Preparation of vascular endothelial growth factor receptor 2/Integrinαv β3 dual‐targeted contrast ultrasound agent and its ultrasound imaging features and targeted ability :a in vitro study
10.3760/cma.j.issn.1004‐4477.2019.03.014
- VernacularTitle:血管内皮生长因子受体2及整合素双靶向超声微泡的制备及其体外成像与寻靶能力检测
- Author:
Cuixian LI
1
;
Beijian HUANG
;
Haixia YUAN
;
Cong LI
;
Wenqing WU
;
Wenping WANG
Author Information
1. 复旦大学附属中山医院超声科
- Keywords:
Ultrasonography;
Contrast agent;
Target;
Vascular endothelial grow th factor;
Integrin
- From:
Chinese Journal of Ultrasonography
2019;28(3):261-266
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare the vascular endothelial grow th factor receptor 2 ( VEGFR2 )/Integrinαv β3 dual‐targeted contrast ultrasound agent ,and further evaluating the physical properties ,imaging characteristics and targeted ability in v itro . Methods VEGFR2 targeted microbubble ( MBV ) ,Integrinαv β3 targeted microbubble ( MBI) and VEGFR2/Integrinαv β3 dual targeted microbubble ( MBD) were prepared by attaching VEGFR2 antibody ,Integrinαv β3 antibody and both of VEGFR2/Integrinαv β3 antibody with no targeted USphere LA respectively , using biotin‐avidin linkage method . USphere LA with no antibody attached were used as non‐targeted microbubble( MBN ) . M icrobubble′s physical properties were observed , and its stability was detected by caculating bubble′s concentration at different time points . MBD′s ultrasound imaging characteristics were evaluated by comparing its grey level of ultrasound imaging with SonoVue at the time of preparation and 3 days after preparation . To detect the targeted ability of microbubble ,different types of microbubble were added into Hepa 1‐6 and C3H10 cells ,respectively . T he above procedure was repeated using the pre‐antibody blocking test in Hepa 1‐6 cell . Results MBD had a mean size of 1 256 nm .T he concentration of microbubble in the first duration of three days was declining slowly ,and its speed was accelerated after five days of preparation . T he grey level of new prepared MBD was similar to that of SonoVue in the same concentration ( P ＝0 .113) ,while the level was higher than that of SonoVue within 3 days after preparation( P ＜0 .001) . T he number of microbubble binding to Hepea1‐6 led a tendency of MBD＞ single targeted microbubble ＞ MBN ( all P ＜0 .05 ) . T he number of C3H10 and pre‐blocked Hepa1‐6 cell attached to each group of microbubble had no statistical difference ( all P ＞0 .05) . In addition ,following the pre‐blocked precedure ,the number of Hepa 1‐6 cell attached to each group of microbubble had no statistical difference either ( all P ＞ 0 .05 ) . Conclusions VEGFR2/Integrinαv β3 dual‐targeted contrast ultrasound agent is a stable microbubble and it has excellent ultrasound imaging and targeting ability in v itro .
