The lower-generation polypropylenimine dendrimers are effective gene-transfer agents
BH Zinselmeyer, SP Mackay, AG Schatzlein… - Pharmaceutical …, 2002 - Springer
BH Zinselmeyer, SP Mackay, AG Schatzlein, IF Uchegbu
Pharmaceutical research, 2002•SpringerObjective. To evaluate polypropylenimine dendrimers (generations 1-5: DAB 4, DAB 8, DAB
16, DAB 32, and DAB 64) as gene delivery systems. Methods. DNA binding was evaluated
by measuring the reduced fluorescence of ethidium bromide, and molecular modelling of
dendrimer-DNA complexes also was performed. Cell cytotoxicity was evaluated against the
A431 cell line using the MTT assay. In vitro transfection was evaluated against the A431 cell
line using the β-galactosidase reporter gene and N-[1-(2, 3-dioleoyloxy) propyl]-N, N, N …
16, DAB 32, and DAB 64) as gene delivery systems. Methods. DNA binding was evaluated
by measuring the reduced fluorescence of ethidium bromide, and molecular modelling of
dendrimer-DNA complexes also was performed. Cell cytotoxicity was evaluated against the
A431 cell line using the MTT assay. In vitro transfection was evaluated against the A431 cell
line using the β-galactosidase reporter gene and N-[1-(2, 3-dioleoyloxy) propyl]-N, N, N …
Abstract
Objective. To evaluate polypropylenimine dendrimers (generations 1-5: DAB 4, DAB 8, DAB 16, DAB 32, and DAB 64) as gene delivery systems.
Methods. DNA binding was evaluated by measuring the reduced fluorescence of ethidium bromide, and molecular modelling of dendrimer-DNA complexes also was performed. Cell cytotoxicity was evaluated against the A431 cell line using the MTT assay. In vitro transfection was evaluated against the A431 cell line using the β-galactosidase reporter gene and N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methylsulphate (DOTAP) served as a positive control.
Results. Molecular modeling and experimental data revealed that DNA binding increased with dendrimer generation. Cell cytotoxicity was largely generation dependent, and cytotoxicity followed the trend DAB 64 > DAB 32 > DAB 16 > DOTAP > DAB 4 > DAB 8, whereas transfection efficacy followed the trend DAB 8 = DOTAP = DAB 16 > DAB 4 > DAB 32 = DAB 64.
Conclusion. The generation 2 polypropylenimine dendrimer combines a sufficient level of DNA binding with a low level of cell cytoxicity to give it optimum in vitro gene transfer activity.
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