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PubMed2004

Two related low molecular mass polypeptide isoforms of amelogenin have distinct activities in mouse tooth germ differentiation in vitro.

UNLABELLED: Embryonic mouse tooth germs were cultured in vitro in the presence of two related amelogenin isoforms to determine their effects on tooth development. Our results show that these individual proteins have specific but quite different effects on epithelial-derived ameloblasts versus mesenchymal-derived odontoblasts. INTRODUCTION: Amelogenins, the main protein components of enamel matrix, have been shown to have signaling activity. Amelogenin isoforms differing only by the presence or exclusion of exon 4, designated [A+4] (composed of exons 2, 3, 4, 5, 6d, and 7) and [A-4] (composed of exons 2, 3, 5, 6d, and 7), showed similar, but different, effects both in vitro and in vivo on postnatal teeth. MATERIALS AND METHODS: Lower first molar tooth germs of E15/16 CD1 mice were microdissected and cultured in vitro in a semisolid media containing either 20% FBS, 2% FBS, or 2% FBS with either 1.5 nM [A+4], [A-4], or both for 6 days. Tooth germs were analyzed by H&E staining and immunohistochemistry for collagen I, dentin matrix protein 2, and DAPI nuclear staining. RESULTS: Teeth cultured in media containing 20% FBS showed normal development with polarized ameloblasts, and odontoblasts producing dentin matrix, and DMP2 expression in odontoblasts and pre-ameloblasts. Culture in 2% FBS media resulted in no ameloblast polarization and modest odontoblast differentiation with scant dentin matrix. Tooth germs cultured with [A+4] in 2% FBS media had well-polarized odontoblasts with robust dentin production and concomitant ameloblast polarization. DMP2 expression was equal to or greater than seen in the 20% FBS culture condition. In cultures with [A-4] in 2% FBS media, odontoblast polarization and dentin production was reduced compared with [A+4]. However, the pre-ameloblast layer was disorganized, with no ameloblast polarization occurring along the dentin surface. DMP2 expression was reduced in the odontoblasts compared with the 20% FBS and [A+4] conditions and was almost completely abrogated in the pre-ameloblasts. CONCLUSION: These data show different signaling activities of these closely related amelogenin isoforms on tooth development. Here we make the novel observation that [A-4] has an inhibitory effect on ameloblast development, whereas [A+4] strongly stimulates odontoblast development. We show for the first time that specific amelogenin isoforms have effects on embryonic tooth development in vitro and also hypothesize that DMP2 may play a role in the terminal differentiation of both ameloblasts and odontoblasts.

Amelogenin↗
PubMed1988

Ultrastructure of odontogenic cells during enameloid matrix synthesis in tooth buds from an elasmobranch, Raja erinacae.

The ultrastructure of the inner dental epithelial cells (IDE) and odontoblasts in elasmobranch (Raja erinacae) tooth buds was investigated by transmission electron microscopy to determine what contribution each cell type makes to the forming enameloid matrix. Row II, early stage, IDE cells contained few organelles associated with protein synthesis, whereas preodontoblasts appeared competent to initiate extracellular matrix production. Row III IDE cells are also devoid of organelles related to secretory protein synthesis, although these IDE cells accumulated large pools of intracellular glycogen. The glycogen appeared to be packaged into vesicles and exocytosed into the lateral extracellular space toward the forming enameloid matrix. Row III odontoblasts had a morphology consistent with an active protein secretory cell. No procollagen granules were present within the odontoblasts, however, nor were many collagen fibers observed in the enameloid matrix. Instead, non-collagenous "giant" fibers having 17.5-nm periodic cross striations were associated with the invaginations of odontoblast cell processes. Giant fibers, which spanned a clear zone adjacent to the odontoblasts, terminated within the enameloid matrix. Smaller 25-nm-wide "unit" fibers emanated from the giant fiber tips to form the bulk of the enameloid matrix. The clear zone, which separated the odontoblasts from the enameloid matrix at early stages, diminished in size at later stages until the odontoblast processes were completely embedded in the enameloid matrix. Nascent enameloid crystallites were observed only after a layer of unmineralized predentin was deposited beneath fully formed enameloid matrix. The results suggest that the major constituent of the enameloid matrix in skates is a non-collagenous protein derived from the odontoblasts. The inner dental epithelial cells appear to contribute large quantities of carbohydrates to the forming enameloid matrix.

