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Focal c-fos expression in developing rat molars: correlations with subsequent intradental and epithelial sensory innervation.

The purpose of this study was to analyze the temporal and spatial patterns of expression of the inducible transcription protein Fos and the Fos-related antigens (Fra) in developing rat teeth. Immunoreactivity (IR) for Fos/Fra was analyzed at postnatal age 1-35 days. A transient gradient of Fos/Fra-IR was found in all the molars in the coronal odontoblasts along sites of dentinogenesis, with numerous cells and intense staining near the pulp horn tip, and fewer cells and less staining in mid-crown and cervical pulp. This gradient was well established in first molars of the one day old rats; it was first seen in second molars in the 2 day old rats; and it was found in third molars at 10 days. The Fos/Fra-IR was transient and faded after a few days. Rat molars have a tilted orientation so that maxillary molar crown cusps point in a posterior direction and mandibular crowns in the anterior direction. In each set of molars dentinogenesis was initiated along the side of each pulp horn closest to the gingival surface, i.e. anterior for maxillary crowns and posterior for mandibular crowns; and the Fos/Fra-IR first appeared next to those asymmetric sites. As the wave of dentinogenesis spread around the crown, it was accompanied by odontoblastic expression of Fos/Fra-IR that had decreasing intensity in mid- and cervical crown. Molar root pulp lacked Fos/Fra-IR, and incisor teeth only had odontoblastic and ameloblastic immunoreactivity in the 1 day old rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Polyamine depletion-mediated effects on murine odontogenesis are dependent on tooth developmental stage and culture conditions.

Polyamines are known to play a central role in processes such as growth and development. Virtually nothing is known about their importance in tooth development, an attractive and frequently used experimental model for studies of developmental processes. A polyamine-depleted state was created in tooth cells in an organ culture system. First lower molar germs from 16-and 17.5-day old mouse fetuses were used. alpha-difluoromethylornithine (DFMO) and methylglyoxal bis-(guanylhydrazone) (MGBG) were used to deplete the cells from their polyamine content. Polyamine interconversion and catabolism were prevented by aminoguanidine sulfate (AG). In day-16 germs cultured in serum-containing medium, DFMO reduced the frequency of cycling cells as shown by [3H]thymidine incorporation, and induced a delay of odontoblast differentiation of about 24 h. Under the same conditions, MGBG induced an arrest of histo-morphogenesis, correlated to a significant decrease in the rate of cell proliferation. Addition of polyamines prevented DFMO- and MGBG-induced delay of tooth differentiation. Interestingly, MGBG did not delay the terminal differentiation of odontoblasts and ameloblasts in cultured day-17.5 molars; in these, cells at the tip of the cusps are only a few hours before their withdrawal from the cell cycle. In serum-deprived medium, dental cytodifferentiations did not occur. Addition of putrescine or spermidine to serum-free media, however, allowed for tooth morphogenesis and cytodifferentiation. Tooth explants in a serum-deprived medium reacted to DFMO in a cytocidal fashion, whereas MGBG showed only a mild toxicity in some cell types. Addition of putrescine to DFMO-containing medium prevented its cytotoxic effect. Addition of spermidine to MGBG-containing medium not only prevented its mild toxicity but also allowed for predentin secretion by differentiated odontoblasts. The results are discussed with regard to the well-established developmental events of tooth germs cultured in vitro and with respect to present knowledge of polyamine metabolism and their involvement in cellular processes.

Animals↗

The quantification of tertiary dentine formation in response to materials commonly placed in deep cavities in general practice in the UK.

PURPOSE OF STUDY: To determine the rate of formation of tertiary dentine in dogs' teeth, in response to five commonly used lining materials. Also, to record the resulting differences in quality of the formed dentine. MATERIALS AND METHODS: Representatives of five types of materials commonly used in general practice in the UK were placed in deep standardised cavities with a remaining dentine thickness of 400-500 microns, and sealed to the cavo-surface with zinc oxide-eugenol. The daily rate of tertiary dentine formed in response to each material was measured over a period of 119 days using a tetracycline staining method. A concomitant study to show the quality of the formed dentine was undertaken using the same materials and experimental conditions. RESULTS: There was a significant difference between the rates of tertiary dentine formation for all materials except zinc oxide-eugenol and Ledermix paste. For each material there was a increased rate of tertiary dentine formation in the second period (29-49 days) compared to the first period (0-28 days). This is considered to be due, in part, to a lag period when damaged odontoblasts are differentiating from mesenchymal elements. The rate of tertiary dentine formation in response to all the test materials was always greater than that of normal physiological dentine formation in unprepared teeth. Qualitatively, there was difference in the formed tertiary dentine beneath each material. All the materials caused odontoblast destruction, altered ground substance production and alterations in the calcification process. CONCLUSION: None of the materials routinely used at present can be considered to be 'ideal' as odontoblast destruction, altered ground substance production and altered calcification occur beneath zinc oxide-eugenol preparations, calcium hydroxide preparations as well as in response to corticosteroid-containing materials such as Ledermix paste and cement.

