Establishing protein expression profiles involved in tooth development using a proteomic approach.

Various growth and transcription factors are involved in tooth development and developmental abnormalities; however, the protein dynamics do not always match the mRNA expression level. Using a proteomic approach, this study comprehensively analyzed protein expression in epithelial and mesenchymal tissues of the tooth germ during development. First molar tooth germs from embryonic day 14 and 16 Crlj:CD1 (ICR) mouse embryos were collected and separated into epithelial and mesenchymal tissues by laser microdissection. Mass spectrometry of the resulting proteins was carried out, and three types of highly expressed proteins [ATP synthase subunit beta (ATP5B), receptor of activated protein C kinase 1 (RACK1), and calreticulin (CALR)] were selected for immunohistochemical analysis. The expression profiles of these proteins were subsequently evaluated during all stages of amelogenesis using the continuously growing incisors of 3-week-old male ICR mice. Interestingly, these three proteins were specifically expressed depending on the stage of amelogenesis. RACK1 was highly expressed in dental epithelial and mesenchymal tissues during the proliferation and differentiation stages of odontogenesis, except for the pigmentation stage, whereas ATP5B and CALR immunoreactivity was weak in the enamel organ during the early stages, but became intense during the maturation and pigmentation stages, although the timing of the increased protein expression was different between the two. Overall, RACK1 plays an important role in maintaining the cell proliferation and differentiation in the apical end of incisors. In contrast, ATP5B and CALR are involved in the transport of minerals and the removal of organic materials as well as matrix deposition for CALR.

Tooth recognition of 32 tooth types by branched single shot multibox detector and integration processing in panoramic radiographs.

Purpose: The purpose of our study was to analyze dental panoramic radiographs and contribute to dentists' diagnosis by automatically extracting the information necessary for reading them. As the initial step, we detected teeth and classified their tooth types in this study. Approach: We propose single-shot multibox detector (SSD) networks with a side branch for 1-class detection without distinguishing the tooth type and for 16-class detection (i.e., the central incisor, lateral incisor, canine, first premolar, second premolar, first molar, second molar, and third molar, distinguished by the upper and lower jaws). In addition, post-processing was conducted to integrate the results of the two networks and categorize them into 32 classes, differentiating between the left and right teeth. The proposed method was applied to 950 dental panoramic radiographs obtained at multiple facilities, including a university hospital and dental clinics. Results: The recognition performance of the SSD with a side branch was better than that of the original SSD. In addition, the detection rate was improved by the integration process. As a result, the detection rate was 99.03%, the number of false detections was 0.29 per image, and the classification rate was 96.79% for 32 tooth types. Conclusions: We propose a method for tooth recognition using object detection and post-processing. The results show the effectiveness of network branching on the recognition performance and the usefulness of post-processing for neural network output.

Three-dimensional configuration of apical epithelial compartments including stem cell niches in guinea pig cheek teeth.

OBJECTIVES: Continuously growing rodent incisors have an apically located epithelial stem cell compartment, known as an "apical bud" (AB). Few studies have described the morphological features of ABs and stem cell niches in continuously growing premolars/molars. We attempted to clarify the relationship between the three-dimensional configuration of ABs and the stem cell niches in guinea pig cheek teeth. METHODS: We perfusion-fixed four-week-old guinea pigs, then decalcified their premolars/molars to produce serial paraffin sections, which we immunostained for Sox2. We reconstructed the serial sections using image processing and analysis software. We processed undecalcified samples for scanning electron microscopy by KOH digestion. RESULTS: Two types of epithelia with M and Δ shapes surrounded the S-shaped dental papilla in the apical region of the premolars/molars, and there were three Sox2-positive epithelial bulges above the M- and Δ-shaped epithelia. Sox2-positive epithelial stem cell niches were restricted to the apical side, and cell proliferation and differentiation immediately proceeded in the crown-analogue dentin. The Sox2-positive epithelial stem cell niches were sparsely distributed and extended to the occlusal side. We also detected continuously proliferating cells in the cervical loop and Hertwig's epithelial root sheath of the root-analogue dentin. CONCLUSIONS: Our findings suggest that guinea pig cheek teeth have three ABs, and the complex configuration of these types of teeth may be attributed to the prompt formation of crown-analogue dentin followed by the long-term formation of root-analogue dentin.