Animals↗
PubMed1991

Interodontoblastic collagen (von Korff fibers) and circumpulpal dentin formation: an ultrathin serial section study in the cat.

The collagenous fibers of von Korff pass from the dentin matrix between the odontoblasts into the dental pulp. Although collagen fibrils are known to be present between odontoblasts, the existence of von Korff fibers has remained controversial. This may be because their continuity between the dentin matrix and the pulp has not been demonstrated ultrastructurally. In this study we have examined the odontoblast layer in the middle to apical regions of perfusion-fixed permanent canine teeth of cats by using transmission electron microscopy. Ultrathin sections of demineralized specimens revealed frequent bundles of collagen fibrils 1) entering the odontoblast layer from the predentin, 2) present between odontoblast cell bodies, and 3) passing from between the odontoblasts into the pulp. The question of continuity of these bundles from the predentin, across the odontoblast layer into the pulp was examined in ultrathin serial sections. Unbroken continuity of a collagen bundle from the predentin between the odontoblasts into the pulp was established in a reconstruction of one series of 22 serial sections and was very strongly suggested by a number of other series in which the numbers of available sections restricted their full visibility. This investigation has shown, therefore, that classical von Korff fibers are present and that these fibers are present in fully erupted teeth with closed apices, i.e., at a time when secondary circumpulpal dentinogenesis is in progress. The findings call for a reexamination of the question of von Korff fibers during mantle dentinogenesis and primary circumpulpal dentinogenesis. Resolution of their existence at the earlier stages of dentinogenesis should be possible by using the ultrathin serial-sectioning technique.

Animals↗
PubMed2005

PTHrP and PTH/PTHrP receptor 1 expression in odontogenic cells of normal and HHM model rat incisors.

Parathyroid hormone related peptide (PTHrP) was discovered as a causative factor of humoral hypercalcemia of malignancy (HHM). We examined PTHrP and its receptor (PTHR1) expression patterns in odontogenic cells in normal and HHM model rat incisors. Nontreated nude rats serving as the normal control and HHM model rats produced by implantation of PTHrP-expressing tumor (LC-6) cells were prepared. HHM rats fractured its incisor, and histopathologically, restrict population of odontoblasts showed findings classified as "shortening of high columnar odontoblasts" and "dentin niche." The incisors were immunostained against PTHrP and PTHR1. In normal rats, PTHrP and PTHR1 colocalized in ameloblasts, cementoblasts, and odontoblastic cells from mesenchymal cells to columnar odontoblasts. In high columnar odontoblasts, PTHrP solely expressed. In the HHM animals, although the expression patterns were identical to those of the normal rats in normal area, the shortened high columnar odontoblasts maintained PTHR1 expression and dentin niche comprising odontoblastic cells expressed both proteins. In the HHM model, the protein expression patterns changed in the odontoblastic cells with histological anomalies, and thus direct relations between the anomalies and PTHrP/PTHR1 axis are suggested.

Animals↗
PubMedDate not supplied

Electron microscope comparison of freeze-substitution and conventional chemical fixation of undecalificied human dentin.

Within the stated limits of cell fine structure preservation and within the observed anatomical limits specified in the Results and Discussion sections, a freeze-substitution method using an ethylene glycol-Hank's solution eutectic mixture with a glutaraldehyde additive can be used to effectively prepare undecalcified human dentin for electron microscopy. The ultrastructual appearance of the odontoblast cell body and the odontoblastic process subjected to freeze-substitution differs from that seen with conventional chemical fixation. Artifacts produced by freeze-substitution differ in appearance and frequency of occurrence from those produced by glutaraldehyde-osmium tetroxide sequential double fixation. The cellular component of dentin shows greater structural preservation of protein when it is subjected to freeze-substitution that when it is prepared by conventional chemical fixation. The absence of ice crystal defects in the odontoblastic process in calcified dentin and the presence of ice crystal defects in the odontoblast cell body suggest that intracellular water in the odontoblastic process in the calcified dentin may exist in a more highly structured state than intracellular water in the odontoblast cell body. If intracellular water exists in a more highly structured state in the odontoblastic process of the calcified dentin than in the cell body, the ratio of protein molecules to cytoplasmic volume may be greater in the odontoblastic process than in the cell body. After glutaraldehyde-osmium tetroxide sequential double fixation, the use of graded alcohol dehydration obtained cell fine structure preservation and artifact control superior to that obtained by use of ethylene glycol cryodehydration. Further refinements of the freeze-substitution technique, as it applies to the preparation of undecalcified human dentin, are necessary to increase the amount of cellular preservation, to decrease the number of ice crystal artifacts, and to improve the overall quality of cell fine structure preservation.

