Roloff Frank, Strauß Sarah, Vogt Peter M, Bicker Gerd, Radtke Christine
Division of Cell Biology, University of Veterinary Medicine Hannover, Bischofsholer Damm15/102, 30173 Hannover, Germany.
Department of Plastic, Hand and Reconstructive Surgery, Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Germany.
Biomed Res Int. 2014;2014:906819. doi: 10.1155/2014/906819. Epub 2014 May 18.
Over the last years, a number of therapeutic strategies have emerged to promote axonal regeneration. An attractive strategy is the implantation of biodegradable and nonimmunogenic artificial scaffolds into injured peripheral nerves. In previous studies, transplantation of decellularized veins filled with spider silk for bridging critical size nerve defects resulted in axonal regeneration and remyelination by invading endogenous Schwann cells. Detailed interaction of elongating neurons and the spider silk as guidance material is unknown. To visualize direct cellular interactions between spider silk and neurons in vitro, we developed an in vitro crossed silk fiber array. Here, we describe in detail for the first time that human (NT2) model neurons attach to silk scaffolds. Extending neurites can bridge gaps between single silk fibers and elongate afterwards on the neighboring fiber. Culturing human neurons on the silk arrays led to an increasing migration and adhesion of neuronal cell bodies to the spider silk fibers. Within three to four weeks, clustered somata and extending neurites formed ganglion-like cell structures. Microscopic imaging of human neurons on the crossed fiber arrays in vitro will allow for a more efficient development of methods to maximize cell adhesion and neurite growth on spider silk prior to transplantation studies.
在过去几年中，出现了许多促进轴突再生的治疗策略。一种有吸引力的策略是将可生物降解且无免疫原性的人工支架植入受损的周围神经。在先前的研究中，植入填充有蜘蛛丝的脱细胞静脉以桥接临界尺寸的神经缺损，可通过内源性施万细胞的侵入实现轴突再生和髓鞘再生。伸长的神经元与作为引导材料的蜘蛛丝之间的详细相互作用尚不清楚。为了在体外可视化蜘蛛丝与神经元之间的直接细胞相互作用，我们开发了一种体外交叉丝纤维阵列。在此，我们首次详细描述了人类（NT2）模型神经元附着于丝支架。伸出的神经突可以跨越单根丝纤维之间的间隙，然后在相邻纤维上延伸。在丝阵列上培养人类神经元导致神经元细胞体向蜘蛛丝纤维的迁移和粘附增加。在三到四周内，聚集的胞体和伸出的神经突形成神经节样细胞结构。体外对交叉纤维阵列上的人类神经元进行显微成像，将有助于在移植研究之前更有效地开发方法，以最大限度地促进细胞在蜘蛛丝上的粘附和神经突生长。
