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T. Yamashita, S. Sonoda, K. Tachibana, R. Suzuki, T. Sakamoto; Gene Delivery by Combination of Bubble Liposome and Ultrasound for Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2714.
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© ARVO (1962-2015); The Authors (2016-present)
We reported gene transfer methods mediated by microbubble-enhanced sonoporation (Sonoda et al. IOVS 2006). The purpose of this study is to examine the efficacy and safety of a newly developed bubble liposome (BL) with sonoporation using corneal epithelial cells.
A new BL is a polyethyleneglycol modified liposomes containing prefluoropropane (Suzuki et al. J Cont. Release in press). Rat corneal epithelial cells (RC-1, 2x10-5 cells/well) were cultured in 24-well dishes. Plasmids containing cDNA of green fluorescent protein (GFP) were added to each culture followed by sonoporation with BL (BL-enhanced sonoporation group, N=40). To evaluate its efficacy, the similar experiments were performed in the following groups; sonoporation alone group (n=15), sonoporation and conventional microbubble group (n=15). The gene transfer efficacy was evaluated by immunofluorescent microscopy. Cell damage was evaluated by MTS-assay (N=12).
BL-enhanced sonoporation significantly increased the gene transfer efficacy (BL-enhaced sonoparation, 27%; Sonoporation alone, 2%; sonoporation and conventional microbubble, 11%: P<0.05: ANNOVA test). An equivalent gene transfer was achieved with less duration of ultrasound exposure in BL-enhanced sonoporation group. No apparent cell damage was found in BL-enhanced sonoporation.
A new BL-enhanced sonoporation can transfer the gene of interest to cultured corneal epithelial cells more effectively without damaging cells. This method will have a merit for future gene therapy for cornea without using viral-vector.
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