Analysis of Variance↗

Early and intermediate time response of the dental pulp to an acid etch technique in vivo.

PURPOSE: To present the ultrastructural features of the pulpal responses, following the application of All-Bond 2 to acid-conditioned, deep, unexposed coronal dentin and exposed pulps in human teeth. MATERIALS AND METHODS: Cylindrical Class V cavities were prepared in human premolars. In the non-exposure group an attempt was made to prepare the floor of the cavity to +/- 0.5 mm from the pulp. In the exposure group, the pulps were intentionally exposed. After hemostasis, the preparation was etched with 10% phosphoric acid. The teeth were restored with All-Bond 2. Histological evaluation was done at 0-7, 28-35, and > 90 days. RESULTS: A typical connective tissue response to injury was observed in the majority of the specimens. Irreversible injury to the odontoblasts closest to the site of cavity preparations resulted in the death of these cells. This was followed by an early neutrophilic response, a subsequent macrophage response and a fibroblastic response that led to the deposition of either reparative dentin or calcific bridge formation by odontoblast-like cells. However, another notable feature was the consistent observation of resin particulates within the dentin-pulp complex. These resin particulates could have been indirectly introduced into the pulp through the pertubation of the junctional complexes or the death of the odontoblasts. They may also enter the pulp directly through a pulpal exposure. In some specimens, the presence of these resin particulates appeared to have triggered a foreign body response, characterized by the presence of a mononuclear inflammatory infiltrate as well as the appearance of multinuclear giant cells. The persistence of unresolved chronic inflammation was associated with the lack of calcific bridge formation in these specimens.

Acid Etching, Dental↗

Metabolic unit of pulp-dentinal complex of human tooth.

The change of dentin depends on odontoblast's metabolism (synthesis and secretion), and the result of those cells activity is apposition of dentin: physiological or reactive. Odontoblasts, dentinal tubuli with odontoblastic processus and peritubular zone, they all make metabolic unit of dentin, and they are the major part of circumpulpal dentin structure. Functional activities of the tooth are the base for constant changes during the entire life of circumpulpal dentin. The apposition, and also the resorption of dentinal tissue, can be found as the respond to trauma, pathological conditions, or after the therapeutic interventions.

Dental Pulp↗

Restricted expression of Mov13 mutant alpha 1(I) collagen gene in osteoblasts and its consequences for bone development.

Cell type-specific differences in the transcriptional control of the mouse gene coding for the alpha 1 chain of collagen type I (Col1a1) have been revealed previously with the help of the Mov13 mouse strain which carries a retroviral insert in the first intron of the gene. Transcription of this mutant Col1a1 allele is completely blocked in all mesodermal cell types tested so far, with the exception of the odontoblast where it is expressed at an apparently normal rate (Kratochwil et al. [1989] Cell 57:807-816). To define the tissue specificity of the mutant allele more precisely, we have now studied its expression in osteoblasts, another skeletogenic cell type which, like odontoblasts, produces high amounts of collagen I. Evidence for transcription of the Mov13 allele was obtained by in situ hybridization in homozygous (M/M) and heterozygous (M/+) bone tissue, in grafts as well as in vivo. The presence of mouse collagen I and the development of bone tissue were demonstrated in M/M skeletal elements grown on the chick chorioallantoic membrane (CAM). Further support for expression of the mutant gene was obtained from two 16 day M/M fetuses in vivo. Bone tissue of diverse embryological origin (vertebrae and ribs of somitic origin, long bones derived from lateral plate, calvariae from head paraxial mesoderm, and mandibulae from head neural crest) expresses the mutant allele. However, in situ hybridization experiments indicate that only a subpopulation of osteoblasts is capable of transcribing it at a high rate, resulting in severe impairment of bone development in grafts and in vivo. Therefore, osteoblasts, in comparison to odontoblasts and fibroblast-like cells, assume an intermediate position with respect to transcription of the Mov13 allele. We suggest that this diversity in the utilization of the mutant collagen gene reflects cell type-specific differences in the transcriptional regulation of the wild type (wt) Col1a1 gene.

