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PubMed2004

Detection of dentin sialoprotein in rat periodontium.

Cloning and sequencing of the cDNA indicates that dentin sialophosphoprotein (DSPP) is a precursor of both dentin sialoprotein (DSP) and dentin phosphoprotein (DPP). Dentin sialophosphoprotein must be proteolytically processed to form these two extracellular matrix (ECM) proteins. Numerous studies led us to conclude that DSP (and DSPP) are exclusively expressed by odontoblasts and preameloblasts. However, recent observations suggest a wider distribution. To test this hypothesis, we conducted systematic studies on rat first molar during root formation with immunohistochemical techniques using specific anti-DSP polyclonal and monoclonal antibodies. We also performed in situ hybridization, using high-stringency RNA probes to detect DSP transcripts. Immunohistochemical studies demonstrated that DSP is not only localized in odontoblasts, dentin ECM and preameloblasts, but also in alveolar bone, cellular cementum, osteocytes, cementocytes, and their matrices. The results of in situ hybridization were consistent with those from immunohistochemistry, showing the expression of DSP transcripts in osteoblasts of alveolar bone, fibroblasts in periodontal ligament and cementoblasts in cellular cementum. Together, these observations suggest that DSP is involved in formation of the periodontium as well as tooth structures.

Alveolar Process↗
PubMed2005

Microhardness changes in dentine after neonatal capsaicin application.

AIM: To determine if desensitization of the nociceptive innervation in the dental pulp has an effect on odontoblast function in the rat. METHODOLOGY: Neonatal systemic application of capsaicin was used to selectively eliminate nociceptive innervation. 12 capsaicin-treated rats were intravitally perfused at 150 days of life with 4% formaldehyde and jaws were prepared for Vicker's microhardness (VMH) measurement. As a control, 12 rats were injected with vehicle on the 3rd day of life and intravital perfusion was carried out exactly as those used for the experimental group. Immunohistological labeling of CGRP was carried out in both groups to assure the efficiency of desensitization in the experimental group. The VMH was measured in the incisors of each animal for a quantitative analysis of dentine quality. RESULTS: Vicker's microhardness was significantly higher in the control rats compared with the capsaicin-treated rats (P < 0.001). CONCLUSIONS: Neonatal systemic application of capsaicin produces changes in the quality of dentine in the rat over time and therefore it is suggestive that selective elimination of the nociceptive innervation in pulpal tissue may effect odontoblast function.

Analgesics, Non-Narcotic↗
PubMed1977

Influence of different endodontic methods of treatment upon the process of apical closure of immature pulpless human teeth and the structure of the newly formed calcified tissue in apical opening.

A description is given of the pathohistological structure of the calcified tissue in the apical opening of four extracted immature pulpless human teeth treated by various endodontic methods. It is concluded that in the tooth treated by mortal extirpation there was neither histological healing nor any tendency for further apex formation. In both the tooth where vital extirpation was performed and in one of the teeth treated as apical periodontitis, the apices were formed from conglomerates of different calcified tissues, identical to the tissue which was formed under comparable conditions in experimental dogs. In the fourth human tooth also treated as apical periodontitis after treatment the apical opening closed with irregular dentine tissue. In this study consideration was given to the problem of the origin of the odontoblasts which co-exist with the hard destructive process present in some pulpless teeth. It is concluded that in the case presented in this study the irregular dentine was formed by the same odontoblasts which took part in the process of normal odontogenesis. This fact, should be respected in the course of endodontic treatment of immature pulpless teeth.

Animals↗
PubMed1993

Periodontal cell migration into the apical pulp during the repair process after pulpectomy in immature teeth: an autoradiographic study.

