一项关于由新型DSPP突变引起的牙本质生成不全II型的家系研究以及对人脱落乳牙中分离出的干细胞的研究。

A family study of dentinogenesis imperfecta shields type II caused by a novel DSPP mutation and investigations on the isolated stem cells from human exfoliated deciduous teeth.

作者信息

Gao Qianhua, Yue Ning, Liu Kehong, Deng Zhongren, Yang Ling, Zou Jing, Du Qin

机构信息

Department of Stomatology, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, No.32, Section 2, The First Ring Road West, Chengdu, 610072, Sichuan, China.

State Key Laboratory of Oral Diseases, Department of Pediatric Dentistry, West China Hospital of Stomatology, National Clinical Research Center for Oral Diseases, Sichuan University, No. 14 Section 3, Renmin South Road, Chengdu, 610041, China.

出版信息

BMC Oral Health. 2025 Apr 7;25(1):503. doi: 10.1186/s12903-025-05912-8.

DOI:10.1186/s12903-025-05912-8

PMID:40197225

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11978159/

Abstract

OBJECTIVE

This study aims to analyze the clinical features and genetic mutation characteristics of a family with Dentinogenesis Imperfecta Shields type II (DGI-II) and to observe the behavior of the stem cells from human exfoliated deciduous teeth (SHED) to explore the relationship between the locus of dentin sialophosphoprotein (DSPP) mutations and family clinical manifestations.

MATERIALS AND METHODS

After collecting clinical data from the family, Whole Genome Sequencing (WGS) followed by Sanger sequencing was used to identify pathogenic genes sites. The physical characteristics of the patient's teeth were examined using Micro-CT, scanning electron microscopy (SEM), and microhardness analysis. The behavior of SHEDs was studied through flow cytometry, adipogenic and osteogenic differentiation, quantitative real-time PCR (qRT-PCR), Western blotting, CCK-8 proliferation assays, colony formation, and cell migration experiments.

RESULTS

A novel frameshift mutation, DSPP c.2695delA.N899fs, was identified in the family. Micro-CT showed significant wear in the patient's teeth. SEM results revealed reduced and irregular dentinal tubules. Microhardness analysis showed significantly lower hardness in the patient's teeth. CCK-8, colony formation, and migration assays demonstrated reduced proliferation and migration capacities in the patient's SHEDs. qRT-PCR and Western blot results showed lower expression of DSPP, RUNX2, OCN, and ALP compared to controls, but higher DSPP protein level in the patient's SHEDs. Osteogenic differentiation tests indicated reduced mineralization capacity of the patient's SHEDs.

CONCLUSION

This study identified a novel frameshift mutation, DSPP c.2695delA.N899fs, in a DGI-II family and demonstrated its impact on SHED proliferation, migration, and mineralization. The findings demonstrated that this novel variant disturbs dentinal characteristics and cell behavior of SHED.

摘要

目的

本研究旨在分析Ⅱ型牙本质发育不全（DGI-II）一家系的临床特征和基因突变特点，并观察人脱落乳牙干细胞（SHED）的行为，以探讨牙本质涎磷蛋白（DSPP）基因突变位点与家系临床表现之间的关系。

材料与方法

收集该家系的临床资料后，采用全基因组测序（WGS）联合桑格测序来鉴定致病基因位点。使用显微CT、扫描电子显微镜（SEM）和显微硬度分析检查患者牙齿的物理特征。通过流式细胞术、成脂和成骨分化、定量实时聚合酶链反应（qRT-PCR）、蛋白质免疫印迹法、CCK-8增殖检测、集落形成和细胞迁移实验研究SHED的行为。

结果

在该家系中鉴定出一种新的移码突变，DSPP c.2695delA.N899fs。显微CT显示患者牙齿有明显磨损。SEM结果显示牙本质小管减少且不规则。显微硬度分析显示患者牙齿的硬度明显较低。CCK-8、集落形成和迁移检测表明患者的SHED增殖和迁移能力降低。qRT-PCR和蛋白质免疫印迹结果显示，与对照组相比，患者的SHED中DSPP、RUNX2、OCN和ALP的表达较低，但DSPP蛋白水平较高。成骨分化试验表明患者的SHED矿化能力降低。