中国组织工程研究2016，Vol.20Issue(30)：4425-4433,9.DOI:10.3969/j.issn.2095-4344.2016.30.003
构建叶酸修饰的乳腺癌靶向纳米超声造影微粒
Construction of folate-modified nanoparticles as ultrasound contrast agent targeting breast cancer
摘要
Abstract
BACKGROUND:Studies have testified that nano-ultrasound contrast agents have a strong permeability, making it possible to image the targeted tissues outside blood vessels and overcome the limitation that micron contrast agents are only available for the blood pool imaging. OBJECTIVE:To construct the folate-modified nanoparticles targeting breast cancer as ultrasound contrast agents, as wel as to observe their ability to specifical y bind to cel s and imaging effect in vitro. METHODS:Both contrast agents, pegylated lactic acid-glycolic acid copolymer wrapping liquid fluorocarbon formed nanoparticles (mPP/PFOB) and folate modified pegylated lactic acid-glycolic acid wrapping liquid fluorocarbon formed nanoparticles (mPPF/PFOB), were constructed by phacoemulsification-evaporation method. (1)Biocompatibility detection:HFF-1 and MCF-7 cel s in the logarithmic phase were cultivated with various concentrations (0, 0.005, 0.01, 0.02, 0.05, 0.1, 0.2 and 1 g/L) of mPP/PFOB or mPPF/PFOB for 24 hours respectively, and then the cel viability was measured. (2)Targeting ability detection in vitro:HFF-1 and MCF-7 cel s in the logarithmic phase were divided into three groups. Cy5-labled mPP/PFOB and mPPF/PFOB were added into groups A and B, respectively;the cel s in group C were pretreated with folate for 2 hours, and sequential y Cy5-labled mPPF/PFOB was added into group C. Fluorescence intensity was detected by flow cytometry after 0.5 hours of culture. The distribution of contrast agents in cel s was observed using confocal microscopy after 20 minutes of culture. (3)Ultrasound imaging in vitro:there were three groups:saline was as group A;the suspension of saline and mPPF/PFOB nanoparticles was prepared as group B;MCF-7 cel s were resuspended with the mixture of saline and mPPF/PFOB nanoparticles to prepare the suspension of nanoparticles and cel s as group C. In each group, the suspension was added into latex gloves, that were then tightened and immersed in water. Final y, the ultrasound was use to detect the ultrasound imaging effect in vitro. RESULTS AND CONCLUSION:Neither nanoparticles were with significant cytotoxicity. The flow cytometry showed that the mean fluorescence intensity in MCF-7 cel s of group B was significantly higher than that of groups A and C. But there were no significant differences in the mean fluorescence intensity in HFF-1 cel s among the three groups. It was observed that mPPF/PFOB mainly gathered around the MCF-7 cel membrane, while mPP/PFOB randomly distributed in the cytoplasm. After mPPF/PFOB binding to MCF-7 cel s, they could enhance ultrasound echo in vitro. These findings indicate that the targeted nanoparticles mPPF/PFOB have good biocompatibility and can specifical y bind to breast cancer MCF-7 cel s in vitro and enhance the imaging capability.关键词
生物材料/纳米材料/纳米粒/聚乙二醇/聚乳酸-羟基乙酸共聚物/叶酸受体/超声诊断/靶向对比剂/乳腺癌/国家自然科学基金分类
医药卫生引用本文复制引用
陈园园,徐枫,杨辉,刘婷,周建桥,蔡晨蕾,叶园园,刘培峰,韩宝三..构建叶酸修饰的乳腺癌靶向纳米超声造影微粒[J].中国组织工程研究,2016,20(30):4425-4433,9.基金项目
国家自然科学基金(81172078)，宁波市社会发展重大择优科技攻关项目(2012C5013) （81172078）
