The Bioeffect Of Focused Ultrasoud Near Feild On ECV304 In Vitro

BackgroundFocused ultrasound is a non-invasive technical that has been widely applied in clinical therapy,especially in the treatment of refractory and high recurrence rate diseases,such as the effective rate of vulvar white lesion treatment reaches to 94.7%.Relative to the biological effect or risk studies,that is,the biophysical mechanisms by which ultrasound affects biological materials,ultrasound-induced biological effects are generally separated into thermal and non-thermal mechanisms.The near field of focused ultrasound acoustic we disscussed here has simple far field and complicated near field with inhomogeneous acoustic intensity,which indicates complexed biological effect.In clinical study,the number of endothelial cells,microvessel density increasing and microvessel remodeling were found in lesion tissues by tissue biopsy after focused ultrasound treatment.From another point of view for this,it indicates most of endothelial cells in tissue were treated with complicated near field in focused ultrasound acoustic channel and proliferated and generated vessal remolding in secondary tissuse reconstruction.But the reason is not clear.Based on the effects of focused ultrasound near field with vulvar white lesion clincal treatment parameters on relative survial ratio of endothial cells in vitro,combined with proteomics method was performed in the biology effect study to identified proteins which were participated in.Objective1. The effect of focused ultrasound near field on ECV304 proliferation in vitro.2. The effect of focused ultrasound near field on ECV304 proteins in vitro.Methods1. The effect of focused ultrasound near field on ECV304 proliferation.Groups in Experiment: irradiation power:4.3,5 W;irradiation frequency:10.1,11.2 MHz; irradiation times:5,10,15,20,25,30 s.Experiment method:MTT method was performed in detecting the proliferation of ECV304 treated with focused ultrasound near field under different parameters.2. The selection of the optimized focused ultrasound near field irradation parameters. The optimized irradiation parameters was selected by analyzed ECV304 relative survival ratios(OD treatment group/OD normal group) with SAS statistical software.3. The effect of focused ultrasound near field on proteins of ECV304.Groups in experiment:normal group,focused ultrasound near field irradation group(irradation parameters:11.2 MHz,4.3 W,10 s).Experiment method: one-dimensional SDS-PAGE electrophoresis(chromatography) liquid chromatography-electrospray ironzation-iron trap two-stage mass spectrometry (LC-ESI-IT MS/MS) was identified differential proteins of ECV304 pre and post focused ultrasound near field irradation.The expression of Actin of ECV304 was analyzed by western blot.Results1. ECV304 proliferation after focused ultrasound near field irradation depends on irradation power,frequency and times.Comparied with the other control groups,the degree of proliferation was biggest with irradation power 4.3 W,frequency 11.2 MHz,time 10 s.The ECV304 growing morphology was closed to subcircular and circular.2. Combined with relative references and reality, irradation power 4.3 W,frequency 11.2 MHz,time 10 s was considered applying in secondary experiment. 3. Total proteins of ECV304 pre and post focused ultrasound near field irradation was significant different in the area which relative molecular weight was at 95~130 kD through one-dimensional SDS-PAGE electrophoresis.4. 7 differential proteins were identified by liquid chromatography-electrospray ironzation-iron trap two-stage mass spectrometry (LC-ESI-IT MS/MS).Most of them were paticipated in cell signal transduction,29%,cytoskeletal protein,28%,paticipated motility.5. The expression of Actin of ECV304 pre and post focused ultrasound near field was analyzed by western blot,agreement with LC-ESI-IT MS/MS.Conclusions1. The proliferation and morphology of ECV304 were both affected by focused ultrasound near field irradation.Irradation power 4.3 W,frequency 11.2 MHz,time 10 s were the optimal parameters benefit for the proliferation.2. The composition of proteins in ECV304 was changed after focused ultrasound near field irradation,which was the foundation for cell morphology and function adaptability change.