Qiu Wei, Guo Ruiming, Yu Hongwen, Chen Xiaoxin, Chen Zehao, Ding Dian, Zhong Jindou, Yang Yumeng, Fang Fuchun
Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
J Adv Res. 2025 Jun;72:287-301. doi: 10.1016/j.jare.2024.07.028. Epub 2024 Jul 30.
Exaggerated neutrophil recruitment and activation are the major features of pathological alterations in periodontitis, in which neutrophil extracellular traps (NETs) are considered to be responsible for inflammatory periodontal lesions. Despite the critical role of NETs in the development and progression of periodontitis, their specific functions and mechanisms remain unclear.
To demonstrate the important functions and specific mechanisms of NETs involved in periodontal immunopathology.
We performed single-cell RNA sequencing on gingival tissues from both healthy individuals and patients diagnosed with periodontitis. High-dimensional weighted gene co-expression network analysis and pseudotime analysis were then applied to characterize the heterogeneity of neutrophils. Animal models of periodontitis were treated with NETs inhibitors to investigate the effects of NETs in severe periodontitis. Additionally, we established a periodontitis prediction model based on NETs-related genes using six types of machine learning methods. Cell-cell communication analysis was used to identify ligand-receptor pairs among the major cell groups within the immune microenvironment.
We constructed a single-cell atlas of the periodontal microenvironment and obtained nine major cell populations. We further identified a NETs-related subgroup (NrNeu) in neutrophils. An in vivo inhibition experiment confirmed the involvement of NETs in gingival inflammatory infiltration and alveolar bone absorption in severe periodontitis. We further screened three key NETs-related genes (PTGS2, MME and SLC2A3) and verified that they have the potential to predict periodontitis. Moreover, our findings revealed that gingival fibroblasts had the most interactions with NrNeu and that they might facilitate the production of NETs through the MIF-CD74/CXCR4 axis in periodontitis.
This study highlights the pathogenic role of NETs in periodontal immunity and elucidates the specific regulatory relationship by which gingival fibroblasts activate NETs, which provides new insights into the clinical diagnosis and treatment of periodontitis.
中性粒细胞的过度募集和激活是牙周炎病理改变的主要特征，其中中性粒细胞胞外诱捕网（NETs）被认为是炎症性牙周病变的罪魁祸首。尽管NETs在牙周炎的发生发展中起关键作用，但其具体功能和机制仍不清楚。
阐明NETs在牙周免疫病理中的重要功能和具体机制。
我们对健康个体和诊断为牙周炎患者的牙龈组织进行了单细胞RNA测序。然后应用高维加权基因共表达网络分析和拟时间分析来表征中性粒细胞的异质性。用NETs抑制剂处理牙周炎动物模型，以研究NETs在重度牙周炎中的作用。此外，我们使用六种机器学习方法建立了基于NETs相关基因的牙周炎预测模型。通过细胞间通讯分析确定免疫微环境中主要细胞群之间的配体-受体对。
我们构建了牙周微环境的单细胞图谱，并获得了九个主要细胞群。我们进一步在中性粒细胞中鉴定出一个NETs相关亚群（NrNeu）。体内抑制实验证实NETs参与了重度牙周炎的牙龈炎症浸润和牙槽骨吸收。我们进一步筛选出三个关键的NETs相关基因（PTGS2、MME和SLC2A3），并验证它们具有预测牙周炎的潜力。此外，我们的研究结果表明，牙龈成纤维细胞与NrNeu的相互作用最多，并且它们可能在牙周炎中通过MIF-CD74/CXCR4轴促进NETs的产生。
本研究突出了NETs在牙周免疫中的致病作用，并阐明了牙龈成纤维细胞激活NETs的具体调控关系，为牙周炎的临床诊断和治疗提供了新的见解。
