ABSTRACT

Background: Porphyromonas gingivalis (P. gingivalis) is a gram-negative bacterium

that have lipopolysaccharide (LPS) in the outer surface. LPS induces pro-
inflammatory cytokines, such as tumor necrosis factor- (TNF-), interleukin-6 (IL-
6), and IL-8, which induce periodontal tissue and alveolar bone destruction. TNF-
activates the immunologic transcription of nuclear factor-kappa B (NF-κB) signaling
pathway. Activating of NF-κB signaling pathway can affect the ability of
mesenchymal stem cell (MSC) differentiation. Dental pulp stem cells (DPSCs) are a
type of MSC and play an important role in alveolar bone regeneration and are
expected to have future applications in cellular therapies for periodontitis. However,
no studies have examined the effects of LPS on DPSCs through NF-κB signaling
pathway. The aim of this study was to investigate how LPS affects the bone nodule
formation in DPSCs through NF-κB signaling pathway. Methods: DPSCs derived
from extracted molar 3 were used in this study. DPSCs were cultured and treated
with/without osteogenic inductors and/or LPS for 2 hours to detect NF-κB activity, or
for 3 weeks to detact bone nodule formation. NF-κB activity detection was performed
with flourometric cell-base ELISA, meanwhile bone nodule formation was assayed
by staining with Alizarin Red. Resulted NF-κB activities were statistically analyzed.
Meanwhile, bone nodule formations were documanted under inverted light
microscope. Result: DMEM and osteogenic medium with LPS (10 mg/mL) group
showed significant increasing NF-κB activation compared to the group without LPS
(p=0.00) and the calcified nodules stained with Alizarin Red was not formed.
Conclusions: This study provides the first findings that LPS from P. gingivalis
inhibits bone nodule formation in DPSCs through NF-κB signaling pathway. These
findings will help clarify the relationship between LPS and periodontal tissue
regeneration using DPSCs.

Keywords: Lipopolysaccharides, Nuclear factor-kappa B, Osteoblasts, Dental pulp

stem cells.