Azithromycin Promotes the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells after Stimulation with TNF-α. (31st October 2018)

• Record Type: Journal Article
• Authors: Meng, Tingting; Zhou, Ying; Li, Jingkun; Hu, Meilin; Li, Xiaomeng; Wang, Pingting; Jia, Zhi; Li, Liyu; Liu, Dayong
• Other Names: Zanvit Peter Academic Editor.
• Is Part Of: Stem cells international. Volume 2018(2018)
• Publication Date: 2018-10-31
• Subjects: Stem Cells -- Periodicals; Stem Cells -- Therapeutic use -- Periodicals; Stem Cells -- Transplantation -- Periodicals; 616.0277405
• DOI: 10.1155/2018/7961962
• Languages: English
• ISSNs: 1687-966X

Abstract

Abstract : Background and Objective . This study investigated the effects and underlying mechanisms of azithromycin (AZM) treatment on the osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) after their stimulation with TNF- α in vitro. Methods . PDLSCs were isolated from periodontal ligaments from extracted teeth, and MTS assay was used to evaluate whether AZM and TNF- α had toxic effects on PDLSCs viability and proliferation. After stimulating PDLSCs with TNF- α and AZM, we analyzed alkaline phosphatase staining, alkaline phosphatase activity, and alizarin red staining to detect osteogenic differentiation. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis was performed to detect the mRNA expression of osteogenic-related genes, including RUNX2, OCN, and BSP . Western blotting was used to measure the NF- κ B signaling pathway proteins p65, phosphorylated p65, I κ B- α, phosphorylated I κ B- α, and β -catenin as well as the apoptosis-related proteins caspase-8 and caspase-3. Annexin V assay was used to detect PDLSCs apoptosis. Results . TNF- α stimulation of PDLSCs decreased alkaline phosphatase and alizarin red staining, alkaline phosphatase activity, and mRNA expression of RUNX2, OCN, and BSP in osteogenic-conditioned medium. AZM enhanced the osteogenic differentiation of PDLSCs that were stimulated with TNF- α . Western blot analysis showed that β -catenin, phosphorated p65, and phosphorylated I κ B- α protein expression decreased in PDLSCs treated with AZM. In addition, pretreatment of PDLSCs with AZM (10 μ g/ml, 20 μ g/ml) prevented TNF- α -induced apoptosis by decreasing caspase-8 and caspase-3 expression. Conclusions . Our results showed that AZM promotes PDLSCs osteogenic differentiation in an inflammatory microenvironment by inhibiting the WNT and NF- κ B signaling pathways and by suppressing TNF- α -induced apoptosis. This suggests that AZM has potential as a clinical therapeutic for periodontitis.