CAJ | 학술논문

目的体外评价"亲和素-生物素"法制备靶向整合素αvβ3的微泡(MBαvβ3)及其黏附人脐静脉内皮细胞(HUVECs)的效能.方法 "亲和素-生物素"法桥连αvβ3抗体于磷脂微泡,体外荧光法鉴定其表面"生物素-亲和素-生物素"结构,应用平行平板流动腔(PPFC)技术评价MBαvβ3特异性黏附HUVECs的效能.结果加入荧光标记的亲和素后,生物素化微泡(MBB)表面可见明亮荧光,普通微泡未见荧光;MBB加入荧光标记的蛋白A也未见荧光;MBB结合亲和素后,再加入荧光标记的生物素或蛋白A,前者表面可见明亮荧光,后者未见荧光.PPFC内,MBαvβ3组和普通微泡组结合数分别为(9.9±3.1)微泡/HUVEC和(0.8±0.3)微泡/HUVEC(P<0.05).结论 "生物素-亲和素"法能成功制备MBαvβ3,具有特异黏附血管内皮细胞的能力.
목적 체외평개"친화소-생물소"법제비파향정합소αvβ3적미포(MBαvβ3)급기점부인제정맥내피세포(HUVECs)적효능.방법 "친화소-생물소"법교련αvβ3항체우린지미포,체외형광법감정기표면"생물소-친화소-생물소"결구,응용평행평판류동강(PPFC)기술평개MBαvβ3특이성점부HUVECs적효능.결과 가입형광표기적친화소후,생물소화미포(MBB)표면가견명량형광,보통미포미견형광;MBB가입형광표기적단백A야미견형광;MBB결합친화소후,재가입형광표기적생물소혹단백A,전자표면가견명량형광,후자미견형광.PPFC내,MBαvβ3조화보통미포조결합수분별위(9.9±3.1)미포/HUVEC화(0.8±0.3)미포/HUVEC(P<0.05).결론 "생물소-친화소"법능성공제비MBαvβ3,구유특이점부혈관내피세포적능력.
Objective To identify microbubbles targeted (MBt) to alpha(v)beta(3) (αvβ3) via biotin-avidin bridge and evaluate the adhesion to human umbilical vein endothelial cells (HUVECs) in vitro.Methods MBt produced via biotin-avidin bridge were validated using fluorescence in vitro.Adhesion of αvβ3-integrin targeted MBt (MBαvβ3) to HUVECs was tested using the parallel plate flow chamber (PPFC) test.Results Bright green fluorescence was observed on the biotinylated microbubbles(MBB) incubated with fluorescein isothiocyanate labeled streptavidin (FITC-SA) and on MBB-SA incubated with FITC labeled biotin.There was no fluorescence seen on non-targeted control microbubbles,MBB incubated with FITC labeled protein A and MBB-SA incubated with FITC labeled protein A. The adherent rate of MBαvβ3 was significantly higher than MBt with non-specific antibody (MBN) in PPFC test,with 9.9±3.1 of MBαvβ3 and 0.8±0.3 of MBN adhered to HUVECs,respectively(P<0.05).Conclusions Avβ3 targeted microbubbles using biotin-avidin bridging method is highly efficient and reliable for HUVECs.