The migration of dental papilla cells into the periodontium during the process of root development may occur as part of the process involved in the formation of the periodontal tissues. The question posed is whether such cells under pathological conditions could retromigrate from periodontium into dental pulp and together with other apical pulp cells of immature teeth, take part in the production of additional dental tissue, e.g. 1) the tertiary/dentine under deep carious lesion where odontoblasts had been destroyed 2) the dentine bridge on an amputation wound and 3) calcified tissue which closes an apex during the apexification process in immature teeth. The migration of periodontal cells locally marked by H3 thymidine immediately after partial pulpectomy in immature dog's teeth was analysed at observation periods of 2, 24 and 50 h and also without H3 thymidine labelling of periodontal cells 8 weeks after pulpectomy. The marked cells were found in the early observation periods after pulpectomy just in the places where the hard tissues were formed in the later observation period of 8 weeks. They were found in large numbers just around the coagulated necrotic foci. The finding supports the assumption that firm necrotic masses are a very important stimulative factor in the reparation process in pulp and periodontium. The experiment also corroborated the existence of periodontal cell retromigration into apical dental papilla of immature teeth. Future research should assess the possible role of the pathological condition in the determination of undifferentiated odontogenic ectomesenchymal periodontal cells into odontoblasts.

Animals↗
PubMed1983

Neutral morphogenetic activity of epithelium in heterologous tissue recombinations.

To assess the existence of specific and nonspecific epithelial instructions for mesenchymal cell differentiation we compared homospecific and heterospecific mouse and quail tissue recombinations. In heterospecific recombinants between trypsin-dissociated mouse molar mesenchyme and quail epithelia neither odontoblasts nor chondrocytes differentiated. Cartilage appeared if the quail epithelium was contaminated with homologous limb mesenchyme and odontoblasts differentiated if the mouse dental epithelium was contaminated with dental papilla cells.

Animals↗
PubMed1978

Electron microscopy of cells displaced into the dentinal tubules due to dry cavity preparation.

Using dogs' teeth, cells displaced into the dentinal tubules due to dry cavity preparation were observed using electron microscopy. In order to obtain adequate fixation a new local-perfusion method was employed. Most of the cells which moved into the dentinal tubules were odontoblast nuclei and partly neutrophilic leukocytes and erythrocytes. Not only odontoblast nuclei but also cytoplasmic organelles, such as mitochondria, rough endoplasmic reticulum, free ribosomes, and lysosomes were displaced into the tubules. The weavings of dentinal fibers in the tubules were observed in some areas.

Animals↗
PubMed1992

Growth hormone regulates nucleolar organizer regions during odontogenesis in the rat.

Nucleolar organizers are major sites of ribosomal RNA synthesis and provide an index of transcriptional activity. In order to further define growth hormone actions on nucleolar organizer regions in tooth forming cells, hypophysectomized rats treated with growth hormone for 4 and 24 h, hypophysectomized and sham-operated animals were used. After demineralization and standard paraffin embedding, longitudinal sections of maxillary incisors were stained by a silver stain technique to reveal nucleolar organizer regions. The area of these regions per nucleus was measured using a modified microdensitometer. Analyses of variance of the resulting data showed that preameloblasts and preodontoblasts have greater silver stained nucleolar organizer region values than ameloblasts and odontoblasts. Hypophysectomy reduced and growth hormone partly restored the level of nucleolar organizer regions in preameloblasts and preodontoblasts, but not in mature ameloblasts or odontoblasts. In the case of the younger preameloblasts and preodontoblasts, the effect of growth hormone was seen within 4 h of growth hormone injection. In conclusion, rRNA synthesis, as revealed by the specific silver staining of nucleolar organizer regions in tooth forming cells, appears to be regulated by growth hormone over a relatively short time frame.

Ameloblasts↗
PubMed1978

Histochemical characterization of alkaline phosphatase in developing rat teeth and bone.

