Dental Stem Cell Biology Research Unit, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
Office of Research Affairs, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
Int Endod J. 2023 Mar;56(3):369-384. doi: 10.1111/iej.13877. Epub 2022 Dec 16.
Tideglusib is a small molecule agonist of the canonical Wnt pathway. The present study investigated the influence of Tideglusib on human dental pulp stem cell (hDPSC) proliferation, apoptosis, migration and odonto/osteogenic differentiation.
hDPSCs were treated with 50, 100 nM or 200 nM Tideglusib. β-catenin accumulation was detected by immunofluorescence staining. Colony-forming unit ability was assessed by staining with Coomassie blue. Cell cycle progression and cell apoptosis were investigated using flow cytometry. Cell migration was examined using an in vitro wound-healing assay. Osteogenic differentiation was examined using alkaline phosphatase (ALP) staining, alizarin red S staining and osteogenic-related gene expression. The gene expression profile was examined using a high-throughput RNA sequencing technique. All experiments were repeated using cells derived from at least four different donors (n = 4). The Mann-Whitney U-test was used to identify significant differences between two independent group comparisons. For three or more group comparisons, statistical differences were assessed using the Kruskal-Wallis test followed by pairwise comparison. The significance level was set at 5% (p < .05).
Tideglusib activated the Wnt signalling pathway in hDPSCs as demonstrated by an increase in cytoplasmic β-catenin accumulation and nuclear translocation. Tideglusib did not affect hDPSC proliferation, cell cycle progression, cell apoptosis or cell migration. In contrast, 50 and 100 nM Tideglusib significantly enhanced mineralization and osteogenic marker gene expression (RUNX2, ALP, BMP2 and DSPP; p < .05).
Tideglusib enhanced the odonto/osteogenic differentiation of hDPSCs. Therefore, incorporating this bioactive molecule in a pulp-capping material could be a promising strategy to promote dentine repair.
Tideglusib 是经典 Wnt 通路的小分子激动剂。本研究探讨了 Tideglusib 对人牙髓干细胞（hDPSC）增殖、凋亡、迁移和牙向/成骨分化的影响。
用 50、100 nM 或 200 nM Tideglusib 处理 hDPSCs。用免疫荧光染色检测β-连环蛋白积累。用考马斯亮蓝染色评估集落形成单位能力。用流式细胞术研究细胞周期进程和细胞凋亡。用体外划痕愈合试验检测细胞迁移。用碱性磷酸酶（ALP）染色、茜素红 S 染色和成骨相关基因表达检测成骨分化。用高通量 RNA 测序技术检测基因表达谱。所有实验均使用至少 4 个不同供体来源的细胞（n=4）重复进行。采用 Mann-Whitney U 检验比较两组间的差异。对于 3 组或更多组间的比较，采用 Kruskal-Wallis 检验，然后进行两两比较。显著性水平设定为 5%（p<.05）。
Tideglusib 激活了 hDPSCs 中的 Wnt 信号通路，表现为细胞质β-连环蛋白积累增加和核转位。Tideglusib 不影响 hDPSC 的增殖、细胞周期进程、细胞凋亡或细胞迁移。相反，50 和 100 nM Tideglusib 显著增强了矿化和成骨标记基因的表达（RUNX2、ALP、BMP2 和 DSPP；p<.05）。
Tideglusib 增强了 hDPSC 的牙向/成骨分化。因此，将这种生物活性分子纳入牙髓盖髓材料中可能是促进牙本质修复的一种有前途的策略。
