GELATIN-CHONDROITIN-HYALURONAN TRI-COPOLYMER SCAFFOLD SEEDED WITH DENTAL BUD CELLS FOR ODONTOGENESIS: AN EX VIVO STUDY ON NUDE MICE

A biologically regenerated tooth may provide a new treatment for tooth loss. In this study, a tissue engineering approach was applied to demonstrate the tooth regeneration. The dental buds of the second molar tooth from 1.5-month-old miniature pigs were harvested by surgical operation before eruption. The dental bud tissues were cultured and expanded in vitro for three weeks to obtain dental bud cells (DBCs). The phenotypes of DBCs were identified with a flowcytometry, and the DBCs were seeded into a gelatin-chondroitin-hyaluronan tri-copolymer (GCHT) scaffold. The DBCs/GCHT scaffold constructs were implanted under dermis of nude mice's thoracic dorsum. Mice were sacrificed at predetermined intervals, and the developing tooth-like tissues were harvested for histological examinations. The present results of flowcytometry showed that the DBCs expressed specific surface markers of mesenchymal stem cells. Animal study revealed that the tooth-like structures expressed cytokeratin 14 at 4, 8, and 12 weeks postoperatively. The vascular endothelial growth factor was expressed on 12 weeks. Dentin-like mineralized tissue and dentin genetic-like cells were generated that expressed dentin martrix protein-1 on 16 and 20 weeks. Osteocytes were formed on 24 weeks and expressed osteopontin. This study reveals that the DBCs combined with an appropriate scaffold regenerated tooth-like structure with specific proteins for odontogenesis in nude mice.