人骨髓间充质干细胞靶向纳米基因载体制备及体外细胞磁共振成像

发布时间：2018-01-24 16:25

本文关键词： 骨髓间充质干细胞磁共振成像靶向基因载体纳米医学　出处：《中国医学影像技术》2017年10期 　论文类型：期刊论文

【摘要】：目的探讨靶向纳米基因载体单链神经节苷脂抗体-聚乙二醇-聚乙烯亚胺-超顺磁性氧化铁(scAbGD2-PEGg-PEI-SPION)转染人骨髓间充质干细胞(hBMSCs)的可行性、效率及体外细胞MR显像能力。方法合成scAbGD2-PEG-g-PEI-SPION后,采用凝胶阻滞实验评估其复合外源性基因的能力;动态光散射法测量scAbGD2-PEG-g-PEI-SPION/pDNA纳米复合物的粒径大小及表面电位;体外细胞毒性实验检测其对hBMSCs的细胞毒性。采用流式细胞仪检测scAbGD2-PEG-g-PEI-SPION靶向转染hBMSCs的效率,并设置PEG-g-PEI-SPION组、cAbGD2-PEG-g-PEI-SPION组、抗体竞争抑制(scAbGD2-PEG-g-PEI-SPION+free AbGD2)组和同型抗体(scAbIgG2a-PEG-g-PEI-SPION)组,通过激光共聚焦显微镜及普鲁士蓝染色观察hBMSCs对纳米复合物的摄入。通过体外细胞MR扫描验证scAbGD2-PEG-g-PEI-SPION的MR成像功能。结果 scAbGD2-PEG-g-PEI-SPION细胞毒性小,复合外源性基因后能够形成稳定的纳米复合物,粒径80~100nm。在相同的N/P比值下,scAbGD2-PEG-g-PEI-SPION组的转染率明显高于其他组(P0.001)。N/P=20时,靶向组具有最高转染率[(59.60±4.50)%]。同时,scAbGD2-PEG-g-PEI-SPION中的SPION可有效标记hBMSCs,在MR T2/T2*加权图像上呈低信号。结论 scAbGD2-PEG-g-PEI-SPION是一种MRI可视的、可有效转染hBMSCs的靶向纳米基因载体。
[Abstract]:Objective to investigate the single chain ganglioside antibody (PEG-PEGg-PEI-SPION) targeting nanometer-sized gene vector, Poly (ethylene glycol) polyethyleneimide-superparamagnetic ferric oxide (scAbGD2-PEGg-PEI-SPION). The feasibility of transfection of human bone marrow mesenchymal stem cells (hBMSCs). Methods after scAbGD2-PEG-g-PEI-SPION was synthesized, the ability of compound exogenous gene was evaluated by gel block assay. The particle size and surface potential of scAbGD2-PEG-g-PEI-SPION/pDNA nanocomposites were measured by dynamic light scattering method. In vitro cytotoxicity test was used to detect its cytotoxicity to hBMSCs. Flow cytometry was used to detect the efficiency of scAbGD2-PEG-g-PEI-SPION targeted transfection of hBMSCs. CAbGD2-PEG-g-PEI-SPION group was set up in PEG-g-PEI-SPION group. Antibody competitive inhibition (scAbGD2-PEG-g-PEI-SPION free AbGD2) and the same type of antibody (GD2-PEG-PEI-SPION free AbGD2). ScAbIgG2a-PEG-g-PEI-SPION) group. The uptake of nanocomposites by hBMSCs was observed by laser confocal microscopy and Prussian blue staining. ScAbGD2-PEG-g-PEI-SPION was verified by in vitro cell Mr scanning. Mr imaging function of the. The cytotoxicity of scAbGD2-PEG-g-PEI-SPION was small. The composite of exogenous genes could form a stable nano-complex with a particle size of 80 ~ 100nm. under the same N / P ratio. The transfection efficiency of scAbGD2-PEG-g-PEI-SPION group was significantly higher than that of other groups (P 0.001N / P = 20:00), and the target group had the highest transfection efficiency. [At the same time, SPION in scAbGD2-PEG-g-PEI-SPION could effectively label hBMSCs. Conclusion scAbGD2-PEG-g-PEI-SPION is visualized by MRI. It can effectively transfect hBMSCs targeting nanogene vector.
【作者单位】：中山大学附属第五医院介入医学中心;中山大学附属第五医院眼科;
【基金】：国家自然科学基金青年基金(81501561) 广东省自然科学基金(2014A030310043、2017A030313873)
【分类号】：R445.2
【正文快照】： 人骨髓间充质干细胞(human　bone　marrow-de-rived　mesenchymal　stem　cells,hBMSCs)具有向肝样细胞分化的潜能,参与肝损伤的修复[1-2]。随着对治疗机制研究的深入,发现移植入肝脏的hBMSCs仅少数分化为肝样细胞,多数hBMSCs以旁分泌和免疫调节的方式参与肝损伤的修复,极大降

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