Experimental study of gene / mode of drug delivery and biological effects in the ultrasonic field
|School||University of Electronic Science and Technology|
|Keywords||Low-frequency ultrasound Microbubbles Gene therapy Model drug Cytoskeletal|
In recent years, with the progress of the development of ultrasound technology, applications, and developed microbubbles, ultrasound and microbubble contrast agent synergy in gene therapy, drug delivery, thrombolysis bolt, inflammation and tumor targeting developing treatment and other medical fields shows the huge potential of becoming a hot topic in the medical field. In this study of human umbilical vein endothelial cells (HUVECs) and HeLa cells as an in vitro cell model, as the purpose of the exogenous gene EGFP gene, nucleic acid fluorescent dye TO-PROTM-1, PI, DAPI as a model drug, respectively, by the MTT colorimetric analysis inverted fluorescence microscope, flow cytometry technology, MGG staining, Coomassie blue staining of a 20 kHz low-frequency ultrasound alone and in conjunction with microbubble contrast agent-mediated EGFP gene transfected endothelial cells, HeLa cells of efficiency and regularity, ultrasound and liposome jointly mediated EGFP gene transfected HeLa cells, efficiency and regularity of ultrasound-mediated mode of drug delivery efficiency and regularity, as well as ultrasound vascular endothelial cell proliferation activity of the growth curve, migration and cytoskeleton the impact. The results showed that: 20 kHz low-frequency ultrasound-mediated EGFP gene transfection of human umbilical vein endothelial cells and HeLa cells, but low transfection efficiency, generally in the range of 0.2% to 2%; cell types of the transfection efficiency. impact of HeLa cells transfected rate is higher than the efficiency of transfection of endothelial cells; influential plasmid concentration on transfection efficiency, the higher the greater the transfection efficiency of the plasmid concentration within a certain range; 2. ultrasound with liposomes jointly mediated conduction EGFP gene transfected HeLa cells, although the transfection efficiency is reduced with the increase of the irradiation time, but the transfection rate of the increase of the ultrasonic irradiation time increases with liposomes, which may be related to the cytotoxicity of ultrasound; 3. microbubble can significantly enhance the EGFP gene transfection of endothelial cells, and HeLa cells, transfection efficiency increased by an average of about 3 to 4 times, 3.5 times and 4.6 times, respectively, the highest; However, microbubble enhanced ultrasound on cells in a certain extent, will injury; 4. ultrasound mediated model drug delivery efficiency is high, but the law of the transporting efficiency and the ultrasound-mediated gene transfection is slightly different: the first increase with increasing ultrasonic irradiation time, the transfection efficiency, obtained in 6 seconds maximum, and then decreased gradually; drug delivery efficiency is achieved in 9 seconds, and then decreased; 5. excess of ultrasonic radiation will inhibit cell proliferation activity, the growth curve becomes more gentle, significantly reduce the migration ability of cells, so that decomposed skeleton structure, smaller structure become loose arrangement of the loss of orderliness, even broken; cytoplasmic will reduce obvious retraction.