Neuronal cells loaded with PEI-coated Fe3O4 nanoparticles for magnetically guided nerve regeneration
Neuronal cells loaded with PEI-coated Fe3O4 nanoparticles for magnetically guided nerve regeneration
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DOI：
10.1039/c3tb20336k
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发表时间：
2013-01-01
影响因子：
7
通讯作者：
Goya, Gerardo F.
中科院分区：
文献类型：
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作者：
Pilar Calatayud, M.;Riggio, Cristina;Goya, Gerardo F.
我们报告了一种一步合成方案，用于获得为上传神经细胞而设计的聚合物涂层磁性纳米颗粒（MNP）。通过在碱性水介质中用硝酸盐氧化 Fe(OH)(2)，并在合成过程中原位添加 PEI，制备了尺寸为 25 +/- 5 nm 的聚乙烯亚胺包覆的 Fe3O4 纳米粒子 (PEI-MNP)。所得PEI-MNP核表现出整齐的八面体形貌和高结晶度。所得纳米颗粒涂覆有约0.7-0.9 nm的薄聚合物层，并在250 K下表现出饱和磁化强度值M-S = 58 A m(2) kg(-1) (T 10 K时为64 A m(2) kg(-1))。在神经母细胞瘤来源的 SH-SY5Y 细胞系上进行了较宽的时间和 MNP 浓度范围的细胞摄取实验。结果显示，孵育24小时后，细胞活力略有下降（100 μg ml(-1)时细胞活力降至70%），毒性作用随着孵育时间的延长而增加（100 μg ml(-1)孵育7天时细胞存活率为30%）。另一方面，原代神经元细胞对PEI-MNPs表现出更高的敏感性，与对照细胞相比，在与50μg ml(-1)孵育3天后，细胞活力降低了44%。发现 SH-SY5Y 细胞上传的 PEI-MNP 量与浓度呈线性依赖性。通过双光束 (FIB/SEM) 技术在单细胞水平上分析 PEI-MNP 的细胞内分布，揭示了完全掺入的 PEI-MNP 和穿过细胞膜的部分内化的 PEI-MNP 簇的共存。由此产生的 MNP 簇分布开启了使用这些 PEI-MNP 在神经细胞中磁驱动轴突再生长的可能性。
We report a one-step synthesis protocol for obtaining polymer-coated magnetic nanoparticles (MNPs) engineered for uploading neural cells. Polyethyleneimine-coated Fe3O4 nanoparticles (PEI-MNPs) with sizes of 25 +/- 5 nm were prepared by oxidation of Fe(OH)(2) by nitrate in basic aqueous media and adding PEI in situ during synthesis. The obtained PEI-MNP cores displayed a neat octahedral morphology and high crystallinity. The resulting nanoparticles were coated with a thin polymer layer of about 0.7-0.9 nm, and displayed a saturation magnetization value M-S = 58 A m(2) kg(-1) at 250 K (64 A m(2) kg(-1) for T 10 K). Cell uptake experiments on a neuroblastoma-derived SH-SY5Y cell line were undertaken over a wide time and MNP concentration range. The results showed a small decrease in cell viability for 24 h incubation (down to 70% viability for 100 mu g ml(-1)), increasing the toxic effects with incubation time (30% cell survival at 100 mu g ml(-1) for 7 days of incubation). On the other hand, primary neuronal cells displayed higher sensitivity to PEI-MNPs, with a cell viability reduction of 44% of the control cells after 3 days of incubation with 50 mu g ml(-1). The amount of PEI-MNPs uploaded by SH-SY5Y cells was found to have a linear dependence on concentration. The intracellular distribution of the PEI-MNPs analyzed at the single-cell level by the dual-beam (FIB/SEM) technique revealed the coexistence of both fully incorporated PEI-MNPs and partially internalized PEI-MNP-clusters crossing the cell membrane. The resulting MNP-cluster distributions open the possibility of using these PEI-MNPs for magnetically driven axonal re-growth in neural cells.