Study of SonoVue and ultrasound enhance liposome mediated gene transfection to retinal pigment epithelium in vitro

Abstract

Objective To compare the transfection rate and safety between SonoVue + ultrasound and liposome mediated gene transfection to retinal pigment epithelium (RPE) in vitro, and to approach the feasibility of SonoVue and ultrasound enhancing liposome mediated gene transfection to RPE in vitro. Methods RPE were cultured in 24 well dishes with green fluorescent protein plasmid of 2 μg/well. All the cells were divided into 5 groups: control group, ultrasound irradiance group, SonoVue + ultrasound irradiance group(2 W/cm2 and 3 W/cm2),liposome group and liposome + SonoVue + ultrasound irradiance group. There were 10 wells in every group. Gene transfection results were observed under inversion fluorescent microscope. Seventy-two hours later, gene transfer was guantified under flow cytometry. The apoptosis rates of RPE were determined with Annexin Ⅴ-FITC and PI staining by flow cytometry. Results The transfection rate of ultrasound exposure group was (1.06 ± 0.13)%. The transfection rate of SonoVue + ultrasound group was (15.81 ± 1.70)% and (20.86 ± 2.63)%. The transfection rate of liposome group was (30.06 ± 3.49)% ,and the differences between above mentioned sets were significant(P<0.05). The transfection rate of liposome + SonoVue + ultrasound was (44.51 ± 1.28) %,and the differences between above mentioned two sets were significant (P<0.05). Conclusions The transfection rate of liposome is higher than that of SonoVue + ultrasound group. SonoVue and ultrasound can obviously enhance liposome mediated EGFP gene transfection to RPE in vitro. Key words: Ultrasonography; Microbubbles; Liposomes; Transfection; Pigment epithelium of ege