Characterization and comparison of neutrophil extracellular traps in gingival samples of periodontitis and gingivitis: A pilot study

BACKGROUND AND OBJECTIVE: Neutrophil extracellular traps (NETs) are a recently discovered antimicrobial mechanism used by neutrophils that have been proposed as an intervention in the pathogenesis of periodontitis. The objective of our study was to characterize the expression of NETs in gingival tissues with periodontitis and controls and to compare the expression of these traps in gingival tissue samples of patients with gingivitis and periodontitis. MATERIAL AND METHODS: An observational cross-sectional study was conducted on patients with periodontitis, gingivitis, and controls that needed tooth extraction. Gingival tissue biopsies were gathered after clinical examination and tooth extraction. Electron microscopy and immunofluorescence were performed to characterize NETs, comparing periodontitis and control patients. Immunohistochemical analysis was performed to quantify neutrophil extracellular trap expression through extracellular citrullinated histone H3 and myeloperoxidase in biopsies from patients with gingivitis and periodontitis. RESULTS: Thirteen biopsies were gathered from 13 patients: five gingivitis, six periodontitis, and two controls. Electron microscopy and immunofluorescence imaging showed greater expression of neutrophils present in periodontal inflamed tissue compared with controls. Release of nuclear content to the extracellular space was observed, compatible with the formation of NETs. The expression of citrullinated histone H3 was higher in gingivitis samples than periodontitis samples (P = 0.0106). Myeloperoxidase expression was higher in periodontitis than gingivitis, but without achieving statistical significance. CONCLUSION: Neutrophil extracellular traps were found in tissue samples of periodontitis as extracellular components of chromatin, along with neutrophil enzymes, that were not present in healthy controls. The comparison of NETs expression in periodontitis and gingivitis showed higher expression in gingivitis, associating them to acute phases of the periodontal inflammatory process.