Immunohistochemical localization of amelogenin in human odontogenic tumors, using a polyclonal antibody against bovine amelogenin

In the present study, we investigated the localization of amelogenin in odontogenic tumors, using an anti-amelogenin polyclonal antibody. In order to make the antibody, antisera against an amelogenin fraction obtained from the enamel matrix of unerupted bovine tooth was raised in rabbits. By Western blot analysis, a main band of 25 kDa and six minor bands (6.8, 12, 18, 20, 23, and 27 kDa) were detected under nonreducing conditions. Immunoreactivity for the amelogenin was observed in ameloblasts and in the immature enamel matrix of 4-day-old rats. In odontogenic tumors, positive reactions for amelogenin were localized in limited areas in adenomatoid odontogenic tumor, calcifying odontogenic cyst, primary intraosseous carcinoma and odontoma. The strongest immunoreactions were shown in enamel matrices in odontomas. Small mineralized foci in epithelial nests showed positive reactions, and a few reactions were observed in epithelium adjacent to the mineralized foci. In calcifying odontogenic cysts, some ghost cells in the lining epithelium were strongly stained. The results indicate that the present antibody for amelogenin is useful for the determination of odontogenic tumors, especially in those in which small mineralized foci are present in the epithelial nests.     

Pathological evaluation of the effects of intentional disocclusion and overloading occlusion in odontogenesis disorders in N-methylnitrosourea-treated hamsters

This study compares the effects of disocclusion and overloading occlusion on dental lesions. Ten-day-old Syrian hamsters were divided into 4 groups: group I, untreated animals; group II, animals whose hemilateral incisors were disoccluded; group III, N-methylnitrosourea (MNU)-treated animals; and group IV, MNU-treated animals whose hemilateral incisors were disoccluded. The ipsilateral maxillary and mandibular incisors were repetitively cut with diamond discs. The hamster is easier to anesthetize. Animals received a 0.2% solution of MNU (10 mg/kg body weight) intragastrically twice a week for 16 wk. All the cut mandibular incisors and the MNU-treated uncut mandibular incisors showed lack of iron deposition on the enamel surface. The eruption rate was significantly higher in the cut disoccluded incisors of groups II and IV (p < 0.05) and significantly lower in the uncut overloaded incisors of groups II and IV (p < 0.05). In the cut mandibular incisors of group IV, the degree of the disturbance of odontogenesis and the atypical proliferation of odontogenic epithelium were more prominent (p < 0.02), and the dental lesions occurred earlier. Histologically, the disturbed Hertwig's epithelial sheath and the Hertwig's epithelial sheath-like transformed U-shaped part and enamel organ seemed to lead to disturbances of amelogenesis and detinogenesis as well as to atypical proliferation of odontogenic epithelium nests. Thus, this method of disocclusion of the incisors of rodents may represent a useful model for the investigation of the effects of various agents on tooth formation over a short experimental period.     

Histochemical localization of cholinesterase activity in the dental epithelium of guinea pig teeth

Cholinesterase is known for its remarkable diversity in distribution and function. An association of this enzyme with proliferative and morpho-differentiating tissues has been reported in several species. Here we report on the first evidence of the presence of cholinesterase in the enamel organ of continuously erupting incisors and molars of the guinea pig. Frozen sections of the incisors and molars of the guinea pig were incubated for histochemical demonstration of cholinesterase activity by means of the thiocholine method as described by Karnovsky and Root. The cholinesterase activity was observed in several types of cells of the dental epithelium; cells forming the basal portion of the enamel organ, outer enamel epithelium and maturation stage ameloblasts of both the incisors and molars. In the crown analogue side, the outer enamel epithelial cells gained strong reactions for cholinesterase and maintained the reaction throughout the secretory and maturation stages of amelogenesis. In contrast, cholinesterase reactions were lacking in the inner enamel epithelium, pre-ameloblasts, and secretory ameloblasts. In the early stage of enamel maturation, ameloblasts began to show positive reactions for cholinesterase, which was upregulated in the incisal direction. Although both tooth types showed similar reactive patterns for cholinesterase at the growing ends, maturation ameloblasts depicted a different pattern of staining displaying the reactions only sporadically in molars. These data indicate the role of cholinesterase in the enamel organ in tooth morphogenesis and function of guinea pig teeth.     

