郑州大学学报（医学版）2025，Vol.60Issue(2)：167-172,6.DOI:10.13705/j.issn.1671-6825.2024.08.053
外源性肿瘤坏死因子刺激基因6对炎症环境下人牙髓干细胞成牙/成骨分化的影响
Exogenous TSG-6 enhances the odontogenic/osteogenic differentiation of human dental pulp stem cells in inflammatory environment
摘要
Abstract
Aim:To explore the role and possible mechanism of tumor necrosis factor-stimulated gene 6(TSG-6)in the odontogenic/osteogenic differentiation of human dental pulp stem cells(hDPSC)in the inflammatory environment.Meth-ods:The pulps of extracted healthy and impacted wisdom teeth were extirpated,and the cells were isolated and cultured.The hDPSC were grouped after identification.The normal control group was cultured with DMEM containing 10％FBS by volume for 96 hours,the induction group was cultured with mineralized induction solution for 96 hours,the inflammatory fac-tor group was cultured with mineralized induction solution containing 50 μg/L TNF-α for 96 hours,and the inflammatory factor+TSG-6 group was cultured with normal medium containing 20 µg/L TSG-6 for 48 hours,and then cultured with mineralized induction solution containing TNF-α for 48 hours.The neutralizing antibody group was incubated with TSG-6 and CD44 neutralizing antibodies for 48 hours and then cultured with mineralized induction solution containing TNF-α for 48 hours.Western blot was performed to measure the expression levels of odontogenic/osteogenic differentiation markers[DSPP,DMP-1,RUNX2]and P65,p-IKB,IκB,respectively.The nuclear transport level of NF-κB in the nucleus of inflam-matory factor+TSG-6 group and neutralizing antibody group were observed by immunofluorescence staining.Twelve 2-day-old SD rats were divided into 4 groups with 3 rats in each group.The rats were sacrificed after anesthesia,and the mandibles were taken and cultured in culture medium containing DMSO,mineralized induction solution,mineralized induction solution containing TNF-α,mineralized induction solution containing TNF-α and TSG-6,respectively for 10 days.The mineralization ability of teeth in each group was evaluated by HE and Masson tri-color staining.Results:Compared with normal control group,the expression levels of DSPP,DMP-1 and RUNX2 proteins in the mineralized induction group were increased(P＜0.05);compared with the mineralization induction group,the expression levels of the above proteins in the inflammatory fac-tor group were decreased(P＜0.05);compared with the inflammatory factor group,the expression levels of the above pro-teins in the inflammatory factor+TSG-6 group were increased(P＜0.05).Compared with normal control group,the expres-sion levels of P65 and p-IKB proteins in the inflammatory factor group were increased(P＜0.05);compared with the inflam-matory factor group,the expression levels of P65 and p-IκB proteins in the inflammatory factor+TSG-6 group were lower(P＜0.05).Compared with the inflammatory factor+TSG-6 group,the expression levels of DSPP,DMP-1 and RUNX2 pro-teins in the neutralizing antibody group were decreased,while the expression levels of P65 and p-IκB proteins were in-creased,and the number of NF-κB nuclear transport cells was increased.Animal experiment showed that TSG-6 treatment could promote the formation of predentin in the teeth of rats under the inflammatory environment.Conclusion:Exogenous TSG-6 could enhance the odontogenic/osteogenic differentiation of hDPSC in the inflammatory environment,and the mecha-nism may be related with inhibiting the activation of NF-κB signaling pathway.关键词
牙髓干细胞/TNF-α/肿瘤坏死因子刺激基因6/NF-κB信号通路/成牙/成骨分化Key words
dental pulp stem cell/TNF-α/tumor necrosis factor-stimulated gene 6/NF-κB signaling pathway/odonto-genic/osteogenic differentiation分类
医药卫生引用本文复制引用
董稳航,张雪,何巍,李锐,王颖..外源性肿瘤坏死因子刺激基因6对炎症环境下人牙髓干细胞成牙/成骨分化的影响[J].郑州大学学报（医学版）,2025,60(2):167-172,6.基金项目
国家自然科学基金项目(82301061) （82301061）
