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Xenografts of recombined bovine odontogenic tissues and cultured cells to hypothymic mice.

It was demonstrated in this study that recombined tissues of bovine dental papilla and reduced enamel epithelium, when transplanted to the subcapsular kidney site of hypothymic mice, elaborated tissues resembling osteodentin and dentin matrices with which latter tissues were closely associated cells resembling odontoblasts. In xenografts of cultured cell populations of these tissues, osteodentin-like matrices occurred in the absence of dentin-like matrices.

Animals↗

Effect of ProRoot MTA on pulp cell apoptosis and proliferation in vitro.

ProRoot Mineral Trioxide Aggregate (MTA) has been indicated as a pulp capping material. The purpose of this study was to evaluate the effect of tooth-colored (white) MTA on pulp cell apoptosis and cell cycle. Mouse odontoblast-like cells (MDPC-23) and undifferentiated pulp cells (OD-21) were exposed to 0 to 100 mg MTA for 24 h. Propidium iodide staining followed by flow cytometry demonstrated that MTA did not induce apoptosis of MDPC-23 or OD-21 (p > 0.05). Cell cycle analysis showed that MTA induced a modest (but significant) increase in the percentage of MDPC-23 in the S and G2 phases, and OD-21 in the S phase of cell cycle, as compared to untreated controls (p </= 0.05). In conclusion, MTA induced proliferation, and not apoptosis, of pulp cells in vitro. These findings suggest a potential mechanism to explain the regenerative effect observed in the dentin-pulp complex when MTA was used for direct pulp capping.

Aluminum Compounds↗

The upregulation of matrix metalloproteinase-9 in inflamed human dental pulps.

The matrix metalloproteinases (MMPs) participate in a wide variety of extracellular matrix degradation. Detailed knowledge of MMPs may be important for understanding the pathogenesis of pulpal inflammation. The purpose of this study was to compare MMP-9 expression in clinically healthy human pulp and inflamed human pulp tissue specimens. We compared the levels of MMP-9 between clinically healthy pulp and inflamed pulp tissues by using the semi-quantitative reverse-transcriptase polymerase chain reaction analysis. In addition, immunohistochemistry was used to identify the in situ localization of MMP-9 expression in pulp specimens. MMP-9 mRNA gene was found to be increased in inflamed pulps as compared with clinically healthy pulp tissues (p < 0.05). The results from immunohistochemistry demonstrated that MMP-9 expression was significantly higher in the inflamed pulps than clinically healthy pulps (p < 0.05). MMP-9 stain was detected in the odontoblasts, fibroblasts, inflammatory infiltrates, and endothelial cells. Taken together, MMP-9 may play an important role in the pathogenesis of pulpal inflammation.

Coloring Agents↗

Histopathology of the teeth in segmental odontomaxillary dysplasia: new findings.

Histological examination of the deciduous teeth in two cases of segmental odontomaxillary dysplasia (SOMD) showed fibrous enlargement of the pulps, an irregular pulp/dentine interface displaying many pseudoinclusions and pulp stones. There were tubular defects in the coronal dentine from pulp horn to cusp tip, an irregular tubular structure to the circumpulpal dentine of the apical half, a focally deficient odontoblast layer and widespread external resorption. Together with the clinical features of unilateral maxillary enlargement, upper alveolar expansion in the distal segment, increased spacing and delayed eruption of the deciduous molars and absence of premolar teeth, these histological appearances allow distinction of this condition from fibrous dysplasia (FD), segmental hemifacial hypertrophy (SHH) and regional odontodysplasia (ROD).

Alveolar Process↗

Mechanisms controlling secondary initiation of dentinogenesis: a review.

Dentinogenesis can be initiated secondarily as an intrinsic ability of the dental pulp to repair, or after interaction of pulp cells with specific exogenous inductive factors. In the present article the basic developmental aspects, highlighting the mechanism by which dentinogenesis is initiated during tooth development, are discussed. Furthermore, clinical and experimental observations concerning the events taking place during secondary initiation of dentine formation, as part of exposed or non-exposed pulp tissue repair, or as a result of dentine matrix or other chemical-pulp cell interactions, are reviewed. Discussion includes hypotheses relating to the crucial biological steps leading to expression of odontoblastic-like cell phenotype and secondary initiation of dentine histogenesis.

Animals↗

Localization of chondroitin sulphate and dermatan sulphate in human dental pulps--an immunohistochemical study.

