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Effects of PEGylation and acetylation of PAMAM dendrimers on DNA Binding, cytotoxicity and in vitro transfection efficiency

Effects of PEGylation and acetylation of PAMAM dendrimers on DNA Binding, cytotoxicity and in vitro transfection efficiency
Effects of PEGylation and acetylation of PAMAM dendrimers on DNA Binding, cytotoxicity and in vitro transfection efficiency
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.

dendrimer, polyplex, biocompatibility, gene delivery, dna, characterization
1543-8384
1734-1746
Fant, Kristina
507d32c8-7d73-4e06-907f-51b0aecd2b55
Esbjörner, Elin
93fe2c7e-20c2-46dc-8fce-5afb8b59f576
Jenkins, Alan
cd72f4fc-eb55-4e5d-a598-218ada99f099
Grossel, Martin C.
403bf3ff-6364-44e9-ab46-52d84c6f0d56
Lincoln, Per
f1edcf40-411d-4f54-bd94-d13394c011d4
Nordén, Bengt
64d4009e-7456-4490-ac87-8d587334c7e0
Fant, Kristina
507d32c8-7d73-4e06-907f-51b0aecd2b55
Esbjörner, Elin
93fe2c7e-20c2-46dc-8fce-5afb8b59f576
Jenkins, Alan
cd72f4fc-eb55-4e5d-a598-218ada99f099
Grossel, Martin C.
403bf3ff-6364-44e9-ab46-52d84c6f0d56
Lincoln, Per
f1edcf40-411d-4f54-bd94-d13394c011d4
Nordén, Bengt
64d4009e-7456-4490-ac87-8d587334c7e0

Fant, Kristina, Esbjörner, Elin, Jenkins, Alan, Grossel, Martin C., Lincoln, Per and Nordén, Bengt (2010) Effects of PEGylation and acetylation of PAMAM dendrimers on DNA Binding, cytotoxicity and in vitro transfection efficiency. Molecular Pharmaceutics, 7 (5), 1734-1746. (doi:10.1021/mp1001312). (PMID:20695423)

Record type: Article

Abstract

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.

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More information

Published date: 9 August 2010
Keywords: dendrimer, polyplex, biocompatibility, gene delivery, dna, characterization

Identifiers

Local EPrints ID: 178861
URI: http://eprints.soton.ac.uk/id/eprint/178861
ISSN: 1543-8384
PURE UUID: 48f10dd4-0397-4ab5-9a91-6e9f254f8da8
ORCID for Martin C. Grossel: ORCID iD orcid.org/0000-0001-7469-6854

Catalogue record

Date deposited: 28 Mar 2011 15:07
Last modified: 19 Nov 2019 02:00

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