Abstract
Objective This research is aimed at investigating how high glucose affects the proliferation and apoptosis in periodontal ligament stem cells (PDLSCs) in the presence of TNF-α. Methods PDLSCs obtained from periodontal healthy permanent teeth were treated under either high-glucose condition (30 mmol/L, G30 group) or normal glucose condition (5.6 mmol/L, G5.6 group) in the presence or absence of TNF-α (10 ng/ml) for 2 to 6 days. Cell proliferation and cell cycle were evaluated by CCK-8, EdU incorporation assay, and flow cytometry. Cell apoptosis was assessed by annexin V/PI staining. Protein expression was detected by western blotting. Cellular ROS expression was evaluated by CellROX labeling and flow cytometry. Specific antibodies targeting TNFR1 and TNFR2 were used to block TNF-α signaling. Vitamin C was also used to verify if the blockage of ROS can rescue PDLSCs in the presence of high glucose and TNF-α. Results CCK-8 assay showed that high glucose exacerbated TNF-α-induced cell viability inhibition (57.0%, 85.2%, and 100% for the G30+TNF-α group, G5.6+TNF-α group, and control group, respectively) on day 6. High glucose increased protein expression of TNFR1 compared with the control group on day 2 (1.24-fold) and day 6 (1.26-fold). Blocking TNFR1 totally reversed the proliferative inhibition in G30+TNF-α group. The addition of vitamin C or TNFR1 antibody totally reversed the elevation of intracellular ROS expression caused by high glucose and TNF-α. Vitamin C partially restored cell proliferation in the presence of high glucose and TNF-α. Conclusion High glucose exacerbates TNF-α-induced proliferative inhibition in human periodontal ligament stem cells through the upregulation and activation of TNF receptor 1. Inhibition of intracellular ROS expression by vitamin C partially rescues PDLSCs in terms of cell proliferation.
Highlights
Periodontitis, which is caused by a bacterial infection and exacerbated by host immunological reactions, may result in attachment loss, bone resorption, and eventually loss of teeth
periodontal ligament stem cells (PDLSCs) are a subgroup of mesenchymal stem cells in terms of phenotype and multilineage differentiation potentials [2]
We found that protein expression of both TNFR1 and TNFR2 in CD146-positive PDLSCs was 2-fold higher than that of CD146-negative periodontal ligament cells
Summary
Periodontitis, which is caused by a bacterial infection and exacerbated by host immunological reactions, may result in attachment loss, bone resorption, and eventually loss of teeth. Periodontal ligament stem cells (PDLSCs) have shown some promise as seeds for regenerating periodontal tissues [1]. PDLSCs are a subgroup of mesenchymal stem cells in terms of phenotype (express stromal cell surface markers, including CD146, Stro-1, CD105, CD90, and CD73) and multilineage differentiation potentials [2]. In our previous study [3], CD146-positive cells from human periodontal ligament were found to express MSC surface markers (CD105, CD90, CD73, CD44, and Stro-1) and show higher proliferative and osteogenic potential than CD146negative cells. PDLSCs are capable of regenerating periodontal tissues, including periodontal ligaments, alveolar bone, cementum, and peripheral nerves and blood vessels in animal models [4]. Clinical case reports [5] show that autologous periodontal ligament progenitor cell transplantation in intrabony defect of 3 periodontitis patients achieves significant alveolar bone regeneration as well as
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