The distribution of dermatan sulphate and chondroitin sulphate in human dental pulps has been assessed using monoclonal antibodies and immunoperoxidase localization techniques. The pulpal tissues were reacted with specific antibodies following pretreatment of the sections with chondroitinase ACII or chondroitinase ABC. Both the 4- and 6-sulphates isomers of chondroitin sulphate were detected in the tissues studied. Very little dermatan sulphate could be detected. These glycosaminoglycans appeared throughout the pulpal connective tissues with a particularly strong localization to the region adjacent to the odontoblastic and predentine layers. Such distribution strongly implicates chondroitin sulphate in the mineralization process of human dentine.

Chondroitin Sulfates↗

Influence of the pulpal components on human dentine permeability in vitro.

AIM: To examine the influence of the retained pulpal components on permeability of human dentine by monitoring drug diffusion. METHODOLOGY: Twelve intact dentine discs were prepared from freshly extracted human third molars. The dentine surface on the enamel side was etched with 10% polyacrylic acid for 30 s. The drug diffusion test was carried out before and after removal of the retained pulpal components. Each dentine disc was inserted between two plastic chambers; enamel- and pulpal-side chambers, which were filled with 0.05 mol L(-1) naproxen sodium (NA) and phosphate-buffered saline (PBS), respectively. After 10 min, the solution on the pulpal-side chamber was collected to determine the concentration of NA using a spectrophotometer. To remove the retained pulpal components and residual NA, the pulp chamber of each disc was washed out with PBS and placed in an ultrasonic cleaner. After removal of these components and the residual NA, the drug diffusion test was repeated. The inner surface of the pulp chamber was observed using scanning electron microscopy (SEM) before and after the removal of the retained pulpal components. RESULTS: The amount of NA that diffused through dentine into the pulp was significantly higher after the pulp chamber was washed out with PBS (paired t-test, P < 0.05). SEM observation demonstrated the presence of the retained pulpal components, odontoblastic layer and some parts of subodontoblastic zone, covering the surface of predentine. These components were removed after the pulp chamber was washed out with PBS followed by cleaning in an ultrasonic cleaner. CONCLUSIONS: The presence of retained pulpal components had a significant influence on drug diffusion through dentine discs.

Anti-Inflammatory Agents, Non-Steroidal↗

Analysis of the calcium distribution in predentine by EELS and of the early crystal formation in dentine by ESI and ESD.

Predentine is a collagen-rich extracellular matrix between the odontoblasts and the dentine with a width of about 15-20 microns. Electron energy-loss spectroscopy of rat incisors shows a significantly higher calcium content in the predentine at the predentine-dentine border than in the middle region of the predentine. At the predentine-dentine border in the dentine, the calcium and the phosphate groups combine to form apatite crystallites. Electron spectroscopic diffraction with zero-loss filtering revealed that the earliest crystallites contain only Debye-Scherrer rings of apatite, which are fewer in number and more diffuse than the diffraction rings from the mature crystallites. We therefore conclude that the early crystallites still contain lattice defects, which are annealed out to some degree with crystal growth. Electron spectroscopic imaging with zero-loss filtering also showed that the earliest crystallites are chains of dots (or small islands); they build up strands composed of islands, which rapidly acquire a needle-like character and coalesce laterally to form ribbon-or plate-like crystallites. The parallel strands sometimes appear to reinforce the macroperiod of the collagen microfibrils (67 nm) by tiny holes without any crystal-substance lined up perpendicular to the parallel strands of the crystallites.

Animals↗

Protein dynamics of bovine dentin phosphophoryn.

Bovine dentin phosphophoryn (BDP), a protein rich in aspartyl (Asp) and o-phosphoseryl [Ser(P)] residues, is synthesized by odontoblasts and believed to be involved in matrix-mediated biomineralization of dentin. The elucidation of the structure-function relationship of phosphophoryn has been a challenge because of its high-molecular weight, high negative charge, repetitive sequence, and lability. We have used the dynamic behavior of the (1)H NMR signal at 600 MHz to provide insight into the molecular dynamics of phosphophoryn. Our results indicate that phosphophoryn is a molecule of uniformly high mobility, thus belonging to a recently identified class of intrinsically disordered proteins that are characterized by sequences of low complexity and rich in polar and charged residues. The significance of our results is that phosphophoryn, because of its uniform nature has the potential to be replaced by biomimetic synthetic peptide analogs that together with amorphous calcium phosphate may lead to the development of novel, nontoxic, apatite-based dental restorative materials.