A method for sampling the stages of amelogenesis on mandibular rat incisors using the molars as a reference for dissection

A method for locating specific stages of amelogenesis on continuously erupting incisors was devised for rats weighing 101 +/- 5 g (n = 32). The technique is based on reflecting reference lines from the mandibular molars as perpendiculars to the labial surface of mandibular incisors. From these reference lines additional measurements are then made along the midline of the labial surface of the incisor in an apical or incisal direction to find the site desired for sampling. Histological studies on 24 decalcified incisors split into segments by using such reference lines and reconstructed by morphometry indicated that a reference line reflected from the contact point between the 2nd and 3rd molars crossed the enamel organ and adjacent enamel at 3,181 +/- 329 microns incisal to the start of the secretory zone of amelogenesis. A reference line from the 2nd and 1st molars crossed the enamel organ and enamel at 1,238 +/- 424 microns incisal to the start of the maturation zone of amelogenesis, while a reference line from the mesial side of the 1st molar crossed the enamel organ and enamel almost exactly where the enamel becomes completely soluble following prolonged decalcification in EDTA. Although reference lines were reproducible within a group of male rats having similar body weights, the linear distance between the apical end of the incisor and the point at which they crossed the tooth increased at a rate of 1 mm per 159 g for rats between 50 and 300 g body weight. This suggests that molars do not maintain a fixed relationship to incisors over time, and extreme care must be taken to standardize an experiment to a specific body weight when using this method.     

Natal teeth: a review

The incidence of natal teeth is approximately 1:2,000 to 1:3,000 live births. The most commonly affected teeth are the lower primary central incisors. Natal teeth usually occur in pairs. The eruption of more than two natal teeth is rare. The majority of natal teeth represent the early eruption of normal primary deciduous dentition. Less than 10% of natal teeth are supernumerary. Natal teeth might resemble normal primary dentition in size and shape; however, the teeth are often smaller, conical and yellowish, and have hypoplastic enamel and dentin with poor or absent root formation. Complications include discomfort during suckling, sublingual ulceration, laceration of the mother's breasts and aspiration of the teeth. A dental roentgenogram is indicated to differentiate the premature eruption of a primary tooth from a supernumerary tooth. Tooth extraction is indicated if the tooth is supernumerary or excessively mobile. If the tooth does not interfere with breastfeeding and is otherwise asymptomatic, no treatment is necessary.     

Immunohistochemical localization of connexin 43 in the developing tooth germ of rat

Distribution of gap junction protein in maxillary tooth germs of 1-day-old rats was examined by immunohistochemistry, using an affinity-purified antibody specific to residues 360–376 of rat connexin (CX) 43. In 1-day-old rats, the maxillary second molar formed the shape of the cusp, but neither dentine nor enamel was formed between the cells of the dental papilla and the inner enamel epithelium. In the tooth germ, CX 43 was expressed in the cells of the stratum intermedium and the inner enamel epithelium. Labelling in the stratum inter-medium was extensive and showed an increasing gradient from peripheral to cuspal regions. CX 43 detected in the inner enamel epithelium was at cell surfaces facing the interface between the dental papilla and the inner enamel epithelium. The cells of the dental papilla and the inner enamel epithelium began differentiation as odontoblasts and secretory ameloblasts respectively, in the cusps of the first molars, where predentine and dentine were formed but enamel matrix was not secreted. CX 43 was present in the stratum intermedium, inner enamel epithelium, preodontoblasts, odontoblasts and subodontoblasts. The incisors showed the most advanced stage of development, where the enamel matrix and calcified dentine were formed in the labial part of the teeth. The CX 43 epitope was seen in the stratum intermedium, inner enamel epithelium, preameloblasts, preodontoblasts, odontoblasts, and subodontoblasts. Immunolabelling was more extensive in the stratum intermedium and subodontoblasts than in preameloblasts, preodontoblasts, and odontoblasts. The immunolabelling in preameloblasts and preodontoblasts was accumulated at cell surfaces facing the predentine. Further, the labelling in preameloblasts and preodontoblasts disappeared or was reduced at the initiation of enamel matrix secretion and calcification of dentine matrix.The present results suggest that gap junctional cell communication has important roles in tooth development. Further, the extensive CX 43 expression in the stratum intermedium and the subodontoblast layer suggests that gap junctions have an important role in amelogenesis and dentinogenesis.     

