Effect of trichostatin A on the osteogenic differentiation potential of periodontal ligament stem cells in inflammatory microenvironment induced by tumor necrosis factor-α stimulation
10.3760/cma.j.issn.1002-0098.2016.04.010
- VernacularTitle:曲古抑菌素A对肿瘤坏死因子α诱导的炎症微环境下牙周膜干细胞成骨分化能力的影响
- Author:
Hong WANG
1
;
Qi CHEN
;
Wenjia LIU
;
Zhenhua YANG
;
Dong LI
;
Fang JIN
Author Information
1. 第四军医大学口腔医学院正畸科 军事口腔医学国家重点实验室
- Keywords:
Periodontal ligament;
Stem cells;
Histone deacetylases;
Trichostatin A;
Osteogenic differentiation
- From:
Chinese Journal of Stomatology
2016;51(4):235-241
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the expression of histone deacetylase(HDAC)1-11 of humanperiodontal ligament stem cells(PDLSC) in normal and inflammatory microenvironments,and to investigate the effect of histone deacetylase inhibitor trichostatin A(TSA) on the osteogenic differentiation potential of PDLSC in inflammatory microenvironment induced by tumor necrosis factor-α(TNF-α) stimulation.Methods PDLSC were isolated from periodontal ligament tissues obtained from the surgically extracted human teeth and cultured by single-colony selection.The expression of HDAC1-11 in cells with or without TNF-α(10 μg/L) stimulation was evaluated by quantitative real time-PCR(RT-PCR).The effect of TSA on cell proliferation was investigated by methyl thiazolyl tetrazolium(MTT) assay.The influence of TSA on osteogenic differentiation of PDLSC in inflammatory microenvironment with TNF-α stimulation was assessed by alizarin red staining,qoantitative RT-PCR and Western blotting,respectively.Results The expression of HDAC in PDLSC with TNF-α stimulation was significantly higher than that in normal PDLSC(P＜0.05) (except HDAC7,P=0.243).TSA had no significant effect on PDLSC proliferation at the concentration of 50 nmol/L(P=0.232).The alizarin red staining showed that PDLSC in TNF-α group generated less mineralized nodule than the control group,while the cell matrix mineralization in TSA group was improved obviously.TNF-α had an inhibitory effect on the expression of osteogenesis related genes,runt-related transcription factor-2(RUNX2) and alkaline phosphatase(ALP),with relative gene expression ratio (experimental/control) decreased to 0.17 ±0.02 and 0.32±0.03,while TSA could significantly increase the genes' expression to 0.67±0.03 and 0.89±0.02(P＜0.01).Western blotting test showed that in TNF-α group the expression of osteogenesis related proteins was obviously reduced,and compared with the TNF-α group,TSA could significantly promote the expression of proteinsin inflammatory microenvironment.Conclusions PDLSC in inflammatory microenvironment by TNF-α stimulation had a higher expression of HDAC than that in normal conditions.TSA,as a histone deacetylase inhibitor,could significantly promote the osteogenic differentiation potential of PDLSC in inflammatory microenvironment by suppressing HDAC.
