Expression of KLF5 in odontoblastic differentiation of dental pulp cells during in vitro odontoblastic induction and in vivo dental repair
Expression of KLF5 in odontoblastic differentiation of dental pulp cells during in vitro odontoblastic induction and in vivo dental repair
复制标题
KLF5在体外成牙本质细胞诱导和体内牙齿修复过程中牙髓细胞成牙本质细胞分化中的表达
DOI：
10.1111/iej.12672
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发表时间：
2017-07-01
影响因子：
5
通讯作者：
Zhang, Q.
中科院分区：
文献类型：
--
作者：
Han, N.;Chen, Z.;Zhang, Q.
目的探讨修复性牙本质形成过程中Kruppel-like factor5(KLF5)是否参与成牙本质细胞的分化。采用茜素红染色、碱性磷酸酶(ALPase)活性、实时荧光定量聚合酶链式反应(RT-PCR)和免疫印迹法检测成牙本质细胞体外分化情况。用实时定量聚合酶链式反应和Western Blot方法检测KLF5在成牙本质细胞分化过程中的表达。通过慢病毒介导的shRNA敲除KLF5，以确定KLF5在成牙本质细胞分化中的作用。结果碱性磷酸酶(ALP；P=0.004)活性显著升高，牙本质涎磷蛋白(DSPP；P=0.004)、牙本质基质蛋白-1(DMP-1；P=0.001)等成牙本质细胞分化相关基因表达上调，证实了成牙本质细胞分化。在体外培养的DPC成牙本质细胞分化过程中，KLF5的表达显著上调(P=0.0002)。KLF5基因敲除对成牙本质细胞分化的影响。直接盖髓后，穿孔部位下方形成牙本质桥样钙化组织。结论KLF5可能参与了修复性牙本质形成过程中成牙本质细胞的分化过程。
Aim To identify whether Kruppel-like factor 5 (KLF5) was involved in odontoblastic differentiation during reparative dentine formation.Methodology Human Dental pulp cells (DPCs) were isolated from healthy human dental pulp tissue and induced for odontoblastic differentiation. Alizarin Red staining, alkaline phosphatase (ALPase) activity, quantitative real-time PCR and Western Blot were performed to evaluate in vitro odontoblastic differentiation. The expression profile of KLF5 during the in vitro odontoblastic differentiation was determined by quantitative real-time PCR and Western Blot. Knock-down of KLF5 by lentivirus-mediated shRNA was performed to determine the function of KLF5 in odontoblastic differentiation. After direct pulp capping with MTA, the maxillary first molar segments dissected from male Wistar rats were prepared for histology analysis and immunohistochemistry staining.Results Odontoblastic differentiation was confirmed by significantly increased alkaline phosphatase (ALP; P = 0.004) activity and upregulated odontoblastic differentiation-related genes including dentine sialophosphoprotein (DSPP; P = 0.004) and dentine matrix protein-1 (DMP-1; P = < 0.001). The expression of KLF5 was significantly upregulated during odontoblastic differentiation of in vitro cultured DPCs (P = 0.0002). KLF5 knock-down impaired odontoblastic differentiation. After direct pulp capping, dentine bridge-like calcified tissues were formed under the perforation sites. KLF5 was expressed in odontoblast-like cells and DPCs beneath the perforation sites during reparative dentine formation.Conclusions KLF5 might be involved in the process of odontoblastic differentiation during reparative dentine formation.