Lipopolysaccharide inhibits osteogenic differentiation of periodontal ligament stem cells partially through toll-like receptor 4-mediated ephrinB2 downregulation
Lipopolysaccharide inhibits osteogenic differentiation of periodontal ligament stem cells partially through toll-like receptor 4-mediated ephrinB2 downregulation
复制标题
脂多糖部分通过Toll样受体4介导的ephrinB2下调抑制牙周膜干细胞的成骨分化
DOI：
10.1007/s00784-020-03211-w
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发表时间：
2020-10-01
影响因子：
3.4
通讯作者：
Wang, Penglai
中科院分区：
文献类型：
--
作者：
Wang, Wen;Yuan, Changyong;Wang, Penglai
目的探讨LPS/TLR 4和ephrinB 2信号通路在牙周膜干细胞（hPDLSCs）成骨分化中的相互作用。进行qPCR、碱性磷酸酶（ALP）染色和茜素红S染色，以评估成骨基因表达、ALP活性和矿化结节形成。LPS处理后，EphrinB 2 mRNA和蛋白表达水平也进行了测定。为了探讨ephrinB 2在LPS抑制hPDLSCs成骨分化中的作用，分别用ephrinB 2-Fc刺激hPDLSCs或用ephrinB 2慢病毒转染hPDLSCs，观察hPDLSCs的成骨分化能力。阻断TLR 4可部分逆转LPS诱导的ephrinB 2表达下降。EphrinB 2-Fc促进矿化结节形成，并增加hPDLSC中ALP、骨钙素（OCN）和骨形态发生蛋白2（BMP 2）的表达。EphrinB 2-overexpressing hPDLSCs与LPS处理后表达更高的ALP和BMP 2 mRNA和更高的ALP活性，并表现出更多的矿化结节的形成，当与宽型hPDLSCs treated with LPS.ConclusionsOur data suggest，LPS降低hPDLSCs的成骨分化能力部分通过下调ephrinB 2的表达通过LPS/TLR 4信号转导。ephrinB 2的上调部分逆转了LPS诱导的hPDLSCs成骨分化能力的受损。临床意义我们的研究结果为LPS介导的PDLSCs成骨分化抑制机制提供了新的见解，并阐明了牙周炎治疗的潜在靶点。
ObjectivesThis study aimed to investigate the possible crosstalk between LPS/toll-like receptor 4 (TLR4) and ephrinB2 signaling in mediating osteogenic differentiation of PDLSCs.Materials and methodsHuman periodontal ligament stem cells (hPDLSCs) were harvested and treated with different concentrations of LPS under osteogenic induction. qPCR, alkaline phosphatase (ALP) staining, and Alizarin Red S staining were performed to assess osteogenic gene expression, ALP activity, and mineralized nodule formation. EphrinB2 mRNA and protein expressions after LPS treatment were also determined. To explore the role of ephrinB2 in LPS-impaired osteogenic differentiation of hPDLSCs, hPDLSCs were stimulated with ephrinB2-Fc or transfected with ephrinB2 lentivirus, and then, the osteogenic differentiation capacity was evaluated.ResultsLPS inhibited osteogenic differentiation of hPDLSCs and downregulated ephrinB2 expression in hPDLSCs during osteogenic differentiation. Blockage of TLR4 partially reversed LPS-induced decrease in ephrinB2 expression. EphrinB2-Fc promoted mineralized nodule formation and increased the expression of ALP, osteocalcin (OCN), and bone morphogenetic protein 2 (BMP2) in hPDLSCs. EphrinB2-overexpressing hPDLSCs treated with LPS expressed higher ALP and BMP2 mRNA and higher ALP activity and showed more mineralized nodule formation, when compared with wide-type hPDLSCs treated with LPS.ConclusionsOur data suggested that LPS decreased the osteogenic differentiation capacity of hPDLSCs partially through downregulation of ephrinB2 expression via LPS/TLR4 signaling. Upregulation of ephrinB2 partially reversed the impaired osteogenic potential of hPDLSCs induced by LPS.Clinical relevanceOur results provided a new insight of mechanism underling LPS-mediated osteogenic differentiation inhibition of PDLSCs and clarified a potential target for the management of periodontitis.