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Ultrasound-mediated lipid microbubble destruction enhances pEGFP-N1 expression in human periodontal ligament fibroblasts cell

ZHANG Yun-yan1,LI You-qiang2,LUO Shu-mei1,ZHONG Xiao-bo1,WANG Liu3,WANG Zhi-gang4(1.Department of Conservative Dentistry And Endodontics,the Affiliated Hospital of Stomatology,Chongqing Medical University,Chongqing 400015,China;2.China Faculty of Laboratory Medicine,Chongqing Medical University,Chongqing 400016,China;3.Department of Stomatology,Chongqing Maternal and Child Health Hospital,Chongqing 400012,China;4.Institute of Ultrasonic Image,Chongqing Medical University,Chongqing 400010,China)  
Objective To investigate whether ultrasound-mediated microbubble destruction can safely and effectively deliver pEGFP-N1 plasmid to human periodontal ligament fibroblasts.Methods The primary cultured human periodontal ligament fibroblasts were divided into five groups.Experiment groups were ultrasound+microbubble+plasmid.Based on different conditions of transfection,experiment groups were further divided into sub groups.Control groups included naked plasmid,ultrasound+plasmid,microbubble+plasmid and liposome+plasmid.After 48 h,GFP expression in the human periodontal ligament fibroblasts was detected using phase-contrast fluorescence microscopy.MTT was adopted to measure the cells vitality of every group.Results The transfection efficiency of ultrasound+microbubble+plasmid group was similar to that of the liposome+plasmid group,which were both higher than that of the other control groups,but the cells vitality of the ultrasound+microbubble+plasmid group was higher than that of the liposome+plasmid group.Conclusion Under some specific conditions,ultrasound mediated microbubble destruction method can safely and effectively enhance the reporter gene transfection and expression.
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