The aims of this study were: 1) to compare the levels of cytokines between healthy and diseased sites, in patients with untreated periodontitis; 2) to correlate cytokine levels with each other and with key periodontal pathogens, in healthy and diseased sites.
Paired gingival crevicular fluid (GCF) samples were obtained from two healthy (probing depth (PD) and clinical attachment level (CAL) ≤3 mm without bleeding) and two diseased sites (PD and CAL ≥5 mm with bleeding on probing [BoP]) of patients with generalized stage III/IV grade B/C periodontitis. GCF levels of eighteen cytokines and subgingival levels of seven periodontal pathogens were assessed by multiplex immunoassay and qPCR, respectively.
A total of 112 subjects and 448 GCF samples were analyzed. The GCF levels of GM-CSF, IL-17, IL-1β, IL-2, IL-21, IL-23 and TGF-β were significantly higher in the diseased than in the healthy sites (p < 0.05). Levels of IL-8 and MIP-1α were significantly higher in the healthy than in the diseased sites (p < 0.05). In the healthy sites, IL-8 and MIP-1α formed an independent cluster of cytokines and, MIP-1α positively correlated with Porphyromonas gingivalis (p < 0.05). In deep sites, smoking negatively associated with GM-CSF, IL-10, IL-17, IL-23, IL-5, IL-6, IL-7, IL-8 and MIP-1α levels (p < 0.05).
Diseased sites exhibited increased levels of T helper 17-related cytokines and TGF-β while healthy sites presented increased levels of the chemokines, IL-8 and MIP-1α. Patients with periodontitis may not only have inflammation in diseased deep sites, but also present significant hidden subclinical inflammation in their shallow clinically healthy sites.

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