携RGD肽靶向高分子造影剂的制备及体外靶向实验

Preparation of RGD-PLGA and in vitro targeting study

目的制备携RGD肽的PLGA造影剂,并观察其在体外的靶向能力。方法采用双乳化法制备PLGA-COOH造影剂,利用碳二亚胺法将RGD肽与PLGA-COOH造影剂共价结合,制备出RGD-PLGA的靶向造影剂。用激光共聚焦检测FITC-RGD与PLGA造影剂的连接情况,并在光镜及荧光显微镜下对比观察RGD-PLGA与普通PLGA靶向SKOV-3细胞的能力,而后利用流式细胞检测技术对比RGD-PLGA与普通PLGA的靶向能力。结果连接FITC-RGD的PLGA显示出明显的绿光,体外靶向实验显示RGD-PLGA较普通PLGA更多地聚集在SKOV-3细胞的周围,流式细胞检测显示RGD-PLGA较普通PLGA有更好的靶向性。结论成功制备出RGD-PLGA,且在体外实验中对高表达整合素αvβ3的SKOV-3细胞有较强的结合能力。

Objective To synthesize RGD-PLGA and further explore its targeting ability in vitro.Methods High molecular polymer PLGA-COOH was prepared using double emulsion technique.Targeted high molecular polymer ultrasound contrast agents RGD-PLGA was synthesized by combining PLGA-COOH ultrasound contrast agents with RGD peptides using covalent binding.The conjugation of FITC-RGD and PLGA was detected by laser scanning confocal microscope,the affinity of RGD-PLGA and PLGA for SKOV-3 cells was investigated with light microscope and fluorescence microscope.Then the targeting ability between RGD-PLGA and control PLGA was compared by flow cytometry technique.Results FITC-RGD-PLGA showed clear green light by laser scanning confocal microscope.It was showed that many RGD-PLGA attached to SKOV-3 tightly in vitro experiment by light microscope and fluorescence microscope.It was also showed that RGD-PLGA has stronger targeting ability to SKOV-3 cells by flow cytometry technique.Conclusion RGD-PLGA was successfully synthesized and proved having strong targeting ability to SKOV-3 cells expressing integrin αvβ3.