The effect of kaempferol on the osteogenic/dentinogenic differentiation of stem cells from the human apical pa-pilla under inflammation and its related mechanism
Objective:To study the effect of kaempferol on the osteogenic/dentinogenic differentiation of stem cells from the human apical papilla(SCAP)under inflammation and the regulation of Wnt/β-catenin signaling pathway.Methods:SCAP was obtained by enzymatic digestion combined with tissue block method,and cell surface markers were detected by flow cytometry.After 2 weeks of osteogenic induction,they were divided into blank control group,TNF-α group,TNF-α+XAV939 group,TNF-α+kaempferol group,TNF-α+kaempferol group+XAV939 group.Alkaline phosphatase(ALP)staining activity as-say,alizarin red staining and real-time RT-PCR,Western blot analysis to detect the expression of pathway-related proteins.Results:Compared with the TNF-α group,ALP activity was significantly increased in the TNF-α+0.1 μmol/L kaempferol group,decreased in the TNF-α+XAV939 group(P<0.05).The expression of ALP activity was inhibited in the TNF-α+0.1 μmol/L kaempferol+XAV939 group compared with the TNF-α+0.1 μmol/L kaempferol group(P<0.05).Alizarin red staining was significantly lighter and the number of mineralized nodules formed was small and sparse in the TNF-α+XAV939 group without added kaempferol compared with the TNF-α+0.1 μmol/L kaempferol+XAV939 group.Compared with the TNF-α group,the expression levels of DSPP,OPN,RUNX2,and OCN were significantly increased in the TNF-α+0.1 μmol/L kaempferol group,while they were all decreased in the TNF-α+XAV939 group(P<0.05).The expression levels of DSPP,OPN,RUNX2,and OCN were all reduced in the TNF-α+0.1 μmol/L kaempferol+XAV939 group compared with the TNF-α+0.1 μmol/L kaempferol group(P<0.05).The expression levels of osteogenesisrelated proteins and pathway-related proteinsβ-catenin and p-GSK3β were higher in the TNF-α group than in the TNF-α+XAV939 group(P<0.05).Compared with the TNF-α+0.1 μmol/L kaempferol group,the expression levels of β-catenin and p-GSK3β were reduced in the TNF-α+0.1 μmol/L kaempferol+XAV939 group(P<0.05).And there was no significant difference in GSK3β expression level a-mong the five groups.Conclusion:Kaempferol concentration of 0.1 μmol/L can promote the potential of SCAP osteogenic/dentinogenic differentiation towards differentiation under inflammatory state,and Wnt/β-catenin signaling pathway plays an important role in this process.
Stem cells from the apical papilla(SCAP)Tumor necrosis factor-α(TNF-α)KaempferolWnt/β-catenin
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