Objective: The aim of this study was to investigate the effect of saliva of patients with chronic periodontitis (CPD) on the differentiation, activation, and secretion of osteoclast-maturing mediators of macrophages.
Methods: A total of 40 saliva samples were collected from healthy donors (n=20) and severe periodontitis patients (n=20). Peripheral blood mononuclear cells (PBMCs) and THP-1 monocyte line cells were challenged with 15% saliva for 5 days. The phenotype, surface marker, and phagocytosis of macrophages were analyzed by flow cytometry and microscopy. Osteoclast-maturing mediators were assayed by using enzyme-linked immunosorbent assay (ELISA) kits.
Results: When PBMCs were treated with CPD saliva for 5 days, 61.25%±11.33% of cells were transformed into large granular cells; 86.78%±13.69% of large granular cells were identified as CD14⁺⁺CD16⁺ macrophages. When THP-1 cells were treated with CPD saliva, most cells attached to the bottom of cell culture plates, thereby exhibiting macrophage morphology and releasing additional osteoclast-maturing mediators. Furthermore, the phagocytosis of THP-1 cells considerably increased in the presence of CPD saliva (66.35%±9.67%) compared with medium control (33.33%±7.52%), or healthy saliva (40.71%±3.52%).
Conclusions: Saliva from patients with CPD can induce macrophage differentiation, activate phagocytose microorganisms, and secrete osteoclast-maturing mediators.
目的 探讨慢性牙周炎（CPD）唾液对巨噬细胞分化、活化、吞噬功能和分泌破骨细胞成熟因子的影响。方法 收集20名正常人和20名CPD患者的唾液样品。15%唾液刺激外周血单个核细胞（PBMCs）和人单核细胞系（THP-1）5 d，用流式细胞仪检测淋巴细胞亚群变化，显微镜观察巨噬细胞分化和吞噬活性；酶联免疫吸附测定（ELISA）检测巨噬细胞分泌破骨细胞成熟因子情况。结果 CPD唾液刺激PBMCs 5 d后，61.25%±11.33%的PBMCs分化为大颗粒细胞，其中86.78%±13.69%的大颗粒细胞为CD14⁺⁺CD16⁺巨噬细胞；CPD唾液可诱导THP-1细胞分化为巨噬细胞，表达高水平CD14，分泌高水平破骨细胞成熟因子；CPD唾液诱导后THP-1细胞的吞噬作用（66.35%±9.67%）明显高于正常唾液组（40.71%±3.52%）和阴性对照组（33.33%±7.52%）。结论 CPD唾液能够诱导巨噬细胞的分化和活化，促进巨噬细胞吞噬细菌，分泌破骨细胞成熟因子。.
Keywords: chronic periodontitis; macrophage; saliva.