Intervention with pyrrolidine dithioearbamate stimulates expression of RANKL and OPG in lipopolysaccharide-stimulated periodontal ligament stem cells

doi:10.3969/j.issn.2095-4344.0901

Abstract

Abstract:

BACKGROUND: It is generally believed that various cytokines, such as receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG), which regulate osteoclast and osteoblast formation and differentiation, are mainly secreted by osteoblasts and bone marrow stromal cells. Whereas in the periodontal tissues, homologous periodontal ligament stem cells (PDLSCs) also play a significant role in the regulation of inflammation and immunity. Thus, virulence factors are very likely to mediate alveolar bone resorption by involvement in the regulation of RANKL and OPG.
OBJECTIVE: To explore whether the inhibition of nuclear factor-κB activity by pyrrolidine dithiocarbamate (PDTC) can relieve the imbalance of RANKL/RANK/OPG system in PDLSCs in inflammatory state.
METHODS: After isolation, culture and differentiation in vitro, passage 4 PDLSCs were divided into four group: control group, cultured in the DMEM medium containing 10% fetal bovine serum; lipopolysaccharide (LPS) group, cultured in the DMEM medium containing 10 mg/L LPS; 10 μmol/L PDTC group, pretreated with 10 μmol/L PDTC for 30 minutes followed by removal of the medium and addition of the medium containing 10 mg/L LPS; 100 μmol/L PDTC group, pretreated with 100 μmol/L PDTC for 30 minutes followed by removal of the medium and addition of the medium containing 10 mg/L LPS. In the latter two groups, cell morphology was observed under inverted microscope at 30 minutes after PDTC pretreatment. After 24 hours of culture, expression of OPG and RANKL mRNA and the ratio of RANKL/OPG in cells were detected in each group.
RESULTS AND CONCLUSION: (1) PDTC pretreatment at a concentration of 100 μmol/L could cause cell morphological changes: some cells were adherent and retracted, and became round with the cell body shrinking in size. (2) Compared with the control group, the expression of RANKL in the PDLSCs was significantly increased in the LPS group (P < 0.05), and the expression of RANKL in the PDLSCs pretreated with 10 μmol/L PDTC had no significant change (P > 0.05), while compared with the LPS group, the expression of RANKL in the PDLSCs pretreated with 100 μmol/L PDTC was significantly lowered (P < 0.05). (3) Compared with the control group, the expression of OPG in the PDLSCs was slightly lowered in the LPS group (P > 0.05). After pretreatment with 10 μmol/L PDTC, the expression of OPG in the PDLSCs was significantly increased (P <0.05) compared with the LPS group, while the expression of OPG in the PDLSCs pretreated with 100 μmol/L PDTC had no significant difference from that in the LPS group (P > 0.05). (4) Compared with the control group, the ratios of RANKL/OPG in the other groups were significantly increased (P < 0.05), especially in the LPS group. With the increase of PDTC concentration, the ratio of RANKL/OPG decreased gradually(P < 0.05). These results reveal that PDTC can relieve the imbalance of RANK/RANKL/OPG system in the PDLSC in inflammatory state by inhibiting nuclear factor-κB activity.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Periodontal Ligament, Stem Cells, Osteoprotegerin, RANK Ligand, Pyrrolidines, Tissue Engineering

CLC Number:

R394.2

Wang Ping-ting, Zhou Pan, Zheng Guo-ying, Liu Da-yong. Intervention with pyrrolidine dithioearbamate stimulates expression of RANKL and OPG in lipopolysaccharide-stimulated periodontal ligament stem cells[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(21): 3365-3370.

Figures/Tables 2

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