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PubMed1998

Transforming growth factor-beta superfamily members expressed in rat incisor pulp.

The transforming growth factor (TGF)-beta superfamily comprises more than 35 structurally related genes that have been implicated in embryonic induction and morphogenesis. Different superfamily members may have distinct regulatory roles in tooth development and maintenance. Degenerate primer sets derived from the highly conserved carboxy terminal region of the TGF-beta superfamily were used for reverse transcriptase polymerase with poly(A)+ RNA from the rat incisor pulp as a template. TGF-beta superfamily members expressed in the pulp with known potential to differentiate into odontoblasts and to form dentine were identified. Nucleotide-sequence analysis of the amplified cDNAs identified those encoding activin-betaB; bone morphogenic protein (BMP)-2, -4, -7 and -8; growth/differentiation factor (GDF)-1, -5 and -6; and glial cell line-derived neurotrophic factor. In addition, Northern blot analysis detected TGF-beta1 -beta2 and -beta3; activin-betaA; BMP-6 and GDF-7 mRNA transcripts in the pulp. Coordinated expression of TGF-beta superfamily members in pulp may be critical in tooth development and repair.

Activins↗
PubMed1999

Immunohistochemical demonstration of exocytosis-regulating proteins within rat molar dentinal tubules.

No morphologically defined synaptic structures have so far been detected between nerve terminals and the dentine-producing odontoblasts. Recent studies of the molecular mechanisms in neuronal exocytosis have identified several proteins that participate in synaptic-vesicle exocytosis. By localizing these proteins with immunohistochemical methods, information about the capacity for synaptic exocytosis should be obtained. Here, antibodies directed against some of the exocytosis-related proteins were used to investigate whether they are present in nerve fibers within the dentinal tubules in rat molars. Antibodies against synaptosome-associated protein of 25 kDa, Rab 3, synaptotagmin and synapsin all produced a punctuate staining pattern, suggesting that the proteins are accumulated in bouton-like elements. The results demonstrate that a set of exocytosis-related proteins is accumulated in the dentinal tubules, most probably within the intradentinal nerves. This finding is consistent with the hypothesis that intradentinal nerves can mediate efferent signals.

Animals↗
PubMed1999

Differences in the pattern of lanthanum diffusion into predentine and dentine in mouse incisors and molars.

Lanthanum nitrate was either perfused intravascularly or segments of mouse tooth were immersed in a fixative solution containing the tracer. The tracer deposits were examined in young (8-day-old) and older (8-week-old) mouse incisors and molars, demineralized or undemineralized. Lanthanum passed the distal junctional complex of odontoblasts and appeared in the predentine of incisors as large electron-dense stellate aggregates, 40-70 nm in diameter, and in molars as round, 20-40 nm dots. In dentine, tracer deposits were detected at three locations. Near the predentine dentine junction, the tracer densely stained a band 0.5-2.5 microm in width, also termed metadentine; in the inner circumpulpal dentine, the staining was weaker or lacking in an area extending 5-7 microm from the predentine-dentine junction; in outer circumpulpal dentine, lateral diffusion had occurred in porosities of intertubular dentine. Lanthanum impregnated the walls of dentine tubules and a peritubular-like dentine. In contrast, the mantle dentine was never stained. These differences in the pattern of diffusion prove that lanthanum staining is age-dependent and varies between mouse incisors and molars, independently of tissue processing. Architectural properties and driving flux are involved in the transport and localization of lanthanum in predentine and dentine.

Age Factors↗
PubMed1999

Stimulation of proliferation and differentiation of dog dental pulp cells in serum-free culture medium by insulin-like growth factor.

Insulin, insulin-like growth factors (IGF) I and II are considered to play an important part in the growth and differentiation of dental pulp cells. The present study examined the effects of these factors on pulp cells in serum-free culture conditions. The DNA content and alkaline phosphatase (ALPase) activity of dog pulp cells increased when they were cultured in a serum-free medium supplemented with transferrin, yolk lipoprotein and basic fibrobrast growth factor (TYF medium). The pulp cells produced type I collagen but not type III, suggesting that they might proliferate and differentiate into odontoblast-like cells in a serum-free culture. Both IGF-I and IGF-II enhanced the ALPase activity of pulp cells cultured in TYF medium to an equivalent level, but a higher concentration of IGF-II was necessary to produce a similar effect on DNA synthesis to that of IGF-I. Insulin dose-dependently enhanced DNA synthesis and increased ALPase activity, but its effects were weaker than those of the IGFs. These findings suggest that IGF-I might have a primary role in the growth and differentiation of pulp cells.

