Developmental changes in primary cilia in the mouse tooth germ and oral cavity.

The primary cilium, a sensory apparatus, functions as both a chemical and mechanical sensor to receive environmental stimuli. The present study focused on the primary cilia in the epithelialmesenchymal interaction during tooth development. We examined the localization and direction of projection of primary cilia in the tooth germ and oral cavity of mice by immunohistochemical observation. Adenylyl cyclase 3 (ACIII)-immunolabeled cilia were visible in the inner/outer enamel epithelium of molars at the fetal stage and then conspicuously developed in the odontoblast layer postnatally. The primary cilia in ameloblasts and odontoblasts-shown by the double staining of acetylated tubulin and γ-tubulin-were regularly arranged from postnatal Day12, projecting apart from each other. The periodontal ligament possessed ACIII-positive cilia, which gathered on both sides of the dentin/cement and alveolar bone in postnatal days. In the oral cavity, numerous long primary cilia immunoreactive for ACIII were condensed at subepithelial stromal cells in the oral processes in fetuses, while postnatally a small number of short cilia were dispersed throughout the stroma of the oral cavity. These findings suggest that the primary cilia showing stage- and regionspecific morphology are involved in the epithelial-mesenchymal interaction during tooth development via mechano- and/or chemoreception for growth factors.

A systematic analysis for localization of predominant growth factors and their receptors involved in murine tooth germ differentiation using in situ hybridization technique.

Tooth development is regulated by various growth factors and their receptors. However, the overall mechanism of growth factor-mediated odontogenesis remains to be elucidated. The present study examined expression sites and intensities of major growth factors and receptors in the tooth germ of murine fetuses and neonates. Signals of TGF-ß and CTGF in fetuses were released from the enamel epithelium, while their neonatal signals arose in odontoblasts. Moreover, BMP/Smad signaling may affect the differentiation of ameloblasts, in contrast to PDGFα whose signals may cause odontoblast differentiation. Growth factors associated with the formation of the periodontium were IGF1, IGF2, IGFBP3, CTGF, and PDGFα. Concerning cusp formation, the enamel knot selectively expressed FGF4, BMP2, and BMP4 with an expression of PDGFα in the enamel-free area. It is concluded that many molecules play critical roles in the epithelium-mesenchyme interaction of tooth germ differentiation, and their expressions are precisely controlled.

Morphological factors of mandibular edentulous alveolar ridges influencing the movement of dentures calculated using finite element analysis.

PURPOSE: The aim of this study was to determine the importance of each morphological factor of edentulous alveolar ridges according to its influence on the movement of complete dentures. METHODS: The shapes of casts and waxed complete dentures were digitized. The determined shapes of the ridges were uniformly divided circumferentially and radially. Principal component (PC) analysis was performed using the coordinates of the points on the grid as the variables (morphological PC). The denture movement under bilateral and unilateral loads was analyzed using a finite element (FE) model constructed from the digitized shape, following PC analysis of the displacement of representative points on the denture (displacement PC). The effects of the morphological PCs were evaluated by means of stepwise multiple regression analysis with displacement PC as a dependent variable. RESULTS: The ridge height, clearance between the ridge and the occlusal plane, and various inclinations, were significantly selected as independent variables where the dependent variable was the displacement PC under a bilateral load. Under a unilateral load, the displacement PC was mainly influenced by the ridge height. The influence of morphological PCs of the non-loaded side tended to be larger than that of loaded side. CONCLUSION: Under a bilateral load, ridge height, clearance to the occlusal plane, and inclination of the ridge are considered to account for denture movement. To evaluate the effect of the ridge morphology on denture movement under a unilateral load, it is effective to determine the partitioned shape together with the height in general.

Factors differentiating the morphology of mandibular edentulous alveolar ridges: a pilot study.

The aim of this study was to determine the morphologic factors that characterize mandibular edentulous alveolar ridges. The shapes of casts and waxed complete dentures were digitally scanned and the extracted shapes of the ridges were uniformly divided both mesiodistally and buccolingually. Principal component analysis was performed using the coordinates of the points on the grid as variables. Over 82% of the variables could be expressed using seven principal components. However, some of these were not taken into account by the existing criteria. Therefore, the influence of each principal component should be investigated. Int J Prosthodont 2010;23:53-55.

Finite element model based on a mandibular cast and a waxed complete denture: evaluation of the accuracy and the reproducibility of analysis.

