Dong Rui, Li Meihui, Gu Xiu Feng, Gao Haiting, Wei Ziyi, Qi Houbao, Zhang Jun, Feng Qiang
Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong Key Laboratory of Oral Tissue Regeneration and Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Shandong University, Jinan, China.
Department of Human Microbiome, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Research Center of Dental Materials and Oral Tissue Regeneration & Shandong Provincial Clinical Research Center for Oral Diseases, Shandong University, Jinan, China.
J Transl Med. 2025 May 14;23(1):540. doi: 10.1186/s12967-025-06569-1.
Periodontitis is a chronic inflammatory disease that significantly impacts periodontal bone regeneration, yet the distinct biological features of osteoblasts in this condition remain poorly understood. This study aims to elucidate the cellular and molecular mechanisms underlying osteoblast dysfunction in periodontitis, with a focus on the role of Fusobacterium nucleatum (Fn) and its effector protein, D-galactose-binding periplasmic protein (Gbp).
Single-cell RNA sequencing (scRNA-seq) data from human gingival tissues of periodontitis patients (PD) and healthy controls (HC) were analyzed to identify cellular heterogeneity and molecular pathways. An experimental periodontitis model in mice and primary osteoblast cultures were used to investigate the effects of Fn and Gbp on osteogenic differentiation. Transcriptomic analysis, gene set enrichment analysis (GSEA), and protein-protein interaction (PPI) networks were employed to explore the underlying mechanisms.
scRNA-seq revealed a reduction in mesenchymal stem cells (MSCs) and osteoblastic lineage cells in PD tissues, with significant downregulation of osteogenic pathways such as Wnt signaling. Fn infection induced alveolar bone destruction in vivo and inhibited osteoblast proliferation, differentiation, and mineralization in vitro. Gbp, an Fn adhesin, similarly impaired osteogenic differentiation by downregulating key osteogenic genes and pathways. Transcriptomic analysis identified shared inflammatory and osteogenic pathways affected by Fn and Gbp, with NF-κB signaling activated and Wnt/β-catenin signaling inhibited. Mechanistically, Gbp interacted with the host protein ANXA2, disrupting the ANXA2/GSK3β complex and inhibiting Wnt/β-catenin signaling, a pivotal route for osteoblast differentiation. ANXA2 knockdown mitigated the Fn/Gbp-induced suppression of osteogenic activity, emphasizing its role in Fn-induced bone loss.
This study demonstrates that Fn and its effector Gbp disrupt osteogenic differentiation by inactivating the Wnt/β-catenin pathway binding to ANXA2.
牙周炎是一种慢性炎症性疾病，对牙周骨再生有显著影响，但在这种情况下成骨细胞独特的生物学特性仍知之甚少。本研究旨在阐明牙周炎中成骨细胞功能障碍的细胞和分子机制，重点关注具核梭杆菌（Fn）及其效应蛋白D - 半乳糖结合周质蛋白（Gbp）的作用。
分析来自牙周炎患者（PD）和健康对照（HC）的人牙龈组织的单细胞RNA测序（scRNA - seq）数据，以确定细胞异质性和分子途径。使用小鼠实验性牙周炎模型和原代成骨细胞培养物来研究Fn和Gbp对成骨分化的影响。采用转录组分析、基因集富集分析（GSEA）和蛋白质 - 蛋白质相互作用（PPI）网络来探索潜在机制。
scRNA - seq显示PD组织中间充质干细胞（MSC）和成骨谱系细胞减少，成骨途径如Wnt信号通路显著下调。Fn感染在体内诱导牙槽骨破坏，在体外抑制成骨细胞增殖、分化和矿化。Gbp作为Fn粘附素，同样通过下调关键成骨基因和途径损害成骨分化。转录组分析确定了受Fn和Gbp影响的共同炎症和成骨途径，NF - κB信号通路被激活，Wnt/β - 连环蛋白信号通路被抑制。机制上，Gbp与宿主蛋白膜联蛋白A2（ANXA2）相互作用，破坏ANXA2/糖原合成酶激酶3β（GSK3β）复合物并抑制Wnt/β - 连环蛋白信号通路，这是成骨细胞分化的关键途径。敲低ANXA2减轻了Fn/Gbp诱导的成骨活性抑制，强调了其在Fn诱导的骨质流失中的作用。
本研究表明，Fn及其效应蛋白Gbp通过与ANXA2结合使Wnt/β - 连环蛋白途径失活，从而破坏成骨分化。
