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骨形态发生蛋白 9 在牙髓细胞牙源性分化中的潜在作用。

Potential Roles of Bone Morphogenetic Protein 9 in the Odontogenic Differentiation of Dental Pulp Cells.

机构信息

State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China; Department of Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, China.

State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China; Department of Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, China.

出版信息

J Endod. 2021 Mar;47(3):436-443. doi: 10.1016/j.joen.2020.10.018. Epub 2020 Oct 28.

Abstract

INTRODUCTION

The differentiation of dental pulp cells (DPCs) plays an important role in the repair of dental pulp injury. Bone morphogenetic protein 9 (BMP9) is one of the most effective BMPs to induce the differentiation of stem cells. However, the role of BMP9 in promoting the odontogenic differentiation of DPCs and dentinogenesis is worth knowing.

METHODS

Fluorescence in situ hybridization and immunohistochemistry staining were performed to detect the BMP9 expression in human dental pulp. BMP9 was overexpressed in human DPCs (hDPCs), and the mineralization of hDPCs was tested by alkaline phosphatase staining and alizarin red staining. The expression of odontogenic differentiation-related genes was examined by quantitative real-time polymerase chain reaction and western blotting. The subcutaneous transplantation experiment was performed to test the odonto-induction ability of BMP9 in vivo. The rat direct pulp-capping experiment was performed to test the function of BMP9 in promoting dentin formation.

RESULTS

BMP9 showed an increased expression in odontoblast layer at both the mRNA and protein levels. BMP9 enhanced the mineralization and induced the expression of odontogenic differentiation-related genes in hDPCs. More mineralized nodules, and increased expression of dentin sialophosphoprotein (DSPP) and dentin matrix protein-1 (DMP1) were detected in the beta-tricalcium phosphate scaffold/cells composites of BMP9 group compared with the control group. Meanwhile, there was thicker reparative dentin formation in the BMP9 group in the rat pulp exposure experiment.

CONCLUSIONS

BMP9 participates in the process of DPC differentiation and promotes DPC mineralization and dentinogenesis. BMP9 might be a potential therapeutic target in the repair of dental pulp injury.

摘要

简介

牙髓细胞(DPCs)的分化在牙髓损伤修复中起着重要作用。骨形态发生蛋白 9(BMP9)是诱导干细胞分化最有效的 BMP 之一。然而,BMP9 在促进 DPC 成牙分化和牙本质形成中的作用值得了解。

方法

通过荧光原位杂交和免疫组织化学染色检测人牙髓中 BMP9 的表达。在人牙髓细胞(hDPCs)中过表达 BMP9,通过碱性磷酸酶染色和茜素红染色检测 hDPCs 的矿化情况。通过定量实时聚合酶链反应和蛋白质印迹检测牙源性分化相关基因的表达。进行皮下移植实验以测试 BMP9 在体内的成牙诱导能力。进行大鼠直接盖髓实验以测试 BMP9 促进牙本质形成的功能。

结果

BMP9 在 mRNA 和蛋白质水平上均在成牙本质细胞层表达增加。BMP9 增强了 hDPCs 的矿化并诱导了牙源性分化相关基因的表达。与对照组相比,在 BMP9 组的β-磷酸三钙支架/细胞复合材料中检测到更多的矿化结节,以及牙本质涎磷蛋白(DSPP)和牙本质基质蛋白-1(DMP1)的表达增加。此外,在大鼠牙髓暴露实验中,BMP9 组形成了更厚的修复性牙本质。

结论

BMP9 参与 DPC 分化过程,促进 DPC 矿化和牙本质形成。BMP9 可能是牙髓损伤修复的潜在治疗靶点。

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