Author:
Dorris Annie C,Douglas Kimberly L,Tabrizian Maryam,Barrett Christopher J
Abstract
Nanoparticles coated with DNA and poly(L-lysine) (PLL) were prepared using a layer-by-layer assembly technique under various solution conditions. Both the level of DNA loading into the materials, the rate and the extent of DNA released from films upon degradation were controlled by varying the pH of polyelectrolyte assembly solutions. To determine optimal conditions for DNA loading, analogous films assembled on flat surfaces were prepared under various pH conditions. Using UV–vis spectroscopy and thin-film ellipsometry, it was found that the extent of DNA incorporation could be varied by more than a factor of two, and that the highest loading was obtained for films built using a combination of DNA and poly(L-lysine) solutions of pH = 4 and pH = 7, respectively. The layers coated onto the silica nanoparticles permitted the surface charge to be characterized by zeta potential electrophoresis. Furthermore, the acid–base dissociation constant measured for PLL on the outermost layer of the DNA/PLL film showed that the pKaof PLL can be shifted by more than three units. Film degradability was investigated via the exposure of films assembled under different pH conditions to α-chymotrypsin. The fraction of DNA released from degraded films can also be increased by a factor of three when films are built under conditions of pH = 4 for the DNA solution. The resultant effect on the transfection ability of pEGFP-N1/PLL coated particles was then measured, and results suggest that the control achieved over the bulk film properties also extends to a strong influence on cell uptake and transfection.Key words: polyelectrolyte multilayer, DNA incorporation, enzymatic degradation, nanoparticles, gene therapy.
Publisher
Canadian Science Publishing
Subject
Organic Chemistry,General Chemistry,Catalysis
Cited by
10 articles.
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