). Equally importantly, the lipid coating removed the necessity of introducing excess free PEI chains into the transfection solution for higher efficiency, generating lipopolyplexes with no signs of cytotoxicity. CONCLUSION: Noncovalent modification of polyplexes with lysinylated and histidylated cholesterol lipids can simultaneously improve efficiency and reduce the toxicity of gene delivery under serum conditions, showing great promise for genetic modification of bone marrow stem cells.

SUBMITTER: Song H

PROVIDER: S-EPMC3428250 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Cationic lipid-coated PEI/DNA polyplexes with improved efficiency and reduced cytotoxicity for gene delivery into mesenchymal stem cells.

Song Hongmei H Wang Gang G He Bin B Li Li L Li Caixia C Lai Yusi Y Xu Xianghui X Gu Zhongwei Z

International journal of nanomedicine 20120822

<h4>Background</h4>Effective gene transfection without serum deprivation is a prerequisite for successful stem cell-based gene therapy. Polyethylenimine (PEI) is an efficient nonviral gene vector, but its application has been hindered by serum sensitivity and severe cytotoxicity.<h4>Methods</h4>To solve this problem, a new family of lipopolyplexes was developed by coating PEI/DNA polyplexes with three serum-resistant cationic lipids, namely, lysinylated, histidylated, and arginylated cholesterol ...[more]

PMID: 22942645

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Project description:The effective delivery of DNA locally could increase the applicability of gene therapy in tissue regeneration and therapeutic angiogenesis. One promising approach is through use of porous hydrogel scaffolds that incorporate and deliver DNA in the form of nanoparticles to the affected sites. While we have previously reported on caged nanoparticle encapsulation (CnE) to load DNA polyplexes within hydrogels at high concentrations without aggregation, frequent issues with limited polyplex release following CnE have been encountered. In this study, we report two alternative approaches to polyplex presentation for decreasing aggregation in porous hydrogels. The first approach reduces polyplex aggregation by utilizing polyethylene glycol modification of the gene carrier polymer polyethyleneimine (sPEG-PEI) to mitigate charge-charge interactions between polyplexes and the scaffold during gelation. The second approach electrostatically presents polyplexes on the surfaces of scaffold pores as opposed to an encapsulated presentation. The sPEG-PEI polymer formed a smaller, less toxic, and more stable polyplex that exhibited less aggregation within HA gels when compared to the traditionally used linear PEI (LPEI) polymer. Surface-coated polyplexes also resulted in a more homogenous distribution of polyplexes in hydrogels. Furthermore, sPEG-PEI polyplexes retained transfection abilities comparable to LPEI in 3D surface-coated transfections. These results demonstrate a significant improvement in scaffold-mediated gene delivery and show promise in applications to multi-gene delivery systems.Statement of significanceA promising gene delivery approach for regenerative medicine is implanting porous hydrogel scaffolds loaded with DNA nanoparticles for delivery to affected sites. However, loading DNA polyplexes at high concentrations within hydrogels results in significant aggregation. Here, we describe two methods for decreasing aggregation of DNA polyplexes in porous gels. First, the gene carrier polymer polyethyleneimine (PEI) was modified with polyethylene glycol (sPEG-PEI) to mitigate the electrostatic interactions between polyplexes and scaffold polymer to in turn decrease aggregation. Second, polyplexes were presented along the surfaces of the pores of the hydrogel instead of being encapsulated within the gel. These methods allow for highly tunable and sustained transgene expression from scaffold-mediated gene delivery while avoiding polyplex aggregation.

Dataset Information

Cationic lipid-coated PEI/DNA polyplexes with improved efficiency and reduced cytotoxicity for gene delivery into mesenchymal stem cells.

Publications

Cationic lipid-coated PEI/DNA polyplexes with improved efficiency and reduced cytotoxicity for gene delivery into mesenchymal stem cells.

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