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Histological evaluation of dog permanent teeth after traumatic intrusion of their primary predecessors.

The aims of this study were to analyze the histomorphology of developing permanent teeth whose primary teeth had suffered traumatic intrusion, as well as to compare the influence of immediate extraction of the intruded tooth to passive re-eruption. Nine dogs from 45 to 50 days old were submitted to the intrusion of the maxillary central and lateral primary incisors using a force applicator adapted to the teeth incisal cuspids. The right side intruded teeth were kept in their sockets and the ones on the left side were extracted 30 min later. After a postoperatory periods of 30 and 60 days, four (group 1) and five (group 2) dogs, respectively, were killed by perfusion. The histological evaluations showed that, in group 1, alterations had occurred in the odontoblastic layer and deposition of the enamel matrix had taken place in some specimens while in group 2, a portion of non-mineralized matrix was observed. We concluded that the morphological changes were because of the immediate trauma of intrusion. No differences were found between the groups where the primary tooth was immediately extracted or left to passively re-erupt.

Ameloblasts↗

Tooth survival and periodontal tissues healing of allogenic-transplanted teeth in the mice.

OBJECTIVE: For the successful organ transplantation, immune rejection has to be considered. Autogenic transplantation of human teeth is generally carried out free of clinical difficulty because of the lack of immune reaction, whereas allogenic tooth transplantations easily induce host immune rejection to donor tissues. The aim of the present study was to evaluate the replacement of donor tissue by host cells after allogenic tooth transplantation. MATERIALS AND METHODS: First molars extracted from lacZ transgenic ROSA26 mice were transplanted into the alveolar socket and the tongue of host wildtype mice, where the first molar had existed. RESULTS: Donor cells from lacZ transgenic mice were not detected in the periodontal ligament space, but rather in the pulp chamber of the donor tooth. Furthermore, if the pulp chamber was widely open to an affluent blood supply, odontoblasts and fibroblasts in the donor tissue survived in the dental pulp. CONCLUSIONS: Our experimental models using lacZ transgenic ROSA26 mice clearly demonstrate that donor periodontal tissue cells are replaced by host cells and that periodontal tissue can regenerate after allogenic tooth transplantation. Furthermore, our models suggest that donor pulpal cells can survive if the vascular supply into the pulp chamber is sufficient.

Animals↗

Analysis of developmental potentials of dental pulp in vitro using GFP transgenes.

BACKGROUND: In recent years there has been increasing progress in identifying stem cells from adult tissues and their potential application for tooth replacement/regeneration. Our previous in vivo studies show that pOBCol3.6GFP and pOBCol2.3GFP transgenic animals provide a unique model to gain insight into progenitor/stem cells in the dental pulp capable of giving rise to odontoblasts. OBJECTIVES: To characterize the behavior of dental pulp cells derived from pOBCol3.6GFP animals in vitro. EXPERIMENTAL DESIGN: Primary cultures were established from the coronal portions of the pulps isolated first molars from 5-day-old pOBCol3.6GFP heterozygous mice and grown for 21 days. In these cultures proliferation, clonogenic capacity, activation of 3.6-GFP and mineralization were examined. RESULTS: Our observations show that dental pulp cells derived from 3.6-GFP contain a population of proliferative, clonogenic cells with the ability to mineralize. We also show the stage specific activation/upregulation of 3.6-GFP in primary cultures derived from dental pulp. In these cultures, expression of Col1a1-3.6-GFP occurs prior to the appearance of mineralized nodules and is unregulated in mineralized nodules. CONCLUSIONS: Col1a1-GFP transgenes appear to fulfill many of the requirements of a marker gene for cell lineage studies in intact tooth and primary cultures derived from dental pulp.

Animals↗

Total etch technique and cavity isolation.

In the total etch technique for chemically adhesive composite restorations, the phosphoric acid penetrates only 10 microns or less into the vital dentin with the dentinal tubules being filled with the odontoblast processes. The acid is completely removed by subsequent air-water jet spray washing. The tubule apertures are perfectly sealed by the protective bonding agent layer with the resin tags adhering to the tubule walls and the resin-impregnated dentin surface. Isolation of the cavity from moisture contamination is required for only less than a few seconds after drying the etched cavity until the bonding agent coating and after this coating until the composite resin placement. Such a short time for isolation is quite easy even without a rubber dam when a trained assistant is cooperating.

