In vitro assessment of axonal growth using dorsal root ganglia explants in a novel three-dimensional collagen matrix
In vitro assessment of axonal growth using dorsal root ganglia explants in a novel three-dimensional collagen matrix
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DOI：
10.1089/ten.2007.0116
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发表时间：
2007-12-01
影响因子：
--
通讯作者：
Pallua, Norbert
中科院分区：
文献类型：
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作者：
Bozkurt, Ahmet;Brook, Gary A.;Pallua, Norbert
本研究的目的是开发一种生物人工神经向导来诱导周围神经系统（PNS）的轴突再生。在这项体外研究中，研究了一种新型的、三维的、高度定向的、交联的猪胶原蛋白支架促进定向轴突生长的能力。采用专利的单向冷冻工艺，经过一系列化学和机械处理，制作出具有纵向导向通道的胶原神经导向器，然后冷冻干燥（孔径20-50 μ m）。将半切的大鼠背根神经节（DRG）定位，使神经和非神经成分可以迁移到胶原支架中。21天后，s100阳性的雪旺细胞（SCs）迁移到支架中，并在引导通道内以柱状排列，类似于“Bungner带”。DRG神经元的神经丝阳性轴突（平均长度+/- 756 μ m +/- 318 μ m，最大1496 μ m）进入支架，引导通道内的生长与定向SCs密切相关。本研究证实了SCs在再生过程中的重要性（神经营养理论）。SCs在引导通道内的排列支持定向轴突生长（接触引导理论）。支架的微观结构特性（开放、多孔、纵向孔道）和DRG加载后的体外数据（沿着迁移的柱状排列的SCs进行轴突再生，类似于“Bungner带”）表明，这种新型定向3D胶原支架可作为未来PNS实验再生研究的基础。
The goal of this study was the development of a bioartificial nerve guide to induce axonal regeneration in the peripheral nervous system (PNS). In this in vitro study, the ability of a novel, 3-dimensional (3D), highly oriented, cross-linked porcine collagen scaffold to promote directed axonal growth has been studied. Collagen nerve guides with longitudinal guidance channels were manufactured using a series of chemical and mechanical treatments with a patented unidirectional freezing process, followed by freeze-drying (pore sizes 20-50 mu m). Hemisected rat dorsal root ganglia (DRG) were positioned such that neural and non-neural elements could migrate into the collagen scaffold. After 21 days, S100-positive Schwann cells (SCs) migrated into the scaffold and aligned within the guidance channels in a columnar fashion, resembling "Bands of Bungner." Neurofilament-positive axons (mean length +/- SD 756 mu m +/- 318 mu m, maximum 1496 mu m) from DRG neurons entered the scaffold where the growth within the guidance channels was closely associated with the oriented SCs. This study confirmed the importance of SCs in the regeneration process (neurotrophic theory). The alignment of SCs within the guidance channels supported directional axonal growth (contact guidance theory). The microstructural properties of the scaffold (open, porous, longitudinal pore channels) and the in vitro data after DRG loading (axonal regeneration along migrated and columnar-aligned SCs resembling "Band of Bungner") suggest that this novel oriented 3D collagen scaffold serves as a basis for future experimental regeneration studies in the PNS.