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At least 919 records · Page 51Linked to original sources
PubMed1992

Anti-peptide antibodies reactive with epitopic domains of porcine amelogenins at the C-terminus.

This was an immunological investigation of the processing of porcine amelogenins in situ. Rabbit and rat anti-peptide sera reacted specifically with the hydrophilic segment of the intact amelogenins at the C-terminus. The immunogens used were the synthetic peptides: (a) C13 composed of PATDKTKREEVDC and (b) C25 composed of MQSLLPDLPLEAWPATDKTKREEVD. These peptides correspond to the C-terminal 12- and 25-residue segments of porcine amelogenin, respectively. Cystine was introduced at the C-terminus of C12 for KLH-binding (C13). Western blot analysis disclosed that: (i) both rabbit and rat anti-C13 sera reacted selectively with the 25-kDa porcine amelogenin and three other minor components (27, 22 and 18 kDa); (ii) anti-C25 peptide sera, additionally, reacted with the 23-kDa amelogenins (a degradation derivative of the 25-kDa protein, lacking the 12-residue segment at the C-terminus) and as trace components, 20-, 16- and 14-kDa moieties. Importantly, all the proteins reactive with the anti-C13 serum were concentrated in the outer secretory enamel adjacent to the ameloblasts, decreasing significantly in the underlying inner secretory enamel. Immunohistochemical studies applying the anti-peptide sera to the developing tooth germs of a minipig also confirmed the localization of reactivity in the outer secretory region. Neither anti-peptide serum reacted with porcine non-amelogenins, serum proteins nor dentine matrix proteins at the dilutions tested. however, it was found that both the anti-C13 and C25 sera reacted with human keratin.(ABSTRACT TRUNCATED AT 250 WORDS)

Amelogenin↗
PubMedDate not supplied

Epithelial-mesenchymal interactions: effects of a dental biomatrix on odontoblasts.

The functional differentiation of odontoblasts requires specific interactions between these cells and the extracellular matrix. To further analyze these phenomena we studied the effects of a "dental papillae biomatrix" on isolated dental papillae cultured in vitro. The dental papillae biomatrix was extracted from EDTA-dissociated day-18 mouse dental papillae by homogenization, NaCl and enzymatic treatments, and deposited on Millipore filters. This biomatrix was studied by means of transmission electron microscopy and indirect immunofluorescence: it contained collagen fibrils, type IV collagen, fibronectin and laminin; cellular residues were also observed. The dental papillae were isolated by trypsin treatment of homologous tooth germs and cultured on uncoated (control) and coated filters. As shown by histological and cytological data, odontoblast-like cells never differentiated in control cultures. In presence of biomatrix and serum, polarized functional cells were observed. The functional state of these cells was enhanced by the addition of ascorbic acid to the culture media. Study of the incorporation of 3H-proline in cultured dental papillae and in macromolecules secreted into the culture media corroborated the morphological findings.

Animals↗
PubMed1978

Histochemical and electron probe analysis of secretory ameloblasts of developing rat molar teeth.

Calcium was not found in secretory ameloblasts and stratum intermedium cells when treated with OsO4-pyroantimonate or when surfaces prepared by fracturing fresh, rapidly frozen, developing molar tooth germs were subject to electron probe X-ray analysis. Pyroantimonate reaction product, considered to be calcium, was found in mitochondria of enamel organ cells which were first placed in a bath containing calcium and potassium. The plasma membrane was disrupted in cells ehich showed mitochondrial localization of reaction product. The results provide no data which indicates that enamel organ cells have a direct, active role in the movement of calcium into the enamel. Rather, it is suggested that the secretory enamel organ might serve as a selective barrier in regulating the initial mineralization of enamel.

Ameloblasts↗
PubMed1980

Epithelio-mesenchymal morphology in ameloblastic fibro-odontoma: a light and electron microscopic study.

Five cases of ameloblastic fibro-odontoma were examined by light and electron microscopy. The ultrastructure of the epithelio-mesenchymal interface was compared with the morphology of the normally developing tooth germ. It was noted that the ultrastructure of the tumors was identical with the normally developing odontogenic tissues up to but not including the differentiation of mesenchymal cells into tall columnar odontoblasts. The absence of odontoblasts in the ameloblastic fibro-odontomas results in an absence of tubular dentin in place of which a homogeneous collagen-containing material is synthesized. The failure of mesenchymal cells to acquire the morphology of tall columnar odontoblasts probably is due to the absence of the normally occurring induction of the mesenchyme by the odontogenic epithelium.

Ameloblasts↗
PubMed1984

Crystal growth in calcifying fronts during dentinogenesis.

