The aim of this article is to reveal the influence of aligned/random poly(l-lactic acid) (PLLA) nanofibers on PC12 cell differentiation from the perspective of metabolic level. First, three materials—PLLA aligned nanofibers (PLLA AF), PLLA random nanofibers (PLLA RF) and PLLA films (control)—were prepared by electrospinning and spin coating. Their surface morphologies were characterized. Subsequently, the cell viability, cell morphology and neurite length of PC12 cells on the surface of the three materials were evaluated, indicating more neurites in the PLLA RF groups but the longer average neurite length in the PLLA AF groups. Next, the metabolite profiles of PC12 cells cultured on the surface of the three nanofibers after 12 h, 24 h and 36 h showed that, compared with the control, 51, 48 and 31 types of differential metabolites were detected at the three time points among the AF groups, respectively; and 56, 45 and 41 types among the RF groups, respectively. Furthermore, the bioinformatics analysis of differential metabolites identified two pathways and three metabolites critical to PC12 cell differentiation influenced by the nanofibers. In addition, the verification experiment on critical metabolites and metabolic pathways were performed. The integrative analysis combining cytology, metabolomics and bioinformatics approaches revealed that though both PLLA AF and RF were capable of stimulating the synthesis of neurotransmitters, the PLLA AF were more beneficial for PC12 cell differentiation, whereas the PLLA RF were less effective.

中文翻译：

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聚(l-乳酸)纳米纤维对PC12细胞分化的代谢组学分析

本文旨在从代谢水平的角度揭示排列/随机聚乳酸（PLLA）纳米纤维对PC12细胞分化的影响。首先，通过静电纺丝和旋涂制备三种材料——PLLA 排列纳米纤维（PLLA AF）、PLLA 无规纳米纤维（PLLA RF）和 PLLA 薄膜（对照）。表征了它们的表面形态。随后，评估了三种材料表面PC12细胞的细胞活力、细胞形态和神经突长度，表明PLLA RF组的神经突较多，而PLLA AF组的平均神经突长度较长。接下来，在 12 小时、24 小时和 36 小时后，在三种纳米纤维表面培养的 PC12 细胞的代谢物谱表明，与对照相比，51，AF组3个时间点分别检测到48种和31种差异代谢物；RF 组中分别有 56、45 和 41 种类型。此外，差异代谢物的生物信息学分析确定了受纳米纤维影响的对 PC12 细胞分化至关重要的两条途径和三种代谢物。此外，还进行了关键代谢物和代谢途径的验证实验。结合细胞学、代谢组学和生物信息学方法的综合分析表明，虽然 PLLA AF 和 RF 都能够刺激神经递质的合成，但 PLLA AF 对 PC12 细胞分化更有利，而 PLLA RF 效果较差。RF组中分别有45种和41种。此外，差异代谢物的生物信息学分析确定了受纳米纤维影响的对 PC12 细胞分化至关重要的两条途径和三种代谢物。此外，还进行了关键代谢物和代谢途径的验证实验。结合细胞学、代谢组学和生物信息学方法的综合分析表明，虽然 PLLA AF 和 RF 都能够刺激神经递质的合成，但 PLLA AF 对 PC12 细胞分化更有利，而 PLLA RF 效果较差。RF组中分别有45种和41种。此外，差异代谢物的生物信息学分析确定了受纳米纤维影响的对 PC12 细胞分化至关重要的两条途径和三种代谢物。此外，还进行了关键代谢物和代谢途径的验证实验。结合细胞学、代谢组学和生物信息学方法的综合分析表明，虽然 PLLA AF 和 RF 都能够刺激神经递质的合成，但 PLLA AF 对 PC12 细胞分化更有利，而 PLLA RF 效果较差。进行了关键代谢物和代谢途径的验证实验。结合细胞学、代谢组学和生物信息学方法的综合分析表明，虽然 PLLA AF 和 RF 都能够刺激神经递质的合成，但 PLLA AF 对 PC12 细胞分化更有利，而 PLLA RF 效果较差。进行了关键代谢物和代谢途径的验证实验。结合细胞学、代谢组学和生物信息学方法的综合分析表明，虽然 PLLA AF 和 RF 都能够刺激神经递质的合成，但 PLLA AF 对 PC12 细胞分化更有利，而 PLLA RF 效果较差。