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Enamel Structure Defects in Kdf1 Missense Mutation Knock-in Mice

The Keratinocyte differentiation factor 1 (KDF1) is reported to take part in tooth formation in humans, but the dental phenotype of Kdf1 mutant mice has not been understood. Additionally, the role of the KDF1 gene in dental hard tissue development is rarely known. In this study, we constructed a Kdf...

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Autores principales: Li, Pei, Zeng, Binghui, Xie, Weihong, Xiao, Xue, Lin, Ling, Yu, Dongsheng, Zhao, Wei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953722/
https://www.ncbi.nlm.nih.gov/pubmed/36831017
http://dx.doi.org/10.3390/biomedicines11020482
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author Li, Pei
Zeng, Binghui
Xie, Weihong
Xiao, Xue
Lin, Ling
Yu, Dongsheng
Zhao, Wei
author_facet Li, Pei
Zeng, Binghui
Xie, Weihong
Xiao, Xue
Lin, Ling
Yu, Dongsheng
Zhao, Wei
author_sort Li, Pei
collection PubMed
description The Keratinocyte differentiation factor 1 (KDF1) is reported to take part in tooth formation in humans, but the dental phenotype of Kdf1 mutant mice has not been understood. Additionally, the role of the KDF1 gene in dental hard tissue development is rarely known. In this study, we constructed a Kdf1 missense mutation knock-in mouse model through CRISPR/Cas9 gene-editing technology. Enamel samples from wildtypes (WT) and Kdf1 homozygous mutants (HO) were examined using micro-computed tomography (micro-CT), scanning electron microscopy (SEM), an atomic force microscope (AFM) and Raman microspectroscopy. The results showed that a novel Kdf1 missense mutation (c. 908G>C, p.R303P) knock-in mice model was constructed successfully. The enamel of HO mice incisors appeared chalky and defective, exposing the rough interior of the inner enamel and dentin. Micro-CT showed that HO mice had lower volume and mineral density in their tooth enamel. In addition, declined thickness was found in the unerupted enamel layer of incisors in the HO mice. Using SEM and AFM, it was found that enamel prisms in HO mice enamel were abnormally and variously shaped with loose decussating crystal arrangement, meanwhile the enamel rods were partially fused and collapsed, accompanied by large gaps. Furthermore, misshapen nanofibrous apatites were disorderly combined with each other. Raman microspectroscopy revealed a compromised degree of order within the crystals in the enamel after the Kdf1 mutation. To conclude, we identified enamel structure defects in the Kdf1 missense mutation knock-in mice, which displayed fragmentary appearance, abnormally shaped prism structure, decreased mineral density, altered crystal ordering degree and chemical composition of the enamel layer. This may support the potential role of the KDF1 gene in the natural development of enamel.
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spelling pubmed-99537222023-02-25 Enamel Structure Defects in Kdf1 Missense Mutation Knock-in Mice Li, Pei Zeng, Binghui Xie, Weihong Xiao, Xue Lin, Ling Yu, Dongsheng Zhao, Wei Biomedicines Article The Keratinocyte differentiation factor 1 (KDF1) is reported to take part in tooth formation in humans, but the dental phenotype of Kdf1 mutant mice has not been understood. Additionally, the role of the KDF1 gene in dental hard tissue development is rarely known. In this study, we constructed a Kdf1 missense mutation knock-in mouse model through CRISPR/Cas9 gene-editing technology. Enamel samples from wildtypes (WT) and Kdf1 homozygous mutants (HO) were examined using micro-computed tomography (micro-CT), scanning electron microscopy (SEM), an atomic force microscope (AFM) and Raman microspectroscopy. The results showed that a novel Kdf1 missense mutation (c. 908G>C, p.R303P) knock-in mice model was constructed successfully. The enamel of HO mice incisors appeared chalky and defective, exposing the rough interior of the inner enamel and dentin. Micro-CT showed that HO mice had lower volume and mineral density in their tooth enamel. In addition, declined thickness was found in the unerupted enamel layer of incisors in the HO mice. Using SEM and AFM, it was found that enamel prisms in HO mice enamel were abnormally and variously shaped with loose decussating crystal arrangement, meanwhile the enamel rods were partially fused and collapsed, accompanied by large gaps. Furthermore, misshapen nanofibrous apatites were disorderly combined with each other. Raman microspectroscopy revealed a compromised degree of order within the crystals in the enamel after the Kdf1 mutation. To conclude, we identified enamel structure defects in the Kdf1 missense mutation knock-in mice, which displayed fragmentary appearance, abnormally shaped prism structure, decreased mineral density, altered crystal ordering degree and chemical composition of the enamel layer. This may support the potential role of the KDF1 gene in the natural development of enamel. MDPI 2023-02-07 /pmc/articles/PMC9953722/ /pubmed/36831017 http://dx.doi.org/10.3390/biomedicines11020482 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Li, Pei
Zeng, Binghui
Xie, Weihong
Xiao, Xue
Lin, Ling
Yu, Dongsheng
Zhao, Wei
Enamel Structure Defects in Kdf1 Missense Mutation Knock-in Mice
title Enamel Structure Defects in Kdf1 Missense Mutation Knock-in Mice
title_full Enamel Structure Defects in Kdf1 Missense Mutation Knock-in Mice
title_fullStr Enamel Structure Defects in Kdf1 Missense Mutation Knock-in Mice
title_full_unstemmed Enamel Structure Defects in Kdf1 Missense Mutation Knock-in Mice
title_short Enamel Structure Defects in Kdf1 Missense Mutation Knock-in Mice
title_sort enamel structure defects in kdf1 missense mutation knock-in mice
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953722/
https://www.ncbi.nlm.nih.gov/pubmed/36831017
http://dx.doi.org/10.3390/biomedicines11020482
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