576019General Dentistry Dep, School and Hospital of Stomatology, China Medical University, Liaoning Provincial Key Laboratory of Oral Diseases, Shenyang 110001, China.
71123Metallic Nano-Materials Division, Shenyang National Laboratory for Materials Science, Institute of Metal Research, Chinese Academy Sciences, Shenyang 110016, China.
J Biomater Appl. 2023 Jan;37(6):1086-1101. doi: 10.1177/08853282221125036. Epub 2022 Sep 5.
Titanium (Ti) and Ti-based alloy materials are ideal brackets that restore bone defect, and the mechanism of related genes inducing bone mesenchymal stem cells (BMSCs) to osteogenic differentiation is currently a hot research topic. In order to screen key genes of BMSCs during the osteogenic expression process, we acquired data sets (GSE37237 and GSE84500) which were in the database Gene Expression Omnibus (GEO). Investigations on differentially expressed genes (DEGs) and their enrichment of functions were conducted. We constructed relative protein-protein interaction (PPI) network by using Search Tool for the Retrieval of Interacting Genes (STRING) and visualized the expression of DEGs with Cytoscape. A total of 279 DEGs were discerned, which could be divided into 177 down regulated genes and 102 up regulated genes. In addition, the DEGs' enrichment and pathways included regulation of actin cytoskeleton, inflammatory mediator regulation of transient receptor potential (TRP) channels, peroxisome proliferator-activated receptors (PPAR) pathway, cell cycle, Rheumatoid arthritis, mitogen-activated protein kinases (MAPK) signaling pathway and Ras signaling pathway ect. It showed that 10 notable up regulated genes were mainly in AMP-activated protein kinase (AMPK) pathway. Then we used a technology named surface mechanical attrition treatment (SMAT) to prepare gradient nanostructured (GNS) surface Ti and seeded well-growing BMSCs on the surface of SMAT Ti and native pure Ti. Cell Counting Kits-8 (CCK-8), apoptosis experiment, immunofluorescence technology and staining experiments for alka-line phosphatase (ALP) and alizarin red staining (ARS) were used to research the proliferation, adhesion and differentiation ability of BMSCs seeded on SMAT Ti compared with native pure Ti. We used quantitative real-time PCR (qRT-PCR) technology so as to verify the expression of the most significant 5 genes. In summary, these results indicated novel point of views into candidate genes and potential mechanism for the further study of BMSCs' behaviors seeded on SMAT Ti.
钛(Ti)和钛基合金材料是修复骨缺损的理想支架,相关基因诱导骨髓间充质干细胞(BMSCs)成骨分化的机制是目前的研究热点。为了筛选成骨表达过程中 BMSCs 的关键基因,我们从基因表达综合数据库(GEO)中获取了数据集(GSE37237 和 GSE84500)。对差异表达基因(DEGs)及其功能富集进行了研究。我们使用搜索工具检索基因交互(STRING)构建了相关的蛋白质-蛋白质相互作用(PPI)网络,并使用 Cytoscape 可视化了 DEGs 的表达。共鉴定出 279 个 DEGs,可分为 177 个下调基因和 102 个上调基因。此外,DEGs 的富集和通路包括肌动蛋白细胞骨架的调节、瞬时受体电位(TRP)通道的炎症介质调节、过氧化物酶体增殖物激活受体(PPAR)途径、细胞周期、类风湿性关节炎、丝裂原激活蛋白激酶(MAPK)信号通路和 Ras 信号通路等。结果表明,10 个显著上调的基因主要集中在 AMP 激活蛋白激酶(AMPK)通路中。然后,我们使用一种名为表面机械磨损处理(SMAT)的技术来制备梯度纳米结构(GNS)表面 Ti,并在 SMAT Ti 和原始纯 Ti 表面接种生长良好的 BMSCs。细胞计数试剂盒-8(CCK-8)、凋亡实验、免疫荧光技术和碱性磷酸酶(ALP)和茜素红染色(ARS)染色实验用于研究 SMAT Ti 表面接种的 BMSCs 与原始纯 Ti 相比的增殖、黏附和分化能力。我们使用定量实时 PCR(qRT-PCR)技术验证了最显著的 5 个基因的表达。总之,这些结果为进一步研究 SMAT Ti 表面接种的 BMSCs 行为提供了候选基因和潜在机制的新观点。