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PubMed1998

S100 protein-immunoreactive trigeminal neurons innervating the rat molar tooth pulp.

S100-immunoreactivity (ir) was examined in tooth pulp primary neurons of the rat. An immunofluorescence method demonstrated that the molar tooth pulp contained S100-immunoreactive (ir) nerve fibers. In the root pulp, pulp horn and roof of the pulp chamber, S100-ir smooth and varicose fibers ramified and formed subodontoblastic nerve plexuses. All the fibers became varicose at the base of the odontoblastic layer and extended to the odontoblastic layer. Some varicose endings could be traced into the dentin. The trigeminal neurons retrogradely labeled with fluorogold (FG) from the first and second maxillary molar tooth pulps exhibited S100- and parvalbumin-ir. Approximately 60% and 24% of the labeled cells were ir for S100 and parvalbumin, respectively. Virtually all parvalbumin-ir FG-labeled cells showed S100-ir, while 40% of S100-ir ones coexpressed parvalbumin-ir. An immunoelectron microscopic method revealed that all myelinated axons and half of the unmyelinated axons in the root pulp contained S100-ir. In the odontoblastic layer, predentin and dentin, S100-ir neurites lost the Schwann cell ensheathment and made close contact with cell bodies and processes of odontoblasts. The odontoblastic layer also contained parvalbumin-ir neurites. These neurites were devoid of the Schwann cell ensheathment and in close apposition to cell bodies and processes of odontoblasts. S100-ir pulpal axons seemed to be insensitive to repeated neonatal capsaicin treatment. This study suggests that S100-ir tooth pulp primary neurons are mostly myelinated and that S100-ir unmyelinated axons in the root pulp are preterminal segments of myelinated stem axons.

Animals↗
PubMed1998

Reciprocal temporospatial patterns of Msx2 and Osteocalcin gene expression during murine odontogenesis.

Msx2 is a homeodomain transcription factor that regulates craniofacial development in vivo and osteocalcin (Osc) promoter activity in vitro. Msx2 is expressed in many craniofacial structures prior to embryonic day (E) E14 but is expressed at later stages in a restricted pattern, primarily in developing teeth and the calvarium. We examine Osc expression by in situ hybridization during murine development, detailing temporospatial relationships with Msx2 expression during preappositional and appositional odontogenesis and calvarial osteogenesis. Osc expression at E14-14.5 is very low, limited to a few perichondrial osteoblasts in the dorsal aspect of developing ribs. At E16.5 and E18.5, Osc expression is much higher, widely expressed in skeletal osteoblasts, including calvarial osteoblasts that do not express Msx2. No Osc is detected in early preappositional teeth that express Msx2. In incisors studied at an early appositional phase, Msx2 is widely expressed in the tooth, primarily in ovoid preodontoblasts and subjacent dental papilla cells. Osc is detected only in a small number of maturing odontoblasts that also express alpha1(I) collagen (Colla1) and that are postproliferative (do not express histone H4). Msx2 expression greatly overlaps both histone H4 and Colla1 expression in ovoid preodontoblasts and dental papilla cells. By the late appositional phases of E18.5 and neonatal teeth, Osc mRNA is highly expressed in mature columnar odontoblasts adjacent to accumulating dentin. In appositional bell-stage molars, reciprocal patterns of Msx2 and Osc are observed in adjacent preodontoblasts and odontoblasts within the same tooth. Osc is expressed in mature columnar odontoblasts, while Msx2 is expressed in adjacent immature ovoid preodontoblasts. In less mature teeth populated only by immature ovoid preodontoblasts, only Msx2 is expressed-no Osc is detected. Thus, Msx2 and Osc are expressed in reciprocal patterns during craniofacial development in vivo, and Msx2 expression in preodontoblasts clearly precedes Osc expression in odontoblasts. In functional studies using MC3T3-E1 calvarial osteoblasts, Msx2 suppresses endogenous Osc, but not osteopontin, mRNA accumulation. In toto, these data suggest that Msr2 suppresses Osc expression in the craniofacial skeleton at stages immediately preceding odontoblast and osteoblast terminal differentiation.

Animals↗
PubMed2003

Analysis of the odontogenic and osteogenic potentials of dental pulp in vivo using a Col1a1-2.3-GFP transgene.

