Effects of PEGylation and acetylation of PAMAM dendrimers on DNA binding, cytotoxicity and in vitro transfection efficiency
| Autor: | Per Lincoln, Elin K. Esbjörner, Alan Jenkins, Bengt Nordén, Kristina Fant, Martin C. Grossel |
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| Rok vydání: | 2010 |
| Předmět: |
Dendrimers
Genetic Vectors Pharmaceutical Science Biocompatible Materials CHO Cells Gene delivery DNA condensation Transfection Biophysical Phenomena Polyethylene Glycols chemistry.chemical_compound Cricetulus Drug Delivery Systems Dendrimer Cricetinae Drug Discovery Materials Testing Animals Cytotoxicity Chemistry Gene Transfer Techniques Acetylation DNA Biochemistry Biophysics PEGylation Molecular Medicine |
| Zdroj: | Molecular pharmaceutics. 7(5) |
| ISSN: | 1543-8392 |
| Popis: | Poly(amidoamine) (PAMAM) dendrimers are promising multipotent gene delivery vectors, providing favorable DNA condensation properties also in combination with the possibility of conjugation of different targeting ligands to their surface. They have been used for transfection both in vitro and in vivo, but their application is currently somewhat limited due to inherent cytotoxicity. In this work we investigate how two types of surface modification, acetylation and PEGylation, affect the DNA binding characteristics, the cytotoxicity and the in vitro transfection efficiency of generation 4 and 5 PAMAM dendrimers. Particularly, we address how the morphology of DNA-dendrimer complexes, formed under low salt conditions, changes upon dilution in cell growth medium, an event that inevitably occurs before the complexes reach the cell surface in any transfection experiment. We find that acetylation and PEGylation essentially eliminates the inherent dendrimer cytotoxicity. However, the transfection efficiency of the modified dendrimers is lower than that of the corresponding unmodified dendrimers, which can be rationally understood by our observations that DNA is less condensed when complexed with these modified dendrimers. Although small DNA-dendrimer particles are formed, the availability for ethidium intercalation and nuclease degradation is significantly higher in the modified DNA-dendrimer complexes than in unmodified ones. Dilution in cell growth medium has a drastic effect on these electrostatically assembled complexes, resulting in increase in size and DNA availability. Our results strongly add to the notion that it is of importance to perform a biophysical characterization under conditions as close to the transfection situation as possible, to enable conclusions regarding structure-activity relations of gene delivery vectors. |
| Databáze: | OpenAIRE |
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