Alleles↗

Nuclear uptake of 1,25-dihydroxyvitamin D3 in developing rodent teeth: an autoradiographic study.

Target cells for 1,25(OH)2 vitamin D3 metabolites are identified in developing rodent teeth by the use of thaw-mount autoradiography. Following the injection of [26, 27-3H]-1,25(OH)2 vitamin D3 into 18-day- and 20-day-old fetal rats and neonatal mice, nuclear concentration of radioactivity is found in different cell types. In incisors of both animal groups, strong nuclear labeling is present predominantly in pulp cells, while relatively weakly labeled cells are found in the layers of odontoblasts, ameloblasts, and stratum intermedium. In molars, nuclear labeling is absent in fetal rats, but is present in 2-day-old neonates in pulp cells and cells in the layers of stratum intermedium of the first molars, but not in the second molars. The absence of labeled pulp cells in the progenitor regions of incisors and in molars of 20-day-old fetal rats, and differential ontogenic appearance of labeled pulp cells in molars, indicates that there is a critical period of receptor emergence. The finding that labeled pulp cells exist in the regions of incisors and molars where secretory odontoblasts are present suggests that nuclear uptake of 1,25(OH)2 vitamin D3 is related to cell maturation and differentiation, and topographically related to the formation of dentin. The results further suggest that, in contrast to bone, the predominant effect of 1,25(OH)2 vitamin D3 is not on tooth cells which are directly involved in the formation of calcified tissue, i.e., ameloblasts and odontoblasts, but rather on supporting tissues such as pulp cells and stratum intermedium.

Animals↗

Acute and chronic reactions of dental sensory nerve fibers to cavities and desiccation in rat molars.

We have studied the effects of air drying of exposed, acid etched dentin on the sensory innervation of rat molars. In the acute series of experiments, trigeminal nerve fibers were labeled by axonal transport of radioactive protein prior to the dentin exposure and desiccation, the anesthetized rats were fixed by aldehyde perfusion 10 min later, and the teeth were prepared for autoradiography. The results confirmed the hydrodynamic theory by showing outward movement of labeled nerve material in response to dentinal drilling and desiccation. It also showed that some odontoblasts could be separated from the dentinal nerve fibers. In the chronic series, teeth were injured 25 h, 5-7 days, or 21 days prior to fixation and nerves were labeled during the last 24 hours; the surviving vital nerve fibers were evident because of their axonal transport of the radioactive label. In that series, sensory nerve fibers were found to have been lost from areas with newly-formed reparative dentin, or from dentinal tubules that had lost their odontoblasts. In the teeth injured 25 h, 5-7 days, or 21 days earlier, an abnormal nonneuronal labeling occurred 0.2-0.3 mm into injured dentin. Our results are discussed in relation to the hydrodynamic theory, nerve-odontoblast interactions, differences between shallow and deep cavity injuries, altered nerve location in response to pulpal or dentinal injury, and characteristics of the pulp-dentin border.

Animals↗

Calcification capacity of dental papilla mesenchymal cells transplanted in the isogenic mouse spleen.

The capacity of the dental pulp to form calcified tissue was examined in papilla cells dissociated from first molar tooth germs of the neonatal mouse and isografted in the spleen for up to 7 days. To obtain papilla cell populations without odontoblasts, pulpal mesenchyme was isolated mechanically from the enamel organ after 0.1% trypsin treatment and rolled on a membrane filter. On day 3 after transplantation, the grafted papilla cells had changed into large, spindle-shaped cells, and initial calcification with needle-like crystals began in association with the collagenous matrix surrounding those cells. On day 7 after transplantation, the spindle cells transformed into odontoblast-like cells containing well-developed secretory organelles, and irregular, but nontubular, calcified tissues were commonly observed surrounding the extracellular collagenous matrix. The calcified tissue matrix with cellular inclusions displayed a structure similar to that of osteodentin. During this period, an intense positive reaction for alkaline phosphatase (ALPase) activity was demonstrated along the cell membranes of the odontoblast-like cells aligned at the periphery of forming calcified tissue. Enzymatic activity could not be detected on the cells incorporated completely into osteodentin-like matrix. The present results show that the papilla cell population transplanted into the spleen formed osteodentin-like material, thus demonstrating the capacity of papilla cells to produce calcified tissue.