Recently, transgenic mice that carry a Green Fluorescent Protein (GFP) reporter gene fused to 2.3 kb fragment of rat Col1a1 regulatory sequences (pOBCol2.3GFPemd) were generated. In the present study, we have examined the patterns of expression of Col1a1-2.3-GFP during odontoblast differentiation in this transgenic line. We report that Col1a1-2.3-GFP is expressed in newly differentiated odontoblasts secreting predentin and fully differentiated odontoblasts. The pattern of expression of Col1a1-2.3-GFP in odontoblasts is correlated with that of dentin sialophosphoprotein (DSPP). Col1a1-2.3-GFP is also expressed in the osteoblasts and osteocytes of alveolar bone. The pattern of expression of Col1a1-2.3-GFP in osteocytes is correlated with the expression of Dmp1. These observations indicate the 2.3 kb rat Col1a1 promoter fragment has sufficient strength and specificity to monitor the stage-specific changes during both odontoblast and osteoblast differentiation. We also used coronal pulp tissues isolated from postnatal pOBCol2.3GFPemd transgenic animals to follow their differentiation after transplantation under the kidney capsule. Our observations provide direct evidence that the dental pulp contains competent progenitor cells capable of differentiating into new generations of odontoblast-like cells which express high levels of Col1a1-2.3-GFP and DSPP and secrete tubular containing reparative dentin. We also report that the dental pulp is capable of giving rise to atubular bone-like tissue containing osteocytes expressing high levels of Col1a1-2.3-GFP and Dmp1. Our studies indicate that pOBCol2.3GFPemd transgenic animals provide a powerful tool for direct examination of the underlying mechanisms and the signaling pathways involved in dentin regeneration and repair, stem cell properties and heterogeneity of the dental pulp.

Animals↗
PubMed1995

Is rabbit dentine innervated? A fine-structural study of the pulpal innervation in the cheek teeth of the rabbit.

The pulpal innervation of rabbit premolars and molars has been studied in transverse sections of perfusion-fixed, demineralised specimens using light microscopy and transmission electron microscopy. A mixed population of small myelinated and unmyelinated axons enters the apical foramen to supply the mesial and distal laminae of these continuously growing teeth. The nerve fibres are remote from the preodontoblasts and odontoblasts near the apical end, but in their passage to the occlusal end the pulp becomes progressively narrower and nerve fibres come to lie subjacent to the odontoblasts and postodontoblasts. Counts of myelinated fibres near the apical end and in the occlusal pulp suggested that the myelin is shed near the occlusal end. Most of the dentine in these teeth is tubular and migrates occlusally with supporting odontoblasts. Near the occlusal end, postodontoblasts deposit an atubular tissue which closes the pulpal ends of the tubules. Nearer the occlusal tip the pulpal contents degenerate and become embedded in the forming atubular tissue. Evidence of axon profiles was found near the occlusal end in the pulp, passing through the odontoblast layer and in the dentine tubules adjacent to odontoblast processes. However, many of the tubules contained an odontoblast process only and the atubular tissue was not innervated. Since innervated tubules eventually become closed by atubular tissue it is assumed that the nerve fibres retract from the tubules before their closure. In common with other teeth the function of the pulpal nerve supply is likely to be mostly nociceptive.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

Combinatorial expression patterns of the connexins 26, 32, and 43 during development, homeostasis, and regeneration of rat teeth.

Gap junctions permit the exchange of regulatory molecules between cells and play important roles during organogenesis. The expression pattern of the gap junction proteins connexin 26, 32, and 43 was studied by immunohistochemistry in the developing, adult, and injured rat teeth. Connexins 32 and 43, but not the connexin 26, were detected during the late stages of embryonic tooth development (bell stage). Expression of connexin 32 was predominant in epithelial cells, whereas connexin 43 was more widely distributed and found in both epithelial and mesenchymal cells. During cytodifferentiation (early postnatal stages), both connexin 32 and 43 were expressed in the epithelial-derived ameloblasts, synthesizing and secreting the enamel matrix proteins. In mesenchyme, connexin 32 was observed only in differentiating odontoblasts, while connexin 43 was expressed in both differentiating and functional odontoblasts, which secrete the dentin matrix. In adult rat teeth, connexin 26 and 43 were expressed in the odontoblastic layer at low and high levels, respectively, while connexin 32 was absent from odontoblasts. Electron microscopy showed that connexin 43 was distributed exclusively at sites of contacts between odontoblasts. However, double immunostaining combined with confocal microscopy suggested an occasional overlap between odontoblasts and calcitonin gene-related peptide-positive nerve fibers. Denervation experiments showed that the expression of connexins in dental pulp was independent of innervation, whereas in injured teeth connexin 43 was upregulated in pulpal fibroblasts. Finally, cultured dental epithelial cells expressed both connexin 32 and 43, and connexin 43 was detected in cultured pulp fibroblasts in vitro, thus mimicking the in vivo distribution pattern of connexins. These results demonstrate that connexins are involved in tooth development and suggest that a given connexin may have distinct roles during odontogenesis and tooth homeostasis.

