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首页> 外文期刊>Acta biomaterialia >The design of electrospun PLLA nanofiber scaffolds compatible with serum-free growth of primary motor and sensory neurons.
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The design of electrospun PLLA nanofiber scaffolds compatible with serum-free growth of primary motor and sensory neurons.

机译：电纺PLLA纳米纤维支架的设计与原代运动和感觉神经元的无血清生长相容。

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Aligned electrospun nanofibers direct neurite growth and may prove effective for repair throughout the nervous system. Applying nanofiber scaffolds to different nervous system regions will require prior in vitro testing of scaffold designs with specific neuronal and glial cell types. This would be best accomplished using primary neurons in serum-free media; however, such growth on nanofiber substrates has not yet been achieved. Here we report the development of poly(l-lactic acid) (PLLA) nanofiber substrates that support serum-free growth of primary motor and sensory neurons at low plating densities. In our study, we first compared materials used to anchor fibers to glass to keep cells submerged and maintain fiber alignment. We found that poly(lactic-co-glycolic acid) (PLGA) anchors fibers to glass and is less toxic to primary neurons than bandage and glue used in other studies. We then designed a substrate produced by electrospinning PLLA nanofibers directly on cover slips pre-coated with PLGA. This substrate retains fiber alignment even when the fiber bundle detaches from the cover slip and keeps cells in the same focal plane. To see if increasing wettability improves motor neuron survival, some fibers were plasma etched before cell plating. Survival on etched fibers was reduced at the lower plating density. Finally, the alignment of neurons grown on this substrate was equal to nanofiber alignment and surpassed the alignment of neurites from explants tested in a previous study. This substrate should facilitate investigating the behavior of many neuronal types on electrospun fibers in serum-free conditions.
机译：对齐的电纺纳米纤维可引导神经突生长，并可能证明对整个神经系统的修复有效。将纳米纤维支架应用于不同的神经系统区域将需要事先对具有特定神经元和神经胶质细胞类型的支架设计进行体外测试。使用无血清培养基中的原代神经元将是最好的方法。然而，在纳米纤维基底上的这种生长尚未实现。在这里，我们报告了聚（l-乳酸）（PLLA）纳米纤维基质的发展，该基质在低电镀密度下支持无血清生长的主要运动和感觉神经元。在我们的研究中，我们首先比较了将纤维固定在玻璃上以保持细胞浸没并保持纤维排列的材料。我们发现，聚乳酸-乙醇酸（PLGA）可以将纤维锚固到玻璃上，并且与其他研究中使用的绷带和胶水相比，对初级神经元的毒性较小。然后，我们设计了一种将PLLA纳米纤维直接静电纺在预涂PLGA的盖玻片上制成的基材。即使纤维束从盖玻片上分离下来，该基材仍可以保持纤维排列，并使细胞保持在同一焦平面上。为了观察增加的润湿性是否能改善运动神经元的存活率，在细胞铺板前对某些纤维进行了等离子体蚀刻。在较低的镀层密度下，蚀刻纤维的存活率降低。最终，在该基质上生长的神经元的排列与纳米纤维排列相等，并且超过了先前研究中测试的外植体中的神经突排列。该基质应有助于在无血清条件下研究电纺纤维上许多神经元类型的行为。

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