Acid Etching, Dental↗

Pulp response in primary teeth with deep residual caries treated with silver fluoride and glass ionomer cement ('atraumatic' technique)

Histological assessment of the dental pulps of 55 carious primary teeth was carried out 3 to 58 months after treatment by the 'atraumatic' technique involving application of 40 per cent silver fluoride to residual caries followed by restoration with glass ionomer cement. Fifty of the 55 teeth examined showed a favourable pulpal response, inducing presence of abundant reparative dentine and a wide odontoblast layer. Histological comparisons were made between these teeth and others not treated with silver fluoride but restored with glass ionomer cement, amalgam or zinc oxide and eugenol. Possible mechanisms of the action of silver fluoride in arresting residual caries are discussed. The question of whether or not treatment of carious dentine with silver fluoride represents a biologically acceptable clinical procedure cannot be answered on the basis of pulpal histology alone. The very high concentration of fluoride in commercial preparations of silver fluoride raises several questions concerning its clinical safety.

Adolescent↗

Electron microscopy: attachment sites between connective tissue cells.

Regions of attachment have been observed between connective tissue cells from four different structures: fibroblasts in embryonic and fetal tendons, fibroblasts in fetal ligamentum nuchae, odontoblasts, and osteoblasts. Morpho logically these sites appear to be focal and to consist of an approximation of the plasma membranes of adjacent cells to within approximately 200 A. In the region of approximation both the extracellular space and the cytoplasm adjacent to the plasmalemma are increased in density. We have postulated a role for these sites in the maintenance of structural integrity.

Animals↗

Tissue- and stage-specific activation of an endogenous provirus after transcription through its integration site in the opposite orientation.

Endogenous proviruses of the Moloney murine leukemia retrovirus (Mo-MuLV) are transcriptionally blocked in early embryos and in general remain silent even when the tissues have become permissive to the expression of newly integrated copies. Eventually, activation in presumably very few cells initiates rapid superinfection leading to viremia and leukemia, but the processes leading to provirus activation are unknown. Differences in the onset and development of viremia between several mouse strains carrying an endogenous Mo-MuLV (Mov lines) are attributed to a chromosomal position effect, but neither cell type nor stage of provirus activation is known for any strain. We have now monitored the appearance of viral transcripts and particles in the Mov13 strain, which carries the Mo-MuLV provirus in inverted orientation in the first intron of a collagen gene (Col1a1) with well-characterized transcriptional activity. We report obligatory tissue- and stage-specific virus activation in osteoblasts and odontoblasts. The significance of this activation pattern is indicated by the fact that of the great variety of cells expressing the wild-type collagen gene, only these two cell types can also transcribe the mutant allele including its viral insert. We propose that this transcription of the proviral genome, albeit in the opposite direction, leads to the activation of the viral promoter.

Animals↗

The future role of a molecular approach to pulp-dentinal regeneration.

The ultimate goal of a regenerative pulp treatment strategy is to reconstitute normal tissue continuum at the pulp-dentin border, regulating tissue-specific processes of tertiary dentinogenesis. Experimental investigations in mature teeth have shown that a network of extracellular matrix molecules and growth factors signal tertiary dentinogenesis. Application of dentin matrix components or growth factors in deep dentinal cavities stimulated up-regulation of biosynthetic activity of primary odontoblasts (reactionary dentin formation). Pulp-capping studies with a broad spectrum of biological agents, including growth factors and extracellular matrix molecules, showed formation of osteodentin and/or tertiary dentinogenesis (reparative dentin formation). Promising biologically active substances should be subjected to careful evaluation in well-designed preclinical investigations as well as in long-term clinical trials before their introduction in clinical practice.

Animals↗

Developmental arrest of mouse tooth germs in vitro by 6-diazo-5-oxonorleucine (DON).