Effect of Down syndrome on the dimensions of dental crowns and tissues

Abnormal growth in Down syndrome (DS) is reflected by variable reduction in size and simplification in form of many physical traits. This study aimed to compare the thickness of enamel and dentine in deciduous and permanent mandibular incisor teeth between DS and non‐DS individuals and to clarify how these tissues contribute to altered tooth size in DS. Sample groups comprised 61 mandibular incisors (29 permanent and 32 deciduous) from DS individuals and 55 mandibular incisors (29 permanent and 26 deciduous) from non‐DS individuals. Maximum mesiodistal and labiolingual crown dimensions were measured initially, then the crowns were sectioned midsagittally and photographed using a stereomicroscope. Linear measurements of enamel and dentine thickness were obtained on the labial and lingual surfaces of the crowns, together with enamel and dentine–pulp areas and lengths of the dentino‐enamel junction. Reduced permanent crown size in DS was associated with a reduction in both enamel and dentine thickness. After adjustments were made for tooth size, DS permanent incisors had significantly thinner enamel than non‐DS permanent teeth. The DS permanent teeth also exhibited significant differences in shape and greater variability in dimensions than the non‐DS permanent teeth. Crown dimensions of deciduous incisors were similar in size or larger in DS compared with non‐DS deciduous teeth. Enamel and dentine thicknesses of the deciduous teeth were similar in DS and non‐DS individuals. The findings indicate that growth retardation in DS reduces both enamel and dentine deposition in the permanent incisors but not in the earlier‐forming deciduous predecessors. The results are also consistent with the concept of amplified developmental instability for dental traits in DS. Am. J. Hum. Biol. 13:690–698, 2001. © 2001 Wiley‐Liss, Inc.     

The Distribution of Cellular Retinoic Acid-Binding Protein I (CRABPI) and Cellular Retinol-Binding Protein I (CRBPI) during Molar Tooth Development and Eruption in the Rat

The distribution of cellular retinoic acid-binding protein (CRABPI) and cellular retinol binding protein (CRBPI) was studied in a series of prenatal and early postnatal rats, covering the main stages of development and eruption of the molar teeth. CRABPI positive cells were found in the mesenchymal cells of the dental follicle from the cap stage and in the dental papilla from the early bell stage. In the dental papilla, CRABPI positive cells were situated adjacent to the enamel organ in the cervical loop region and in the subodontoblastic region. Newly formed odontoblasts were CRABPI positive for a short period of time. The enamel organ was CRBPI and CRABPI negative, except for the presence of CRABPI positive cells in the internal enamel epithelium over the tip of cusps and in parts of the stratum intermedium. During root formation, CRABPI positive cells were found in the developing periodontal ligament, in the dental papilla adjacent to the epithelial root sheath and in the subodontoblastic zone. During crown formation, CRBPI positive cells were mainly localized to the mesenchymal cells of the dental papilla during the cap stage of crown development. The periosteum of the developing mandible contained CRABPI positive cells while some osteoclasts appeared to show a weak but positive reaction to CRBPI. The findings were considered in terms of the possible significance of retinoid-binding proteins during tooth and bone development.     

Histological and three dimensional organization of the odontogenic organ in the lower incisor of 100 gram rats

A three dimensional reconstruction of the epithelial tissue at the apical end of the lower rat incisor was made from serial 1 μm thick cross sections. This tissue formed an elongated structure, called the odontogenic organ, which was composed of a bulbous and a “U”-shaped part. Both parts were joined to one another at the posterior aspect of the apical foramen. The bulbous part of the odontogenic organ was situated at the lingual side of the “U”-shaped part and protruded anteriorly over the pulp. It was formed by cells of the outer dental epithelium and stellate reticulum whose organization suggested that the bulbous part was important in the production of cells for renewal of all the epithelia of the incisor. The “U”-shaped part of the odontogenic organ was apparently derived from the bulbous part and delineated the pulp by forming the lateral, mesial and labial sidewalls around the apical foramen. It was composed of all the epithelial cell types recognizable as precursors to (a) cells of the enamel organ which form the enamel, and (b) Hertwig's epithelial root sheath, a part of the odontogenic organ which induces the formation of dentin on the lingual aspect of the incisor.     