Alkaline Phosphatase↗

Temporospatial gene expression and protein localization of matrix metalloproteinases and their inhibitors during mouse molar tooth development.

The gene expression and protein distribution of matrix metalloproteinase (MMP) -2, -9, membrane type-1 MMP (MT1-MMP), as well as of TIMP-1, -2, and -3 were analyzed during mouse molar development. Immunohistochemical data demonstrated that all the MMPs investigated were expressed in the dental epithelium and mesenchyme. In contrast, gene and protein expression analysis for TIMPs showed that they were differentially expressed. TIMP-1 was expressed in the dental epithelium and mesenchyme between E13 and E16 and was transiently up-regulated at E14, the cap stage. TIMP-1 expression was also detected in differentiating odontoblasts. TIMP-2 RNA transcripts were found in the peridental and dental mesenchyme, odontoblasts, and ameloblasts. Protein analysis revealed high expression on the lingual side of the dental epithelium and underlying mesenchyme together with transient expression in the enamel knot at E14 and expression in the gingival tissue and enamel matrix postnatally. TIMP-3 RNA transcripts were found in discrete regions of the dental epithelium, including at high levels in the cervical loop at E16. Expression was also detected in preodontoblasts at E16 and transiently during ameloblast differentiation. Analysis of the protein distribution revealed a lower level of TIMP-3 on the lingual side of the dental epithelium at E14. MT1-MMP was expressed in the dental mesenchyme between E13 and E16, at relatively high levels in the cervical loop at E14, and in the odontoblasts and ameloblasts. The distinct temporospatial distribution patterns of the TIMPs suggest that these inhibitors play several intrinsic roles during tooth development.

Animals↗

Matrix proteins of the teeth of the sea urchin Lytechinus variegatus.

The teeth of the sea urchin Lytechinus variegatus grow continuously. The mineral phase, a high magnesium calcite, grows into single crystals within numerous compartments bounded by an organic matrix deposited by the odontoblasts. Electron microscopic examination of glutaraldehyde-fixed Ethylene Diamine Tetra acetic acid (EDTA) demineralized teeth shows the compartment walls to be organized from multiple layers of cell membrane which might contain cytoplasmic protein inclusions. Proteins extracted during demineralization of unfixed teeth were examined by gel electrophoresis, high performance liquid chromatography, and amino acid analysis. The tooth proteins were acidic, they contained phosphoserine, and they were rich in aspartic acid. By contrast, the proteins of similarly extracted mineralized Aristotle's lantern skeletal elements were nonphosphorylated and were rich in glutamic acid. Vertebrate tooth and bone matrix proteins show similar differences. Surprisingly, an antibody to the principle rat incisor phosphoprotein showed a significant cross-reactivity with the urchin tooth protein, by dot-blot and enzyme-linked immunosorbent assay procedures. Thus, the urchin tooth proteins contain epitope regions similar to those which are phenotypic markers of vertebrate odontoblasts. Whether this is an expression of convergent or divergent evolutionary processes, it is likely that the matrix proteins play a similar role in matrix mineralization. The sea urchin tooth may thus be an excellent model for the study of odontoblast-mediated mineral-matrix relationships.

Amino Acids↗

An ultrastructural study of dentinogenesis and amelogenesis in rat molar tooth germs cultured in vitro.

Molar tooth germs from three-day-old rats were cultured successfully for fourteen days, permitting the study of the development in vitro of both extracellular matrix and cellular elements such as odontoblasts and ameloblasts. The ultrastructure of the cultured tooth germs was compared with the ultrastructure of tooth germs in vivo at a comparable developmental stage. Progenitor cells of odontoblasts and ameloblasts were found to differentiate in vitro. Odontoblasts seemed to contain more lysosome-like bodies and fewer secretory granules than in vivo. They formed normally mineralizing dentine or a thick layer of dense, unmineralized predentine with incidentally some amorphous, extracellular material. Enamel was exclusively present opposite well developed dentine. It was often hyper- or hypomineralized and enamel rods were not as regularly shaped as in vivo. In places where no enamel formation had taken place, large amounts of amorphous extracellular material were sometimes seen. From these observations it can be concluded that cellular development in cultured tooth germs appeared more or less normal, but extracellular matrix formation and mineralization were sometimes disturbed.

