VCAM-1靶向双模态光声/超声纳米级分子探针的制备及其体外寻靶实验

Preparation of VCAM-1 receptor-targeted dual-modal photoacoustic/ultrasound nanoscale molecular probe with gold nanorods and liquid fluorocarbon and their targeted ability in vitro

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.