Additional dentinogenesis in calcifying fronts of permanent tooth germs in kittens was examined using a transmission electron microscope. The irregular island-like structures were observed in calcifying fronts. These islands, increased in size, formed the coalescence. Pairs of fine crystals ran parallel in the marginal zone of these islands. At the inner part of the islands, long and wide crystals were abundant. Furthermore, a three-dimensional visualization of crystals revealed that dentin crystals were of needle-like shape. These findings suggest that pairs of fine, needle-like crystals first appear and additional calcification progresses due to fusions between fine crystals, and maturation in crystallization occurs along the long axis following these fusions.

Animals↗
PubMed1980

Ultrastructure of the human enamel organ. I. External enamel epithelium, stellate reticulum, and stratum intermedium.

The fine structure of external enamel epithelium, stellate reticulum and stratum intermedium in primary tooth germs (bell stage) from four human foetuses was investigated. Characteristically, the cells of the differentiated external enamel epithelium, stellate reticulum and stratum intermedium exhibit many free ribosomes, few rough endoplasmic reticulum cisterns, well-developed Golgi complexes, many coated and smooth vesicles, often in relation to the cell membranes, and many bundles of tonofilaments. The cells are connected by numerous desmosomes and gap junctions. A parallel differentiation of stratum intermedium - external enamel epithelium, and the ameloblast layer is demonstrated. The morphology of the cells of the three layers indicates that these have secretory, transport and supporting functions.

Cell Membrane↗
PubMed1988

Membrane-cytoskeleton interactions: inhibition of odontoblast differentiation by a monoclonal antibody directed against a membrane protein.

It is known that high-molecular-weight (HMW) membrane proteins mediate interactions with constituents of the extracellular matrix and/or with cytoskeletal elements. To study participation of HMW membrane proteins in odontoblast or ameloblast differentiation, an immunological approach has been adopted. Antibodies directed against membrane proteins (Mr, 110-190) from mouse embryos have been produced by the hybridoma technique. Supernatants of hybridoma cultures were screened for their ability to stain dental tissues and also tested for their biological activities on dental cells in primary culture or on developing tooth germs in organ culture. An IgM monoclonal antibody, MC16A16, directed against a 165-kDa antigen present in plasma membrane preparations, reacted strongly with the dental epithelium and weakly with the mesenchyme. MC16A16 also reacted with the cell surface of nonpermeabilized cultured dental cells and could detach epithelial cells cultured on glass, but not mesenchymal cells which maintained vinculin-containing focal contacts. This antibody, which affected the organization of dental-cell microfilaments in primary culture, also inhibited the polarization of odontoblasts, but not that of ameloblasts.

Animals↗
PubMed1982

Ultrastructural localization and gradient of activity of alkaline phosphatase activity during rodent odontogenesis.

The ultrastructural localization and gradient of activity of alkaline phosphatase were studied with respect to cell differentiation, matrix synthesis, and matrix mineralization in the incisor and molar teeth of 4-day-old Sprague-Dawley rats. The animals were perfused intracardially at room temperature with 2.5% glutaraldehyde in 0.1M sodium cacodylate (pH 7.4) with 3-4% sucrose. The jaws were dissected, immersion-fixed for 24 h, and the incisor and molar tooth germs removed. These were determined in 10% EDTA in NaOH (pH 7.4) with 7% sucrose. After reactivation of the enzyme with 0.1M MgCl in Tris-maleate buffer (pH 7.4) at 4 degrees C, the medium consisting of 6 ml 3% sodium beta-glycerophosphate, 4 ml 0.2M Tris-HCl buffer (pH 9.2), 3 ml 1.6% MgSO4, 12 ml 0.5% lead citrate (pH congruent to 12), and 2.1 g sucrose. The pH was adjusted to 9.2 with 0.2M HCl, the volume made up to 30 ml, and the solution centrifuged for 10 min at 5000 rpm. Control teeth were incubated in medium minus the substrate. Finally, the specimens were routinely post-fixed and embedded for sectioning and examination with a Philips 300 electron microscopy. A gradient of alkaline phosphatase activity was mapped along the developing teeth in the cells of the stratum intermedium, the proximal borders of the ameloblasts, the early dentine matrix, the predentine-dentine border, matrix vesicles, and the plasma membranes of odontoblasts and subodontoblast cells. The gradient of alkaline phosphatase activity was evident in the forming tooth from the cervical loop to the crown apex and was related to the cellular events, matrix synthesis, and matrix mineralization occurring during odontogenesis.

Alkaline Phosphatase↗
PubMed1985

Immunohistochemical studies with a monoclonal antibody on the distribution of phosphophoryn in predentin and dentin.

