Int J Stem Cells.  2024 Nov;17(4):437-448. 10.15283/ijsc23205.

The Characterization and Regulation of Schwann Cells in the Tooth Germ Development and Odontogenic Differentiation

• He J
• Wang T
• Liu D
• Yang J
• He Y
• Zhao S
• Ju Y

• KMID: 2562363
• DOI: http://doi.org/10.15283/ijsc23205

Abstract

Schwann cells (SCs), a type of glial cell in the peripheral nervous system, can serve as a source of mesenchymal stem cells (MSCs) to repair injured pulp. This study aimed to investigate the role of SCs in tooth germ development and repair of pulp injury. We performed RNA-seq and immunofluorescent staining on tooth germs at different developmental stages. The effect of L-type calcium channel (LTCC) blocker nimodipine on SCs odontogenic differentiation was analyzed by real-time polymerase chain reaction and Alizarin Red S staining. We used the PLP1-CreERT2/ Rosa26-GFP tracing mice model to examine the role of SCs and Ca_v 1.2 in self-repair after pulp injury. SC-specific markers expressed in rat tooth germs at different developmental stages. Nimodipine treatment enhanced mRNA levels of osteogenic markers (DSPP, DMP1, and Runx2) but decreased calcium nodule formation. SCs-derived cells increased following pulp injury and Ca v 1.2 showed a similar response pattern as SCs. The different SCs phenotypes are coordinated in the whole process to ensure tooth development. Blocking the LTCC with nimodipine promoted SCs odontogenic differentiation. Moreover, SCs participate in the process of injured dental pulp repair as a source of MSCs, and Ca_v 1.2 may regulate this process.

Keyword

Schwann cells; Tooth development; Ca_v 1.2; Odontogenic differentiation; Pulp injuries

Figure

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  Fig. 1 Protein-protein interactions network and RNA-seq analysis. (A) Protein-protein interaction network of the differentially expressed genes of rat nervous system of tooth germ between E14.5 and P7 by RNA-seq. (B) Protein-protein interaction network of the differentially expressed genes associated with specific markers of the different phenotypes of Schwann cells between E14.5 and P7. (C) Differential expression of rat neural development marker of tooth germ by RNA-seq. *p<0.05.

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  Fig. 2 Expression of rat neural development markers (GAP43, MBP101, S100β) in tooth germ at different stages. (A, F) Neural development markers expression on E14.5, amplification times is 400×. (B, G) Expression of neural development markers on E16.5, amplification times is 400×. (C, H) Expression of neural development markers on E18.5, amplification times is 200×. (D, I) Expression of neural development markers at P1. (E, J) Expression of rat neural development markers in tooth germ on P7. DAPI in blue. S100: S100β, OE: outer enamel, EM: ectodermal mesenchyme, OEE: outer enamel epithelium, SR: stellate reticulum, IEE: inner enamel epithelium, DP: dental papilla, DF: dental follicle, SI: stratum intermedium, E: enamel, D: dentin, P: dental pulp. *Co-stained.

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  Fig. 3 Effects of L-type calcium channel on the odontogenic differentiation of Schwann cells (SCs) in vitro. (A) Immunofluorescence staining of DSP in SCs after odontogenic differentiation. Scale bar=50 μm. (B) The mRNA expression of DSPP, DMP-1 and Runx2 in the three groups was measured at 14 days by real-time polymerase chain reaction. (i) Control: SCs in Schwann cell medium. (ii) OD: SCs in odontogenic differentiation medium (ODM). (iii) OD/Nimo(＋): SCs in ODM supplemented with 1μM nimodipine. Error bar: ±SD (n=3); *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 OD group with or without nimodipine versus control. (C) Images of Alizarin Red S staining at 28 days of odontogenic differentiation.

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  Fig. 4 SCs-derived cells and Cav1.2 participate in the regeneration of incisors after injury. (A-C) represent the expression of GFP＋ cells (green) and Cav1.2 (red) at the dental cusp, middle teeth and apical area of control and injured incisors with recovery for 0, 12, 48, 72 hours, 5 days, and 1 week, respectively. Boxed areas are shown magnified in (D) correspondingly. (D) The possible route of SCs-derived cells migrating to the odontoblast layer. The asterisk highlights GFP＋ cells. (E) Co-expression of GFP＋ cells and Cav1.2. Boxed areas are shown magnified to the right. Scale bar=50 μm. d: dentin, p: dental pulp, CL: cervical loop.

Reference

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