DNA and polylysine adsorption and multilayer construction onto cationic lipid-coated microbubbles.
We report on a novel application of the layer-by-layer (LbL) assembly technique to attach multiple layers of DNA and poly-l-lysine (PLL) onto preformed lipid-coated microbubbles to increase the DNA loading capacity. We first measured the effects of the cationic lipid fraction and salt concentration on the microbubble stability. Microbubble production and stability were robust up to a cationic lipid fraction of 40 mol % in 10 mM NaCl. DNA adsorption was heterogeneous over the microbubble shell and occurred primarily on the condensed phase domains. The amount of adsorbed DNA, and subsequently adsorbed PLL, increased linearly with the fraction of cationic lipid in the shell. DNA loading was further enhanced by the LbL assembly method to construct polyelectrolyte multilayers (PEMs) of DNA and PLL. PEM buildup was demonstrated by experimental results from zeta potential analysis, fluorescence microscopy, UV spectroscopy, and flow cytometry. The PEMs exhibited two growth stages and were heterogeneously distributed over the microbubble surface. The DNA loading capacity onto the microbubbles was enhanced by over 10-fold by using five paired layers. However, the PEM shell did not prevent oscillation or destruction during ultrasound insonification. These results suggest that the surface can be compartmentalized to make multifunctional, high-payload ultrasound contrast agents for targeted gene therapy.
Biomedical Engineering Department, University of California, Davis, California 95616, USA. mb2910@columbia.edu
Mark A
MA
Borden
Charles F
CF
Caskey
Erika
E
Little
Robert J
RJ
Gillies
Katherine W
KW
Ferrara
D000327:Adsorption; D000818:Animals; D002412:Cations / Q000737:chemistry; D003102:Colloids / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004573:Electrolytes; D008055:Lipids / Q000737:chemistry*; D008297:Male; D008854:Microscopy, Electron; D009994:Osmolar Concentration; D011107:Polylysine / Q000737:chemistry*; D012474:Salmon; D013094:Spermatozoa
D002412:Cations; D003102:Colloids; D004573:Electrolytes; D008055:Lipids; D011107:Polylysine; D004247:DNA