Abstract
The present study aimed to investigate whether gingival fibroblasts (GFs) of patients with aggressive periodontitis (AgP) are more sensitive to lipopolysaccharide (LPS) stimulation than GFs of control subjects. AgP causes rapid periodontal destruction, including the production of cytokines [i.e. interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α] and matrix metalloproteinases (MMP)-1, -3 and -9 in AgP GFs. LPS upregulates IL-1β, IL-6, TNF-α, MMP-1, MMP-3, MMP-9 and mitochondrial reactive oxygen species (mtROS). Fibroblasts are known to be associated with immune responses to bacterial virulence factors, but the precise mechanisms underlying this severe periodontal disease are unclear. In the present study, primary human GFs of four patients with AgP and four healthy subjects were challenged in vitro with LPS from Porphyromonas gingivalis (P. gingivalis). The generation of mtROS in GFs was assessed using MitoSOX Red. The expression of genes encoding inflammatory cytokines and MMPs in GFs was analyzed using reverse transcription-quantitative polymerase chain reaction, and the expression of proteins was analyzed using ELISA and Western blotting. Human GFs of patients with AgP exhibited higher levels of mtROS, and higher mRNA and protein expression levels of proinflammatory cytokines, including IL-1β, IL-6, MMP-1, MMP-3 and MMP-9 compared with healthy human GFs following stimulation with LPS from P. gingivalis. In the present study, it was demonstrated that GFs of patients with AgP display hyperreactivity when challenged with LPS.