Optimization of PAM scaffolds for neural tissue engineering: Preliminary study on an SH-SY5Y cell line
Optimization of PAM scaffolds for neural tissue engineering: Preliminary study on an SH-SY5Y cell line
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DOI：
10.1089/ten.tea.2007.0163
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发表时间：
2008-06-01
影响因子：
4.1
通讯作者：
Domenici, Claudio
中科院分区：
文献类型：
--
作者：
Kullenberg, Johanna;Rosatini, Federica;Domenici, Claudio
神经组织工程化是组织工程学最具挑战性的目标之一。神经组织高度复杂，具有组织功能所必需的有组织的三维(3D)分布。组织工程的最佳支架必须提供这种分布，直到细胞能够激活其正常功能并与宿主组织发展神经连接。不同的策略，如基因治疗和细胞移植，特别是在视网膜组织中已经进行了测试，但到目前为止，它们只在动物身上诱导了视网膜退化。这项工作的目的是研究神经细胞组装作为支架特征和表面化学的函数，以应用于视网膜组织工程中，使用具有明确定义的几何形状的微型制造图案。由于视网膜神经元呈六角形排列，因此利用压力辅助微量注射器(PAM)系统制备了聚(DL-丙交酯-乙交酯)酸的六角形支架。一个模型细胞，神经母细胞瘤起源于人类视网膜(SH-SY5Y)，在种植到支架上后，分析了其行为，测量了细胞密度随支架线宽和长度的函数，以确定最佳的六边形几何形状。我们还用四甲基偶氮唑盐比色法评价了支架对细胞代谢和轴突伸展的影响。就二维支架而言，结果表明，尽管细胞的代谢活性保持不变，但就细胞密度而言，边长为500微米、线宽为20+/-5微米的六边形对神经细胞的黏附效果最好。在3D支架上，细胞代谢率约为对照组的三倍，支架中的最佳层数为三到四层。
Engineering neural tissue is one of the most challenging goals of tissue engineering. Neural tissue is highly complex and possesses an organized three-dimensional (3D) distribution that is essential for tissue function. An optimal scaffold for tissue engineering has to provide this distribution until the cells are able to activate their normal functions and develop neural connections with the host tissue. Different strategies such as gene therapy and cell transplantation particularly in retinal tissue have been tested, but so far they have only induced retinal degeneration in animals. The objective of this work was to study neural cell assembly as a function of scaffold features and surface chemistry for application in retinal tissue engineering using microfabricated patterns with a well-defined geometry. Because retinal neurons are known to be arranged in hexagonal arrays, hexagonal scaffolds of poly(DL-lactide-co-glycolide) acid were fabricated using a pressure-assisted microsyringe (PAM) system. The behavior of a model cell, neuroblastoma originating from human retina (SH-SY5Y), was analyzed after seeding on the scaffolds, measuring cell density as a function of line width and length of the scaffold to identify the optimal hexagonal geometry. We also evaluated the influence of scaffold on cell metabolism using the methyl thiazolyl tetrazolium assay and on neurite extension. As far as two-dimensional scaffolds are concerned, the results show that although metabolic activity per cell remains constant, hexagons with sides of 500 mu m and line widths of 20 +/- 5 mu m are optimum for neural cell adhesion in terms of cell density. On 3D scaffolds, cell metabolism is about three times higher than controls, and the optimum number of layers in the scaffold is three or four.