The development of enamel structure in rat incisors as compared to the teeth of monkey and man

The rat incisor is an excellent model system in which to study amelgenesis. However, the information obtained has not been extrapolated to the human because of alleged structural differences between the teeth. The obvious differences include continuous eruption in rat incisors and an enamel rod pattern in rats which seemingly differs from the keyhole pattern of human enamel. A comprehensive analysis was made of those features of enamel structure considered fundamental to the understanding of its formation. This was done by applying the knowledge of amelogenesis obtained in rat incisors to the teeth of monkey and man. The following points of basic similarity were established between these species: (1) Interrod enamel is secreted first. It forms the side walls of cavities which are initially occupied by Tomes' processes. (2) The formation of interrod cavities is followed by deposition of enamel rods within these spaces. (3) The rods conform to the shape of the cavities and are secreted from one surface of Tomes' process. (4) At the initial site of rod deposition its enamel is continuous with the interrod enamel wall. (5) Growth of the rod compresses the process to one side of the cavity resulting in an arcade-shaped "space" between the rod and the remaining interrod walls. This study demonstrates that it is no longer necessary to postulate a keyhole structure for primate enamel, and it has established that a fundamental similarity exists in the basic structure and in the mode of formation of enamel in all three species.     

Interproximal contact hypoplasia in primary teeth: A new enamel defect with anthropological and clinical relevance

This study reports the prevalence, distribution, and expression of enamel defects in a sample of primary teeth (n = 225) from a prehistoric site in western India (1400–700 BC). Five enamel surfaces of individual, isolated primary teeth were observed for surface defects using a binocular stereomicroscope with variable power of magnification (8–20×). Standards for evaluating dental enamel defects (DDE) recommended by the Fédération Dentaire International (FDI) were employed. Details of defect expression were also recorded, including size, shape, and surface of tooth crown affected. Hypoplastic enamel defects were observed in 28% of teeth, but the distribution and expression of defects was not random. More than 50% of canine teeth had hypoplastic defects (HD); incisors and molar teeth exhibited far fewer HD. The buccal surface of canines was the most commonly affected crown surface. Areas of missing enamel were also common on the mesial and distal surfaces of canines and incisors and on the mesial surface of molar teeth. The high frequency of enamel defects found on interproximal crown surfaces warrants a label, and the name interproximal contact hypoplasia (IPCH) is proposed. Linear enamel hypoplasia (LEH) was absent from this primary dental sample. IPCH is more frequent in mandibular than in maxillary teeth, but no side preference was detected. In canine teeth, buccal hypoplasias (localized hypoplasia of primary canines; LHPC) were not positively correlated with interproximal hypoplastic defects. The etiology of IPCH may involve mesial compaction of developing teeth due to slow longitudinal growth of the jaws. Episodic bone remodeling results in ephemeral fenestrae in the mesial and distal walls of the dental crypt permitting tooth–tooth contact and disruption of amelogenesis. IPCH prevalence decreases across the subsistence transition from sedentary Early Jorwe agriculturalists to seminomadic Late Jorwe hunters and foragers, but the difference is not statistically significant. This may be due to underrepresentation of mandibular teeth in the sample. Am. J. Hum. Biol. 11:718–734, 1999. © 1999 Wiley-Liss, Inc.     

PrPc is temporospatially expressed in molar development of rats

Odontogenesis, tooth development, is derived from two tissue components: ectoderm and neural crest‐derived mesenchyme. Cyto‐differentiation of odontogenic cells during development involves time‐dependent and sequential regulation of genetic programs. This study was conducted to detect molecules implicated in cyto‐differentiation of developing molar germs of rats. Differential display‐PCR revealed that PrP^c was differentially expressed between cap/early bell‐staged germs (maxillary 3rd molar germs) and root formation‐staged germs (maxillary 2nd molar germs) at postnatal day 9. Both levels of PrP^c mRNA and protein expression were higher in the root formation stage than the cap/early bell stage and increased in a time‐dependent manner. Immunofluorescence revealed for the first time that PrP^c was not localized in the enamel organ, but localized in dental follicular cells for the development of the periodontal ligament and cementum as well as odontoblasts, both of which are of neural crest origin. These results suggest that the physiological functions of the PrP^c in tooth development may be implicated in the differentiation of neural crest‐derived mesenchyme including the periodontal tissues for root formation rather than epithelial tissue. Anat Rec, 296:1929–1935, 2013. © 2013 Wiley Periodicals, Inc.     

