lipofectamine and perflutren

Albumin microbubbles have been intensively studied for their application in gene delivery. However, with negative surface potential, albumin microbubbles hardly bind plasmid DNA, which might contribute to their low transgene efficiency. In this study, we developed polyethylenimine (PEI) coated albumin microbubbles (PAMB) which were prepared by sonicating the mixture of human albumin, PEI, polyethylene glycol and glucose. CHO cells, COS cells and 293T cells were transfected with PEI, PEI+albumin, PAMB and Lipofectamine 2000, respectively. Our results showed that the surface potential was elevated and PAMB could bind plasmid DNA. The transgene efficiency of PAMB was higher than PEI and PEI+albumin (P<0.05), and PAMB performed the same transgene effect as Lipofectamine 2000 did but with lower cytotoxicity than Lipofectamine 2000. Albumin microbubbles modified by PEI has high transgene efficiency and low cytotoxicity even without ultrasound medication, making it a useful non-virus gene delivery method in vitro.

Topics: Albumins; Animals; Cell Proliferation; Chlorocebus aethiops; CHO Cells; COS Cells; Cricetinae; Cricetulus; DNA; Fluorocarbons; Genetic Vectors; HEK293 Cells; Humans; Lipids; Microbubbles; Plasmids; Polyethylene Glycols; Polyethyleneimine; Sonication; Transfection; Transgenes

Small interfering RNA (siRNA) is expected to be a novel therapeutic tool, however, its utilization has been limited by inefficient delivery systems. Recently, we have developed novel polyethyleneglycol modified liposomes (Bubble liposomes; BL) entrapping an ultrasound (US) imaging gas, which can work as a gene delivery tool with US exposure. In this study, we investigated whether the BL were suitable for the delivery of siRNA. BL efficiently delivered siRNA with only 10 s of exposure to US in vitro. Specific gene silencing effects could be achieved well even in the presence of serum or with the disruption of endocytosis. We suggest that siRNA is directly introduced into the cytoplasm by the BL and US and the mechanism enables effective transfection within a short time and in the presence of high serum. Transfection of siRNA into the tibialis muscles with BL and US was also performed. The gene-silencing effect could be sustained for more than 3 weeks. Thus, BL could be a useful siRNA delivery tool in vitro and in vivo.

Topics: 1,2-Dipalmitoylphosphatidylcholine; Animals; Cell Line; Chlorocebus aethiops; Chloroquine; COS Cells; Cytoplasm; Fluorocarbons; Gene Expression; Gene Transfer Techniques; Kidney; Lipids; Liposomes; Luciferases, Firefly; Mice; Microbubbles; Muscle, Skeletal; NIH 3T3 Cells; Phosphatidylethanolamines; Polyethylene Glycols; RNA, Small Interfering; Serum; Skin; Transfection; Ultrasonics