Animals↗
PubMed2000

Human dentin production in vitro.

The main hard tissues of teeth are composed of dentin and enamel, synthesized by the mesenchyme-derived odontoblasts and the epithelial-derived ameloblasts, respectively. Odontoblasts are highly differentiated post-mitotic cells secreting the organic matrix of dentin throughout the life of the animal. Pathological conditions such as carious 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. In this study we reproduced this physiological event in an in vitro culture system using pulps of human third molars. Pulp cells cultured in presence of beta-glycerophosphate formed mineralization nodules, which grew all over the culture period. The immunohistochemical study revealed that, as odontoblasts, pulp cells contributing to the nodule formation express type I collagen, osteonectin, and nestin. By the exception of nestin, these proteins are also detected in the nodules. The composition of the nodules was also analyzed by Fourier transform infrared microspectroscopy. The spectra obtained showed that both the organic and the mineral composition of the nodules have the characteristics of the human dentin and differ from those of enamel and bone. Taken together, these results show that both the molecular and the mineral characteristics of the human dentin matrix are respected in the in vitro culture conditions.

Adolescent↗
PubMed1985

Unusual collagen aggregates induced in rat incisor dentin after vinblastine administration.

Young male Wistar rats weighing 100 g were injected intravenously with 0.2 mg of vinblastine sulfate (VBL). At 24, 27, and 36 hours and 2, 4, and 6 days after introduction of the drug, the rats were fixed by means of perfusion and the upper incisors were removed for electron microscopy. Many secretion granules had accumulated in the young odontoblasts 24 hours after VBL administration. Newly formed predentin located between odontoblasts contained small amounts of an unusual collagen aggregate characterized by a fine, symmetrically striated structure, and paired, dense bands, which were present at both ends. After 27 hours, following the disappearance of the secretion granules in the odontoblasts, unusual collagen aggregates had increased in amount. Two days after administration, the predentin layer, which was close to the odontoblasts, was composed largely of unusual collagen aggregates. During dentinogenesis, the layer of unusual collagen aggregates gradually shifted into the dentin, where, 4 days after VBL administration, it was, to a large degree, located. Mineralization of the unusual collagen aggregates began with the deposition of fine, filamentous crystals that later grew into needle-shaped crystals 40 A thick. These findings suggest that unusual collagen aggregates are produced as a result of the secreting function of odontoblasts and are mineralized, as are normal collagen fibrils in dentin.

Animals↗
PubMed1999

Appearance and distribution of dendritic cells and macrophages in dental pulp during early postnatal morphogenesis of mouse mandibular first molars.

Dendritic cells and macrophages were examined in dental pulp during the postnatal development of mouse mandibular first molars, by immuno- and enzyme histochemistry. F4/80 antibody against dendritic cells and macrophages demonstrated labeled cells predominantly in and around the odontoblastic layer during tooth development from postnatal day 0 (PN0) to PN5. Labeling with Mac-1, Mac-2, and MOMA-2 antibodies against macrophages showed varied distribution patterns. Mac-1-positive cells were not detected in the dental pulp. Mac-2-positive cells appeared in the dental pulp at PN0, but not in or around the odontoblastic layer, and disappeared by PN3. A few MOMA-2-positive cells were detected in the dental pulp during the period examined. The F4/80-positive cells in and around the odontoblastic layer did not exhibit acid phosphatase or non-specific esterase activities. In addition, the F4/80-positive cells showed continued expression of Fcgamma receptor, but not class II major histocompatibility complex (MHC). Other antibodies against dendritic cells (NLDC-145, MIDC-8, and 33D1) did not label the F4/80-positive cells. We concluded that the F4/80-positive and class II MHC-negative cells in and around the odontoblastic layer may be immature dendritic cells in the early stages before eruption, weaning, and crucial exposure to antigenic stimuli. They may not only act primarily as immunosurveillance cells, but also play a role in a regulatory function and differentiation of odontoblasts.

Acid Phosphatase↗
PubMed2005

Expression of osteocalcin and Jun D in the early period during reactionary dentin formation after tooth preparation in rat molars.

