Yu Lintong, Yang Feng, Li Chenxi, Ruan Jing, Wang Jun
Department of Pediatric Dentistry, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China; College of Stomatology, Shanghai Jiao Tong University, Shanghai, China; National Center for Stomatology, Shanghai, China; National Clinical Research Center for Oral Diseases, Shanghai, China; Shanghai Key Laboratory of Stomatology, Shanghai, China; Shanghai Research Institute of Stomatology, Shanghai, China.
Department of Ophthalmology, Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, Shanghai, China.
J Endod. 2025 Jul;51(7):939-947. doi: 10.1016/j.joen.2025.04.007. Epub 2025 Apr 22.
Although cell-free DNA (cfDNA) is involved in several inflammation-related diseases, its role in pulp inflammation remains unclear. The aims of this study were to explore the role of cfDNA in the development of irreversible pulpitis and to identify a potential therapeutic strategy.
Pulpitis plasma samples were collected from patients diagnosed with irreversible pulpitis, and the cfDNA concentration associated with pulpitis was measured using a Quant-iT PicoGreen dsDNA kit. Dental pulp stem cells and Tohoku Hospital Pediatrics -1 macrophages were stimulated with pulpitis plasma, and the expression of inflammatory factors was investigated via real-time reverse transcription PCR and ELISA. Toll-like receptor 9 expression in pulpitis blood-derived cells was analyzed by flow cytometry. Finally, the effects of cationic poly(amidoamine) (PAMAM) on the treatment of inflammation in vitro and in a rat pulpitis model were evaluated.
We first revealed that pulpitis-derived plasma had elevated cfDNA levels, which caused M1-like macrophage polarization through the toll-like receptor 9 signaling pathway and then induced an inflammatory response in dental pulp stem cells. M1-like macrophage polarization affected the pulp inflammatory microenvironment and could be reversed via a synergistic effect of irrigation and cationic PAMAM for cfDNA scavenging. PAMAM has shown potential for inhibiting inflammatory M1-like macrophage polarization and alleviating pulpitis at the cellular and organism levels.
This study elucidated the importance of cfDNA in pulpitis and provides a potential therapeutic strategy. cfDNA scavenging could alleviate the inflammatory response in pulpitis and potentially preserve more vital pulp for teeth with irreversible pulpitis.
尽管游离DNA(cfDNA)参与了多种炎症相关疾病,但它在牙髓炎症中的作用仍不清楚。本研究的目的是探讨cfDNA在不可逆性牙髓炎发展中的作用,并确定一种潜在的治疗策略。
收集诊断为不可逆性牙髓炎患者的牙髓炎血浆样本,使用Quant-iT PicoGreen双链DNA试剂盒测量与牙髓炎相关的cfDNA浓度。用牙髓炎血浆刺激牙髓干细胞和东北医院儿科-1巨噬细胞,通过实时逆转录PCR和酶联免疫吸附测定法研究炎症因子的表达。通过流式细胞术分析牙髓炎血液来源细胞中Toll样受体9的表达。最后,评估阳离子聚酰胺胺(PAMAM)在体外和大鼠牙髓炎模型中对炎症治疗的效果。
我们首先发现,牙髓炎来源的血浆中cfDNA水平升高,其通过Toll样受体9信号通路导致M1样巨噬细胞极化,进而在牙髓干细胞中诱导炎症反应。M1样巨噬细胞极化影响牙髓炎症微环境,通过冲洗和阳离子PAMAM清除cfDNA的协同作用可使其逆转。PAMAM已显示出在细胞和机体水平上抑制炎症性M1样巨噬细胞极化和减轻牙髓炎的潜力。
本研究阐明了cfDNA在牙髓炎中的重要性,并提供了一种潜在的治疗策略。清除cfDNA可减轻牙髓炎中的炎症反应,并可能为患有不可逆性牙髓炎的牙齿保留更多有活力牙髓。