Institute of Pharmacology and Toxicology, Biomedical Center, University of Bonn, Sigmund-Freud-Strasse 25, 53105 Bonn, Germany.
Pharm Res. 2012 May;29(5):1255-69. doi: 10.1007/s11095-011-0660-x. Epub 2012 Jan 25.
Targeting of specific cells and tissues is of great interest for clinical relevant gene- and cell-based therapies. We use magnetic nanoparticles (MNPs) with a ferrimagnetic core (Fe(3)O(4)) with different coatings to optimize MNP-assisted lentiviral gene transfer with focus on different endothelial cell lines.
Lentiviral vector (LV)/MNP binding was characterized for various MNPs by different methods (e.g. magnetic responsiveness measurement). Transduced cells were analyzed by flow cytometry, fluorescence microscopy and iron recovery. Cell transduction and cell positioning under physiological flow conditions were performed using different in vitro and ex vivo systems.
Analysis of diverse MNPs with different coatings resulted in identification of nanoparticles with improved LV association and enhanced transduction properties of complexes in several endothelial cell lines. The magnetic moments of LV/MNP complexes are high enough to achieve local gene targeting of perfused endothelial cells. Perfusion of a mouse aorta with LV/MNP transduced cells under clinically relevant flow conditions led to local cell attachment at the intima of the vessel.
MNP-guided lentiviral transduction of endothelial cells can be significantly enhanced and localized by using optimized MNPs.
针对特定的细胞和组织对于临床相关的基因和细胞治疗具有重要意义。我们使用具有不同涂层的具有铁磁性内核（Fe(3)O(4)）的磁性纳米颗粒（MNPs）来优化基于 MNPs 的慢病毒基因转移，重点关注不同的内皮细胞系。
通过不同的方法（例如磁性响应性测量）来表征不同 MNPs 中的慢病毒载体（LV）/MNP 结合。通过流式细胞术、荧光显微镜和铁回收来分析转导细胞。使用不同的体外和体内系统进行生理流动条件下的细胞转导和细胞定位。
对具有不同涂层的多种 MNPs 进行分析，结果鉴定出与 LV 结合能力提高且在几种内皮细胞系中增强复合物转导特性的纳米颗粒。LV/MNP 复合物的磁矩足以实现灌注的内皮细胞的局部基因靶向。在临床相关的流动条件下，用转导细胞灌注小鼠主动脉，导致细胞在血管内膜局部附着。
通过使用优化的 MNPs，可以显著增强和定位基于 MNP 的慢病毒转导的内皮细胞。