We examined, in rats, the expression of osteocalcin and Jun D in the early stage of reactionary dentin formation after tooth preparation and the accompanying morphological changes. Reverse transcription/polymerase chain reaction analysis revealed strong expression of osteocalcin mRNA in pulp tissue at 2 and 3 days post-preparation compared with that in control teeth. Light microscopy demonstrated that, at the dentin-pulp interface, damaged odontoblasts were detached from the dentin matrix immediately after preparation, with neutrophils lining the dental surface after 1 day. After 2-3 days, differentiated odontoblasts appeared at the interface. Reactionary dentin with tubular structures was formed under the cavity after 10 days. Immunoelectron microscopy showed that trace amounts of osteocalcin were expressed in odontoblasts at 2 days post-preparation, and abundant osteocalcin was found in the highly developed Golgi apparatus and granules at 3 days post-preparation. Osteocalcin was also found on type I collagen fibrils in newly formed predentin. The existing dentinal tubules were filled with osteocalcin-coated type I collagen fibrils. We observed, by immunohistochemistry, that Jun D was temporally expressed in the nuclei of the odontoblasts at 1 and 2 days post-preparation. However, no Jun D was found in the dental pulp cells at any other time or in control teeth. Thus, osteocalcin expression is correlated with reactionary dentin formation, and Jun D is associated with osteocalcin expression in odontoblasts. Osteocalcin may also serve as an obturator of the dentinal tubules to protect dental pulp vitality against external irritants after preparation.

Animals↗
PubMed1982

Dental sensory receptor structure in human teeth.

Electron microscopy was used to study normal human extracted teeth in order to define the junctions between sensory nerve endings and other cells in external pulp and inner dentin at the crown tip. Two sets of associated cells were found: (1) Connective tissue cells. The pulpal fibroblast newtork made occasional desmosome junctions with the odontoblast newtork, and the cells of each network formed many gap junctions and desmosomes with one another. (2) Nerve endings. The terminal axons formed a succession of appositions with each other or with Schwann cells in the plexus of Raschkow and the cell-free zone, possibly with fibroblasts in the cell-free zone and odontoblast layer, and with odontoblasts in the odontoblast layer, predentin and dentin. The appositions between nerve endings and their companion cells at all levels usually maintained a regular intercellular spacing of at least 15-20 nm. In predentin and dentin, axons could be easily identified by their distinctive vesicles and mitochondria, and they often occurred within clusters of adjacent dentinal tubules; in the odontoblast layer axon identification was much more difficult. Axo-axonic appositions were found in the plexus of Raschkow, the cell-free zone, predentin and dentin; in many cases, bare axons were separated from each other only by a 5-10 nm extracellular space. Dental sensory mechanisms are discussed in relation to these observations.

Adolescent↗
PubMed2006

Peripherin- and CGRP-immunoreactive nerve fibers in rat molars have different locations and developmental timing.

UNLABELLED: Developing rat molars gain mature sensitivity to electric stimulation at 4-5 weeks after eruption, but the related mechanisms are incompletely understood. Preliminary studies showed weak co-localization of calcitonin gene-related peptide (CGRP) immunoreactivity (IR) with peripherin (PER) or neurofilament protein (NF) in rat molar nerve fibers, while the latter two co-localized extensively. OBJECTIVE: Our goal was to compare timing and location of PER-IR and CGRP-IR innervation in rat first molars during tooth maturation. METHODS: We used single and double immunocytochemistry to study molars of rats aged 10 days to 1 year. Neural patterns were compared with odontoblast maturation stages, dentinogenesis, formation of cell-free and cell-rich zones, and root closure. RESULTS: Spatial and temporal patterns showed that most CGRP-IR and PER-IR have different terminal domains in teeth. PER-IR fibers were well established among immature odontoblasts prior to tooth eruption, but CGRP-IR fibers were absent. Two weeks after eruption of first molars, many CGRP-IR beaded fibers entered dentin, the larger PER-IR fibers began shifting away from odontoblasts towards the pulp, and the symmetrical PER-IR pulpal pattern was being established. The CGRP-IR fibers continued to increase their asymmetric dentinal innervation until root growth was completed, during which time odontoblasts matured, the cell-free and cell-rich zones appeared, and roots closed. CONCLUSIONS: Sensory maturation of rat molars coincides with closed root apices, extensive innervation of dentin by CGRP-IR nerve fibers, and the appearance of the mature avascular odontoblast layer next to cell-free and cell-rich zones in the pulp horns.

Animals↗
PubMed2006

Rescue of odontogenesis in Dmp1-deficient mice by targeted re-expression of DMP1 reveals roles for DMP1 in early odontogenesis and dentin apposition in vivo.