Alkaline phosphatase (EC 3.1.3.1.) in developing teeth and in bone has been studied. Prior to hard tissue function a rather high enzyme activity was noted in differentiating odontoblasts, stratum intermedium, and outer enamel epithelium. A lower activity was observed in the cells of the dental papilla and stellate reticulum. After the onset of hard tissue formation the alkaline phosphatase activity was generally increased. Enzyme activity was also found in the proximal part of tall, secretory ameloblasts. In the short postsecretory ameloblasts a high enzyme activity was noted. At the onset of dentin mineralization there was an increase in enzyme activity in the cells of the subodontoblastic layer. In bone the highest alkaline phosphatase activity was found in osteoblasts. A difference was noted between the alkaline phosphatase of hard and soft tissues by means of the addition of inhibitors to the incubation media. Within the hard tissues it was possible to distinguish between two alkaline phosphatases after pretreatment with heat (56 degrees C) or the addition of specific inhibitors (sodium metavandate, ortho-and pyrophosphate). An isoenzyme which was sensitive to these procedures was demonstrated in odontoblasts and in the pulpal connective tissue. Another alkaline phosphatase isoenzyme, which was resistant to pretreatment with heat or the addition of vanadate or phosphate, was demonstrated in the subodontoblastic cell layer, stratum intermedium and the outer cells of the reduced enamel epithelium.

Alkaline Phosphatase↗
PubMed1980

Autoradiographic investigation of dentin production in rat incisors after vincristine administration.

0.7 mg/kg VCR was administered intravenously to nine rats in three experimental groups. One hour after VCR injection the animals were injected with 3H-proline. The animals were sacrificed 1.5, 5 and 24 h after the VCR administration, and by comparison with nine control animals similarly injected with sodium chloride solution and 3H-proline, it was shown that VCR causes an early, intracellular retention of labeled proline in the odontoblasts, an abnormal intracellular distribution of labeled material and a generally decreased secretion of dentinal matrix from the odontoblasts in the maxillary incisors. The severity of the effects were shown to vary in different parts of the incisors.

Animals↗
PubMed1988

Production of monoclonal antibodies against murine dental papilla.

The mesenchymal component of the embryonic tooth, the dental papilla, shows unique cellular behavior. Only the dental papilla cells are capable of differentiating into odontoblasts. The dental papilla also directs the morphogenesis of the tooth. Our aim is to develop suitable markers for further studies on the molecular mechanisms behind the determination and differentiation of the dental mesenchymal cells. We have produced monoclonal antibodies against this embryonal cell population. Altogether 1114 enzymatically separated mesenchymes from dissected molar tooth germs of 17-day-old mouse embryos were fixed in paraformaldehyde (PFA) and sonicated in saline. A rat was immunized and hybridomas were produced by standard methods. The supernatants were screened by immunohistology, using both frozen sections and sections of PFA-fixed paraffin-embedded mouse molar teeth. Altogether, 19 wells produced antibodies reacting with dental tissues. One monoclonal antibody recognizes a 67,000 dalton intracellular antigen enriched in secretory odontoblasts and ameloblasts. The antigen is also found in osteoblasts and chondrocytes in the developing jaw, and in some cultured murine cells. The method described here appears to be successful for generating and screening monoclonal antibodies against the dental papilla.

Ameloblasts↗
PubMed1995

Effects of essential fatty acid deficiency on rat molar pulp cells.

In rats fed an essential fatty acid deficient (EFAD) diet, either during pregnancy (DN) or for 4 wk postnatally (ND), the cell density in the central part of the pulp increased about two- and threefold, respectively, of that in rats who had received a conventional diet containing sunflower oil. Cells were especially numerous around capillaries. The cell density was also increased twofold in the subodontoblastic layer in the outer part of the pulp, cells being smaller in ND compared with DN. In contrast, the odontoblasts were reduced in height, and the Höhl cells formed a thin layer in EFAD rats. This emphasizes some aspects of pulp specificity which reacted differently from odontoblasts. We suggest that the function of killer cells which normally destroy cells at the periphery of the pulp may be impaired by the diet, leading to cell accumulation.

Animals↗
PubMed1997

Effects of actinomycin D on developing hamster molar tooth germs in vitro.

