Investigation of Proliferative Activity in the Developing Human Tooth Using Ki-67 Immunostaining

Abstract

Objective: The aim of this study was to investigate the proliferation of the developing human tooth germ and its surrounding tissues using Ki-67 immunostaining. Materials and Methods: Sections of mandibular dental arch tissues collected from 4 cadaveric human fetuses of 13, 16, 21 and 30 weeks of gestation were used. The immunoreactivity of Ki-67 in the tissue sections was assessed visually under a light microscope. Immunohistochemical controls were performed by replacing the primary antibody with phosphate-buffered saline or normal rabbit lgG. Results: The control sections did not display Ki-67 immunoactivity. Specimens of 13 weeks of gestation revealed intense Ki-67 immunostaining throughout the entire developing mandibular primary molars. At 16 weeks of gestation, immunostaining was observed in the inner enamel epithelium and dental papilla, in conjunction with the dental lamina showing decreased immunostaining. At 21 weeks, Ki-67 immunostaining was observed only in the inner enamel epithelium and dental papilla. The immunoreactivity of active ameloblasts and odontoblasts decreased, along with the proliferation capacity of the dental lamina. At 30 weeks, both enamel and dentin formation was observed along the cusped aspect of the tooth germ. Ameloblasts and odontoblasts were no longer immunoreactive in this region, while both types of cells were immunoreactive at the cervical regions of the crown. Dental lamina cells showed disintegration and were totally Ki-67-negative at 30 weeks of gestation. Conclusion: The Ki-67 immunoreactivity of the dental lamina decreased during intrauterine tooth development. Positive immunostaining was observed at specific sites in the enamel organ and dental papilla during the cap and bell stages.