Abstract
ObjectivesDental pulp regeneration is considered an ideal approach for treating dental pulp disease. Because pulp is composed of various cells, determining the proper seed cells is critical. We explored the potential of human umbilical cord mesenchymal stem cells (hUCMSCs) as seed cells for dental pulp regeneration.MethodsLiquid extract of human treated dentin matrix (LE-TDM) was acquired to culture hUCMSCs. Odontoblast-specific markers were detected by western blot, qRT-PCR, and immunofluorescence assays. Endothelial differentiation of hUCMSCs was examined according to VEGF induction by western blot, qRT-PCR, and Matrigel assays. hUCMSCs and VEGF-induced hUCMSCs (V-hUCMSCs) were also cocultured in vivo for the Matrigel plug assay and in vitro for RNA-sequencing (RNA-seq). Finally, encapsulated mono-cultured hUCMSCs or cocultured hUCMSCs and V-hUCMSCs in scaffolds were injected into the root segments and transplanted into immunodeficient mice for dental pulp regeneration.ResultsUnder LE-TDM induction, hUCMSCs expressed specific odontoblast markers (DSPP, DMP-1, DSP). Under VEGF induction, hUCMSCs expressed functional endothelial markers (CD31, eNOs, vWF). In vivo, the Matrigel plug assay indicated that cocultured hUCMSCs and V-hUCMSCs formed extensive vessel-like structures. RNA-seq results indicated that cocultured V-hUCMSCs exhibited high Hif-1 signaling pathway activity. Both the hUCMSCs mono-culture and coculture groups showed pulp-like tissue regeneration. The cocultured group showed more extracellular matrix and vascularization than the mono-cultured group in vivo.ConclusionhUCMSCs can differentiate into odontoblast-like cells and functional endothelial cells. Cocultured hUCMSCs and V-hUCMSCs formed vessel-like structures and regenerated dental pulp-like tissue. Therefore, hUCMSCs can be used as an alternative seed cell source for angiogenesis and dental pulp regeneration.
Highlights
The vitality of teeth is mainly maintained by dental pulp, which is responsible for the nerve supply, nutritional supply, and dentin production of teeth
We first evaluated whether human umbilical cord mesenchymal stem cells (hUCMSCs) have the capacity to differentiate into odontoblast-like cells under the microenvironment induced by Liquid extract of human treated dentin matrix (LE-TDM) in vitro, to confirm their angiogenic potential in vivo
The results indicated that VC exhibited high hypoxia-inducible factor 1 (Hif-1) signaling pathway activity, suggesting that hUCMSCs could activate the Hif-1 signaling pathway in VC (Figure 6C)
Summary
The vitality of teeth is mainly maintained by dental pulp, which is responsible for the nerve supply, nutritional supply, and dentin production of teeth. Conventional root canal therapy replaces pulp tissue with artificial materials, without restoring the function of the dental pulp. The loss of dental pulp in young permanent teeth will attenuate root development and can even lead to tooth loss (Huang, 2008; Kim et al, 2010). Restoring the function of pulp tissue through tissue engineering technology can solve these problems (Huang, 2008, 2009; Xuan and Li, 2018). A major challenge in regenerating functional dental pulp is the reconstruction of a complex, highly organized histological tissue structure, which contains several types of cells in different regions and areas of vascularization (e.g., odontoblasts in the peripheral lining against the dentinal wall and the microvasculature in the central region). The key to the regeneration of dental pulp is identifying the most suitable seed cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