Alkaline Phosphatase↗
PubMed1997

Temperature rise in pulpal chamber during fabrication of provisional resinous crowns.

STATEMENT OF PROBLEM: The heat generated during the exothermic polymerization reaction of autopolymerizing resinous materials and the heat generated by ultraviolet lamps during irradiation of photopolymerizing resinous materials could cause pulpal damage when a direct technique is used to fabricate provisional restorations. This could occur if temperature elevations overcome the physiological heat dissipating mechanisms of the dental-periodontal system. PURPOSE: This in vitro study compared the rise in temperatures in the pulpal chamber during fabrication of provisional complete veneer crowns by direct method with different autopolymerizing and photopolymerizing resins. The effect of curing resinous crowns in different matrices, such as a polyvinyl siloxane impression and a vaccuum-formed polypropylene sheet, was also evaluated. RESULTS: The results demonstrated that the amount of heat generated during resin polymerization and transmitted to the pulpal chamber could be damaging to pulpal tissues including odontoblasts. When curing of provisional resinous crowns was performed in the polyvinyl siloxane impression, significantly lower temperatures were recorded compared with curing in the vacuum-formed polypropylene sheet. CONCLUSIONS: To prevent pulpal damage, effective cooling procedures are strongly recommended when directly fabricating resinous provisional crowns.

Analysis of Variance↗
PubMed1999

Immunohistochemical localization of the differentiation marker E11 in dental development of rats.

E11 antigen, originally characterized in a rat osteosarcoma cell line, is known to be expressed during late stages of the osteogenic cell lineage both in vitro and in vivo. The aim of the present study was to monitor the occurrence and distribution patterns of the E11 antigen using monoclonal antibodies (mAb E11 and MEP-1) during different stages of tooth germ development of new-born rats by means of immunohistochemistry. Both antibodies strongly bound to plasma membranes of ameloblasts in presecretory and secretory stages. In addition, odontoblasts and cells of the periodontium were immunoreactive for E11 and MEP-1. During maturation, the immunoreactivity of ameloblast plasma membranes decreased significantly. Our data suggest that E11 and MEP-1 might be important as markers for cell differentiation and mineralization processes during tooth germ development.

Ameloblasts↗
PubMed1992

Autonomic nerve endings in the odontoblast/predentin border and predentin of the canine teeth of dogs.

Autonomic nerve endings in the odontoblast/predentin border and the predentin were observed by electron microscopy in the canine teeth of dogs. Adrenergic nerve endings with a number of very electron-dense granular vesicles and a minority of agranular vesicles of different sizes were clearly observed, but seen rarely, at the odontoblast/predentin border, in the predentin adjacent to the odontoblast processes, and as free endings in the middle part of the predentin. Cholinergic nerve endings were also observed with a number of agranular vesicles of different sizes without any visible exocytosis, in the odontoblast/predentin border, and in the predentin. Near the areas of collagen formation, extensive exocytosis of granular vesicles of different sizes was observed in adrenergic nerve endings located as free nerve endings in the predentin.

Adrenergic Fibers↗
PubMed1997

Osteocalcin expression in young and aged dental pulps as determined by RT-PCR.

Dental pulps were obtained from third molars of young adults (17-25 yr) or from molar teeth of individuals > 50 yr of age and examined for the expression of osteocalcin (OC) mRNA by RT-PCR. OC was selected as a determinant of pulp vitality, because it has long been associated with the production of hard tissue matrix in teeth and bone. For comparative purposes, the expression of OC in each pulp was normalized relative to its housekeeping gene-product GAPDH by the establishment of a OC/GAPDH ratio. This study demonstrated that OC expression, presumably by cells of odontoblast lineage, does not diminish relative to the extant cell population. Our findings suggest, despite a reduction in volume and cell numbers, that the pulps of aging teeth retain a capacity for dentin deposition and a potential for caries and trauma resistance.