Adolescent↗
PubMed1992

Developmental expression of a 53 KD dentin sialoprotein in rat tooth organs.

Rat dentin contains a major sialic acid-rich glycoprotein, DSP, with an overall composition similar to that of bone sialoproteins but whose biological role in dentinogenesis is unknown. Using polyclonal affinity-purified antibodies to rat DSP and four immunohistochemical methods of detection, we studied the cell and tissue localization of DSP and the time course of its appearance during odontoblast differentiation. DSP first appeared within young odontoblasts concomitant with early secretion of pre-dentin matrix and before the onset of mineralization but was absent in pre-odontoblasts. DSP immunostaining also localized within secretory odontoblasts and was intense in odontoblastic processes. Early pre-dentin stained positive for DSP, in contrast to more mature pre-dentin, where immunoreactivity was less intense and more restricted to odontoblastic processes. In the zone of mineralized dentin matrix, a moderate and uniform staining pattern was evident. Intense immunostaining was also seen within the cells and matrix of dental pulp during dentinogenesis. Other cells and tissues within the tooth organ and those surrounding it were non-reactive. These findings suggest that DSP is developmentally expressed in cells of the odontoblastic lineage and may be a biochemical marker of odontoblastic activity.

Aging↗
PubMed1993

Bovine bone morphogenetic protein-induced dentinogenesis.

Differentiation of odontoblasts is important for dentin formation in tooth germs and mature teeth. Although previous reports have indicated that there may be a kind of inductive agent that could induce mesenchymal cells in dental pulps to differentiate into odontoblasts, and secrete dentin matrix, the primary inductive factor of odontoblasts has not been found. Bone morphogenetic protein (BMP), which induces the formation of cartilage and bone when implanted in muscle tissue, is found in dentin matrix. The relationship between the differentiation of odontoblasts and BMP was observed by means of immunohistochemical staining with monoclonal antibody (MAb) against BMP in dental pulp tissue and cell culture; [3H]thymidine incorporation; and measurement of alkaline phosphatase activity. The conclusions are: (1) BMP exists in odontoblasts, ameloblasts, and dentin matrix (the positive reaction in ameloblasts appeared earlier and remained stronger); (2) BMP promotes incorporation of [3H]thymidine and increases the activity of alkaline phosphatase in cultured dental pulp cells; (3) BMP-induced dental pulp cells in dental pulp tissue cultures differentiate from mesenchymal to odontoblast-like cells; and (4) BMP induces formation of osteodentin and tubular dentin when used as a dental capping agent of dogs' teeth. Bone morphogenetic protein plays an important role in differentiation of odontoblasts and might be one of the inductive agents of odontoblasts. Further investigations of BMP as a biologic dental capping agent are warranted.

Animals↗
PubMed1992

Immuno-electron-microscopic localization of laminin and collagen type IV in normal and denervated tooth pulp of the cat.

The distribution of laminin-like immunoreactivity in adult normal and denervated cat mandibular tooth pulps was studied by the use of fluorescence microscopy and pre-embedding immunogold electron microscopy. Immunoreactivity to collagen IV was also assessed in order to distinguish basement membranes. In normal pulps, light-microscope laminin-like immunoreactivity was strong along blood vessels and Schwann cell sheaths, and a faint immunoreactivity was seen also in the odontoblast layer. Electron microscopy confirmed the laminin-like immunoreactivity of endothelial and Schwann cell basement membranes at all pulpal levels. In the odontoblast layer and the predentine, nerve-like structures lacking basement membranes but possessing strong membrane laminin-like immunoreactivity were encountered. In addition, a clear-cut laminin-like immunoreactivity of plasma membranes of the somata and processes of odontoblasts was seen. Observations on denervated pulps as well as pulps in which nerve regeneration had taken place did not reveal any changes in the pattern of laminin-immunoreactivity in basement membranes or odontoblasts. Distribution of collagen IV-like immunoreactivity was very similar to laminin-like immunoreactivity in basement membranes of blood vessels and Schwann cells, and appeared unaffected by denervation. The odontoblasts and nerve-like profiles in the odontoblast layer were devoid of collagen IV-like immunoreactivity. We propose that odontoblast-associated laminin could be of significance as guidance for regenerating terminal pulpal nerve fibers to appropriate targets.

