摘要
目的制备一种靶向炎症内皮细胞的双模态纳米级分子探针,并对其进行体外寻靶研究。方法采用双乳化法制备包裹金纳米棒(Gold nanorods)-液态氟碳全氟己烷(PFH)的高分子聚合物纳米粒(GNPs)、只包裹PFH的纳米粒(NPs)和只包裹金纳米棒的纳米粒(Au-NPs);采用碳二亚胺法制备靶向血管内皮粘附因子(VCAM-1)靶向连接的纳米粒(VCAM-1-GNPs);于光镜、电镜下对GNPs基本性质进行检测;用激光辐照仪进行辐照观察GNPs、NPs和Au-NPs的光致相变情况;采用光镜和流式细胞仪观察GNPs与靶向抗体连接情况;用不同浓度肿瘤坏死因子(TNF-α)作用于人脐静脉内皮细胞(HUVECs)后,采用蛋白质印迹法(Western Blot)测量HUVECs膜上VCAM-1的表达量;观察靶向组和非靶向组与VCAM-1-GNPs的结合情况。结果成功制备包裹金纳米棒-液态氟碳双模态靶向纳米粒,平均粒径(463.67±8.23)nm;激光辐照GNPs相变明显,而NPs和Au-NPs未见明显相变;流式仪检测GNPs和VCAM-1抗体的连接率为99.87%;10ng/ml浓度的TNF-α作用于细胞后可见VCAM-1明显表达;光镜下VCAM-1-GNPs特异性与靶向组HUVECs结合,非靶向组则未见明显结合。结论成功制备包裹金纳米棒-液态氟碳双模态靶向纳米粒,GNPs光致相变效果明显,靶向性好。
Objective To prepare vascular cell adhesion molecule-1(VCAM-1)-targeted polymer ultrasound/photoacoustic dual-modality photoacoustic/ultrasound nanoscale molecular probe enfloding gold nanorods and liquid perflurocarbon,and explore their targeting ability in vitro.Methods The polymer PLGA nanoparticles(GNPs)loaded gold nanorods and liquid perflurocarbon(Perfluorohexane,PFH),only PFH nanoparticles(NPs)and only gold nanorods(Au-NPs)were synthesized by double emulsion and the VCAM-1-GNPs were accomplished via carbodiimide method.The basic physical characteristics of GNPs were observed with light microscope and electron microscopy.The optical droplet vaporization(ODV)of GNPs,NPs and Au-NPs were motivated by laser apparatus and observed by light micriscope.The HUVECs was simulated by different concentration of TNF-αand the product-VCAM-1was detected by Western Blot.The coupling efficiency and imaging of GNPs with VCAM-1 were detected.The targeted GNPs and none targeted GNPs with HUVECs were observed.Results The GNPs were synthesized successfully,and the average diameter was(463.67±8.23)nm.The GNPs could be motivated by laser and the ODV imaging was observed obviously,while NPs and Au-NPs were not clear.The coupling efficiency of GNPs with VCAM-1was 99.87%.The VCAM-1expressed on HUVECs markedly via the effect of TNF-α(10ng/ml).The VCAM-1-GNPs with HUVECs in the targeted group had excellent connection while none-targeted had poor.Conclusion The polymer nanoparticles coated gold nanorods and perflurocarbon is prepared.The ODV is obvious.The targeted GNPs has excellent coupling efficiency with HUVECs in vitro.
出处
《中国医学影像技术》
CSCD
北大核心
2016年第3期333-337,共5页
Chinese Journal of Medical Imaging Technology
基金
国家自然科学基金(81471713)
关键词
金纳米棒
液气相变
VCAM-1抗体
光声成像
Gold nanorods
Liquid-gas phase transition
VCAM-1antibody
Photoacoustic imaging