Shamosi Atefeh, Mahmoudi Elaheh, Kermanian Fatemeh
Department of Anatomy, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran.
Department of Mycology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran.
Iran J Biotechnol. 2022 Apr 1;20(2):e3179. doi: 10.30498/ijb.2022.310552.3179. eCollection 2022 Apr.
One of the challenges in using stem cells to neural repair is to induce their differentiation into neurons and lack of glial formation.
Mesenchymal stem cells have revealed great potential for neural reorganization and renewal by taking advantage of differentiation capabilities. Here we explored the potential use of olibanum extract in freeze-dried scaffolds for induction of stem cells differentiation.
In this study, gelatin/ collagen/olibanum/ graphene oxide (GEL/COL/OL/GO) freeze-dried scaffolds were synthesized and then adult rat bone marrow mesenchymal stem cells (BMMSCs) were seeded on scaffolds. The viability of cells was evaluated using MTT test on days 1, 3 and 5. The morphology of the cells seeded on scaffolds was studied using SEM and specific protein expression detected by immunohistochemical analysis. Real-time PCR was applied to detect the expression of Chat, Pax6, Hb-9, Nestin, Islet-1, and neurofilament-H (NF-H). The data were analyzed using Tukey test and one-way ANOVA and the means difference was considered significant at P<0.05, P<0.01, and P<0.001.
Showed that the pore size is increased in GEL/COL/OL/GO scaffolds compared with GO-free scaffolds and higher attachment and proliferation of BMMSCs on GEL/COL/OL /1.5% GO scaffolds compared to GEL/COL/OL/3% GO scaffolds. The cell viability results after 5 days of incubation showed the significant biocompatibility of GEL/COL/OL /1.5% GO freeze-dried scaffold. The results of immunohistochemical and PCR analysis revealed positive role of GEL/COL/OL/1.5% GO scaffolds in upregulation of neuron-specific markers.
These results reveal the great potential of GEL/COL/OL/GO scaffolds for nerve regeneration. Our data suggested that both OL extract and GO can regulate the MSCs differentiation into neurons.
利用干细胞进行神经修复面临的挑战之一是诱导其分化为神经元并避免形成神经胶质细胞。
间充质干细胞通过利用其分化能力,在神经重组和更新方面显示出巨大潜力。在此,我们探索了乳香提取物在冻干支架中诱导干细胞分化的潜在用途。
在本研究中,合成了明胶/胶原蛋白/乳香/氧化石墨烯(GEL/COL/OL/GO)冻干支架,然后将成年大鼠骨髓间充质干细胞(BMMSCs)接种于支架上。在第1、3和5天使用MTT试验评估细胞活力。使用扫描电子显微镜研究接种于支架上的细胞形态,并通过免疫组织化学分析检测特定蛋白表达。应用实时聚合酶链反应检测胆碱乙酰转移酶(Chat)、配对盒基因6(Pax6)、同源盒基因9(Hb-9)、巢蛋白(Nestin)、胰岛素基因增强子结合蛋白1(Islet-1)和神经丝-H(NF-H)的表达。使用Tukey检验和单因素方差分析对数据进行分析,均值差异在P<0.05、P<0.01和P<0.001时被认为具有显著性。
显示与不含氧化石墨烯的支架相比,GEL/COL/OL/GO支架的孔径增大,并且与GEL/COL/OL/3%氧化石墨烯支架相比,BMMSCs在GEL/COL/OL/1. %氧化石墨烯支架上的附着和增殖更高。孵育5天后的细胞活力结果显示GEL/COL/OL/1.5%氧化石墨烯冻干支架具有显著的生物相容性。免疫组织化学和聚合酶链反应分析结果显示GEL/COL/OL/1.5%氧化石墨烯支架在上调神经元特异性标志物方面具有积极作用。
这些结果揭示了GEL/COL/OL/GO支架在神经再生方面的巨大潜力。我们的数据表明乳香提取物和氧化石墨烯均可调节间充质干细胞向神经元的分化。