Animals↗
PubMed2006

Pulpal responses to cavity preparation in aged rat molars.

The dentin-pulp complex is capable of repair after tooth injuries including dental procedures. However, few data are available concerning aged changes in pulpal reactions to such injuries. The present study aimed to clarify the capability of defense in aged pulp by investigating the responses of odontoblasts and cells positive for class II major histocompatibility complex (MHC) to cavity preparation in aged rat molars (300-360 days) and by comparing the results with those in young adult rats (100 days). In untreated control teeth, immunoreactivity for intense heat-shock protein (HSP)-25 and nestin was found in odontoblasts, whereas class-II-MHC-positive cells were densely distributed in the periphery of the pulp. Cavity preparation caused two types of pulpal reactions based on the different extent of damage in the aged rats. In the case of severe damage, destruction of the odontoblast layer was conspicuous at the affected site. By 12 h after cavity preparation, numerous class-II-MHC-positive cells appeared along the pulp-dentin border but subsequently disappeared together with HSP-25-immunopositive cells, and finally newly differentiated odontoblast-like cells took the place of the degenerated odontoblasts and acquired immunoreactivity for HSP-25 and nestin by postoperative day 3. In the case of mild damage, no remarkable changes occurred in odontoblasts after operation, and some survived through the experimental stages. These findings indicate that aged pulp tissue still possesses a defense capacity, and that a variety of reactions can occur depending on the difference in the status of dentinal tubules and/or odontoblast processes in individuals.

Aging↗
PubMed2003

Gene expression analysis in cells of the dentine-pulp complex in healthy and carious teeth.

Knowledge of the molecular events that occur in carious disease has so far been constrained due to difficulties in obtaining sufficient quantities of the dental tissues and cells involved. Our histological findings indicate that a pulp-odontoblast cellular complex can be obtained from carious and healthy human teeth when exposed to low-temperatures prior to pulpal extirpation and from rodent teeth processed at room-temperature. In contrast, pulpal tissue extracted from room-temperature processed human teeth and low-temperature processed rodent teeth resulted in the odontoblast layer remaining attached to the pulp chamber. Semi-quantitative RT-PCR (sq-RT-PCR) analysis confirmed that markers previously shown to be preferentially expressed in odontoblasts, namely dentin sialophosphoprotein (DSPP) and Nestin, amplified more readily from the extracted pulp-odontoblast complex, as compared to pulpal tissue alone, in both human and rodent samples. Subsequent gene expression analysis of collagen-1alpha and collagen-3alpha indicated levels were significantly higher in carious pulpal tissue. In addition, analysis characterising the expression of members of the transforming growth factor and bone morphogenic protein families and their receptors indicated in general, that these genes were expressed by healthy odontoblasts and up-regulated in both pulpal cells and odontoblasts in response to carious injury. Use of this temperature-sensitive dental tissue preparation procedure allows detection of differential gene expression in odontoblasts and other pulpal cells in healthy and carious tissue.

Adult↗
PubMed1998

Histogenesis of the chequered pattern of ivory of the African elephant (Loxodonta africana).