Adolescent↗
PubMed2003

In vitro cytotoxicity of five glass-ionomer cements.

To evaluate the cytotoxic effects of five glass-ionomer cements (GICs) on an odontoblast cell line (MDPC-23), disks of every material were prepared and divided into Group 1: Vitrebond, Group 2: Vitremer, Group 3: Fuji II LC, Group 4: Fuji IX GP, Group 5: Ketac-Molar, Group 6: Z-100 (positive control). In Group 7, phosphate-buffered saline solution (negative control) was applied on filter paper. After placing the samples in the bottom of wells, the cells (30,000cells/cm(2)) were plated and incubated for 72h. The cell number was counted, the cell morphology was assessed by scanning electron microscopy and the cell metabolism was evaluated using methyltetrazolium assay. The statistical analysis of Kruskal-Wallis was used to determine if the scores obtained for the cell metabolism and number of cells were different at the 95% confidence level. In groups 1, 2, 3, 4, 5, and 6 the materials decreased the cell number by 74.5%, 75.5%, 45.5%, 29.5%, 32.5%, and 88.5%, respectively. In groups 1, 2, 3, 4, and 5, the experimental GICs reduced the cell metabolism by 79%, 84%, 54%, 40%, and 42.5%, respectively. Despite the fact that all experimental materials were cytotoxic to the MDPC-23 cells, the GICs were the least cytotoxic. On the other hand, the RMGICs caused the highest cytophatic effects.

Animals↗
PubMed1985

Aberrations in pulpal histology and dentinogenesis in transplanted erupting teeth.

Responses of pulpal tissues were assessed histologically in teeth that had been transplanted after root formation was moderately well advanced. Degeneration occurred in the coronal third, but the area was repopulated by cells proliferating from the center of the pulp. Apically, the tissues, including the odontoblasts, remained vital and functional. Widespread intrapulpal formation of osteodentin was prominent in later observation periods. Apical closure occurred prematurely by production of osteodentin and cellular cementum in the form of an apical diaphragm; these mineralized tissues were confluent with a similarly active deposition of cellular cementum apically on the root surface.

Animals↗
PubMed1983

Considerations on the classification of odontogenic tumours.

The histogenetic, organogenetic and embryological aspects of tooth development are used in 4 hypotheses which form a basis for understanding the formal genesis of benign odontogenic tumours. The ectomesenchyme which gives rise to odontoblasts and cementoblasts is distinguished from mesenchymal tissue. Using 4 histogenetic cell groups (epithelial, ectomesenchymal, neuroectodermal and mesenchymal), the various types of odontogenic tumour are discussed. A short analysis is presented of different odontogenic tumour classifications, and some principle problems of terminology and taxonomy are examined.

Ameloblastoma↗
PubMed1998

Expression and localization of membrane type 1 matrix metalloproteinase in tooth tissues.

Matrix metalloproteinases (MMPs) have been detected in forming dental enamel and are thought to play an important role during enamel biomineralization. Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a membrane bound member of the MMP gene family that has previously been shown to be expressed by cells associated with bone and cartilage formation (osteoclasts, osteoblasts and chondrocytes). Thus, we asked if MT1-MMP was also expressed by the cells responsible for the formation of enamel and dentin. A porcine MT1-MMP cDNA composed of 3284 bp was isolated from an enamel organ-specific cDNA library. Multiple tissue Northern blot analysis revealed that the MT1-MMP message was expressed highly in the enamel organ and pulp organ when compared to the expression levels observed in other non-mineralizing tissues. Northern blot analysis of stage-specific enamel organs (early secretory, late secretory, or maturation stage) and their corresponding pulp organs revealed that MT1-MMP expression increased as the dentin matured. In the enamel organs, however, the MT1-MMP message level became reduced only during the late secretory stage. Immunohistochemical analysis showed that MT1-MMP was present on the surface of the cells (ameloblasts and odontoblasts) responsible for dentin and enamel formation. Thus, MT1-MMP is highly expressed in developing tooth tissues and may play a role in the biomineralization of enamel and dentin.