A monoclonal antibody was raised against phosphophoryn, a unique noncollagenous phosphoprotein in dentin. Mouse myeloma NS-I cells were fused with spleen cells obtained from BALB/c mice immunized with phosphophoryn from fetal calf tooth germs. Mice inoculated with the hybridoma produced ascites fluid containing the antibody and this reacted only with a band of phosphophoryn transblotted from polyacrylamide gel. Immunohistochemical studies with the antibody showed that phosphophoryn was present in odontoblasts, odontoblastic processes and dentin, but not in the matrix of predentin, and that the phosphophoryn content of the dentin layer was high at and around the predentin-dentin junction and gradually decreased toward the enamel layer. The area corresponding to mantle dentin was not stained with the antibody.

Animals↗
PubMed1992

The carboxy-terminal extension of the collagen binding domain of fibronectin mediates interaction with a 165 kDa membrane protein involved in odontoblast differentiation.

Terminal differentiation of the odontoblast is characterized by an elongation and a polarization of the cell. The change in the cell shape and the reorganization of the cytoplasm involve the microfilament system. An immunological approach has previously implicated a transmembrane interaction between fibronectin and vinculin in the control of odontoblast differentiation. A 165 kDa protein localized on the cell-surface of odontoblasts mediated this interaction. In order to define the nature of the interaction of the 165 kDa protein with fibronectin, peptides were prepared by proteolytic cleavage of fibronectin with alpha-chymotrypsin. The results indicate that the 165 kDa protein interacted with a 62 kDa peptide located towards the amino-terminal extremity of fibronectin, but not with a 47 kDa related fragment. Both these 62 kDa and 47 kDa peptides included the collagen-binding domain and were retarded on a heparin-Ultrogel column. Microsequences demonstrated that the 62 kDa and 47 kDa fragments had the same amino-terminal extremity and that the larger fragment was extended in the carboxy-terminal direction. This carboxy-terminal extension of the collagen binding domain of fibronectin is implicated in the interaction of this molecule with the 165 kDa protein. On the other hand, odontoblasts differentiated normally when tooth germs were cultured in the presence of GRGDS synthetic peptide, suggesting that RGD-dependent integrins were not involved in odontoblast differentiation. Staining of dental mesenchymal cells in primary culture and of differentiated odontoblasts in situ with antibodies directed against the beta 1-subunit of integrins confirmed previous observations and showed that although beta 1 integrins are involved in the attachment of cultured dental cells, they are not implicated in the process of odontoblast differentiation.

Actin Cytoskeleton↗
PubMed2003

[The expression of Sonic Hedgehog during late bell stage by Western Blot and in situ hybridization].

OBJECTIVE: To investigate the expression and biological effect of Shh during late bell stage by morphological and semi-quantitative analysis. METHODS: Tooth germs were selected from new born Bal b/c mouse (P1, P2, P3, P5, P7). Semi-quality of Shh was measured by Western Blot and the expression place and strength of Shh were observed by in situ hybridization. RESULTS: Shh was expressed in the ameloblast layer during late bell stage; the expression strength was high in secretive period and decreased with development; the active N-section was detectable before P3. CONCLUSION: Shh expresses specially in the ameloblast layer in late bell stage, and expression quality is related to the function of ameloblasts.

Ameloblasts↗
PubMed1981

Collagen type I trimer synthesis by cultured embryonic mouse molars.

Embryonic mouse tooth germs were cultured in vitro and the collagens type I, type III and type I trimer were purified and biochemically characterized. Collagen type I trimer has been identified by means of CM-cellulose chromatography, CNBr peptide analysis, pepsin resistance and molecular sieve chromatography. Already before the odontoblasts were functional, this molecule was found to be a constituent of the dental extracellular matrix. However, the synthesis of collagen type I trimer was considerably increased when odontoblasts polarized and became functional. the incorporation of 5-bromodeoxyuridine into the dental cells inhibited the polarization of odontoblasts as well as the amplification of collagen type I and type I trimer synthesis.

Animals↗
PubMed2005

Dentinal defects in Hyp mice not caused by hypophosphatemia alone.

The Hyp mouse is a murine homolog of human X-linked hypophosphatemic rickets and displays hypo-mineralization in bone and dentin due to a defect of the phosphate-regulating gene with homology to endopeptidase on the X chromosome (Phex) gene. It has long been considered that the bone and dentin defects in Hyp mice are caused by hypophosphatemia alone, however, several recent studies have indicated the possibility that intrinsic defects are present in Hyp mice osteoblasts. Further, we previously found a hyper-expression of osteocalcin (OC) mRNA in Hyp mouse odontoblasts and suggested the possibility of the presence of intrinsic defects. In the present study, we evaluated morphological features and OC mRNA expression levels in tooth germs of Nor mice with a normal phex gene and a low concentration of serum phosphate, and compared them to those in Hyp and wild-type mice. Nor mice exhibited low serum phosphate levels, however, did not show the characteristic features of dentin defects seen in Hyp mice, such as widened predentin and hyper-expression of OC mRNA. These results suggest that the hypo-mineralization of dentin in Hyp mice is not dependent on serum phosphate level, but rather is affected by intrinsic defects in odontoblasts.