Actin-associated proteins in ameloblast differentiation

Focal contacts are systems of adherens junctions of the cell-extracellular matrix type, which allow the transfer of fundamental signals from the extracellular matrix to nuclear compartments, capable of regulating adhesion, proliferation, migration and differentiation of cells. Recently, many authors have concentrated their attention on epitheliomesenchymal interactions which guide organogenesis of dental germ, identifying numerous growth and differentiation factors and having the inner enamel epithelial cells of the enamel organ as a target. Given that the two cellular compartments in their tooth germ are separated by a basal membrane and by an extracellular matrix, which touches it, we wanted to evaluate the presence of focal contacts through the identification of talin and vinculin, proteins of the actin-associated protein complex. In this study we utilized the hemimandibles of young Wistar rats and we extracted the related odontogenic tooth organs present at their apical end. Specimens are processed with antibody against vinculin and talin. Results show that these junctional system proteins are present at the apical poles of both cellular compartments suggesting that putative epithelial-mesenchymal interactions, other than marker molecules, may use focal contacts as a system for transmission of signals.     

Combined surgical-orthodontic therapy for compound odontoma

Odontogenic tumor is a rare condition in dental medicine that mostly proceeds unrecognized until the occurrence of clinical symptoms such as delayed eruption, or is incidentally detected on routine x-ray examination. The exact cause is not known, however, previous dental trauma and infection have been postulated as the potential factors in the development of odontogenic tumor. The earliest possible operative extirpation of the tumorous growth is recommended to eliminate permanent tooth impaction and to enable normal growth of the teeth. In some cases, corticotomy, including complete removal of the bony coat of the tooth, may be needed to additionally facilitate and precipitate its eruption. Orthodontic therapy is also of great importance in correct alignment of the teeth 'n the dental arch as well as in the management of other anomalies that may be associated with odontogenic tumor. A patient with compound odontoma is presented, along with the course of combined surgical-orthodontic therapy. The patient reported previous intrusion trauma that had occurred at the age of 4 years, which may have been the potential factor in the development of odontoma. In this case, there was a massive odontogenic tumor which had compromised the growth of permanent teeth, and the growth impulse was almost at the end since the patient was 11 years old and the apexes of the upper incisors were partially closed. The first operation included complete removal of the tumorous mass that had interrupted spontaneous eruption of the upper permanent incisors. It did not result in immediate spontaneous tooth eruption, so an additional operation was needed. The objective of the second operative procedure was complete removal of the covering bone over the unerupted upper permanent incisors in order to eliminate the physical barrier to tooth growth and eruption. The objective of fixed orthodontic therapy was full eruption of the partially erupted upper incisors. After 16 months, the upper incisors were regularly located in the dental arch. In this case, orthodontic therapy had another objective, i.e. to ensure rotation of the first upper premolar, to provide space for the upper permanent canine eruption and to establish regular intercuspidation after upper second premolar hypodontia. In colclusion, combined operative and orthodontic therapy can be recommended irrespective of the stage of the impacted tooth development because any treatment to precipitate tooth eruption has favorable effects. Impacted teeth should always be provided all treatment options for faster eruption, as demonstrated in our case where a good clinical result was achieved within 2.5 years. The role of regular clinical and x-ray controls for assessment of the impacted tooth eruption should also be emphasized.     

Timing of odontogenic neural crest cell migration and tooth-forming capability in mice.

