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首页> 外文期刊>Biomaterials >Spinal cord organotypic slice cultures for the study of regenerating motor axon interactions with 3D scaffolds
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Spinal cord organotypic slice cultures for the study of regenerating motor axon interactions with 3D scaffolds

机译：脊髓器官型切片培养物，用于再生运动轴突与3D支架的相互作用的研究

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Numerous in-vitro techniques exist for investigating the influence of 3D substrate topography on sensory axon growth. However, simple and cost-effective methods for studying post-natal motor axon interactions with such substrates are lacking. Here, spinal cord organotypic slice cultures (OSC) from post-natal day 7-9 rat pups were presented with spinal nerve roots, or blocks of fibrin hydrogel or 3D microporous collagen scaffolds to investigate motor axon-substrate interactions. By 7-14 days, axons from motor neuronal pools extended into the explanted nerve roots, growing along Schwann cell processes and demonstrating a full range of axon-Schwann cell interactions, from simple ensheathment to concentric wrapping by Schwann cell processes and the formation of compact myelin within a basal lamina sheath. Extensive motor axon regeneration and all stages of axon-Schwann interactions were also supported within the longitudinally orientated microporous framework of the 3D collagen scaffold. In stark contrast, the simple fibrin hydrogel only supported axon growth and cell migration over its surface. The relative ease of demonstrating such motor axon regeneration through the microporous 3D framework by immunofluorescence, two-photon microscopy and transmission electron microscopy strongly supports the adoption of this technique for assaying the influence of substrate topography and functionalization in regenerative bioengineering.
机译：存在许多用于研究3D基底形貌对感觉轴突生长的影响的体外技术。但是，缺乏研究出生后运动轴突与此类底物相互作用的简单且经济有效的方法。在这里，来自产后第7-9天的幼鼠的脊髓器官型切片培养物（OSC）带有脊髓神经根或纤维蛋白水凝胶或3D微孔胶原蛋白支架块，用于研究运动轴突-底物相互作用。到7-14天时，来自运动神经元池的轴突延伸到外植神经根，沿着雪旺氏细胞过程生长，并展示了各种轴突-雪旺氏细胞相互作用，从简单的包膜到雪旺氏细胞过程的同心包裹以及紧密结构的形成基底层鞘内的髓磷脂。 3D胶原蛋白支架的纵向微孔框架内还支持广泛的运动轴突再生和轴突-施万恩相互作用的所有阶段。与之形成鲜明对比的是，简单的纤维蛋白水凝胶仅支持轴突的生长和细胞在其表面的迁移。通过免疫荧光，双光子显微镜和透射电子显微镜通过微孔3D框架来证明这种运动轴突再生的相对容易性，强烈支持了该技术在再生生物工程学中用于分析基质形貌和功能化的影响。

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