Department of Biology, Faculty of Science, Arak University, Arak, Iran; Stem cell Technology Research Center, Tehran, Iran.
Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran.
Mater Sci Eng C Mater Biol Appl. 2018 Dec 1;93:686-703. doi: 10.1016/j.msec.2018.08.023. Epub 2018 Aug 11.
Application of stem cells in combination with nanofibrous substrates is an interesting biomimetic approach for enhanced regeneration of damaged tissues such as bone and cartilage. The investigation of the complex interplay between nanotopographical cues of niche and noncoding RNAs in stem cells fate is an effective tool to find a new strategy for enhancing the induction of osteogenesis. In this study, we investigated the effects of aligned and random orientations of nanofibers as a natural ECM-mimicking environment on the network of noncoding RNA in mesenchymal stem cells. Aligned and randomly oriented Ploy (L-lactide) PLLA scaffolds were fabricated via electrospinning. Human Adipose Tissue-Derived Mesenchymal Stem Cells (hASCs) were isolated from adipose tissue and were cultured on surfaces of these scaffolds. Their capacity to support hMSCs proliferation was also investigated by MTT assay and the expression of c-Myc gene. Then, after 7, 14 and 21 days, the osteogenic commitment of hMSCs and the miRNA regulatory network in BMP signaling pathway were evaluated by measuring alkaline phosphatase (ALP) activity, extracellular calcium deposition, and bone-related gene activation by Real-Time PCR. Furthermore, osteogenic differentiation was evaluated with regard to their noncoding RNA network. Our results for the first time showed an interaction between nanotopographical cues and miRNA activity in hMSCs. We found that the nanotopographical cues could be used to influence the osteogenic differentiation process of hMSCs through the modulation of lncRNAs and miR-125b as negative regulators of osteogenesis as well as the H19 modulator BMP signaling pathway that acts as a miRNA sponge. Moreover, we also demonstrated for the first time that MEG3 as a long noncoding RNA is controlled by miR-125b and microRNA-triggered lncRNA decay mechanism. This strategy seems to be an important tool for controlling stem cell fate in engineered tissues and provide new insights into most biocompatible scaffolds for bone-graft substitutes.
干细胞与纳米纤维基底的联合应用是一种有趣的仿生方法，可增强骨和软骨等受损组织的再生。研究龛位的纳米形貌线索与干细胞命运中非编码 RNA 之间的复杂相互作用是寻找增强成骨诱导新策略的有效工具。在这项研究中，我们研究了纳米纤维的定向和随机取向作为天然细胞外基质模拟环境对间充质干细胞中非编码 RNA 网络的影响。通过静电纺丝制备了定向和随机取向的聚（L-丙交酯） PLLA 支架。从脂肪组织中分离出人脂肪组织来源的间充质干细胞（hASCs），并在这些支架表面培养。通过 MTT 测定和 c-Myc 基因的表达也研究了它们支持 hMSCs 增殖的能力。然后，在第 7、14 和 21 天，通过测量碱性磷酸酶（ALP）活性、细胞外钙沉积和通过实时 PCR 激活与骨相关的基因，评估 hMSCs 的成骨承诺和 BMP 信号通路中的 miRNA 调控网络。此外，还评估了其非编码 RNA 网络的成骨分化。我们的结果首次表明，纳米形貌线索与 hMSCs 中的 miRNA 活性之间存在相互作用。我们发现，纳米形貌线索可以通过调节 lncRNA 和 miR-125b 作为成骨的负调节剂以及作为 miRNA 海绵的 H19 调节 BMP 信号通路来影响 hMSCs 的成骨分化过程。此外，我们还首次证明，MEG3 作为长非编码 RNA 受 miR-125b 控制和 microRNA 触发的 lncRNA 降解机制。这种策略似乎是控制工程组织中干细胞命运的重要工具，并为骨移植物替代品的最生物相容支架提供了新的见解。