Animals↗
PubMed1991

Extraction of 'nylon' teeth and associated abnormalities in Tanzanian children.

The aim of this study is to determine the prevalence of extraction of 'nylon' teeth and its long term adverse effects on the permanent dentition of Tanzanians. A total of 1890 subjects, aged between 3 and 5 years, and 1377 subjects aged between 12 and 19 years, in different parts of the country were examined. The prevalence of missing teeth in the primary dentition due to 'nylon' teeth practice ranged from 0% to 16.9%, with a weighted mean of 9.5%. Canines were the most involved teeth, accounting for 95% of all missing teeth. Mutilated permanent teeth, a result of traditional extraction of deciduous tooth germs, were observed only in Singida district in 8% of the examined subjects. 22.8% of these mutilations presented as malformations, while the remaining 77.2% presented as missing teeth. Most missing teeth and all the malformed teeth were seen in the mandible. The mandibular canines were the most affected, followed by the mandibular lateral incisors. It is suggested that 'nylon' teeth practice in Tanzania may be a recent invention rather than a traditional tribe related custom. The precariously high prevalence of 'nylon' teeth practice in Tanzania, and the people's perception of this malpractice, requires that particular attention be paid to this issue during oral health educational programmes in the affected communities.

Adolescent↗
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↗
PubMed1981

Autoradiographic demonstration of unscheduled DNA synthesis in oral tissues treated with chemical carcinogens in short-term organ culture.

A system in which oral tissues of inbred F344 adult rats and Syrian golden hamster embryos were used in combination with autoradiography was developed for measurement of unscheduled DNA synthesis (UDS). For this, oral mucosa, submandibular gland, tooth germ and mandible in short-term organ cultures were treated with 4-nitroquinoline 1-oxide or N-methyl-N-nitrosourea plus (methyl-3H)thymidine. Significant numbers of silver grains, indicating UDS, were detected over the nuclei of cells of all these tissues except rat salivary gland after treatment with carcinogens. This autoradiographic method is suitable for detection of UDS in oral tissues in conditions mimicking those in vivo. Results obtained in this study indicated a potential use of this system for studies on the mechanism of carcinogenesis at a cellular level comparable to in vivo carcinogenesis studies on oral tissues.

4-Nitroquinoline-1-oxide↗
PubMed1985

Electron microscopic demonstration of non-mineralized and hypomineralized areas in dentin and cementum by silver methenamine staining of collagen.

An electron microscopic study on silver methenamine staining of hard dental tissues was made on a material that comprised human permanent teeth and primary tooth germs from human and porcine fetuses. It was demonstrated that silver-stained material consisted of collagen fibrils. In predentin and precementum all collagen fibrils were stained, while collagen fibrils of dentin and cementum were unstained except for some fibrils of minor special areas such as Owen's contour lines, interglobular dentin, Tomes's granular layer, and in cementum small "interglobular-like" areas. It is concluded that silver methenamine visualizes collagen fibrils of hypo- and unmineralized areas in dental hard tissues and therefore may be used to demonstrate abnormal patterns of mineralization. Finally variations of silver methenamine stainability in relation to differences in material and methods were studied and discussed.

Animals↗
PubMed1980

Tunicamycin inhibits mouse tooth morphogenesis and odontoblast differentiation in vitro.

Tunicamycin (TM), an antibiotic that selectively inhibits dolichol-mediated protein glycosylation, inhibited morphogenesis and differentiation of odontoblasts in the molar tooth germ in vitro. These effects of TM are reversible and dose-dependent, and in advanced teeth the effect of TM was not complete unless the basement membrane was removed prior to culture. TM did not prevent secretion of predentin or enamel when added to the cultures after initiation of predentin secretion. TM dramatically inhibited protein glycosylation and the accumulation of labeled proteoglycans and glycoproteins in the basement membrane. Our previous studies indicated that odontoblast differentiation is triggered by an interaction between the basement membrane and mesenchymal cells. We suggest that TM inhibits odontoblast differentiation by causing alterations in the basement membrane which prevent the necessary cell-matrix interaction required for odontoblast differentiation.

Animals↗