The aim of this study was to evaluate the toxic effects of actinomycin D on the developing hamster tooth germ in organ culture. Hamster tooth germs during early secretory amelogenesis were exposed in vitro for 24 h to 10(-9) M-5 x 10(-5) M actinomycin D. Actinomycin D dose-dependently (> or = 10(-7) M) decreased the tooth germ dry weight but mineralization was affected only by doses > or = 10(-5) M. However, the uptakes of TCA-insoluble 32P and [3H]thymidine were significantly reduced dose-dependently from > or = 10(-8) M actinomycin D, indicating that the drug inhibits the synthesis of phosphate-containing macromolecules as well as DNA synthesis. Histologically, 10(-8) M actinomycin D was the lowest dose which was not toxic to any cell type in the developing tooth germ. At 10(-7) M actinomycin D, the most sensitive cells were the proliferating pre-odontoblasts followed by pre-ameloblasts; the mature secretory ameloblasts and odontoblasts appeared unaffected. Higher doses resulted in increased cytotoxicity to the secretory cells and, eventually, total degeneration of most cells. The data suggest that children treated for cancer during tooth development using anti-chemotherapy cocktails containing actinomycin D (serum levels > 10(-7) M) may develop defects later on in the mature dentition as a direct consequence of the toxicity of the drug to the tooth organ.

Ameloblasts↗
PubMed1997

Cell proliferation in developing human dental pulp. A combined flow cytometric and immunohistochemical study.

Information concerning cell proliferation and differentiation in dental pulp may be important to understand tooth response to exogenous stimuli. Since few data concerning human dental pulp are available, we have investigated the growth fraction and the localization of proliferating cells in pulp tissue of third molars of young adult human males and females, using flow cytometry and immunohistochemistry. Flow cytometric analysis demonstrates a low proliferative activity of pulp tissue that appears to be confined to radicular pulp, as revealed by immunohistochemical detection of proliferating cells. No polyploid or aneuploid cell populations could be identified, and G2-blocked cells, if any, represented a negligible cell population. Odontoblasts, cells of the sub-odontoblastic layer, and cells of coronal pulp were found to be not proliferating under normal conditions. These data provide the basis for future investigations on proliferative activity and regenerative potentiality of human pulp cells in experimental and clinical situations.

Adolescent↗
PubMed1997

Dentinal and pulpal uptake of mercury from lined and unlined amalgam restorations in minipigs.

The aim of the present investigation was to examine dentin and pulp in amalgam-restored teeth in minipigs with respect to presence of mercury, and to evaluate whether lining of restored cavities with a resin-modified glass ionomer cement had any effect on the penetration of mercury. Class I and V cavities were cut in 49 posterior and canine teeth of 4 minipigs. 35 cavities were restored with amalgam and 14 with composite material. 26 of the amalgam cavities were lined with a resin-modified glass ionomer cement. Following observation periods of 17-28 months, the pigs were killed and the jaws sectioned. After decalcification, the teeth were processed for light or electron microscopy examinations. Mercury was visualized by autometallography. Silver-enhanced mercury was found in all teeth with amalgam, whereas teeth with composites were devoid of mercury. Mercury could be traced in the odontoblast processes, in the body of odontoblasts, and on rare occasions in the nerve tissue of the pulp from lined and from unlined amalgam restorations. The present study thus demonstrates transport of mercury through dentinal tubules to the pulp, and that lining of cavities with a resin-modified glass ionomer cement does not prevent penetration of mercury.

Animals↗
PubMed1998

Transforming growth factor-beta1 (TGF-beta1) in dentine matrix. Ligand activation and receptor expression.

Transforming Growth Factor-beta (TGF-beta) and other members of this family of growth factors have been implicated in tooth development and dental tissue repair. This study aimed to investigate regulatory factors for TGF-beta1 in rabbit incisor dentine matrix and the expression of receptors for TGF-beta in dental tissues better to understand the control of its biological activity. Approximately half of the TGF-beta1 in dentine matrix was present in active form. TGF-beta1 was found in association with latency associated peptide (LAP), betaglycan and decorin in an isolated dentine matrix preparation. Immunohistochemistry showed strong staining for TGF-beta type I and II receptors in odontoblasts with more variable and weaker staining of other pulpal cells. Association of TGF-beta1 with betaglycan, decorin and LAP may regulate the availability and biological activity of this growth factor and influence its presentation to the TGF-beta type I and II receptors on odontoblasts. During dental tissue repair, such control processes will be important in regulating the biological effects of TGF-beta1 on cells of the dentine-pulp complex.