Ameloblasts↗
PubMed2004

Short-term histomorphological effects of Er:YAG laser irradiation to rat coronal dentin-pulp complex.

OBJECTIVE: The objective of the study was to examine the morphological changes of neural elements in dentin-pulp complex ultrastructurally after Er:YAG laser irradiation and elucidate the mechanism of pain reduction in cavity ablation. STUDY DESIGN: The Er:YAG laser was applied at occlusal surfaces of upper and lower first molar cusps of 6 rats, and shallow cavities were ablated. The dentin and pulps were examined with light and electron microscopes at 6 hours after the irradiation. Teeth, without laser irradiation, from three rats were used as controls. RESULTS: Disruption of nerve terminals in the dentinal tubules, degeneration of nerve terminals between odontoblasts, and disruption of the myelin sheath in the pulp core were demonstrated with electron microscope. CONCLUSION: Some Er:YAG laser beams could penetrate to deeper areas than ablated area, and damage of nerve fibers and terminals might be a mechanism of pain reduction in cavity ablation with Er:YAG laser.

Aluminum Silicates↗
PubMed2001

The secretion of amelogenins is associated with the induction of enamel and dentinoid in an ameloblastic fibro-odontoma.

Ameloblastic fibro-odontoma is the unique entity of epithelial-ectomesenchymal odontogenic tumors, which is characterized by enamel formation in addition to dentine. We examined immunohistochemically a case of this tumor in which enamel having prism structures was developed in the absence of odontoblast differentiation but was in contact with mesenchymal matrices. Histological examination showed diverse morphological features of epithelial tumor cells, e.g., cuboidal cells comprising tooth bud-like projections, ameloblast- and stellate reticulum-like cells, and residual cells in forms of extended cords or islands of odontogenic epithelium. Immunostaining with anti-amelogenin sera proved that the intracellular production of amelogenins was initiated at the tooth bud-like stage. The secreted amelogenins were detected almost exclusively in the induced enamel and dentinoid areas, as well as in the core region of cementicle-like spheres deposited in the encapsulating stroma. The results obtained indicate that the odontogenic tumor epithelia and its products, i.e., amelogenins, participate in multifaceted aspects of dental hard tissue formation that takes place during oncogenesis.

Adult↗
PubMed2000

Ultrastructural preservation of rat embryonic dental tissues after rapid fixation and dehydration under microwave irradiation.

Adequate preservation of the cells and matrix of mineralising tissues remains difficult, as organic components and initial mineral deposits may be lost during conventional processing for electron microscopy. In this study, we have reduced significantly the processing time using microwave irradiation. Rat molar tooth germs were fixed in 4% glutaraldehyde + 4% formaldehyde with 0.1 M sodium cacodylate in a laboratory microwave oven for two periods of 20 s with a maximal temperature of 37 degrees C. After conventional washing and post-fixation, specimens were dehydrated in graded ethanols under microwave irradiation for a total of 7 min 20 s. For comparison, some specimens were processed by conventional methods. After embedding, ultrathin sections were examined by electron microscopy. In differentiating ameloblasts and odontoblasts, plasma membranes, mitochondria, rough endoplasmic reticulum, the Golgi complex, together with all other cytoplasmic organelles exhibited excellent preservation. Microtubules, microfilaments and coated vesicles were particularly evident. Crystal-like mineral deposits were conspicuously present in relation to dentine matrix vesicles and collagen fibrils as well as in enamel matrix. The matrix of forming enamel had a globular electron-lucent appearance. It is concluded that this is a rapid method which provides a preserved or even improved morphology.

Ameloblasts↗
PubMed2001

A novel rat dentin mRNA coding only for dentin sialoprotein.