This study aimed to propose a hypothesis on the events which lead to the development of the characteristic chequered pattern of elephant ivory. Twenty fragments of ivory and six elephant tusks were obtained through the National Parks Board of South Africa. Polished surfaces were prepared in sagittal and longitudinal planes and the characteristics of the distinctive chequered pattern described. Light- and electron-microscopical techniques and image analyses were employed to determine the morphological basis of the pattern and to describe the spatial distribution, density and morphology of the dentinal tubules. These investigations showed that the distinctive pattern was the result of the sinusoidal, centripetal course followed by dentinal tubules. The apical, slanted part of the sinusoidal curve is the result of the centripetally moving odontoblast, which, during formation of ivory, progresses towards the centre of the tusk on a decreasing circumference. It is suggested that this leads to cell crowding, increased pressure between odontoblasts and subsequent apical movement of their cell bodies, cell degeneration and fusion. Odontoblastic degeneration and fusion probably relieve the pressure between the crowded odontoblasts by reducing their numbers and the remaining odontoblasts now orientate their centripetal course towards the tip of the tusk, thereby forming the anterior-directed part of the sinusoidal path of the tubule. As odontoblasts progress centripetally the diameter of the pulpal cavity decreases further and the processes of apical movement, fusion and degeneration of odontoblasts are repeated. This occurs until the pulpal cavity is obliterated.

Anatomy, Cross-Sectional↗
PubMed2001

Molecular aspects of tooth pathogenesis and repair: in vivo and in vitro models.

Several growth factors and extracellular matrix molecules, which are expressed during embryonic tooth development, are re-expressed in dental tissues under pathological conditions. Pathological conditions such as caries lesions and dental injuries are often lethal to the odontoblasts, which are then replaced by other pulp cells. These cells are able to differentiate into odontoblast-like cells and produce a reparative dentin. Here we demonstrate the in vivo distribution of several molecules in human permanent teeth under normal and pathological conditions. The intermediate filament protein nestin, which is a marker of young odontoblasts, is absent from old permanent teeth. Similarly, the Notch protein, which is involved in cell fate specification and is localized in the sub-odontoblastic cell layer during odontogenesis, is not detected in adult dental tissues. In carious and injured teeth, nestin is expressed in a selective manner in odontoblasts surrounding the injury site, while Notch is expressed in the sub-odontoblastic layer of cells. We reproduced this physiological event in an in vitro culture system. Pulp cells cultured in the presence of beta-glycerophosphate formed mineralization nodules. As odontoblasts, pulp cells contributing to the nodule formation express type I collagen, osteonectin, dentin sialophosphoprotein, and nestin. In this in vitro assay system, nestin is up-regulated after local application of Bone Morphogenetic Protein 2 and 4. Fourier transform infrared microspectroscopy showed that both the organic and the mineral compositions of the nodules have the characteristics of human dentin and differ from those of enamel and bone. These findings show that both the molecular and the mineral characteristics of the human dentin matrix are respected in the in vitro culture conditions.

Adult↗
PubMed2001

Induction and regulation of crown dentinogenesis: embryonic events as a template for dental tissue repair?

Close regulation of odontoblast differentiation and subsequent secretory activity is critical for dentinogenesis during both embryogenesis and tissue repair. Some dental papilla cells achieve commitment and specific competence, allowing them to respond to epithelially derived inductive signals during the process of odontoblast differentiation. Temporo-spatial regulation of odontoblast differentiation is dependent on matrix-mediated interactions involving the basement membrane (BM). Experimental studies have highlighted the possible roles of growth factors in these processes. Regulation of functional activity of odontoblasts allows for both ordered secretion of the primary dentin matrix and maintenance of vitality and down-regulation of secretory activity throughout secondary dentinogenesis. After injury to the mature tooth, the fate of the odontoblast can vary according to the intensity of the injury. Milder injury can result in up-regulation of functional activity leading to focal secretion of a reactionary dentin matrix, while greater injury can lead to odontoblast cell death. Induction of differentiation of a new generation of odontoblast-like cells can then lead to reparative dentinogenesis. Many similarities exist between development and repair, including matrix-mediation of the cellular processes and the apparent involvement of growth factors as signaling molecules despite the absence of epithelium during repair. While some of the molecular mediators appear to be common to these processes, the close regulation of primary dentinogenesis may be less ordered during tertiary dentinogenic responses.

Basement Membrane↗
PubMed1995

Cytoskeleton of the mesenchymal cells of the rat dental papilla and dental pulp.