The mammalian tooth develops through sequential and reciprocal interactions between cranial neural crest (CNC)- derived ectomesenchymal cells and the stomadial epithelium. Classic tissue recombination studies demonstrated that premigratory CNC cells and CNC-derived ectomesenchymal cells possess odontogenic capacity and can respond to oral epithelial signals to form a tooth, suggesting that the CNC cells contributing to odontogenic tissue are not prespecified. Here we show that, in mice, CNC cells have populated the forming first branchial arch before the 9-somite stage and continue to migrate into the arch by the 13-somite stage. Grafts of the first arch from the 10-somite embryo or earlier yielded membranous bone and cysts but no teeth after subrenal culture. However, grafts of the first arch with its dorsally adjacent tissue containing migrating neural crest cells from the same age embryos gave rise to teeth. In contrast, teeth formed in first arch grafts that do not contain migrating neural crest cells from embryos with 12 or more somites. Interestingly, the acquisition of tooth forming capability in the first arch coincides with the onset of Fgf8 expression in the oral epithelium. These results suggest that there exists a population of odontogenic neural crest cells that migrates into the first arch between the 10- and 12-somite stages. These cells either possess odontogenic potential and are able to initiate tooth development, or can respond to odontogenic signals derived from the oral epithelium to support tooth formation.     

Delayed tooth eruption in membrane type-1 matrix metalloproteinase deficient mice

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is expressed highly in mineralizing tissues including bones and teeth. Mice deficient in MT1-MMP (-/-) display severe defects in skeletal development including dwarfism, osteopenia, and craniofacial abnormalities. Death occurs in these mice by about 3 weeks of age. Since MT1-MMP is expressed by the ameloblasts of the enamel organ and by the odontoblasts of the dental papilla, we asked if the developing teeth were adversely affected in the knockout animals. Molars from MT1-MMP -/- mice and controls were examined by histological, X-ray, and SEM analysis at 4, 18-20, and 25 days of postnatal development. At 4 days of development the molars from the -/- mice appeared histologically normal. At 18-20 days of development, the first molars of the -/- mice had apparently normal tooth crowns with normal dentin and enamel; however, the roots were truncated and the teeth had not yet erupted. In contrast to the -/- mice, the first molars of the 18-20-day control animals had erupted. SEM analysis of a -/- first molar and incisor revealed a normal enamel prism pattern. However, X-ray analysis demonstrated that tooth eruption was delayed by approximately 5 days and that the tooth roots were abnormally short in the knockout animals. Since MT1-MMP-deficient mice have been demonstrated to display a generalized increase in bone resorption, these data suggest that inefficient growth of bone surrounding the tooth root complex causes a delay in tooth eruption.     

Immunohistochemical demonstration of an enamel sheath protein, sheathlin, in odontogenic tumors

Enamel proteins can be useful markers for assessment of the functional differentiation of neoplastic epithelium and the nature of extracellular matrices in odontogenic tumors. In the present study, we examined immunohistochemical localization of sheathlin, a recently cloned enamel sheath protein, in various odontogenic tumors to evaluate functional differentiation of tumor cells and the nature of hyalinous or calcified matrices in odontogenic neoplasms. Distinct immunolocalization of sheathlin was observed in the immature enamel of the tooth germ at the late bell stage. Secretory ameloblasts facing the enamel matrix also showed positive staining in their cytoplasm. Definite localization of sheathlin was demonstrated in the enamel matrix in odontogenic tumors with inductive dental hard tissue formation such as ameloblastic fibroodontomas and odontomas. Immunoexpression of sheathlin was, furthermore, demonstrated in eosinophilic droplets in solid nests of adenomatoid odontogenic tumor (AOT) and ghost cells in the epithelial lining of calcifying odontogenic cyst (COC). In AOT, cells facing the eosinophilic droplets also expressed the protein in their cytoplasm. There was neither intracellular staining for sheathlin in the tumor cells nor extracellular staining in the matrix of ameloblastomas and calcifying epithelial odontogenic tumors. Dentin, dysplastic dentin-like hyaline material and cementum in the tumors examined were negative for sheathlin. These results show that immunodetection of sheathlin is a useful marker for functional differentiation of secretory ameloblasts and enamel matrix, which is often hard to differentiate from other hard tissues in odontogenic tumors. Our findings from the view point of sheathlin expression support that the tumor cells of ameloblastomas do not attain full differentiation into functional ameloblasts. It is very interesting that epithelial cells in odontogenic tumors can differentiate into functional ameloblasts without induction by odontogenic mesenchyme, as shown by immunoexpression of sheathlin in eosinophilic droplets within solid epithelial sheets in AOT and ghost cells in the epithelial lining of COC where inductive participation of mesenchymal cells was most unlikely. Received: 19 May 1999 / Accepted: 27 September 1999     

