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Biodelivery of nerve growth factor and gold nanoparticles encapsulated in chitosan nanoparticles for schwann-like cells differentiation of human adipose-derived stem cells

机译:神经生长因子和金纳米粒细胞的生物次数包封在壳聚糖纳米粒子中,用于施曼样细胞分化的人脂肪衍生的干细胞

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The constant release of neurotrophic factors through a nanomaterial-based delivery system can be an important strategy in medical and pharmaceutical fields for nerve tissue engineering. The present study was aimed at encapsulating NGFand AuNPs in chitosan nanoparticles (NGF-CNPs and AuNPs-CSNPs) and its evaluation on the differentiation potential of human adipose-derived stem cells (h-ADSCs) to Schwann-like cells. The NGF-CNPs were prepared by ionotropic gelation method with tripolyphosphate (TPP) as a crosslinker. After synthesis and characterization of nanoparticles, NGF encapsulation efficiency and release profile were observed by Bradford assay. Next, the effects of NGF-CSNPs and AuNPs-CSNPs on h-ADSCs survival were assessed through MTT assay. Also, the efficacy of Schwann-like cells differentiation was assessed by immunocytochemistry and real-time RT-PCR for S1OOP and MBP markers. NGF encapsulation efficiency was found about 85% and controlled and sustained release of NGF was observed during 7 days in vitro (74.63 ± 2.07%). The findings revealed that these nanoparticles are cytocompatible. The immunocytochemical analysis indicated that NGF-CSNPs and AuNPs-CSNPs could significantly increase the differentiated rate and myelinogenic potential of Schwann-like cells (p < 0.05). Besides, the expression level of GFAP, S100P, and MBP demonstrated significant upregulation in NGF-CSNPs and AuNPs-CSNPs groups compared to the control group (p < 0.05). Hence, it can be proposed that NGF-CNPs and AuNPs-CSNPs are capable of controlled release with improving the ability of h-ADSCs differentiation to Schwann-like cells. Also, the results show the potential future application of this differentiation in nerve tissue regeneration.
机译:通过纳米材料的递送系统持续释放神经营养因子可以是神经组织工程的医学和药物的重要策略。本研究旨在将NGFAND AUNP的壳聚糖纳米颗粒(NGF-CNPS和AUNPS-CSNP)包封,并评估人脂肪衍生的干细胞(H-ADSCs)的分化潜力,类似于施旺的细胞。 NGF-CNP通过与三聚磷酸盐(TPP)作为交联剂来制备NGF-CNP。在纳米颗粒的合成和表征后,通过Bradford测定观察NGF包封效率和释放曲线。接下来,通过MTT测定评估NGF-CSNPS和AUNPS-CSNP对H-ADSCs存活的影响。此外,通过用于S1OOP和MBP标记的免疫细胞化学和实时RT-PCR评估Schwann样细胞分化的疗效。发现NGF包封效率在体外7天期间观察到NGF的约85%,并观察到NGF的持续释放(74.63±2.07%)。结果表明,这些纳米颗粒是细胞偶联的。免疫细胞化学分析表明,NGF-CSNP和AUNPS-CSNP可以显着提高施曼样细胞的分化率和敏素源性潜力(P <0.05)。此外,与对照组相比,GFAP,S100P和MBP的表达水平在NGF-CSNP和AUNPS-CSNPS组中表现出显着上调(P <0.05)。因此,可以提出NGF-CNP和AUNPS-CSNPS能够控制释放,并改善H-ADSCs分化与施旺的细胞的能力。此外,结果表明这种分化在神经组织再生中的潜在未来应用。

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