Nanoengineering neural stem cells on biomimetic substrates using magnetofection technology
Nanoengineering neural stem cells on biomimetic substrates using magnetofection technology
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DOI：
10.1039/c6nr05244d
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发表时间：
2016-11-07
期刊：
影响因子：
6.7
通讯作者：
Chari, Divya M.
中科院分区：
文献类型：
--
作者：
Adams, Christopher F.;Dickson, Andrew W.;Chari, Divya M.
组织工程研究正在见证一种重大的范式转变，即在仿生材料上进行细胞培养，这种材料复制细胞来源的天然组织特征。在这方面，对神经细胞的研究很少，特别是在纳米医学方面。例如，磁性纳米颗粒(MNPs)等平台已被证明是神经移植群体细胞跟踪和基因工程的多功能工具。然而，据我们所知，目前所有的研究都是使用在非神经模拟底物上培养的神经细胞进行的，这些底物不能代表大脑和脊髓微环境的机械弹性特性。因此，可以预测，与神经模拟环境中生长的细胞相比，这些数据在翻译和生理上的相关性较小。因此，我们对磁感应技术(利用具有显著治疗应用潜力的外加磁场增强MNP的输送)及其在基因工程神经干细胞(NSCs；具有高度临床相关性的群体)中的应用进行了首次测试。我们证明，施加磁场可以安全地增强MNP介导的神经干细胞在胶原中生长为3D球状结构的转染，与常规使用的硬基质相比，它更接近于复制神经组织的内在机械和结构特性。此外，众所周知，MNP的摄取是通过内吞作用来调节的，我们还研究了生长在软硬基质上的NSC膜的活性。利用高分辨率扫描电子显微镜，我们能够证明神经干细胞在软基质上比在硬基质上表现出更低的膜活性，这一发现可能对在生理相关系统中繁殖的细胞的MNP介导的工程策略产生特别的影响。
Tissue engineering studies are witnessing a major paradigm shift to cell culture on biomimetic materials that replicate native tissue features from which the cells are derived. Few studies have been performed in this regard for neural cells, particularly in nanomedicine. For example, platforms such as magnetic nanoparticles (MNPs) have proven efficient as multifunctional tools for cell tracking and genetic engineering of neural transplant populations. However, as far as we are aware, all current studies have been conducted using neural cells propagated on non-neuromimetic substrates that fail to represent the mechanoelastic properties of brain and spinal cord microenvironments. Accordingly, it can be predicted that such data is of less translational and physiological relevance than that derived from cells grown in neuromimetic environments. Therefore, we have performed the first test of magnetofection technology (enhancing MNP delivery using applied magnetic fields with significant potential for therapeutic application) and its utility in genetically engineering neural stem cells (NSCs; a population of high clinical relevance) propagated in biomimetic hydrogels. We demonstrate magnetic field application safely enhances MNP mediated transfection of NSCs grown as 3D spheroid structures in collagen which more closely replicates the intrinsic mechanical and structural properties of neural tissue than routinely used hard substrates. Further, as it is well known that MNP uptake is mediated by endocytosis we also investigated NSC membrane activity grown on both soft and hard substrates. Using high resolution scanning electron microscopy we were able to prove that NSCs display lower levels of membrane activity on soft substrates compared to hard, a finding which could have particular impact on MNP mediated engineering strategies of cells propagated in physiologically relevant systems.