Exploration of the role of the subodontoblastic layer in odontoblast-like cell differentiation after tooth drilling using Nestin-enhanced green fluorescent protein transgenic mice
Exploration of the role of the subodontoblastic layer in odontoblast-like cell differentiation after tooth drilling using Nestin-enhanced green fluorescent protein transgenic mice
复制标题
DOI：
10.1016/j.job.2022.01.001
复制
发表时间：
2022-03-14
影响因子：
2.4
通讯作者：
Ohshima, Hayato
中科院分区：
文献类型：
--
作者：
Imai, Chihiro;Sano, Hiroto;Ohshima, Hayato
目的：原始成牙本质细胞和再生成牙本质细胞样细胞（OBLCs）对Nestin表达的调控可能存在差异。本研究旨在利用巢蛋白增强的绿色绿色荧光蛋白（GFP）反应性来探讨亚成牙本质层（SOBL）在巢蛋白增强的GFP转基因小鼠钻牙后OBC分化过程中的作用。方法：在深度麻醉下，在5或6周龄小鼠上颌第一磨牙近中面制备凹槽状空腔。结果：成牙本质细胞表达Nestin蛋白和mRNA，而对照组冠状面SOBL细胞表达Nestin-GFP。损伤后1- 2d，成牙本质细胞Nestin免疫反应性降低，NestinmRNA表达减少; 3- 5d，新分化的OBLCs沿着牙髓牙本质边缘排列，Nestin表达明显增强，NestinmRNA表达增强，形成修复性牙本质。牙髓-牙本质边缘的巢蛋白-GFP阳性细胞在第1天和第2天的数量显著增加。结论：Nestin-GFP阳性细胞向Nestin阳性OBLCs定向分化，提示外源性刺激解除了静止状态下SOBL细胞内源性Nestin蛋白和mRNA表达的限制，使其沿牙髓-牙本质边缘沿着迁移，并向OBLCs分化。(C)2022日本口腔生物学协会。Elsevier B. V.出版，保留所有权利。
Objectives: Original odontoblasts and regenerated odontoblast-like cells (OBLCs) may differently regulate Nestin expression. This study aimed to investigate the role of the subodontoblastic layer (SOBL) using green fluorescent protein (GFP) reactivity in the process of OBLC differentiation after tooth drilling in Nestin-enhanced GFP transgenic mice.Methods: A groove-shaped cavity was prepared on the mesial surface of the maxillary first molars of 5-or 6-week-old mice under deep anesthesia. Immunohistochemical staining for Nestin and GFP and Nestin in situ hybridization were conducted on the sections obtained at 1-14 days postoperative.Results: Odontoblasts showed intense endogenous Nestin protein and mRNA expression, whereas the coronal SOBL cells showed a Nestin-GFP-positive reaction in the control groups. The injured odontoblasts had significantly decreased Nestin immunoreactivity as well as decreased expression of Nestin mRNA 1-2 days after the injury; subsequently, newly differentiated OBLCs were arranged along the pulpdentin border, with significantly increased Nestin expression as well as increased expression of Nestin mRNA on days 3-5 to form reparative dentin. Nestin-GFP-positive cells at the pulp-dentin border significantly increased in number on days 1 and 2. GFP(+)/Nestin(+) and GFP(-)/Nestin(+) cells were intermingled in the newly differentiated OBLCs.Conclusions: The commitment of Nestin-GFP-positive cells into Nestin-positive OBLCs suggests that the restriction of endogenous Nestin protein and mRNA expression in the static SOBL cells was removed by exogenous stimuli, resulting in their migration along the pulp-dentin border and their differentiation into OBLCs. (C) 2022 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved.