The influence of microbubble's concentration on bioeffects induced by ultrasound-mediated microbubble destruction / 中华超声影像学杂志

ABSTRACT

Objective To investigate the effect on bioeffects such as sonoporation and cell killing that induced by domestic ultrasound contrast agent(perfluoropropane-albumin microsphere)and diagnostic ultrasound by microbubble concentration.Methods Suspensions of hepatocyte with microbubbules in different concentrations were exposed to diagnostic ultrasound.The study included blank contrast group(no microbubble,no ultrasound),exposed group(no microbubble,exposed to ultrasound)and 6 microbubble groups in different concentration(with different microbubbule/cell ratios for 1,5,10,50,100 and 200,exposed to ultrasound for 10min).The uptake of fluorescein isothiocyanatedextran(FD500)by hepatocyte was observed and the percentages of sonoporation cells were counted,the cell viability was determined by trypan blue stain immediately after exposure,and apoptosis of ceils were detected by flow cytometry,with double staining of fluorescein isothiocyanate(FITC)-labeled Annexin V/propidium iodide(PI).Results Fluorescence stain results:compared with blank contrast group or exposed group,the sonoporatin of groups with microbubble/cell ratio of 5 to 200 increased significantly(P＜0.05).Cell killing effectscompared with the blank contrast group,only the cell death percentages of microbubble groups with microbubble/cell ratio of 50,100 and 200 increased significantly(P＜0.05).Compared with the exposed group,only that of groups with microbubble/cell ratio of 100 and 200 increased significantly(P＜0.05).There was no significant different between blank contrast group and exposed group for all these results.Conclusions Sonoporation and cell killing effect can be induced by diagnostic ultrasound in HL-7702 with domestic ultrasound contrast agent(perfluoropropane-albumin microsphere).For diagnosis,the ratio of microbubbule/cell should better be under 10.While for gene transfection,the ratio should better be 50 and could be 100 if it is necessary.