Study of preparation of the Annexin V-nanoscale ultrasound contrast agents and targeting ultrasound imaging in vitro / 中华超声影像学杂志

ABSTRACT

Objective To research the Annexin V-nanoscale ultrasound contrast agents'preparation, ultrasound imaging and the ability to binding apoptosis cells of tumor in vitro.Methods The nanoscale bubble (Nanobubbles,NBs ) packaged the octaflouropropane (C3 F8 ) gas was prepared by thin film hydration.The Annexin V-Nanobubbles (AVNBs ) solutions was acquired through conjugating the biotinylated-Annexin V to the surface of the NBs by biotin-streptavidin bridging chemistry.The size and zeta potential of AVNBs were measured by NanoPlus-3 zeta/nano particle analyzer.The shift in size distribution of AVNBs bubbles was analyzed for the stability,after it was stored at 4 ℃ for different time. AVNB's shape were measured by scanning electron microscopy.The AVNBs bubble was measured using an ultrasound system for echogenicity in vitro,and SonoVue was for control.Finally,the ability of AVNBs binding with apoptosis cells of tumor in vitro was determine via the fluorescence microscope.Results AVNBs has a size distribution of (640.2±32.1 )nm,and a mean zeta potential of (-23.30 ±5.71 )mV.Its size remained relatively constant and appeared to show less size variation within the 24 h analysis period. AVNBs solutions were visible milky white and slightly suspension liquid with the naked eye.Under scanning electron microscopy,AVNBs were uniform hollow sperical cavitation bubble with small size and larger dispersibility in solution.The AVNBs and SonoVue solution had the same higher grayscale signal intensity by ultrasonic imaging.The AVNBs binded well with apoptosis cells of tumor in vitro,and the rate of binding was (97.55 ± 1 .30 )%.Conclusions The AVNBs particles prepared by method of thin film hydration have a nanoscale size,good stability and echogenicity.It can be targeted binding with the apoptosis cells of tumor in vitro.