Effect of EDTA on TGF-β1 released from the dentin matrix and its influence on dental pulp stem cell migration
Effect of EDTA on TGF-β1 released from the dentin matrix and its influence on dental pulp stem cell migration
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DOI：
10.1590/1807-3107bor-2016.vol30.0131
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发表时间：
2016-01-01
影响因子：
2.5
通讯作者：
SAKAI, Vivien Thiemy
中科院分区：
文献类型：
--
作者：
GONÇALVES, Lidiany Freitas;FERNANDES, Ana Paula;SAKAI, Vivien Thiemy
储存在牙本质中的生物活性分子，如转化生长因子β 1（TGF-β 1），可能参与与牙齿组织修复相关的信号事件。作者进行了一项体外评价，评价了用10%乙二胺四乙酸（EDTA）、2.5%次氯酸钠（NaOCl）或磷酸盐缓冲盐水（PBS）处理后牙本质切片释放的TGF-β 1的量，以及这种生长因子对人类脱落乳牙（SHED）干细胞迁移的影响。制备60个1 mm厚的牙片（有或没有前牙本质层），并用10%EDTA处理1分钟，2.5%NaOCl处理5天或保存在PBS中。制备牙齿切片条件培养基并用于TGF-β 1 ELISA和迁移测定。还通过迁移试验检测了含有不同浓度重组人TGF-β 1（0.5、1.0、5.0或10.0 ng/mL）的培养基。资料以单因素变异数分析及Tukey检定进行评估。对应于由含有或不含有前牙本质层的牙齿切片调节并用10%EDTA处理的培养基的光密度值在统计学上大于其他组，接近1 ng/mL。观察到向由含有前牙本质层的牙齿切片调节并用PBS、10%EDTA或2.5%NaOCl处理的介质迁移的速率增加。重组人TGF-β 1也刺激迁移的SHED，无论使用的浓度。EDTA可以被认为是从牙本质基质中提取TGF-β 1的有效提取剂。然而，它不影响SHED迁移，表明其他组分可能导致细胞迁移。
Bioactive molecules stored in dentin, such as transforming growth factor beta1 (TGF-beta 1), may be involved in the signaling events related to dental tissue repair. The authors conducted an in vitro evaluation of the amount of TGF-beta 1 released from dentin slices after treatment with 10% ethylenediaminetetraacetic acid (EDTA), 2.5% sodium hypochlorite (NaOCl) or phosphate-buffered saline (PBS), and the effect of this growth factor on stem cell migration from human exfoliated deciduous teeth (SHED). Sixty 1-mm-thick tooth slices were prepared with or without the predentin layer, and treated with either 10% EDTA for 1 minute, 2.5% NaOCl for 5 days or kept in PBS. Tooth slice conditioned media were prepared and used for TGF-beta 1 ELISA and migration assays. Culture medium with different concentrations of recombinant human TGF-beta 1 (0.5, 1.0, 5.0 or 10.0 ng/mL) was also tested by migration assay. The data were evaluated by ANOVA and Tukey's test. Optical density values corresponding to media conditioned by tooth slices either containing or not containing the predentin layer and treated with 10% EDTA were statistically greater than the other groups and close to 1 ng/mL. Increased rates of migration toward media conditioned by tooth slices containing the predentin layer and treated with PBS, 10% EDTA or 2.5% NaOCl were observed. Recombinant human TGF-beta 1 also stimulated migration of SHED, irrespective of the concentration used. EDTA may be considered an effective extractant of TGF-beta 1 from the dentin matrix. However, it does not impact SHED migration, suggesting that other components may account for the cell migration.