Ameloblasts↗

Ca-ATPase and ALPase activities at the initial calcification sites of dentin and enamel in the rat incisor.

Enzymatic activities of calcium-magnesium dependent adenosine triphosphatase (Ca-ATPase) and nonspecific alkaline phosphatase (ALPase) were localized at the initial calcification sites of dentin and enamel of rat incisor teeth using electron-microscopic cytochemistry. Ca-ATPase was localized in the Golgi cisternae, cytoplasmic vesicles and along the outer surface of the presecretory and secretory ameloblasts, whereas it was totally absent from the odontoblasts in the pulp. Inversely, ALPase reaction was localized along the outer surface of the odontoblasts, but almost completely absent from the ameloblasts. Diffuse extracellular reactions of both enzymes were distributed throughout the unmineralized fibrous matrix of mantle dentin in which a large number of matrix vesicles were scattered. Both Ca-ATPase and ALPase reactions, which appeared in the matrix vesicles in the process of formation of mantle dentin, became most conspicuous at the site of initial dentin calcification. At this stage, an intense Ca-ATPase reaction also appeared along some of the collagen fibrils adjacent to the reactive matrix vesicles. No ALPase reaction was localized along these Ca-ATPase reactive collagen fibrils. Our observations suggest strongly that Ca-ATPase in the matrix vesicles originates from the inner enamel epithelium and/or preameloblasts whereas ALPase originates from the odontoblasts in the pulp. The importance of the coexistence of both enzymes for the control of initial calcification of dental hard tissues is suggested.

Alkaline Phosphatase↗

Localization of NADPH-diaphorase activity in the dental pulp, periodontium and alveolar bone of the rat.

In this study we examined the presence and localization of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the dental pulp, periodontal tissues and alveolar bone of the rat. The presence of NADPH-d activity was also examined in cat pulp. The rat histochemical analysis revealed the presence of prominent NADPH-d activity both in cells of the sub-odontoblastic cell layer and in the odontoblasts, in the root as well as in the coronal pulp regions. In the pulpal horns, odontoblasts often had long processes with a high level of labelling indicating NADPH-d activity extending through the predentin and dentin. Moreover, endothelial cells of pulpal blood vessels were positive for NADPH-d in both species. However, no clearcut examples were found of pulpal nerve fibres positive for NADPH-d in the rat or cat and denervation performed in rats did not alter the enzyme staining patterns. In the periodontal tissue, NADPH-d activity was localized to cells on the alveolar bone surface of the periodontal ligament and, in addition, alveolar bone marrow crypts were filled with intensely labelled cells. In the gingival papillae, NADPH-d activity was observed in the basal cell layer of the epithelium. Endothelial cells of periodontal and gingival blood vessels showing positive staining for NADPH-d were occasionally noted.

Alveolar Process↗

Dentin sialoprotein: biosynthesis and developmental appearance in rat tooth germs in comparison with amelogenins, osteocalcin and collagen type-I.

A non-collagenous protein, extracted from rat incisor dentin, is a dentin sialoprotein (DSP). We examined immunohistochemically the developmental appearance and tissue distribution of DSP in 1 to 3-day-old rat molar and incisor tooth germs. The earliest staining for DSP was observed in newly differentiated odontoblasts. In more advanced stages, immunostaining for DSP gradually increased in pre-dentin, odontoblasts and dentin, and appeared in many cells of the dental papilla. In early stages of development before the breakdown of the dental basement membrane, pre-ameloblasts were also positive for DSP. This staining disappeared from the ameloblast cell body soon after deposition of the first layer of mineralized dentin. Radiolabelling of tooth matrix proteins with 14C-serine in vitro followed by immunoprecipitation and fluorography confirmed that DSP was synthesized by tooth-forming cells. The immunolocalization for DSP was different from that of either collagen type-I, osteocalcin or the amelogenins. Whereas collagen type-I and osteocalcin were restricted to the mesenchymal dental tissues, the amelogenins were detectable in both epithelial and mesenchymal dental cells and tissues at the epithelio-mesenchymal interface at early stages of development, prior to the onset of dentin mineralization. We conclude that DSP is expressed in and secreted by odontoblasts and some dental papilla cells from early stages of dentinogenesis onwards, i.e. later than type-I collagen, but before deposition of the first layer of mineralized dentin. In pre-mineralizing stages, some of the matrix proteins may be endocytosed from the pre-dentin by both cell types involved in the epithelio-mesenchymal interaction.

