Effects of the Incorporation of a Hydrophobic Middle Block into a PEG−Polycation Diblock Copolymer on the Physicochemical and Cell Interaction Properties of the Polymer−DNA Complexes
- 22 October 2008
- journal article
- research article
- Published by American Chemical Society (ACS) in Biomacromolecules
- Vol. 9 (11), 3294-3307
- https://doi.org/10.1021/bm800876v
Abstract
One-component homopolymers of cationic monomers (polycations) and diblock copolymers comprising poly(ethylene glycol) (PEG) and a polycation block have been the most widely used types of polymers for the formulation of polymer-based gene delivery systems. In this study, we incorporate a hydrophobic middle block into the conventional PEG−polycation architecture and investigate the effects of this hydrophobic modification on the physicochemical and cell-level biological properties of the polymer−DNA complexes that are relevant to gene delivery applications. The ABC-type triblock copolymer used in this study consists of (A) PEG, (B) hydrophobic poly(n-butyl acrylate) (PnBA), and (C) cationic poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) component polymers. The properties of the triblock copolymer/DNA complexes are compared with those of two other more conventional DNA carriers derived, respectively, using a PDMAEMA homopolymer and a PEG−PDMAEMA diblock copolymer that had comparable molecular weights for individual blocks. In aqueous solution, the PEG−PnBA−PDMAEMA polymer forms positively charged spherical micelles. The electrostatic complexation of these micelles with plasmid DNA molecules results in the formation of stable small-sized DNA particles that are coated with a micelle monolayer, as confirmed by agarose gel electrophoresis, dynamic light scattering (DLS), and cryogenic transmission electron microscopy (cryo-TEM). Proton nuclear magnetic resonance (1H NMR) spectroscopy measurements indicate that the whole micelle−DNA assembly (named “micelleplex” for convenience) is shielded predominantly by the PEG chains. DLS and optical microscopy imaging measurements indicate that compared with PDMAEMA−DNA polyplexes, the micelleplexes have a significantly lower tendency to aggregate under physiological salt concentrations and show reduced interactions with negatively charged components in serum such as albumin and erythrocytes. While the micelleplexes are comparable to the PEG−PDMAEMA-based DNA polyplexes in terms of their stability against aggregation under high salt concentrations and in the presence of the albumin protein, they have a slightly higher tendency to interact with erythrocytes than the diblock copolymer polyplexes. Agarose gel electrophoresis measurements indicate that relative to the PEG−PDMAEMA polyplexes, the micelleplexes provide better protection of the encapsulated DNA from enzymatic degradation and also exhibit greater stability against disintegration induced by polyanionic additives; in these respects, the PDMAEMA homopolymer-based polyplexes show the best performance. In vitro studies in HeLa cells indicate that the PDMAEMA polyplexes show the highest gene transfection efficiency among the three different gene delivery systems. Between the micelleplexes and the PEG−PDMAEMA polyplexes, a higher gene transfection efficiency is observed with the latter system. All three formulations show comparable levels of cytotoxicity in HeLa cells.Keywords
This publication has 54 references indexed in Scilit:
- The Micellar Shuttle: Thermoreversible, Intact Transfer of Block Copolymer Micelles between an Ionic Liquid and WaterJournal of the American Chemical Society, 2006
- Circulating DNA and DNase Activity in Human BloodAnnals of the New York Academy of Sciences, 2006
- Polymers for gene delivery across length scalesNature Materials, 2006
- A PEG‐Based Biocompatible Block Catiomer with High Buffering Capacity for the Construction of Polyplex Micelles Showing Efficient Gene Transfer toward Primary CellsChemMedChem, 2006
- Biophysical Characterization of Complexation of DNA with Block Copolymers of Poly(2-dimethylaminoethyl) Methacrylate, Poly(ethylene oxide), and Poly(propylene oxide)Langmuir, 2005
- Progress and problems with the use of viral vectors for gene therapyNature Reviews Genetics, 2003
- Perspectives on Polymeric Gene DeliveryJournal of Bioactive and Compatible Polymers, 2003
- Observation of DNA–polymer condensate formation in real time at a molecular levelFEBS Letters, 2000
- Effect of serum components on the physico-chemical properties of cationic lipid/oligonucleotide complexes and on their interactions with cellsBiochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism, 1998
- X-ray diffraction studies on cation-collapsed DNAJournal of Molecular Biology, 1984