ABSTRACT

Introduction: Human periodontal ligament stem cells (hPDLSCs) are stem cells found near the tooth periodontal ligament. These cels are involved in the regeneration of the periodontal ligament and alveolar bone during orthodontic treatment and chronic periodontitis.

Objectives: The Homeobox gene HOXA10 regulates the osteogenic differentiation of stem cells. However, the role of HOXA10 in hPDLSCs remains unclear. Therefore, we studied the effects of HOXA10 on human PDLSC osteogenic differentiation in vitro.

Methods: First, hPDLSCs were isolated and characterized. Second, we assessed the effects of overexpression and knockdown of HOXA10 on PDLSC osteogenic differentiation. Finally, the specific Wnt signaling pathway activator lithium chloride (LiCl) and inhibitor ICG-001 were used to investigate the involvement of the Wnt signaling pathway in HOXA10-induced regulation of osteogenic differentiation.

Results: Overexpressing HOXA10 inhibited PDLSC osteogenic differentiation in vitro, shown by ALP and Alizarin Red staining, while HOXA10 knockdown demonstrated the opposite effects. HOXA10 negatively regulated nuclear β-catenin and osteogenic differentiation markers including alkaline phosphatase (ALPL) and integrin-binding sialoprotein (IBSP). Upregulating HOXA10 reduced nuclear β-catenin and increased DKK1 expression. However, HOXA10 knockdown enhanced nuclear β-catenin accumulation and reduced DKK1 expression. These negative effects on osteogenic differentiation by HOXA10 overexpression were restored by the Wnt/β-catenin pathway activator LiCl. The increased osteogenic differentiation effects of HOXA10 knockdown were antagonized by ICG-001, a Wnt pathway inhibitor.

Conclusion: These data demonstrate that HOXA10 inhibits the osteogenic differentiation of periodontal ligament stem cells by regulating β-catenin localization and DKK1.

摘要

简介: 人牙周膜干细胞 (hPDLSCs) 是在牙齿牙周膜附近发现的干细胞。这些细胞在正畸治疗和慢性牙周炎期间参与牙周韧带和牙槽骨的再生。

目的：同源框基因 HOXA10 调节干细胞的成骨分化。然而，HOXA10 在 hPDLSCs 中的作用仍不清楚。因此，我们在体外研究了 HOXA10 对人 PDLSC 成骨分化的影响。

方法：首先，分离和表征hPDLSC。其次，我们评估了 HOXA10 的过表达和敲低对 PDLSC 成骨分化的影响。最后，使用特异性 Wnt 信号通路激活剂氯化锂 (LiCl) 和抑制剂 ICG-001 研究 Wnt 信号通路在 HOXA10 诱导的成骨分化调节中的作用。

结果：过表达 HOXA10在体外抑制 PDLSC 成骨分化，ALP 和茜素红染色显示，而 HOXA10 敲低则显示相反的效果。HOXA10 负调控核 β-连环蛋白和成骨分化标志物，包括碱性磷酸酶 ( ALPL ) 和整合素结合唾液蛋白 ( IBSP )）。上调 HOXA10 可减少核 β-catenin 并增加 DKK1 表达。然而，HOXA10 敲低增强了核 β-连环蛋白的积累并降低了 DKK1 的表达。Wnt/β-连环蛋白通路激活剂 LiCl 恢复了 HOXA10 过表达对成骨分化的这些负面影响。HOXA10 敲低增加的成骨分化作用被 Wnt 通路抑制剂 ICG-001 拮抗。

结论：这些数据表明 HOXA10 通过调节 β-catenin 定位和 DKK1 来抑制牙周膜干细胞的成骨分化。