Amino Acid Motifs↗

Glycosaminoglycans in embryonic mouse teeth and the dissociated dental constituents.

The nature, amounts and distribution of glycosaminoglycans (GAG) before and during odontoblast terminal differentiation were studied. GAG have been isolated from intact mouse tooth germs and from dissociated dental epithelia and dental papillae after labeling with [3H]glucosamine or 35SO2-(4) asd precursor. The kinds and relative amounts of 3H-labeled GAG were analyzed by chromatography on a DEAE-cellulose column and cellulose thin-layer sheets. The amounts of individual GAG relative to total GAG were determined from the elution profiles, whereas their nature was identified by the selective removal of chromatographic peaks after enzymatic or chemical degradation. We found hyaluronate and probably a minute quantity of heparan sulfate in the dental epithelium, while hyaluronate, heparan sulfate, and chondroitin sulfate were the main types of GAG in the dental papilla. The chondroitin sulfate recovered was further fractionated by cellulose thin-layer chromatography into two isomers, namely chondroitin-2-sulfate (the major component) and chondroitin-6-sulfate. Changes in the elution profile from DEAE-cellulose chromatography of tooth GAG extracted from different developmental stages suggest that modifications of GAG occur during odontogenesis. Alcian blue staining localized large amounts of hyaluronate and sulfated GAG along the epithelio-mesenchymal junction. Tissue specificity and changing patterns of GAG were demonstrated during odontogenesis.

Animals↗

Acellular dental matrices promote functional differentiation of ameloblasts.

EDTA treatment of post-natal mouse molars made possible the isolation of cell-free dental matrices composed of basal lamina, predentin, dentin and enamel. Trypsin-isolated dental papillae and enamel organs from embryonic-mouse mandibular molars were combined with isolated matrices and cultured in vitro. In such recombinations, functional odontoblasts were never observed. On the other hand, competent preameloblasts in contact with the epithelial side of occlusal predentin overtly differentiated. Matrices treated with guanidine-EDTA or acetic acid were unable to promote the functional differentiation of ameloblasts. These data are discussed in terms of the epitheliomesenchymal interactions involved in odontogenesis.

Acetates↗

Dental cell interaction with extracellular-matrix constituents: type-I collagen and fibronectin.

It has been suggested that, during odontoblast differentiation, the extracellular matrix present at the epitheliomesenchymal junction modulates the activity of the cytoskeleton by means of membrane constituents (proteins, proteoglycans or gangliosides). To investigate this, we studied the interaction of iodinated fibronectin and type-I collagen with dissociated dental tissues and with membrane proteins prepared from these tissues. Isolated dental papillae and enamel organs were cultured for increasing periods of time in the presence of iodinated proteins. Fibronectin and type-I collagen were preferentially bound to dental papillae; however, after 6 h of incubation, fibronectin no longer interacted with the dental papillae, and the bound radioactivity was released. In the meantime, de novo synthesized fibronectin was deposited in the extracellular matrix of the dental papillae. Membrane proteins were prepared from isolated enamel organs and dental papillae. After sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis, these proteins were transferred to nitrocellulose by electroblotting and then incubated in the presence of either 125I-labelled fibronectin or 125I-labelled type-I collagen. Autoradiography confirmed the preferential interaction of fibronectin with the dental papilla. Fibronectin interacted with three high-molecular-weight proteins (Mr, 145,000, 154,000 and 185,000), which were not detected when membranes were prepared from enamel organs. Under the same conditions, type-I collagen did not interact with membrane proteins. The known interaction of type-I collagen with the plasma membrane of dental-papilla cells might be mediated either by another constituent of the extracellular matrix or by cell-surface-associated proteoglycans.

Animals↗

The histological effects of composite resin materials on the pulps of monkey teeth.

Pulpal response of three bis glycol methacrylate composite resins, Smile, Simulate and Experimental Composite no. 2 were tested on primary and permanent monkey teeth using zinc oxide eugenol (ZOE) and silicate as controls. All materials were placed in Class V cavity preparations in Rhesus monkey teeth and evaluated at 3 days and 5 and 8 weeks. The materials were randomly placed in anterior and posterior teeth utilizing 75 primary and 75 permanent teeth. Following perfusion the teeth were prepared by routine histological procedures. The 3 day response of the composite resins was moderate, characterized by disruption of the odontoblasts, vacuolization and mild inflammatory response underlying the cavity. At 5 weeks the formation of reparative dentin and a decrease in the inflammatory response was similar for all resins observed. At 8 weeks a slight increase in reparative dentin and continued decrease in inflammation was noted when compared to the 5 weeks responses. At all time intervals ZOE produced the least pulpal response while silicate produced the most severe response.

