Tumor necrosis factor-alpha inhibits osteogenic differentiation of periodontal ligament stem cells through activating nuclear factor-kappa B signaling pathway

doi:10.3969/j.issn.2095-4344.1793

Abstract

Abstract:

BACKGROUND: Previous studies have focused on the effects of inflammatory factors-activated nuclear factor-kappa B signaling pathway on bone cells. Little has been reported on the effects of micro-inflammatory environment on osteogenic differentiation and nuclear factor-kappa B signaling pathways of human periodontal ligament stem cells.
OBJECTIVE: To investigate the effect of tumor necrosis factor-alpha on osteogenic differentiation and nuclear factor-kappa B signaling pathway of periodontal ligament stem cells.
METHODS: Human periodontal ligament stem cells were cultured in vitro by enzymatic digestion combined with tissue explant method, and the cell source was identified by immunohistochemical staining. The study protocol was approved by the Ethics Committee of the Affiliated Hospital of Qinghai University, and informed consent was obtained from each patient prior to the enrollment in the study. The cells were cultured in simple osteogenic induction medium (control group) or osteogenic induction medium containing 10 μg/L tumor necrosis factor-alpha (tumor necrosis factor-alpha group). Alizarin red staining and alkaline phosphatase activity were used to detect the effect of tumor necrosis factor-alpha on osteogenic differentiation of periodontal ligament stem cells. Real-time quantitative RT-PCR was used to detect the effect of tumor necrosis factor-alpha on the expression of osteogenic differentiation genes, Osterix and Runx2. Western blot was used to detect the effect of tumor necrosis factor-alpha on nuclear factor-kappa B signaling pathway.
RESULTS AND CONCLUSION: Immunohistochemical staining showed that human periodontal ligament stem cells were positive for anti-vimentin and negative for anti-keratin. After alizarin red staining, mineralized nodules formed in the tumor necrosis factor-alpha group were fewer in number, smaller in scope and lighter in staining than those in the control group. Alkaline phosphatase activity in the tumor necrosis factor-alpha group was significantly lower than that in the control group (P < 0.05). Real-time quantitative RT-PCR results showed that the expression levels of Osterix and Runx2 mRNA in tumor necrosis factor-alpha group were significantly lower than those in the control group (P < 0.05). Western blot assay showed that, compared with the control group, the expression of p-IκBα protein increased significantly in the tumor necrosis factor-alpha group (P < 0.05), while the expression of p65 and IκBα proteins decreased (P < 0.05). These findings indicate that tumor necrosis factor-alpha can inhibit the osteogenic differentiation of human periodontal ligament stem cells by activating the nuclear factor-kappa B signaling pathway.

Key words: human periodontal ligament stem cells, tumor necrosis factor-alpha, osteogenic differentiation, nuclear factor-kappa B signaling pathway, alkaline phosphatase activity, osteogenic gene

CLC Number:

Zhang Yun, Chang Qun’an, Li Shengmei, Zhang Yuan.

Tumor necrosis factor-alpha inhibits osteogenic differentiation of periodontal ligament stem cells through activating nuclear factor-kappa B signaling pathway

[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 3993-3997.

Figures/Tables 2

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