Prepare Of Paclitaxel Loaded HER-2 Targeted Plga-cooh Ultrasound Contrast Agent And In Vitro Experiment

PARTⅠPREPARATION AND TARGETING STUDY OF HERCEPTIN LOADED PLGA-COOH-TARGETED ULTRASOUND CONTRAST AGENTObjective To prepare a kind of Herceptin loaded and breast cancer targeted high molecular polymer ultrasound contrast agent and investigate their affinity for breast cancer in vitro.Methords The high molecular polymer PLGA-COOH ultrasound contrast agents were produced by the double emulsion technique.Herceptin was covalently linked to the PLGA-COOH nanoparticle surface using a carbodiimide technique. Its physical property was determined. The combination of Herceptin with the PLGA-COOH nanoparticle was proved by immunofluorescent assay ,and the PLGA-COOH nanoparticle targeting specifity to breast cancer cells was observed with light microscope and confocal microscope, normal PLGA-COOH nanoparticle was served as control group.Results The diameter range of targeted high molecular polymer ultrasound contrast agents was 466.8-857.6nm,the average diameter was 662.2±69.7nm,85.9% of the diameter was in the range.red punctiform fluorescence was observed by confocal microscope,and the conjugation of the targeted high molecular polymer ultrasound contrast agents with MCF-7 cells was tight while the conjugation in control group was negative.Conclusions The Herceptin loaded PLGA-COOH targeted ultrasound contrast agents was prepared successfully. The targeted ultrasound contrast agents can bind to breast cancer effectively in vitro.PARTⅡPRODUCTION OF PACLITAXEL LOADED PLGA-COOH-TARGETED ULTRASOUND CONTRAST AGENT AND IN VITRO EXPERIMENTObjective To prepare a kind of paclitaxel loaded PLGA-COOH targeted Ultrasound Contrast Agent,and to explore its effect of enhancing ultrasound imaging in vitro.Methods The paclitaxe loaded high molecular polymer PLGA-COOH ultrasound contrast agents were produced by the double emulsion technique. Herceptin was covalently linked to the paclitaxel loaded PLGA-COOH nanoparticle surface by carbodiimide technique. The physical property, entrapment efficiency and the drug-loading amounts of paclitaxel were determined. The antibody activity of Pac-UCA-HER was determined by ELISA.The paclitaxel loaded PLGA-COOH-targeted nanoparticle targeting specifity to breast cancer cells was observed with confocal microscope, and to explore its effect of enhancing ultrasound imaging in vitro.Results The average diameter of paclitaxel loaded PLGA-COOH targeted nanoparticle was (733.4±30.7) nm. The drug entrapment efficiency was (65.08±2.31)% and the drug-loading amounts was (6.51±0.23)%.The rate of antibody activity was (72.8±5.0)%.The conjugation of paclitaxel loaded PLGA-COOH-targeted nanoparticle with MCF-7 cells was tight while the control group was negative.In vitro experiment showed that they can enhance the ultrasound imaging greatly.Conclusions The paclitaxel loaded PLGA-COOH targeted nanoparticle with higher entrapment efficiency can bind to breast cancer effectively in vitro,and can also enhance the ultrasound imaging greatly.