Enamel Tissue Engineering Using Subcultured Enamel Organ Epithelial Cells in Combination with Dental Pulp Cells

Abstract

We describe a strategy for the in vitro engineering of enamel tissue using a novel technique for culturing enamel organ epithelial (EOE) cells isolated from the enamel organ using 3T3-J2 cells as a feeder layer. These subcultured EOE cells retain the capacity to produce enamel structures over a period of extended culture. In brief, enamel organs from 6-month-old porcine third molars were dissociated into single cells and subcultured on 3T3-J2 feeder cell layers. These subcultured EOE cells were then seeded onto a collagen sponge in combination with primary dental pulp cells isolated at an early stage of crown formation, and these constructs were transplanted into athymic rats. After 4 weeks, complex enamel-dentin structures were detected in the implants. These results show that our culture technique maintained ameloblast lineage cells that were able to produce enamel in vivo. This novel subculture technique provides an important tool for tooth tissue engineering.

<b>Induction of neural crest cells from human dental pulp-derived induced pluripotent stem </b><b>cells </b>

“…We have previously shown that dental epithelial cells have the potential to induce chondrocytes from mesenchymal cells (14,15). The extracellular matrix in the regenerated cartilage was strongly stained with alcian blue (14,15,56). To exclude the possibility of cell contamination, we confirmed that only human-origin cells were stained positive for anti-human nuclei antibody, while ratand porcine-origin cells were not stained.…”

“…To assess chondrogenic differentiation potential, the migrated cells were combined with dental epithelium and implanted into the omenta of immunodeficient rats. We have previously shown that dental epithelial cells have the potential to induce chondrocytes from mesenchymal cells (14,15). The extracellular matrix in the regenerated cartilage was strongly stained with alcian blue (14,15,56).…”

“…These approaches are categorized according to whether the cell source derives from post-natal-or embryonic-tooth bud cells (13,16,17,21,40,41,63), and results from these two sources differ considerably. When porcine post-natal tooth buds at late-bell stages are used, tissues are regenerated with structures containing dentin and enamel (14,15,52,63). However, most produced tissues exhibit a disorganized heterogeneous morphology (14,15,52,63).…”

“…When porcine post-natal tooth buds at late-bell stages are used, tissues are regenerated with structures containing dentin and enamel (14,15,52,63). However, most produced tissues exhibit a disorganized heterogeneous morphology (14,15,52,63). On the other hand, our previous study showed that mouse molar tooth bud cells at the cap stage of embryonic day 14 (ED14) are able to form teeth with regular crown shapes (21).…”

Tissue Engineering for Tissue and Organ Regeneration

“…The first one consists in the engineering of dental constituents, such as the periodontium, the pulp/dentin, or the enamel/dentin complexes (Duan et al, 2011;Honda et al, 2009;Huang, 2009;Park et al, 2010). In parallel with these experiments of tissue engineering, attempts are also made to reconstruct a whole tooth (Arany et al, 2009;Hu et al, 2006a;Komine et al, 2007;Nakao et al, 2007;Ohazama et al, 2004).…”