Amelogenin↗

Ultrastructural localization of osteocalcin in rat tooth germs by immunogold staining.

Osteocalcin was localized by indirect immunogold staining of thin frozen sections of rat tooth germs which had been fixed by different methods. Acrolein fixation proved to be satisfactory considering the preservation of fine structure and antigenicity. In odontoblasts, osteocalcin was found to be localized in the cisternae of the rough endoplasmic reticulum and Golgi apparatus. Few positive transport vesicles were found. Staining for osteocalcin in odontoblastic processes was only observed after strong fixation and was intense in odontoblasts engaged in early dentine formation. Predentine was slightly positive in the neighbourhood of positive processes. Matrix vesicles were negative and strong osteocalcin labeling of dentine seemed to appear after the onset of mineralization.

Animals↗

[New evidence for the etiology of so-called radiation caries. Proof for directed radiogenic damage od the enamel-dentin junction].

AIM: In spite of a great number of relevant studies the etiology of radiation induced caries still is discussed in controversy: The assumption of direct radiation induced lesions in the hard tissue is in contrast to an indirect pathogenesis mediated via radio-xerostomia. METHODS: A systematic study is presented, comparing teeth with a manifest radiation caries (group 1, about 60 Gy, long interval to the extraction) and clinically caries free teeth (group 2, about 30 Gy, short interval) with tooth specimens after an experimental enoral (in situ) irradiation (60 Gy, group 3) and after in vitro irradiation (500 to 2,500 Gy, group 4). 60Co was the irradiation source. Sound teeth were used as a standard (group 5). For non destructive visualisation of subsurface histotomograms by confocal laser scanning microscopy (CLSM) teeth were either used as fresh sections or as Technovit embedded thin slices (sawing grinding technique). RESULTS: Tooth samples from radiotherapy patients (cancer therapeutic doses, long interval before extraction; group 1) showed three characteristic changes: 1, rarefcation of the branching (ramification) of odontoblast processes near the junction, 2. dentine tubules end in front of the interface to the hard tissue and 3, in dentine the interface is characterised by an zone (about 10 microns wide) of low intensity of the remitted light. CONCLUSIONS: The obliteration of the dentine tubules, preceded by a degeneration of the odontoblast processes, is obviously the result of a direct radiogenic cell damage with hampered vascularisation and metabolism particularly in the area of the terminations of the odontoblast processes. The deficit in metabolism combined with a latent damage of the parenchyma (hypo-remitting zone) is evidence for the functional symptoms (subsurface caries). The prerequisite for the micromorphological manifestation of this direct irradiation damage is a vital tooth and in consequence cannot be simulated in situ or in vivo.

Cobalt Radioisotopes↗

Microscopy of the dentine of enamel-free areas of rat molar teeth.

Dentine of the enamel-free areas (EFA) of first mandibular molar teeth of rats aged between 20-150 days was examined by scanning electron microscopy. Demineralized sections were stained with Gram-Twort's stain and examined for dental caries. In newly erupted teeth, a few dentinal tubules opened on the EFA surface. As teeth became worn down, extensive areas of EFA showed numerous patent tubules. In primary dentine, no occlusion or partial occlusion of tubules was apparent at any age except at the occlusal surface. It is postulated that this surface narrowing was caused either by dentine debris or salivary deposits. The only other covering of tubule openings appeared to be a thin salivary pellicle which in older animals was plaque-like. Patent tubules in primary dentine contained odontoblast processes which even in worn teeth extended to EFA surfaces. Processes in adjacent enamel-covered dentine also extended to the enamel-dentine junction. Only a few examples of bacterial invasion of EFA dentine were seen: in younger animals bacteria occupied cracks in the EFA surface, but in older animals they also occupied tubules. The EFA dentine does not appear to respond to attrition by infilling of the tubules; the persistence of vital odontoblast processes in worn teeth suggests that dead tract formation is not extensive. Continuous occlusal wear may restrict the progress of caries in EFA but odontoblasts in the rat may be also physiologically and immunologically involved in restricting the ingress of bacteria into otherwise poorly protected EFA.

Age Factors↗