Animals↗

Histological study on the origin of dentinal immunoglobulins and the change in their localization during caries.

At the incipient stage of the caries process prior to destruction, immunoglobulins were first localized in the odontoblast layer in spite of absence of plasma cells in the pulp, and secondly located in the dentinal tubules of the pulpal side of dentin. When the tooth became involved, caries immunoglobulins were localized under the lesion. Bovine serum albumin, which was administered on tooth surfaces, was not observed in the non-decayed area of dentin. When injected into blood vessels, the protein was found in both decayed and unaffected dentin. It is suggested that, at the incipient stage of the caries process, dentinal immunoglobulins come from the pulp, although the possibility of salivary origin at a later stage is not excluded; and that immunoglobulin localization changes during the caries process.

Animals↗

Histological distribution of phosphophoryn in normal and pathological human dentins.

Dentin phosphophoryn is a highly phosphorylated protein which has a hydrophilic character but is not soluble in dilute acetic acid. A histochemical method was developed for staining this protein with Stains-all in situ utilizing those chemical properties. We have succeeded in detecting the presence of this protein in circumpulpal orthodentin of human permanent and deciduous teeth, but not in mantle dentin, secondary dentin and reparative dentin. Phosphophoryn staining was also absent in the dentin of dentinogenesis imperfecta (DI) Type II, a genetic disorder of dentin formation. From these results, it is suggested that phosphophoryn is synthesized and secreted only by physiologically-differentiated odontoblasts and that the mineralization processes of mantle, secondary, reparative and DI dentins may be different from that of circumpulpal orthodentin.

Child↗

Healing of pulp exposures: an ultrastructural study.

The fine structure of tissue changes during the first 14 days following pulp exposure and capping with a hard setting Ca(OH)2 cement has been studied. The early changes included hemorrhage and moderate inflammation which were largely resolved during the first week. During the second week differentiation of cells occurred at the wound surface. These cells had the characteristic features of odontoblasts and formed a predentin-like collagen matrix. The capping material was closely adapted to cellular structures at the wound surface or to the predentin-like matrix at all observation periods. Dentin fragments displaced into the pulp tissue during cavity preparation, acted as sites for pulp stone formation.

Animals↗

Early cellular events in an actinomycin D-created dentin niche in the rat incisor.

Administration of actinomycin D at a dose level of 0.375 microgram/g resulted in the selective disruption of developing odontoblasts at a critical stage of morphogenesis. A dentin niche was formed which was later repaired by cellular reparative dentin. The cellular changes which resulted in dentin niche formation were studied histologically and ultrastructurally in serial longitudinal and transverse sections from tissues obtained 10 h to 80 h following injection of the drug. Five stages were identified: initial destruction (10-20 h), rapid destruction (30-40 h), debris removal (50-60 h), proliferation (60-80 h) and matrix deposition (post 80 h). The cellular changes found in the dental papilla were considerably different from those found in inflammation, resolution and repair of fibrous connective tissue. These early stages were dominated by apoptosis and heterophagy, and after 80 h by disordered dentin matrix formation. The three-dimensional morphology of the defect was reconstructed from serial sections. The shape of the niche was the result of interference by actinomycin D in the patterns of proliferation and migration of the cells in the apical region of the rat incisor tooth.

Acid Phosphatase↗

Development of transplanted pulp tissue containing epithelial sheath into a tooth-like structure.

The aim of these studies was to find out whether intact neonatal pulp tissue containing residual epithelial cells can induce the development of a tooth-like structure in situ. First maxillary neonatal hamster molar pulps containing adhering undifferentiated epithelial cells were transplanted submucosally in the oral cavity of recipient mothers for periods ranging from 2-8 weeks and the tissues were then processed for light microscopy. Developing tooth-like structures containing mineralised tubular dentine, predentine and a vascularised pulp-like chamber lined with functional odontoblast-like cells were observed in the specimens within 2 weeks of transplantation. Enamel and root formation were not observed. These data indicate that neonatal dental pulp tissues containing epithelial cell remnants have the capacity to develop into tooth-like structures and that this could be the explanation for the development of tooth-like structures sometimes observed in infants after extraction of a natal tooth.

Amelogenesis↗