Journal of endodontics · 2018
Human Dental Pulp Cells Express Cellular Markers for Inflammation and Hard Tissue Formation in Response to Bacterial Information.
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Sugiuchi Akina, Sano Yousuke, Furusawa Masahiro, Abe Shu, Muramatsu Takashi
Аннотация
INTRODUCTION: Lipopolysaccharide (LPS) is a major component of the outer membranes of gram-negative bacteria associated with deep dental caries and pulpitis. When bacteria invade dentinal tubes and dentin is continually destroyed, tertiary dentin is formed by preexisting odontoblasts. However, the relationship between LPS and tertiary dentin formation remains unclear. We investigated whether LPS stimulation induces the formation of hard tissue in human dental pulp cells (hDPCs).
METHODS: Immortalized hDPCs were cultured, and Escherichia coli-derived LPS (1 μg/mL) was incorporated into the culture medium. Samples were obtained after 0, 1, 3, 7, 14, and 21 days, and messenger RNA expression of IL-1β, IL-6, Wnt5a, Runx2, ALP, and alkaline phosphatase (ALP) activity was investigated.
RESULTS: Quantitative real-time polymerase chain reaction revealed higher messenger RNA expression levels of IL-1β and IL-6 in the LPS group on 1 day (P < .05). The expression levels of dentinogenesis-related markers including Wnt5a, Runx2, and ALP were higher in the LPS group (2.0-, 4.7- and 10.0-fold, respectively) than that in the control group at 14 days (P < .01). ALP activity was significantly stronger in the LPS group than in the control group at 21 days (P < .01). Treatment of Box5, an antagonist of Wnt5a, showed a decreased expression of Runx2 and ALP (P < .05).
CONCLUSIONS: These results indicate that LPS stimulation induces the gene expression of inflammatory cytokines and hard tissue formation through Wnt5a signaling pathways in hDPCs.
METHODS: Immortalized hDPCs were cultured, and Escherichia coli-derived LPS (1 μg/mL) was incorporated into the culture medium. Samples were obtained after 0, 1, 3, 7, 14, and 21 days, and messenger RNA expression of IL-1β, IL-6, Wnt5a, Runx2, ALP, and alkaline phosphatase (ALP) activity was investigated.
RESULTS: Quantitative real-time polymerase chain reaction revealed higher messenger RNA expression levels of IL-1β and IL-6 in the LPS group on 1 day (P < .05). The expression levels of dentinogenesis-related markers including Wnt5a, Runx2, and ALP were higher in the LPS group (2.0-, 4.7- and 10.0-fold, respectively) than that in the control group at 14 days (P < .01). ALP activity was significantly stronger in the LPS group than in the control group at 21 days (P < .01). Treatment of Box5, an antagonist of Wnt5a, showed a decreased expression of Runx2 and ALP (P < .05).
CONCLUSIONS: These results indicate that LPS stimulation induces the gene expression of inflammatory cytokines and hard tissue formation through Wnt5a signaling pathways in hDPCs.