Mineral trioxide aggregate affects cell viability and induces apoptosis of stem cells from human exfoliated deciduous teeth
Mineral trioxide aggregate affects cell viability and induces apoptosis of stem cells from human exfoliated deciduous teeth
复制标题
DOI：
10.1186/s40360-018-0214-5
复制
发表时间：
2018-05-15
影响因子：
2.9
通讯作者：
Wu, Pei-Chang
中科院分区：
文献类型：
--
作者：
Tsai, Chia-Ling;Ke, Mu-Chan;Wu, Pei-Chang
背景：三氧化矿物聚集体(MTA)广泛应用于恒牙盖髓手术，是牙髓治疗的金标准材料。本研究的目的是研究MTA与人脱皮乳牙(SHED)干细胞直接接触时对细胞存活率和细胞凋亡率的影响。从正常的人乳牙(第3-4代)采集和培养的棚子种植在方形覆盖玻璃上。将带有种子棚的玻璃培养在涂有或不涂有MTA涂层的平板中。分为四组：MTA直接接触组、直接对照组、MTA间接接触组和间接对照组。培养1、2、3d后，观察细胞形态，用WST-1细胞毒试验检测细胞活性。结果：MTA在第1、2、3天对SHADS细胞活力有明显的损伤作用，直接接触作用更严重。与MTA直接接触后，Annexin V和TUNEL染色均呈阳性，可见细胞凋亡。Western印迹分析显示，与MTA接触后，SHEDS细胞中Bcl2和BclxL的表达明显下降。结论：直接接触MTA后1周，SHEDS的存活率明显降低，并诱导细胞凋亡。结果提示，MTA直接接触对牙髓组织有一定的细胞毒作用。由于新鲜混合MTA的强碱性特性，预计会有不同的反应。
Background: Mineral trioxide aggregate (MTA) is widely used for pulp-capping procedures in permanent teeth and as a gold standard material in endodontics. The aim of the study was to investigate the effect of MTA on cell viability and apoptosis when MTA is directly in contact with Stem Cells from Human Exfoliated Deciduous Teeth (SHEDs).Methods: MTA was mixed and coated in the bottom of a 24-well plate. SHEDs collected and cultured from normal exfoliated human deciduous teeth (passages 3-4) were seeded on square cover glasses. The glasses with seeded SHEDs were incubated in the plates with or without MTA coating. They were divided into four groups: MTA direct contact, direct control, MTA indirect contact, and indirect control. After 1, 2 and 3 days of culturing, cell morphology was observed and cell viability was assessed by the WST-1 cell cytotoxicity assay. TUNEL assay, immunofluorescent labeling and western blot analysis were used to study the effects of MTA on SHEDs apoptosis.Results: MTA impaired cell viability of SHEDs in 1, 2 and 3 days, and the effect of direct contact was more severe. Cell apoptosis with positive Annexin V and TUNEL staining was noted when there was direct contact with MTA. Western blot analysis revealed that Bcl-2 and Bcl-xL decreased after SHEDs were in contact with MTA.Conclusions: This study shows that direct contact with 1 week post-set MTA significantly decreases the viability of SHEDs and induced cell apoptosis. The results suggest that there is a possible cytotoxic effect of pulp tissue when there is direct contact with MTA. Different responses would be expected due to the strong alkaline characteristics of fresh mixed MTA.