Abstract
Stem cell-based developmental engineering has been considered as a promising strategy for tissue/organ regeneration. Tooth is formed by sequential reciprocal interactions between epithelium derived from surface ectoderm and mesenchymal cells derived from cranial neural crest. The neural crest cell is an appealing cell source for tooth development and regeneration research. In this study, we investigated the odontogenic differentiation and dentin-pulp complex regeneration potential of neural crest cells. Our results showed that neural crest cells (O9-1 mouse cranial neural crest cell line) can sequentially differentiate into dentin matrix acidic phosphoprotein 1 (DMP-1)-positive odontoblasts within a developing tooth germ in vitro. Moreover, O9-1 cells and induced pluripotent stem cell (iPSC)-derived neural crest-like cells (iNCLCs) can form well-organized vascularized dentin-pulp complex when transplanted in vivo with tooth scaffold. Furthermore, both O9-1 cells and iNCLCs can be differentiated into odontoblast-like cells, positive staining with odontogenic-related markers DMP-1 and dentin sialophosphoprotein (DSPP), under odontogenic induction with the administration of bone morphogenetic protein 4 (BMP-4). These results demonstrated that neural crest cells, especially the unlimited iNCLCs, are a promising cell source for tooth development and dental tissue/tooth organ regeneration studies.
Highlights
Tooth damage or loss, caused by dental carious, periodontitis, or traumatic injury, has a severe impact on our life
To distinguish the epithelium and mesenchyme and trace the biological behavior of O9-1 cells in the developing tooth germ, the dental epithelial tissue isolated from Rosa26 mice showed strong red fluorescence, and the O91 cells labeled by GFP showed strong green fluorescence in the reconstituted complex (Figure 1C)
These results demonstrated that O9-1 cells can differentiate into odontoblasts and contribute to tooth germ formation
Summary
Tooth damage or loss, caused by dental carious, periodontitis, or traumatic injury, has a severe impact on our life. Dental Tissue Regeneration derived from cranial neural crest (Kollar and Fisher, 1980; Chai et al, 2000). The cranial neural crest cells migrate to pharyngeal arches and contribute to a broad variety of derivatives, including craniofacial bone, cartilage, connective tissue, and teeth (Santagati and Rijli, 2003; Noden and Trainor, 2005; Kulesa et al, 2010). Even though there were reports that neural crest stem cells still present in the adult tissues such as gingiva (Zhang Q. et al, 2018), bone marrow (Morikawa et al, 2009; Niibe et al, 2017), and dental periodontal tissues (Ibarretxe et al, 2012), it is quite difficult to isolate plenty of primary NCCs for the research of stem cell-based tooth development and regeneration
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