Dentin matrix protein 1 (DMP1) is expressed in both pulp and odontoblast cells and deletion of the Dmp1 gene leads to defects in odontogenesis and mineralization. The goals of this study were to examine how DMP1 controls dentin mineralization and odontogenesis in vivo. Fluorochrome labeling of dentin in Dmp1-null mice showed a diffuse labeling pattern with a 3-fold reduction in dentin appositional rate compared to controls. Deletion of DMP1 was also associated with abnormalities in the dentinal tubule system and delayed formation of the third molar. Unlike the mineralization defect in Vitamin D receptor-null mice, the mineralization defect in Dmp1-null mice was not rescued by a high calcium and phosphate diet, suggesting a different effect of DMP1 on mineralization. Re-expression of Dmp1 in early and late odontoblasts under control of the Col1a1 promoter rescued the defects in mineralization as well as the defects in the dentinal tubules and third molar development. In contrast, re-expression of Dmp1 in mature odontoblasts, using the Dspp promoter, produced only a partial rescue of the mineralization defects. These data suggest that DMP1 is a key regulator of odontoblast differentiation, formation of the dentin tubular system and mineralization and its expression is required in both early and late odontoblasts for normal odontogenesis to proceed.

Animals↗
PubMed1999

Stimulation of the dentine-pulp complex of rat incisor teeth by transforming growth factor-beta isoforms 1-3 in vitro.

Expression of transforming growth factor-beta (TGF-beta) isoforms by odontoblasts leads to their sequestration within dentine matrix. TGF-beta1 and -beta3 stimulate matrix secretion and also initiate odontoblast cytodifferentiation in vitro and in vivo. Using a recently established organ-culture model, the aim here was to examine the effects of TGF-beta isoforms on the response of the dentine-pulp complex during culture. Agarose beads were soaked in isoforms 1-3, and placed on the odontoblast area of slices of 28-day-old rat incisor tooth. The slices were maintained in Trowel-type cultures for 7 days. Both TGF-beta1 and -beta3 stimulated a local increase in predentine secretion at the site of the bead application when compared to control cultures. Mitogenic effects on the cells of the subodontoblast layer were also seen and occasionally small foci of newly differentiated odontoblast-like cells could be observed a little distant from the application site of TGF-beta3. TGF-beta2 had a minimal effect on the cultured tissues. These results demonstrate that TGF-beta1 and -beta3 can stimulate secretion of extracellular matrix by odontoblasts, are mitogenic to pulp cells, and that TGF-beta3 may have inductive effects on pulpal cells. Such activities might be important during reparative processes in the dentine-pulp complex after tissue injury.

Animals↗
PubMed2000

Stimulation of the rat dentine-pulp complex by bone morphogenetic protein-7 in vitro.

Human recombinant bone morphogenetic protein-7 (BMP-7), when applied to freshly cut dentine in monkey teeth, stimulated tertiary dentine formation, but it is unclear whether this response involved upregulation of the synthetic and secretory activity of existing odontoblasts or the induction of differentiation of new odontoblast-like cells. Using a recently developed organ-culture system for whole tooth slices, the aim here was to examine the effects of BMP-7 on the stimulation and modulation of existing odontoblasts in the absence of tissue injury. Agarose beads were soaked in a 500 ng/ml or 100 ng/ml solution of BMP-7 in culture medium and placed on the odontoblast area of the dentine pulp complex of rat tooth slices. The slices were embedded in a semisolid agar-based medium and cultured at the liquid gas interface for 7 days. Results showed that beads soaked in 500 ng/ml BMP-7 stimulated a localized increase in extracellular matrix secretion by odontoblasts at the site of application, with greater stimulatory effects than from the lower concentration. These effects may be important in the reparative processes after tissue injury within the dentine-pulp complex and may be useful in the therapeutic induction of tertiary dentinogenesis.

Animals↗
PubMed2000

In vitro synchronization of embryonic mouse incisor preodontoblasts and preameloblasts: repercussions on terminal differentiation.

