Oxidative stress in human gingival fibroblasts from periodontitis versus healthy counterparts.

Abstract

Elevated p53 promotes oxidative stress and production of pro-inflammatory cytokines in liposaccharide (LPS)-treated healthy human gingival fibroblasts (HGFs). This study compared oxidative stress, production of inflammatory cytokines, and p53 expression in HGFs from patients with chronic periodontitis (CP) and healthy subjects in vitro upon LPS from Porphyromonas gingivalis challenge. Human gingival fibroblasts were isolated from 6 biopsies-3 from healthy donors and 3 from diseased area in CP (Grade B, Stage III). HGFs were cultured with or without 1μg/ml 24h LPS. Oxidative stress was assessed by analyzing the level of reactive oxygen species (ROS). Mitochondrial membrane potential and respiration were determined by immunofluorescence and respirometry, respectively. Tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β were determined by enzyme-linked immunosorbent assay. P53 expression was monitored by Western blot and immunofluorescence. Human gingival fibroblasts from CP exhibited increased levels of mitochondrial p53, enhanced ROS production, decreased mitochondrial membrane potential, increased mitochondrial oxygen consumption, and increased secretion of TNF-α, IL-6, and IL-1β, as compared to HGFs from healthy donors. Moreover, LPS exacerbated these changes. Human gingival fibroblasts from CP exhibited stronger basal and LPS-inducible oxidative stress and inflammatory response as compared to HGFs from healthy subjects by increased p53 in mitochondria.