Molars and incisors were excised from the mandibles of 15-day-old mouse embryos and explanted to agar-solidified Eagle's basal medium with 10% fetal calf serum and to medium containing 6-diazo-5-oxonorleucine (DON), a glutamine analog which inhibits glycoprotein and glycosaminoglycan synthesis. The concentration of DON ranged from 5-100 microgram/ml; the lowest dose at which consistent results were obtained was 75 microgram/ml. At the end of a 4-day culture period, controls displayed characteristic incisiform or molariform morphologies and possessed odontoblasts and preameloblasts. The growth and development of DON-treated germs, however, was arrested and such germs remained morphologically and histologically similar to the developmental state attained at the time of explantation. DON-treated germs also contained less abundant Alcian blue staining material than controls. Development arrest by DON was prevented by addition of glutamine but not by glucosamine or the purine analog amino imidazol carboximine. Suppressed germs resumed normal development when removed from medium containing 75 microgram/ml of DON to control medium. These results suggest that glycoprotein or glycosaminoglycans play a role in epithelial-mesenchymal inductive interactions.

Ameloblasts↗

Human root caries: histopathology of arrested lesions.

The histopathology of arrested root caries lesions was examined in extracted human teeth. The main structural characteristics of arrested lesions were the completely mineralized surface area and the formation of a distinct sclerosis of the dentinal tubules. Intertubular dentin was, with the exception of the dentinal tubules, fully mineralized up to the surface. Dentinal tubules near the surface were either filled with ghosts of microorganisms or with crystals of different shapes. Sclerosis of the dentinal tubules was characterized by the presence of three different patterns of intratubular mineralization that occur in distinct regions of the zone of tubular sclerosis. The patterns were distinguishable by the type of crystals and their association with organic structures such as collagenous fibrils or odontoblast processes. It is suggested that arrested lesions are based on (1) the formation of an inner barrier that interrupts the diffusion of substrata from the pulp to invaded bacteria, (2) the formation of an outer barrier by a compact, highly mineralized surface region which blocks the diffusion of products of bacterial metabolism into dentin, and (3) an area of mineralization which extends from the outer barrier toward the root canal within demineralized dentin. The present study demonstrates the considerable potential of caries lesions in dentin to become arrested, and subsequently partially remineralized. These phenomena seem to depend on the severity of an active lesion and its location on the root surface. This should be taken into account when diagnosing root caries lesions. The potential of root caries lesions to become arrested indicates that the treatment concept of active root caries lesions should be reconsidered.

Adult↗

Cloning human enamelin cDNA, chromosomal localization, and analysis of expression during tooth development.

Enamelin is the largest protein in the enamel matrix of developing teeth. In the pig, enamelin is secreted as 186-kDa phosphorylated glycoprotein, which is rapidly processed by enamel proteinases into smaller cleavage products. During the secretory stage of enamel formation, enamelin is found among the crystallites in the rod and interrod enamel and comprises roughly 5% of total matrix protein. Although the function of enamelin is unknown, it is thought to participate in enamel crystal nucleation and extension, and the regulation of crystal habit. Here we report the results of enamelin in situ hybridization in a day 1 mouse developing incisor that shows that enamelin is expressed by ameloblasts, but not by odontoblasts or other cells in the dental pulp. The restricted pattern of enamelin expression makes the human enamelin gene a prime candidate in the etiology of amelogenesis imperfecta (AI), a genetic disease in which defects of enamel formation occur in the absence of non-dental symptoms. We have cloned and characterized a full-length human enamelin cDNA and determined by radiation hybrid mapping and fluorescent in situ hybridization (FISH) that the gene is located on chromosome 4q near the ameloblastin gene in a region previously linked to local hypoplastic AI in six families. These findings will facilitate the search for specific mutations in the enamelin gene in kindreds suffering from amelogenesis imperfecta.

Ameloblasts↗

Effects of TGFbeta1 on dental pulp cells in cultured human tooth slices.