Preodontoblasts divide asynchronously and their terminal differentiation occurs gradually. Experimental data suggested that the expression of competence by preodontoblasts to respond to specific epigenetic signals, triggering their overt differentiation, requires a minimal number of cell cycles. The intrinsic timing mechanism could imply division counting and preodontoblasts of juxtaposed cell generations might sequentially withdraw from the last physiological cycle. To test such an hypothesis, embryonic mouse lower incisors were cultured in vitro and treated sequentially with nocodazole in order to induce a transitory synchronization of the dividing preodontoblasts and preameloblasts. This synchronization led to a disorganization of the physiological gradual terminal differentiation of the odontoblasts, giving rise to three distinct domains comprising respectively: 1) odontoblasts with altered polarization and predentin secretion; 2) odontoblasts demonstrating equivalent polarization and predentin deposition; and 3) preodontoblasts-odontoblasts involved in gradual terminal differentiation. These results strongly suggest that the gradient of odontoblast functional differentiation results from sequential withdrawal from asynchronous cell cycles of competent cells able to overtly differentiate.

Ameloblasts↗
PubMed2003

Possible role of immunocompetent cells and the expression of heat shock protein-25 in the process of pulpal regeneration after tooth injury in rat molars.

Recent studies have established that heat shock proteins (HSPs) potentially play a role in immunosurveillance. The purpose of the present study was to clarify the relationship between the chronological changes of immunocompetent cells and the expression of HSP-25 in the process of pulpal regeneration after tooth injury in rat molars by immunocytochemistry for HSP-25 and class II major histocompatibility complex (MHC) antigen. In untreated control teeth, intense HSP-25 immunoreactivity was found in the cell bodies of odontoblasts. Both cavity preparation and tooth replantation caused the degeneration of the odontoblast layer to result in the loss of HSP-25 immunoreactions in the suffered dental pulp at the early stages after tooth injury. Numerous class II MHC-positive cells appeared along the pulp-dentin border and extended their cell processes into the dentinal tubules at 12-24 h after cavity preparation and 3 days after tooth replantation. Newly differentiated odontoblast-like cells with HSP-25 immunoreactivity were arranged at the pulp-dentin border and the class II MHC-positive cells retreated towards the subodontoblastic layer by post-operative days 3-5 after tooth injury. Thus, the common cellular events occur during pulpal regeneration following two different experimental injuries. These findings indicate that the time course of changes in the expression of HSP-25 immunoreactivity reflects the degeneration/regeneration process of odontoblasts and that the temporal appearance of the class II MHC-positive cells at the pulp-dentin border suggests their participation in odontoblast differentiation as well as in initial defence reactions during the pulpal regeneration process.

Animals↗
PubMed2000

bcl-2, p53, and MIB-1 in human adult dental pulp.

Little is known about the renewal of some groups of cells in dental pulp, and the occurrence and significance of physiological cell death in dental pulp is not yet understood. The possibility of odontoblast disappearance by apoptosis has been proposed, and the presence of apoptotic cells in the rat and human odontoblastic and subodontoblastic layers has been recently described. bcl-2 and p53 are proteins involved in the apoptotic pathway, whereas MIB-1 is a proliferating cell marker. The aim of our study was an immunohistochemical evaluation of bcl-2, p53, and MIB-1 in healthy normal pulps of young human subjects. With bcl-2 immunostaining, some positive cells were found in the odontoblastic and subodontoblastic layers, whereas with MIB-1, only a few stromal cells were positive, and all odontoblasts were consistently negative. No cells were positive to p53. The bcl-2 immunoreactivity of the cells of the odontoblastic and subodontoblastic layers could help to explain the presence of apoptotic cells found in these regions.

Adolescent↗
PubMed1998

Dentin-specific proteins in MDPC-23 cell line.

Only four established odontoblast-like cell lines have been reported in the literature (1-6). Of the four, only two synthesize dentin-specific proteins. These studies report that the cell line MO6-G3 synthesizes phosphophoryn (DPP), dentin sialoprotein (DSP) and dentin matrix protein-1 (DMP-1) while MDPC-23 synthesizes DSP, but not DMP-1. The objective of the present study was to determine whether polyclonal antibodies to rat DSP and DPP would label odontoblasts on microscopic sections of day-19 fetal mouse incisor odontoblasts as well as cultured cells of the MDPC-23 cell line. The spontaneously immortalized MDPC-23 cell line was derived from fetal mouse molar papillae, made continuous by the 3T6 method and cloned by dilution. These cultures have been passaged 77 times after cloning, form multilayered nodules, and have high alkaline phosphatase activity. The data show positive reactivity in odontoblasts in 19-d mouse fetal incisors as well as in cultures of MDPC-23 cells by fluorescence and confocal microscopy. In addition, these cultures were characterized by phase microscopy and scanning and transmission electron microscopy. These findings suggest that MDPC-23 cells are of the odontoblast lineage.

Animals↗