This study describes the immunolocalization of actin, cytokeratins and vimentin during differentiation of the dental papilla in the rat. Incisors and first molars were sectioned from mandibles of Wistar rats from embryonic day (E)-14 to (E)-21 and weeks 1, 2, 3, 12 and 104 after birth, fixed in 90% alcohol, decalcified in EDTA, infiltrated with 5% sucrose, frozen in dry ice, and cryosectioned at 10 microns. The sections were immunolabelled using indirect immunofluorescence with a panel of monoclonal antibodies and FITC-phalloidin for F-actin localization. F-actin was present in follicular mesenchyme and odontoblast processes. Vimentin labelled dental papilla fibroblasts, differentiating, functional (secretory) and aged odontoblasts. Vimentin was uniformly localized in the cytoplasm of pre-odontoblasts but was redistributed to the apical pole of these cells during polarization. Of the cytokeratins, only cytokeratin 19 was found in differentiating odontoblasts. It was not present in dental papilla fibroblasts, functional or aged odontoblasts. These results suggest that actin and the redistribution of vimentin may be involved in odontoblast differentiation and odontoblast process formation/support and that these events may be preceded by the expression of cytokeratin 19.

Actins↗
PubMed1982

Epithelial-derived basal lamina regulation of mesenchymal cell differentiation.

The mechanisms by which epithelial-mesenchymal interactions result in differentiation are not known. A number of recombinations between vertebrate tissues associated with epidermal organs (e.g. skin, feather, mammary gland, salivary gland, tooth organ) indicate that regional mesenchymal specificity is instructive for determination and differentiation of epithelial phenotypes. In epidermal organs within which mesenchyme becomes determined and differentiates into a unique phenotype, such as during tooth organogenesis and odontoblast differentiation. Does the epithelial-derived basal lamina regulate mesenchymal differentiation into odontoblasts and the expression of dentine extracellular matrix? Experiments were designed to test the hypothesis that murine or avian epithelial-derived basal lamina possess information which is instructive for determined dental mesenchyme to differentiate into odontoblasts. The strategy was to examine homologous and heterologous tissue recombinants between Theiler stage 25 C57BL/6 molar tooth organs and Hamburger-Hamilton equivalent stage 22-26 Japanese Pharoah quail mandibular processes. Trypsin-dissociated molar epithelium and mesenchyme, reconstituted, secreted a basal lamina within 8 hours and mesenchyme differentiated into odontoblasts and formed dentine matrix within 3 days. Isolated trypsin-dissociated mesenchyme did not differentiate in vitro, whereas heterologous recombinants between odontogenic mesenchyma and quail epithelia resulted in odontoblasts and dentine production. Mouse tooth or quail mandibular epithelia served to regulate odontogenic mesenchyme differentiation. EDTA-dissociated mouse molar mesenchyme, in the absence of epithelium but with adherent basal lamina, routinely differentiated into odontoblasts. Control tooth organs routinely formed both dentine and enamel extracellular matrices within 7-10 days in our serumless, chemically-defined organ culture system. Regulation of determined mesenchymal cells to differentiate into functional and highly specialized odontoblasts appears to be mediated by epithelial-derived basal lamina and is not species or organ-specific.

Animals↗
PubMed2006

Mandibular phenotype of p20C/EBPbeta transgenic mice: Reduced alveolar bone mass and site-specific dentin dysplasia.

CCAAT enhancer binding proteins (C/EBP) comprise a family of basic-leucine zipper transcription factors that regulate cellular differentiation and function. To determine the role of C/EBP transcription factors in osteoblasts and odontoblasts, we generated a transgenic (TG) mouse model with Co1a1 (pOBCol3.6) promoter-targeted expression of a FLAG-tagged dominant negative C/EBP isoform, p20C/EBPbeta (previously LIP). Two of the four transgenic lines presented with abnormalities in the developing incisors, including breakage, overgrowth, and malocclusion. Histological examination revealed that the amount of alveolar bone was reduced in TG compared to wild-type (WT) mice. By microcomputed tomography (microCT), the bone volume fraction of the mandible was reduced at the level of the first and third molars, demonstrating a severe mandibular osteopenia. The lingual dentin morphology of TG incisors differed dramatically from WT. Labial dentin (enamel side) showed normal thickness and tubular dentin structure, whereas the lingual dentin was thinner (25-30% of WT at the alveolar crest) with an amorphous globular structure characteristic of dentin dysplasia. FLAG immunostaining was seen in both lingual and labial odontoblasts, indicating that the site-specific defect was not due to a lack of labial transgene expression. Northern blot analysis demonstrated reduced osteocalcin expression in TG mandibles, while bone sialoprotein was increased, consistent with prior results in calvariae and long bones. Dental sialophosphoprotein, a marker of the odontoblast lineage whose absence causes dentin dysplasia, was modestly reduced in TG mice by Northern blot and in situ hybridization analysis. By fluorescence microscopy, pOBCol2.3-GFP, a marker of the odontoblast lineage, was expressed in both labial and lingual odontoblasts, although GFP-marked lingual odontoblasts were more flattened than WT cells. Moreover, GFP-positive processes in the lingual dentin tubules were truncated and less organized than those in WT dentin. MicroCT analysis showed reduced tissue density in the lingual dentin. These data suggest that C/EBP transcription factors may be involved in the regulation of odontoblast polarization and dentin matrix production.

