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工程 2D 方法纤维平台结合姜黄和聚苯胺纳米粒子,通过 qRT-PCR 预测βIII-微管蛋白和 TREK-1 的表达,以检测 PC12 细胞的神经元分化。

Engineering 2D approaches fibrous platform incorporating turmeric and polyaniline nanoparticles to predict the expression of βIII-Tubulin and TREK-1 through qRT-PCR to detect neuronal differentiation of PC12 cells.

机构信息

Department of Bionanosystem Engineering, Graduate School, Jeonbuk National University, Jeonju 561-756, Republic of Korea.

Department of Bionanosystem Engineering, Graduate School, Jeonbuk National University, Jeonju 561-756, Republic of Korea; Division of Mechanical Design Engineering, Jeonbuk National University, Jeonju 561-756, Republic of Korea.

出版信息

Mater Sci Eng C Mater Biol Appl. 2021 Aug;127:112176. doi: 10.1016/j.msec.2021.112176. Epub 2021 May 24.

Abstract

The bioengineering electroactive construct of a nerve-guided conduit for repairing and restoring injured nerves is an exciting biomedical endeavor that has implications for the treatment of peripheral nerve injury. In this study, we report the development the polycaprolactone (PCL) nanofibrous substrate consisting of turmeric (TUR) and polyaniline nanoparticles (PANINPs) exhibits topological and biological features that mimics the natural extracellular matrix (ECM) for nerve cells. We evaluated the morphology of 2-dimensional (2D) fibrous substrates, and their ability of stem cell adhesion, growth and proliferation rate were influenced by use of various concentrations of turmeric in PCL-TUR substrates. The results showed that 0.62 wt% of TUR and 0.28 wt% of PANINPs in PCL nanofibers substrate exhibited the optimal cellular microenvironment to accelerate PC12 cellular activities. The in vitro experiments revealed that PCL-TUR@PANI substrates significantly stimulated the proliferation, differentiation, and spontaneous outgrowth and extension of neurites from the cells. The substrate has the capacity to respond directly to neuronal markers with significant upregulation of βIII-Tubulin and TREK-1 through myelination, and also trigger neurotrophic protein expression, which was confirmed via immunocytochemistry and quantitative real-time polymerase chain reaction (qRT-PCR) analysis. This study provides a new technique to design substrate of nerve tissue-specific microenvironment for peripheral nerve cell regeneration and could offer promising biomaterials for in vivo peripheral nerve repair.

摘要

用于修复和恢复受损神经的神经引导导管的生物工程电活性构建体是一项令人兴奋的生物医学研究,对周围神经损伤的治疗具有重要意义。在这项研究中,我们报告了聚己内酯(PCL)纳米纤维基质的开发,该基质由姜黄(TUR)和聚苯胺纳米粒子(PANINPs)组成,具有模仿神经细胞天然细胞外基质(ECM)的拓扑和生物学特征。我们评估了二维(2D)纤维基质的形态,以及不同浓度姜黄对 PCL-TUR 基质中干细胞黏附、生长和增殖率的影响。结果表明,PCL 纳米纤维基质中 0.62wt%的 TUR 和 0.28wt%的 PANINPs 表现出最佳的细胞微环境,可加速 PC12 细胞的活性。体外实验表明,PCL-TUR@PANI 基质显著刺激了细胞的增殖、分化以及神经突的自发生长和延伸。该基质具有直接响应神经元标志物的能力,通过髓鞘化显著上调βIII-Tubulin 和 TREK-1,还能触发神经营养蛋白的表达,这通过免疫细胞化学和实时定量聚合酶链反应(qRT-PCR)分析得到了证实。本研究为设计周围神经细胞再生的神经组织特异性微环境的基质提供了一种新技术,并为体内周围神经修复提供了有前途的生物材料。

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