Role of Dental Niche Cells in Tooth Development and Regeneration

Summary

Mammalian teeth develop via an essential mechanism shared by many organs, the inductive epithelial-mesenchymal interaction. The cellular underpinnings of such interactions continue to be a mystery. In this study, they develop a dual-fluorescence model to monitor and examine dental cells from embryonic to postnatal stages, in which Pitx2⁺ epithelium and Msx1⁺ mesenchyme are sufficient for tooth reconstitution. Critical cellular dynamics during the development of the molars, when tooth germs are arranged by the Msx1⁺ Sdc1⁺ dental papilla and surrounding dental niche, were further revealed by single-cell RNA sequencing and spatial mapping. Surprisingly, niche cells are more effective at regenerating teeth and have the ability to directly regenerate papilla cells by interacting with the dental epithelium. Last but not least, they pinpoint a collection of migratory Msx1⁺ Sox9⁺ cells from the dental niche as the potential cell source for the dental papilla. their findings suggest that dental niche cells directly support tooth organogenesis and shed light on the crucial cell makeup required for tooth engineering.

Fig.1 Graphical abstract. (Zhang, 2020)

Research Criteria

This study used single-cell sequencing and bioinformatic techniques combined with the DistMap method to characterize the cellular dynamics of epithelial and mesenchymal cells during molar development from embryonic to postnatal stages (E12.5, Embryonic day 12.5 to P7, Postnatal day 7). Gene locations were identified in E14.5 molars.

Sample Type

Mouse tooth germ was used to isolate epithelium and mesenchyme.

Result—scRNA-Seq Revealed Critical Cellular Dynamics of Molar Development

Single cell transcriptomics analysis was performed on these cells throughout molar development, spanning from E12.5, E14.5, and E16.5 to P1 and P7, in order to elucidate the essential cell composition for tooth reconstitution. The five developmental stages yielded thirteen main representative cell clusters. The top differentially expressed genes for the eight Msx1⁺ mesenchymal cells and the five Pitx2⁺ epithelial cells clusters were listed. Stage-by-stage analysis showed that during embryonic stages, the majority of the cell clusters for both dental epithelial and mesenchymal cells were kept constant.

Fig.2 Global cellular dynamics analysis of tooth germs by scRNA-seq.

Result—Spatial Mapping and Pseudotime Trajectory Analysis Suggested a Central Role of Dental Niche Cells in Tooth Induction

To develop a digitalized gene spatial matrix, in situ hybridization (ISH), immunohistochemistry (IHC), and immunofluorescence (IF) data of E14.5 tooth-associated signature genes were gathered from the public domains. After that, DistMap was used to establish a virtual tooth model. 13 cell clusters with their respective signature genes were identified from E14.5 scRNA-seq data. Then, they mapped the distinctive gene profiles of each cell cluster and visualized their spatial localization using the virtual tooth model. Notably, they discovered that numerous genes displayed distinctive expression patterns in the dental niche in the dental mesenchyme, in addition to identifying Sdc1 and Enpp1 as surface markers that specifically localized to the dental papilla region. Several representative ones, including Runx2, C1qtnf3, Cdkn1c, Postn, and Sox9, were chosen for validation by RNA-scope and immunostaining.

Fig.3 Spatial mapping of single cell transcriptome by DistMap. (Hu, 2022)

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Reference

Hu, H.; et al. Dental niche cells directly contribute to tooth reconstitution and morphogenesis. Cell Reports. 2022, 14(6): 111737.

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