Dentin sialoprotein (DSP) is a major glycoprotein present in the mineralized dentin matrix that is expressed mainly by young and mature odontoblasts. Mutations in the DSP coding regions are linked to Dentinogenesis imperfecta I and II. indicating the importance of DSP in tooth formation. Previous studies have identified multiple mRNA transcripts in dentin that code for both DSP and phosphophoryns (PPs). Using reverse transcriptase-polymerase chain reaction (RT-PCR) to characterize these mRNA transcripts, we have identified a cDNA that codes for DSP, but not PP. This cDNA codes for a protein with 324 amino acids, 303 amino acids being identical to the published rat DSP sequence. However, the subsequent 21 amino acids are unique to this cDNA. Based on the coding sequence, the core protein is predicted to have a pI=4.24, a net charge of -34, and to contain four potential N-glycosylation sites and six potential sites for phosphorylation by casein kinase. That the corresponding mRNA was present in day 5 molar tooth germs was confirmed using RNA protection assays. These data, therefore, identify a novel transcript in rat tooth germs that codes only for DSP (designated as DSPII).

Age Factors↗
PubMed1999

Immunolocalization of Bone Morphogenetic Protein-2 and -3 and Osteogenic Protein-1 during murine tooth root morphogenesis and in other craniofacial structures.

The distribution of Bone Morphogenetic Protein-2, and -3 (BMP-2 and BMP-3) and Osteogenic Protein-1 (OP-1, also known as BMP-7) during root morphogenesis and in other craniofacial structures was examined in sections of 12- to 18-d-old mouse heads using polyclonal and monoclonal antibodies. BMP-3 and OP-1 were localized in alveolar bone, cementum, and periodontal ligament, whereas BMP-2 was only localized in the alveolar bone of periodontium. All three BMPs were localized in predentine, dentine, odontoblasts, osteoblasts, osteocytes, osteoid, cartilage, chondrocytes and spiral limbus. BMP-2 and OP-1 were also localized in spiral ligament and interdentate cells of the cochlea, whilst BMP-3 was restricted to the spiral ganglion. BMP-3 was also localized in ducts of submandibular and sublingual salivary glands, acini of the lacrimal gland, Purkinje cells in the cerebellum, nerve fibres of the cerebellum and brain, afferent cells of the dorsal root ganglia, inferior alveolar nerve, and peripheral processes of the vestibulocochlear nerve. OP-1 was also localized in hair and whisker follicles, sclera of the eye and in ameloblasts. The demonstration of BMP-3 in the nervous system suggests that this protein may be neurotrophic during development and maintenance of the nervous system. The composite expression of BMPs/OPs during periodontal tissue morphogenesis suggests that optimal therapeutic regeneration may entail the combined use of different BMPs/OPs.

Afferent Pathways↗
PubMed2002

Location, arrangement and possible function of interodontoblastic collagen fibres in association with calcium hydroxide-induced hard tissue bridges.

AIM: To assess the location, arrangement and possible function of interodontoblastic collagen fibres in association with calcium hydroxide-induced hard tissue bridges by using light and transmission electron microscopy techniques and immunohistochemical staining localization. METHODOLOGY: Prior to the study, an animal use protocol form was reviewed and approved by the Screening Committee for Animal Research of the Tokyo Medical and Dental University. Exposed monkey pulps were capped with a hard-set calcium hydroxide and histopathologically evaluated at 3, 14, 21, 30 and 90 days, using light microscopy with silver staining and transmission electron microscopy to differentiate structural features of interodontoblastic collagen fibres. In addition, an attempt was made to identify and to differentiate between several types of collagen and fibronectin using immunohistochemical localization techniques. RESULTS: At 14 days, interodontoblastic collagen fibres were observed extending from the original dentine, passing through the odontoblasts, and consisted of two portions: a thick fibril and a thin fibril. At 21 days, interodontoblastic collagen fibres were seen penetrating into the predentine and becoming incorporated into the mineralized dentine. At 30 days, interodontoblastic collagen fibres reached the cell process. Although interodontoblastic collagen fibres were no longer observed near the odontoblastoid cells at the area of the newly formed tubular dentine, interodontoblastic collagen fibres were observed embedded within the primary formed dentine bridge. Immunohistochemical staining demonstrated type I collagen and fibronectin within the interodontoblastic collagen fibres. CONCLUSIONS: Interodontoblastic collagen fibres were routinely detected throughout early dentine bridges. Interodontoblastic collagen fibres are thought to be important for initial dentine bridging to induce and support a dentinogenesis framework.

Animals↗