关键词: electrical stimulation electrospinning scaffold neural differentiation

Mesh : Tissue Scaffolds / chemistry Nerve Regeneration Nanotubes, Carbon / chemistry Aniline Compounds / chemistry Acrylic Resins / chemistry Cell Differentiation Tissue Engineering / methods Biocompatible Materials / chemistry Electric Stimulation Humans Cell Adhesion Microscopy, Electron, Scanning Stem Cells / cytology Tensile Strength Neurons / metabolism cytology Animals Nestin / metabolism

来  源:   DOI:10.1088/2057-1976/ad5e84

Abstract:
Biocompatible polymer-based scaffolds hold great promise for neural repair, especially when they are coupled with electrostimulation to induce neural differentiation. In this study, a combination of polyacrylonitrile/polyaniline (PAN/PANI) and Carbon Nanotubes (CNTs) were used to fabricate three different biomimetic electrospun scaffolds (samples 1, 2 and 3 containing 0.26 wt%, 1 wt% and 2 wt% of CNTs, respectively). These scaffolds underwent thorough characterization for assessing electroconductivity, tensile strength, wettability, degradability, swelling, XRD, and FTIR data. Notably, scanning electron microscopy (SEM) images revealed a three-dimensional scaffold morphology with aligned fibers ranging from 60 nm to 292 nm in diameter. To comprehensively investigate the impact of electrical stimulation on the nervous differentiation of the stem cells seeded on these scaffolds, cell morphology and adhesion were assessed based on SEM images. Additionally, scaffold biocompatibility was studied through MTT assay. Importantly, Real-Time PCR results indicated the expression of neural markers-Nestin,β-tubulin III, and MAP2-by the cells cultured on these samples. In comparison with the control group, samples 1 and 2 exhibited significant increases in Nestin marker expression, indicating early stages of neuronal differentiation, whileβ-tubulin III expression was significantly reduced and MAP2 expression remained statistically unchanged. In contrast, sample 3 did not display a statistically significant upturn in Nestin maker expression, while showcasing remarkable increases in the expression of both MAP2 andβ-tubulin III, as markers of the end stages of differentiation, leading to postmitotic neurons. These results could be attributed to the higher electroconductivity of S3 compared to other samples. Our findings highlight the biomimetic potential of the prepared scaffolds for neural repair, illustrating their effectiveness in guiding stem cell differentiation toward a neural lineage.
摘要:
生物相容性聚合物基支架在神经修复方面有着巨大的前景,特别是当它们与电刺激结合以诱导神经分化时。在这项研究中,聚丙烯腈/聚苯胺(PAN/PANI)和碳纳米管(CNT)的组合用于制造三种不同的仿生电纺支架(样品1、2和3含有0.26wt%,1重量%和2重量%的CNT,分别)。这些支架进行了彻底的表征,以评估电导率,抗拉强度,润湿性,降解性,肿胀,XRD,和FTIR数据。值得注意的是,扫描电子显微镜(SEM)图像显示三维支架形态,排列纤维的直径范围为60nm至292nm。 为了全面研究电刺激对种植在这些支架上的干细胞神经分化的影响,基于SEM图像评估细胞形态和粘附。此外,通过MTT法研究支架的生物相容性。重要的是,实时荧光定量PCR结果显示神经标记物-Nestin的表达,β-微管蛋白III,和MAP2-在这些样品上培养的细胞。与对照组相比,样品1和2表现出Nestin标记表达的显着增加,表明神经元分化的早期阶段,而β-微管蛋白III表达显着降低,而MAP2表达保持统计学不变。相比之下,样本3在Nestin标记表达中没有显示出统计学上显著的上升,同时显示MAP2和β-微管蛋白III的表达显着增加,作为分化末期的标志,导致有丝分裂后神经元。这些结果可归因于S3与其他样品相比更高的电导率。我们的发现强调了制备的支架的仿生潜力,用于神经修复,说明了它们在引导干细胞向神经谱系分化方面的有效性。
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