膀胱癌BIU-87细胞靶向超声纳米脂质微泡造影剂的制备及体外超声成像

Preparation of BIU-87 cells targeting nanobubbles as ultrasound contrast agent and imaging study in vitro

目的制备具有膀胱癌BIU-87细胞靶向结合的超声纳米脂质微泡造影剂,并观察体外靶向能力、检测其对细胞增殖的影响和体外超声成像效果。方法通过机械振动法制备纳米脂质微泡,用生物素-亲和素桥接法将NYZL1连接到纳米脂质微泡的表面制备靶向纳米脂质微泡造影剂,利用倒置显微镜观察其一般特性,Zate检测仪检测粒径大小和表面电荷,使用倒置显微镜来观察其体外靶向效果,用MTT法检测其不同浓度对细胞增殖的影响,使用超声诊断仪观察体外显影效果。结果膀胱癌BIU-87细胞靶向超声纳米脂质微泡呈圆形,分布均匀,无聚集;在体外靶向实验中,靶向纳米脂质微泡造影剂能高效特异性的结合在膀胱癌BIU-87细胞表面并沿细胞表面规则排列;不同浓度下两种微泡对细胞增殖无明显变化(P>0.05);体外超声成像实验中,靶向纳米微泡呈点状细密高回声。结论本实验成功制备膀胱癌BIU-87细胞超声纳米脂质微泡造影剂,能够在体外与膀胱癌BIU-87细胞特异性地靶向结合,两种微泡对细胞生长无明显影响且体外超声显像效果良好。

Objective To prepare nanobubbles specifically targeting BIU-87 cells,and to observe the targeting ability and effect on cell proliferation and ultrasound imaging in vitro.Methods Mechanical vibration method was used to prepare non-targeting nanobubbles.The biotin-avidin bridge method was used to conjugate biotinylated NYZL1,and non-targeting nanobubbles linked to biotin-NYZL1 become targeted nanobubbles.The morphology,concentration and shelf life of nanobubbles were detected with optical microscope,and the particle size and surface charge were measured with Zate detector.The targeting ability was observed with microscope.The proliferation of cells was detected by MTT method.Ultrasound imaging was observed in vitro.Results The targeted nanobubbles were successfully generated,which were round,evenly distributed,without aggregation.In targeted in vitro experiment,the nanobubbles attached to the surface of BIU-87 cells firmly and regularly arranged along the cell surface.There was no significant difference between different concentrations of nanobubbles on cell proliferation（P0.05）.In in vitro ultrasound imaging experiment,the targeted nanobubbles showed fine punctuate hyper echo.Conclusion The targeted nanobubbles are successfully prepared,which can specifically target and attach to BIU-87 cells in vitro and their ultrasound imaging is fine.