Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi'an Jiaotong University, Xi'an, China.
Department of Pediatric Dentistry, College of Stomatology, Xi'an Jiaotong University, Xi'an, China.
Braz J Med Biol Res. 2024 Oct 7;57:e13606. doi: 10.1590/1414-431X2024e13606. eCollection 2024.
This study aimed to illustrate the biological behavior and changes in cell function during the progression of apical periodontitis in deciduous teeth and to explore the underlying molecular mechanism. Deciduous teeth periodontal ligament stem cells (DePDLSCs) were derived and their identity was confirmed. The viability, inflammation, and osteogenic ability of cells were tested by exposing them to various concentrations of lipopolysaccharide (LPS) (0-100 μg/mL) using the cell counting kit-8 (CCK-8) assay, reverse transcription polymerase chain reaction (real-time PCR), alkaline phosphatase (ALP) staining, and ALP activity assay. In addition, osteogenic-induced cells with and without 10 μg/mL LPS were harvested for high-throughput sequencing. Based on sequencing data, proinflammatory factors and ALP expression were measured after interference with the PI3K-AKT signaling pathway activator, 740Y-P. LPS biphasically affected the proliferation and osteogenesis of DePDLSCs. Low concentrations of LPS showed stimulatory effects, whereas inhibitory effects were observed at high concentrations. Sequencing analysis showed that the PI3K-AKT signaling pathway was significantly downregulated when DePDLSCs were treated with 10 μg/mL LPS. The LPS-induced inflammation and osteogenesis inhibition of DePDLSCs were partially rescued by 740Y-P treatment. In conclusion, LPS affected DePDLSCs proliferation and osteogenesis in a biphasic manner. Moderate activation of PI3K-AKT signaling pathway was beneficial for osteogenic differentiation and anti-inflammatory effect in DePDLSCs. This research may provide etiological probes for apical periodontitis and its treatment.
本研究旨在阐明乳牙髓牙周韧带干细胞(DePDLSCs)在根尖周炎进展过程中的生物学行为和细胞功能变化,并探讨其潜在的分子机制。从乳牙髓牙周韧带干细胞(DePDLSCs)中分离并鉴定其特性。通过使用细胞计数试剂盒-8(CCK-8)检测、逆转录聚合酶链反应(实时 PCR)、碱性磷酸酶(ALP)染色和 ALP 活性测定,检测细胞暴露于不同浓度脂多糖(LPS)(0-100 μg/mL)时的活力、炎症和成骨能力。此外,收集有无 10 μg/mL LPS 的成骨诱导细胞进行高通量测序。基于测序数据,用 PI3K-AKT 信号通路激活剂 740Y-P 干扰后,测量促炎因子和 ALP 的表达。LPS 对 DePDLSCs 的增殖和成骨作用呈双相影响。低浓度 LPS 表现出刺激作用,而高浓度 LPS 则表现出抑制作用。测序分析表明,当 DePDLSCs 用 10 μg/mL LPS 处理时,PI3K-AKT 信号通路显著下调。740Y-P 处理部分挽救了 LPS 诱导的 DePDLSCs 炎症和成骨抑制。总之,LPS 以双相方式影响 DePDLSCs 的增殖和成骨作用。适度激活 PI3K-AKT 信号通路有利于 DePDLSCs 的成骨分化和抗炎作用。本研究可为根尖周炎及其治疗提供病因学探针。