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Updated: Aug 20, 2026
Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice
Published on: January 12, 2017
Effect of Notoginsenoside R1 on reparative dentin formation via regulating the function of human dental pulp stem
Xuan He1, Dan Liang2, Xinyi Zhao2
1the Department of Operative Dentistry and Endodontology, College of Stomatology, Hospital of Stomatology, Guangxi Medical University, Guangxi, Nanning, China; Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction, Guangxi, Nanning, China; Guangxi Clinical Research Center for Craniofacial Deformity, Guangxi, Nanning, China.
Objective:
This study investigates the influence of Notoginsenoside R1 (NGR1) on the odontogenic differentiation of human dental pulp stem cells (hDPSCs) and its potential application in direct pulp capping, utilizing both in vitro and in vivo models. The results aim to provide a theoretical basis for expanding the clinical use of NGR1.
Methods:
hDPSCs were isolated, cultured and characterized in vitro. The effects of varying concentrations of NGR1 (0, 50, 100, 200 μg/ml) on cell proliferation and migration were evaluated using the CCK-8 and Transwell assays, respectively. The effect of NGR1 on odontogenic differentiation was evaluated by alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining, as well as RT-PCR to detect the mRNA expression levels of ALP and DSPP. A direct pulp capping model was established in adult dogs. The experimental group was rinsed with 100 μg/ml NGR1 solution before capping, and samples were collected at 1, 4, 8, and 12 weeks. Micro-CT scanning and H&E staining were used to analyze reparative dentin and pulp morphology. Additionally, RNA-Seq of hDPSCs treated with 100 μg/ml NGR1 was conducted, followed by DEG screening, GO/KEGG enrichment, GSEA, and PPI analyses, RT-PCR and IHC were used to verify the DEGs.
Results:
NGR1 promotes hDPSCs proliferation, migration, and odontogenic differentiation. In vitro experiments demonstrated that NGR1 enhanced ALP activity and mineralized nodule formation. Both the 100 μg/ml and 200 μg/ml groups significantly promoted the mineralization of hDPSCs. RT-PCR results showed that mRNA expression levels of ALP and DSPP were significantly upregulated in NGR1-treated groups. Animal experiments revealed the formation of continuous, uniform, and thickened reparative dentin over time in both the Control(0 μg/ml) and NGR1-treated groups. H&E staining showed dentin bridges at 4, 8, and 12 weeks in both Control(0 μg/ml) and NGR1 groups. Histological observations suggested that reparative dentin in the NGR1-treated groups appeared to have numerous, regularly arranged dentinal tubules and a compact architecture, whereas the Control(0 μg/ml) groups showed a relatively less organized structure. RNA-Seq results identified 50 upregulated DEGs, including the dentin-related gene S100A9. These DEGs were enriched in the complement and coagulation cascades, cytokine-cytokine receptor interaction, Staphylococcus aureus infection, and tuberculosis pathways. PPI analysis identified SPRR1A, SPRR1B, and SPRR3 as potential hubs. RT-PCR and IHC confirmed that the expression trends of the differentially expressed genes were consistent with the sequencing results.
Conclusion:
NGR1 promotes the proliferation, migration, and odontogenic differentiation of hDPSCs, with the 100 μg/ml concentration providing an optimal balance between proliferative and pro-differentiation effects. In vivo, NGR1 improves the microstructural quality of reparative dentin. Transcriptomic analysis revealed upregulation of S100A9 and enrichment of repair-related pathways, which may serve as a basis for future mechanistic studies.
