Journal Article

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

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

2025

Overview

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. Significance This study provides insights into the genetic and cellular basis of DGI-II, elucidating the role of a novel DSPP mutation in tooth structure development and stem cell behavior. However, in vivo validation of this DSPP mutation is important to further confirm this conclusion.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Abnormalities / Causes of / Cbfa-1 protein / Cell Differentiation / Cell migration / Cell Movement / Cell Proliferation / Child / Cholecystokinin / Computed tomography / Dental enamel / Dental research / Dentin / Dentinogenesis / Dentinogenesis imperfecta / Dentinogenesis imperfecta (DGI) / Dentinogenesis Imperfecta - genetics / Dentinogenesis Imperfecta - pathology / Dentistry / Diagnosis / Disease / DNA sequencing / DSPP gene / Dspp protein / Epoxy resins / Extracellular Matrix Proteins - genetics / Female / Flow cytometry / Frameshift Mutation / Gene mutations / Genes / Genetic aspects / Genetic disorders / Genomes / Genomics / Genotype & phenotype / Health aspects / Hearing loss / Humans / Investigations / Male / Medicine / Microscopy, Electron, Scanning / Mineralization / Mutation / Nucleotide sequencing / Oral and Maxillofacial Surgery / Pedigree / Phosphoproteins - genetics / Physical characteristics / Polymerase chain reaction / Scanning electron microscopy / Shields type-II / Sialoglycoproteins - genetics / Spectrum analysis / Stem Cells / Stem cells from human exfoliated deciduous teeth (SHED) / Teeth / Tooth, Deciduous - cytology / Tooth, Deciduous - pathology / Tubules / Western blotting / Whole genome sequencing / X-Ray Microtomography