Odontogenic Tissue Generation Derived from Human Induced Pluripotent Stem Cells Using Tissue Engineering Application

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Abstract

<p>Human dental pulp cells significantly contribute to the generation of patient-specific human induced pluripotent stem cells (hiPSCs) because dental pulp is easily accessible and contains high-quality mesenchymal stem cells. This study aimed to generate hiPSCs from deciduous dental pulp cells, using three factors, OCT3/4, SOX2, and KLF4, and to evaluate the feasibility of hiPSCs as substitutes for odontogenic cells. HiPSCs were mixed with heterogeneous cells of porcine third molar tooth germs extracted from the mandibles of six-month-old pigs. The mixed cells were seeded in a disc-shaped poly (D, L-lactic-co-glycolic acid) scaffold. The cell–scaffold complexes were then wrapped around the omentum of immunocompromised rats as recipients to promote vascularization and maturation of the implants; the implants were harvested at 16 weeks after transplantation. The paraffin-embedded sections of the implants were used for histological observation and for immunohistochemical and immunofluorescence analyses. Histologically, several small pieces of odontogenic tissue were observed in the implants. The enamel organ-like structures were observed and the tall and columnar-shaped cells facing the enamel stained positive for anti-human nuclei and amelogenin antibodies. Dentin–pulp complexes with dentinal tubules were observed and the columnar-shaped cells facing the dentin stained positive for anti-human nuclei and dentin sialophosphoprotein antibodies. Dental root-like structures accompanying the Hertwig’s epithelial root sheath (HERS)-like bilayer were observed and cells constituting the HERS-like bilayer stained positive for anti-human nuclei and cytokeratin 14 antibodies and negative for anti-vimentin antibody. The cementum adjacent to the dentin was recognized, and staining for bone sialoprotein (BSP) was observed to be intense at the cementum–dentin border. Cementoblasts and cementocytes stained positive for anti-human nuclei and BSP antibodies. These results suggest that hiPSCs have the potential to differentiate into ameloblasts, odontoblasts, and cementocytes, and are capable of generating odontogenic tissues.</p>

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