Alveolar Bone Loss↗
PubMed1989

Responses of pulpal nerves to cavity preparation in rat molars: an immunohistochemical study using neurofilament protein (NFP) antiserum.

The response of neural elements to a dentin injury was morphologically investigated in rat molars by use of immunostaining for neurofilament protein (NFP). An artificially formed cavity in dentin by drilling rapidly caused the displacement of some odontoblasts into the exposed dentinal tubules, while others were detached from the predentin. The subodontoblastic nerve plexus consisting of NFP-immunoreactive nerves shifted inward together with the separated odontoblasts, while a movement of the nerves into the exposed dentinal tubules was not recognized. The odontoblasts separated from the predentin degenerated and disappeared one day after the cavity preparation; at this time, the subodontoblastic nerve plexus underlying the drilled dentin was remarkably disrupted, presumably losing dentinal sensation of the drilled area. Three days after the cavity preparation, the destroyed odontoblastic layer began to be repaired by newly differentiating odontoblasts; the reparative dentin was produced from 5 to 7 days onward. Numerous NFP-positive nerves, beaded in type, gathered in the odontoblastic layer in accordance with the differentiation of the new odontoblasts. The increased beaded nerve fibers were suggested to represent peptide-containing nerves. In 10-15 days, the reparative dentin accumulated quite remarkably under the cavity area. The NFP-positive subodontoblastic nerve plexus was entirely reconstituted and also regained continuity to its surrounding plexus. The nerve fibers in the reconstituted plexus were mostly non-beaded in type as seen in the control teeth. Since dentinal tubules in the reparative dentin are not normally continuous to the primary dentinal tubules, dentinal sensation may not have been restored.

Animals↗
PubMed1993

Biocompatibility testing of restorative materials influencing dentin and pulp.

A comparative study was carried out using 27 healthy human bicuspids that had been extracted for orthodontic reasons from adolescents 11-14 years old. A microphotometrical analysis of buccal and lingual odontoblast layer thickness was performed using the SAMBA 2000 system and the results were submitted to statistical comparison. The buccal odontoblast layer (BOL) was found to be thicker than the lingual odontoblast layer (LOL). These results were independent of age, sex, and tooth position in the dental arch. Moreover, a strong relationship existed between the layer thicknesses as linear function on a logarithmic scale. Standards for the evaluation of pulp-dentin biocompatibility are based essentially upon qualitative or semiquantitative histological criteria. A method of quantitative evaluation based upon microphotometrical measurements of the thickness of the buccal odontoblast layer, considered variable, and the lingual odontoblast layer, considered stable, was applied to coronal filling materials that had previously been tested by a classic method. Seventy bicuspids extracted from adolescents 11-14 years of age whose class-V cavities had been filled in situ with one of four restorative materials, and whole residual dentin thickness was less than 750 microns, were divided into two groups: an intermediate-term group (A) and a long-term group (B); both groups were subdivided into four experimental series as a function of test materials. A microphotometrical analysis was carried out using an automatic scanning system (SAMBA 2000) for the quantification of buccal and lingual odontoblast layer thicknesses. The results confirmed that due to the stability of the LOL thickness it may legitimately be used as reference in the study of the analogous BOL thickness.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