Animals↗
PubMed1998

The phosphophoryn gene family: identical domain structures at the carboxyl end.

Phosphophoryns (PPs), a family of aspartic acid and phosphoserine rich dentin proteins, are considered to be archetypal regulators of extracellular matrix biomineralization. Their very unusual composition, extensive phosphorylation, and tissue-specific heterogeneity have made their characterization a difficult task. We have recently cloned several rat incisor PP genes from our odontoblast cDNA library. The first clone, designated as DMP2, was 2.4 kb long, with an open reading frame of approximately 700 bp and an untranslated region of approximately 1.7 kb. Northern blot analysis of odontoblast mRNA showed a single message between 5 and 6 kb. When a 5' RACE technique was used to obtain the full length clone, a second approximately 2 kb DNA fragment (DMP3) was also found. Cloning and sequencing of DMP3 showed that the 3' end was highly homologous to the 3' end of DMP2, but the 5' end of this clone had 100% homology to dentin sialoprotein (DSP). DMP3 is not a typical Asp-rich phosphophoryn, but DMP2 contains a domain N-terminal to the common region which has the hallmark Asp- and Ser-rich composition of the phosphophoryns. Both DMP2 and DMP3 are closely localized on mouse chromosome 5q21, corresponding to human chromosome 4q21. The expression of these two genes is regulated differently. Thus, there may be a family of phosphophoryn-related genes coding for a common C-terminal peptide sequence domain, but involving different N-terminal domains with different functions.

Amino Acid Sequence↗
PubMed1998

Changes in the cytoskeleton of cells within the periodontal ligament and dental pulp of the rat first molar tooth during ageing.

The periodontal ligament and dental pulp connective tissues are foetal-like, and their process of ageing may therefore differ from other tissues of mesenchymal origin. Several studies have already highlighted the lack of age changes in the extracellular matrix (ECM), but more needs to be known about cellular changes. For this study, tissues from the first molar teeth of Wistar rats aged 12 wk and 104 wk were compared by immunolocalisation of cytoskeletal components. Tissues from the first molar were immunostained, employing a panel of 16 monoclonal antibodies against cytokeratins, vimentin, F-actin and tubulin. Within the pulp, labelling for vimentin in both odontoblasts and pulpal fibroblasts and F-actin in the cell processes of odontoblasts was detected at both ages but with marked reduction in labelling in the older tissue. Within the periodontal ligament, vimentin labelling was weaker in the aged tissue, especially nearer the cementum. More significantly, the fibroblasts of the aged periodontal ligament expressed cytokeratin 19. In contrast to reports of little age change to the ECM, the cells of the pulp, and particularly the periodontal ligament, show marked changes to their cytoskeletal components.

Aging↗
PubMed1989

Dentin induction by implants of autolyzed antigen-extracted allogeneic dentin on amputated pulps of dogs.

Implantation of autolyzed antigen-extracted allogeneic (AAA) dentin matrix gelatin powder caused homeoinduction on amputated dental pulps. This event began with migration of spindle-shaped mesenchymal cells into the cavity on the amputated pulp. This was followed by proliferation of undifferentiated large cells concomitantly with vascular invasion and attachment of spindle-shaped cells and large cells to the AAA dentin. The adhering cells differentiated into osteodentinoblasts and/or preodontoblasts to form osteodentin matrix and predentin respectively. Osteodentinocytes and odontoblasts, then, formed osteodentin and tubular dentin. The healing of pulp dressed with inactivated AAA dentin using guanidine HCl was delayed. These results suggested that AAA dentin matrix powder may have chemotactic and mitogenic activity for undifferentiated mesenchymal cells and may provide a suitable scaffolding for fixation of these cells. Compartments of microenvironment formed by AAA dentin and the enveloping hard substances may play some role in differentiation into odontoblasts.

Animals↗