Hyperglycemia and xerostomia are key determinants of tooth decay in type 1 diabetic mice

Insulin-dependent type 1 diabetes mellitus (DM) and oral diseases are closely interrelated. Poor metabolic control in diabetics is associated with a high risk of gingivitis, periodontitis and tooth loss. Salivary flow declines in diabetics and patients suffer from xerostomia. Reduced saliva predisposes to enamel hypomineralization and caries formation; however, the mechanisms that initiate and lead to progression of tooth decay and periodontitis in type 1 DM have not been explored. To address this issue, we analyzed tooth morphology in Akita ?/? mice that harbor a point mutation in the Ins2 insulin gene, which leads to progressive hyperglycemia. Mandibles from Akita ?/? and wild-type littermates were analyzed by microCT, scanning EM and histology; teeth were examined for amelogenin (Amel) and ameloblastin (Ambn) expression. Mice were injected with pilocarpine to assess saliva production. As hyperglycemia may alter pulp repair, the effect of high glucose levels on the proliferation/differentiation of cultured MD10-F2 pulp cells was also analyzed. Results showed that Akita ?/? mice at 6 weeks of age showed chalky white incisors that correlated with marked hyperglycemia and impaired saliva production. MicroCT of Akita ?/? teeth revealed excessive enamel wearing and hypomineralization; immunostaining for Amel and Ambn was decreased. A striking feature was invasion of dentinal tubules with Streptococcus mitis and microabcesses that originated in the coronal pulp and progressed to pulp necrosis and periapical periodontitis. High levels of glucose also inhibited MD10-F2 cell proliferation and differentiation. Our findings provide the first evidence that hyperglycemia in combination with reduced saliva in a model of type1 DM leads to decreased enamel mineralization/matrix proteins and predisposes to excessive wearing and decay. Importantly, hyperglycemia adversely affects enamel matrix proteins and pulp repair. Early detection and treatment of hyperglycemia and hyposalivation may provide a useful strategy for preventing the dental complications of diabetes and promoting oral health in this population.     

Localization of RELM‐β/FIZZ2 Is Associated with Cementum Formation

Resistin‐like molecule‐β/found in inflammatory zone 2 (RELM‐β/FIZZ2) is a cysteine‐rich secretory protein that is localized in the epithelium of the gastrointestinal tract and lung alveoli. Previous reports have suggested that this protein regulates glucose metabolism and inflammation. In the present study, to analyze the involvement of RELM‐β/FIZZ2 in tooth development, we immunohistochemically examined the localization of RELM‐β/FIZZ2 in tooth germs of embryonic days (E) 15‐20 and postnatal days (P) 7‐42 rats. RELM‐β/FIZZ2 was hardly detected in the tooth germ at the bud (E15) stage. However, at the cap (E17) and bell (E20) stages, this protein was detectable in the inner enamel epithelium; whereas cells in the other parts of the enamel organ including the outer enamel epithelium and stellate reticulum did not show the immunoreactivity. During the root formation stage (P14‐28), cells in Hertwig's epithelial root sheath (HERS) localized RELM‐β/FIZZ2. Intense immunoreactivity was also seen in the matrix of the root dentin facing the HERS and the dental follicle. This reactivity was not present on the more upwardly located dentin surface. In contrast, cementum matrix positive for osteopontin and bone sialoprotein was observed on the dentin instead of immunoreactivity for RELM‐β/FIZZ2. Osterix‐positive cells, indicating cementoblast progenitors, were also detected in the dental follicle near the matrix positive for RELM‐β/FIZZ2. These results suggest that RELM‐β/FIZZ2 secreted by the inner enamel epithelium was mainly localized in the matrix at the surface of the apical root dentin and might be involved in cementogenesis. Anat Rec, 2017. © 2017 Wiley Periodicals, Inc. Anat Rec, 300:1865–1874, 2017. © 2017 Wiley Periodicals, Inc.