Experimental Study Of Function Of Targeted Nano-lipid Ultrasound Contrast Agent In Vitro And To Evaluate The Rabbit VX2Tumor Imaging

PART IIN VITRO EVALUATION OF TARGETING CAPABILITY OFTARGETED LIPID NANOPARTICLES COMBINING ANTI-ICAM-1MONOCLONE ANTIBODYObjective To construct targeted lipid nanoparticles combining theanti-human ICAM-1moloclone antibodies to the shellvia“avidin-biotin”bridging chemistry and to evaluate its ability of targetingto HUVEC cells.Methods Common lipid nanoparticles and biotined lipid nanoparticleswere prepared by perfluorocarbon gas with several lipids in a certainratio.Then more avidin was added to incorporate to the biotinednanoparticles. After being washed to remove excess avidin, a dose of biotinmouse-anti human ICAM-1monoclone antibodies was added to completethe preparation of targeted lipid nanoparticles. Detect the partical size and electric potential. HUVEC was divided into4groups randomly:①c＋pgroup(given common nanoparticles)②c＋p-Ab group(given targetednanoparticles)③s＋p group（given TNF-α and common nanoparticles）④s＋p-Ab group （given TNF-α and targeted nanoparticles）.The ability oflipid nanoparticles targeting to HUVEC was observed on fluorescentmicroscopy.Results Common nanoparticles partical size were(623.21±91.37）nm,targeted nanoparticles partical size were(760.61±145.02)nm. Both of c＋pand c＋p-Ab groups had little nanoparticles adhere to HUVEC. A greatquantity of green nanoparticles targeted to HUVEC in s＋p-Abgroup,which is higher than s＋p group dramatically.Conclusion This study constructed a type of targeted lipidnanoparticles conjugating mouse-anti human ICAM-1monocloneantibodies on the shell successfully,which has little partical size and wasappropriate for targeting HUVEC in vitro. PART IICOMPARISON OF SELF-MADE TARGETED LIPOSOMENANOPARTICLES ULTRASONIC CONTRAST ANGENTAND GENERAL LIPID MICROBUBBLES INULTRASOUND CONTRAST IMAGING OF RABBIT VX2TUMOR MODELObject To investigate the ultrasound imaging characteristics of theself-made lipid nanoparticles targeted ultrasound contrast agents in VX2rabbit tumor model.Methods To construct targeted lipid nanoparticles combining the antiICAM-1moloclone antibodies to the shell via “avidin-biotin” bridgingchemistry. Inject self-made common lipid nanoparticles and targetednanoparticles ultrasound contrast agents into rabbit ear vein with1ml/kgdose. Use ultrasound imaging diagnostic mode, real-time monitor twodifferent ultrasound contrast agent imaging characteristics of their own.Collect images to evaluate the imagings in different time point respectively.Analyze the peak time and the peak echo intensity with DFY-Ⅱ. Theparameters of each group were compared.Results Control lipid ultrasound contrast agent time to peak was20s and the targeted self-made lipid nanoparticles ultrasound contrast agentwas30s. Peak intensity of the targeted group was107.34±4.40dB which ishigher than that of the control group67.88±4.29dB. In control group, thedifference value of positive peak is63.84±3.58dB in20s and get to thenegative peak-68.54±1.91dB in90s. In targeted group, the difference value of positive peak is94.6±3.07dB in30s and get to the negative peak-68.88±4.71dB in90s.Conclusions Self-made lipid nanoparticles targeted ultrasoundcontrast agents have their own unique processes and characteristics inultrasound imaging, which can enhance the grey value imaging of the livertumor greatly and may be helpful for tumor diagnosis.