M1 Macrophages Drive Cxcr2+ Netosis Via Cxcl8/Cxcr2 In Pulpitis

To elucidate how M1 macrophages exacerbate pulpitis by driving CXCR2+ neutrophil extracellular traps (NETs) formation via the CXCL8/CXCR2 signaling axis. We performed single-cell RNA sequencing (scRNA-seq) on normal and irreversible pulpitis tissues to delineate neutrophil subpopulations driving NETs release and their cell-cell communication networks within the inflammatory microenvironment. Through integration of machine learning algorithms (LASSO, random forest) combined with in vitro functional assays, we identified and validated novel NETs-associated biomarkers in pathological dental pulp. Single-cell RNA sequencing (scRNA-seq) revealed three distinct neutrophil clusters in dental pulp, with histopathological validation identifying CXCR2+Neu as the predominant NETs-releasing subpopulation in inflamed pulp. Mechanistically, cell-cell interaction analysis coupled with in vitro functional assays demonstrated that M1 macrophages drive NETs release via CXCL8/CXCR2-mediated chemotactic signaling. Clinically, pharmacological inhibition of CXCR2 in a murine pulpitis model ameliorated pulpal inflammation through NETs suppression, establishing CXCR2 as a viable therapeutic target CXCR2-positive neutrophils (CXCR2+ Neu) were identified as the predominant source of neutrophil extracellular traps (NETs) in pulpitis, and targeting CXCR2 emerges as a promising therapeutic strategy to mitigate pulpal inflammatory damage.