Human periodontal ligament stem cells (hPDLSCs) are ideal seed cells for periodontal regeneration. A greater understanding of the dynamic protein profiles during osteogenic differentiation contributed to the improvement of periodontal regeneration tissue engineering. Tandem Mass Tag quantitative proteomics was utilized to reveal the temporal protein expression pattern during osteogenic differentiation of hPDLSCs on days 0, 3, 7 and 14. Differentially expressed proteins (DEPs) were clustered and functional annotated by Gene Ontology (GO) terms. Pathway enrichment analysis was performed based on the Kyoto Encyclopedia of Genes and Genomes database, followed by the predicted activation using Ingenuity Pathway Analysis software. Interaction networks of redox-sensitive signalling pathways and oxidative phosphorylation (OXPHOS) were conducted and the hub protein SOD2 was validated with western blotting. A total of 1024 DEPs were identified and clustered in 5 distinctive clusters representing dynamic tendencies. The GO enrichment results indicated that proteins with different tendencies show different functions. Pathway enrichment analysis found that OXPHOS was significantly involved, which further predicted continuous activation. Redox-sensitive signalling pathways with dynamic activation status showed associations with OXPHOS to various degrees, especially the sirtuin signalling pathway. SOD2, an important component of the sirtuin pathway, displays a persistent increase during osteogenesis. Data are available via ProteomeXchange with identifier PXD020908. This is the first in-depth dynamic proteomic analysis of osteogenic differentiation of hPDLSCs. It demonstrated a dynamic regulatory mechanism of hPDLSC osteogenesis and might provide a new perspective for research on periodontal regeneration.

中文翻译：

---

人牙周膜干细胞成骨分化过程中的动态蛋白质组学分析

人牙周膜干细胞（hPDLSC）是牙周再生的理想种子细胞。对成骨分化过程中动态蛋白质谱的深入了解有助于牙周再生组织工程学的改善。利用串联质量标签定量蛋白质组学揭示了hPDLSC在第0、3、7和14天成骨分化过程中的时间蛋白表达模式。差异表达的蛋白（DEP）聚类并用基因本体论（GO）术语进行功能注释。路径富集分析是根据《京都议定书》的基因和基因组百科全书数据库进行的，随后使用Ingenuity Pathway Analysis软件进行了预测的激活。进行了氧化还原敏感信号通路和氧化磷酸化（OXPHOS）的相互作用网络，并通过蛋白质印迹验证了毂蛋白SOD2。共确定了1024个DEP，并将其聚集在5个代表动态趋势的独特集群中。GO富集结果表明具有不同趋势的蛋白质显示出不同的功能。途径富集分析发现OXPHOS参与其中，进一步预测了连续激活。具有动态激活状态的氧化还原敏感信号通路显示了与OXPHOS的不同程度的关联，特别是瑟土因信号通路。SOD2是Sirtuin途径的重要组成部分，在成骨过程中显示出持续的增加。数据可通过ProteomeXchange获得，其标识符为PXD020908。这是hPDLSCs成骨分化的第一个深入的动态蛋白质组学分析。它显示了hPDLSC成骨的动态调节机制，并可能为牙周再生研究提供新的前景。