Periodontal ligament fibroblasts regulate osteoblasts by exosome secretion induced by inflammatory stimuli

Abstract

ObjectivesThis study evaluated the role of human periodontal ligament fibroblasts (hPDLFs)-derived exosomes in periodontitis progression and discovered whether hPDLFs influence bone remodeling activity via exosome secretion. Materials and MethodsExosomes were isolated and quantified from Porphyromonas gingivalis lipopolysaccharide (LPS)-treated primary hPDLFs and evaluated by western blotting, dynamic light scattering, and transmission electron microscopy. GW4869 was used to block exosome secretion in conditioned medium (CM). hPDLFs-derived CM, CM containing GW4869 (CM + GW4869) and exosomes were used to stimulate MG-63 cell lines. The expression levels of proinflammatory mediators, osteogenic genes, and osteoclastogenesis-related genes were measured by quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, western blotting, and alkaline phosphatase staining. ResultsExosome-enriched protein and total exosomal protein levels were higher in the LPS-treated group than in the vehicle controls. hPDLFs-derived exosomes were incorporated into MG-63 osteoblasts and slightly upregulated the expression of Interleukin-6 and tumor necrosis factor-alpha. CM and exosomes inhibited alkaline phosphatase, Collagen-I, Runt-related transcription factor 2, and Osteoprotegerin expression as well as ALP activity, and blocking exosome secretion by GW4869 eliminated the inhibitory effects. ConclusionThese results indicate that LPS-pretreated hPDLFs induce inflammation and inhibit osteogenic activity of osteoblasts through secreting exosomes. This study provides a potential mechanism by which localized periodontal inflammation may influence bone remodeling by release exosomes.