Transforming growth factor-beta1 (TGF beta1) is a potent modulator of tissue repair in various tissues. To analyze its role during human dental repair, we used thick-sliced teeth cultured as described previously (Magloire et al., 1996). The supply of TGF beta1 to the pulp tissue was accomplished by means of a small tube glued onto the dentin. We show that this device allowed the growth factor to diffuse locally through dentinal tubules and to bind to the cells present in the coronal pulp opposite the TGF beta1-delivery tube. The tube was filled with 20 ng/mL TGF beta1, and slices were cultured for 4 days. Results show a preferential accumulation of cells in the odontoblastic and subodontoblastic layers in the vicinity of the tube. Cell proliferation increased in the subodontoblastic layer and in the underlying pulp, and BrdU-positive cells were abundant around the blood vessels. TGF beta1 induced type I collagen production by the odontoblastic/subodontoblastic/pulp cells in the stimulated zone, as demonstrated by in situ hybridization. These results suggest that TGF beta1 could be directly involved in the regulation of cell proliferation, migration, and extracellular matrix production in the human dental pulp and eventually in the repair process occurring after tooth injury.

Adolescent↗

Gelatinase A (MMP-2) in developing tooth tissues and amelogenin hydrolysis.

Matrix metalloproteinases (MMPs) are thought to play important roles during enamel and dentin biomineralization. Previously, membrane type-1 matrix metalloproteinase (MT1-MMP) was localized to the plasma membranes of ameloblasts and odontoblasts of the developing tooth. The best-characterized function of MT1-MMP is to initiate the activation of gelatinase A (MMP-2). Thus, we hypothesized that gelatinase A may also be expressed by developing tooth tissues. A full-length porcine gelatinase A mRNA was isolated by RT-PCR homology cloning of an enamel-organ-specific cDNA library. Northern blot and in situ hybridization analyses demonstrated gelatinase A expression in developing tooth tissues. Immunohistochemical analysis localized gelatinase A close to the plasma membrane of these tissues. Furthermore, recombinant gelatinase A was demonstrated to cleave recombinant amelogenin into several fragments of differing molecular masses. Thus, gelatinase A is expressed by developing tooth tissues along with its activator MT1-MMP and may, therefore, play an important role during tooth development.

Ameloblasts↗

Identification of cis-DNA regions controlling Bmp4 expression during tooth morphogenesis in vivo.

Epithelial-mesenchymal interactions are required for tooth formation. Bone morphogenetic protein 4 (Bmp4) is a crucial signaling molecule during this process. For better understanding of the role of the Bmp4 gene during tooth development, we studied the mechanisms that control its temporal and spatial expression during development. Using a transgenic approach, we determined that the domains which controlled Bmp4 expression in epithelium-derived ameloblasts were located in the region between 0.26 kb and 1.1 kb of the murine Bmp4 promoter. In contrast, the domains controlling Bmp4 expression in mesenchyme-derived odontoblasts and pulp cells existed in other regions of the Bmp4 gene. We have also demonstrated that the domains controlling Bmp4 expression in primordial tooth cells differ from those controlling Bmp4 expression in mature tooth tissues. The determination of unique domains by controlling the expression of the Bmp4 gene suggests that different transcriptional factors regulate the Bmp4 level at different stages during tooth morphogenesis.

Ameloblasts↗

Effect of infusing selected chemical compounds on dentinal fluid movement in the rat.

Intra-arterial infusion of carbamyl phosphate or of carbamyl-DL-aspartic acid into rats on a cariogenic diet greatly stimulated the movement of fluid through the odontoblastic processes. The infusion of sodium cyanate also stimulated fluid movement. Guanidine HCL and L-asparagine were active at higher concentrations. Purifying the urea on a mixed-bed ion exchange resin virtually removed its stimulatory effect on dentinal fluid movement. The action of urea is apparently attributable to contamination with sodium cyanate.

Acriflavine↗

Electron microscopic structure of the two layers of carious dentin.

Two layers of carious dentin from extracted human teeth were observed with an electron microscope. The first layer, which is superficial and fuchsin-stainable, showed degenerated collagen fibers and granular or leaflike inorganic crystals irregularly scattered. The second layer, which is profound and fuchsinunstainable, showed expanded odontoblastic processes, sound collagen fibers, and apatite crystals bound to the fibers like fringes.

Apatites↗