PURPOSE: The purpose of this study was to evaluate a newly developed method to construct customized finite element models from the viewpoints of the accuracy of measurement and the reproducibility of calculated denture movement under loads. METHODS: A cast of an edentulous mandibular alveolar ridge and a waxed complete denture were used. Measurement of the surface was done using a 3D-digitizer. After superposing, they were rotated so that the occlusal plane became level. The border of the alveolar ridge on the cast was decided in each buccolingual section. From a series of cross sections, the surface of the alveolar ridge was made. Based on the surfaces of the mucosa and denture, a finite element model consisting of the denture and underlying mucosa, which was given a uniform thickness, was constructed. This procedure and analysis under bilateral or unilateral loads on an artificial molar region were done repeatedly to evaluate its reproducibility and the error of the measurement of the surface. RESULTS: The standard error of the measured shapes of spheres was estimated to be within 0.1mm. The error caused by superposing was estimated to be within 0.38 mm. The results of analysis showed that the coefficient of variation of the displacement of the denture at selected nodes was approximately 14.1% at most. CONCLUSIONS: We conclude that this method has sufficient measurement accuracy and reproducibility for the calculated movement of dentures.

Cellular distribution of glutamate transporters in the gastrointestinal tract of mice: an immunohistochemical and in situ hybridization approach.

L-Glutamate transport by intestinal epithelial cells is an initial step of the entire glutamate metabolism pathway in the gut mucosa. The present study examined the cellular distribution of glutamate transporters in the digestive tract of adult mice using immunohistochemistry and in situ hybridization technique. Expression of EAAC1 mRNA was more intense in the ileum, where the epithelium in crypts and the basal half of intestinal villi showed high levels of transcripts, suggesting an essential role of EAAC1 in differentiating or premature epithelial cells. Electron-microscopically, EAAC1 immunoreactivity was predominantly localized in the striated border of enterocytes. Immunoreactivity for GLT-1 was found in the lateral membrane of epithelial cells at the bottom of gastric glands and at the intestinal crypts, and also in the lateral membrane of secretory cells at the duodenal gland. GLAST immunoreactivity was restricted to the fundic and pyloric glands, and was especially intense in the neck portion of both glands. However, in situ hybridization analysis failed to confirm the expression of GLT-1 and GLAST at the mRNA level, possibly due to limited sensitivity. The strong and specific luminal localization of EAAC1 in the intestinal epithelium suggests that EAAC1 is a predominant transporter of glutamate, at least in the lower part of the small intestine.

Immunohistochemical demonstration of acidic mammalian chitinase in the mouse salivary gland and gastric mucosa.

Acidic mammalian chitinase (AMCase) is the sole chitinolytic enzyme that has been identified thus far in the gastrointestinal tract of mammals. AMCase mRNA expression has been demonstrated in the salivary gland and stomach of mice and in the stomach of humans, while a bovine homologue of AMCase is produced in the liver and secreted into the blood. The present study using antibody raised against bovine AMCase demonstrates the cellular distribution of AMCase in salivary and gastric secretions at the protein level. Immunostaining using mouse tissues detected intense immunoreactivity for AMCase in serous-type secretory cells of the parotid gland and von Ebner's gland. Gastric chief cells, localized at the bottom of gastric glands, were also immunoreactive for AMCase. Electron-microscopically, the immunoreactivity was localized in granules in the apical cytoplasm of these secretory cells, and not in other structures. Western blot analysis confirmed the existence of AMCase in the parotid gland and stomach, and in their secretions in mice. However, no immunoreactive band was clearly detectable in immunoblots of the human parotid saliva and gastric juice. At least in the mouse, AMCase is secreted into the saliva and gastric juice, and may function as a digestive enzyme or play a defensive role against chitinous pathogens.

Cellular expression of gut chitinase mRNA in the gastrointestinal tract of mice and chickens.

Recently, the second mammalian chitinase, designated acidic mammalian chitinase (AMCase), has been identified in human, mouse, and cow. In contrast to the earlier identified macrophage-derived chitinase (chitotriosidase), this chitinase is richly expressed in the gastrointestinal (GI) tract, suggesting its role in digestion of chitin-containing foods as well as defense against chitin-coated microorganisms and parasites. This in situ hybridization study first revealed cellular localization of the gut-type chitinase in the mouse and chicken. In adult mice, the parotid gland, von Ebner's gland, and gastric chief cells, all of which are exocrine cells of the serous type, expressed the gut chitinase mRNA. In the chicken, oxyntico-peptic cells in glandular stomach (proventriculus) and hepatocytes expressed the chitinase mRNA. Because cattle produce the gut chitinase (chitin-binding protein b04) only in the liver, the gut chitinases in mammals and birds have three major sources of production, i.e., the salivary gland, stomach, and liver. During ontogenetic development, the expression level in the parotid gland and stomach of mice increased to the adult level before weaning, whereas in the stomach of chickens intense signals were detectable in embryos from incubation day 7.