The role of sensory nerves in the development of inflammation of oral tissues

Experimental stimulation and clinical procedures applied on the tooth crown cause vascular reactions in the dental pulp of cats and rats. These reactions depend on the activation of trigeminal afferent nerves and release of neuropeptides. A brief stimulation causes vasodilation, which is mainly mediated by calcitonin gene-related peptide (CGRP). A longer stimulation results in plasma extravasation which is mediated mainly by substance P (SP) and prostaglandins in the pulp. In adjacent oral tissues the mechanisms following stimulation or local irritation are more complex and other mediators are also involved. Nitric oxide (NO) which is instantly produced in the tissues is such a novel mediator. The chemosensitive nature of the nerves involved (capsaicin sensitive) may lead to their activation also by inflammatory mediators released in the tissues. Thus, sensory nerves may modulate the progress of inflammation. Since sensory nerves in oral tissues are often the first structures to be activated during clinical procedures, tissue reactions that occur can be assumed to be initiated and perpetuated by the sensory neuropeptides. Much work is now made to modulate the sensory nerveinduced cascade of events in oral tissues to find new treatment strategies.     

CMAP: a novel cystatin-like gene involved in liver metastasis

The distribution and ultrastructure of class II major histocompatibility complex (MHC)-positive cells were investigated in human dental pulp, employing immunohistochemistry using an anti-human leukocyte antigen (HLA)-DR-monoclonal antibody. HLA-DR-immunopositive cells, appearing spindle-like or dendritic in profile, were densely distributed throughout the dental pulp. Under the electron microscope, these cells exhibited various sizes of vesicles containing clear or opaque contents, multivesicular bodies and characteristic fine tubulovesicular structures in their cytoplasm. Some reactive cells possessed coated pits and vesicles including electron-dense materials, indicating an active endocytosis. At the periphery of the pulp tissue, the HLA-DR-immunopositive cells were predominantly situated in the subodontoblastic layer, with some located in the odontoblast layer and/or predentin and extending their cytoplasmic processes into the dentinal tubules. Cell processes of these cells occasionally made contact with several odontoblast processes in the same way as the nerve fibers in the predentin. These cells never contained the typical phagosomes frequently observed in the HLA-DR-immunoreactive macrophages in the subodontoblastic layer and the pulp core. The results suggest that the HLA-DR-immunopositive cells in the odontoblast layer and/or predentin have some regulatory function on the odontoblasts under physiological conditions, in addition to their involvement in the initial defense reaction after tooth injury.     

Diet and risk of colon cancer in a large prospective study of older women: an analysis stratified on family history (Iowa, United States)

The aim of this study was to determine if diffusible angiogenic growth factors were released in human dental pulp during orthodontic tooth movement. These factors, if diffusible, could induce angiogenesis in other tissues, and may then be isolated and identified. The pulps from 14 premolar teeth treated with straight wire fixed orthodontic appliances for 2 weeks were compared with those of 14 untreated control premolar teeth from the same subjects. Following tooth extraction and sectioning, 1-mm horizontal sections of pulp tissue were embedded in collagen with 1-mm sections of rat aorta and co-cultured in growth media for up to 4 weeks. Sections of rat aorta alone were also cultured. Angiogenic changes in the form of microvessel growth were observed by light microscopy. Microvessel identification was confirmed by electron microscopy and by immunohistochemistry using staining for factor VIII-related antigen marker for endothelial cells. When compared at days 5, 10, and 14 of co-culture, the number of microvessels was significantly greater in the pulps from orthodontically moved teeth than in those from the control teeth. The number of rat aorta microvessels was also significantly greater when co-cultured with pulp from orthodontically moved teeth than with pulp from control teeth and when compared with control cultures of rat aorta alone. There were no significant differences in microvessel numbers between the rat aorta co-cultured with pulp from control teeth and control cultures of rat aorta alone. These results indicate an increase in angiogenic growth factors in the pulp of orthodontically moved teeth, and the enhanced response of the rat aorta when co-cultured with this pulp shows that these factors appear to be diffusible.     

Pulp reactions to restoration of experimentally induced crown fractures

OBJECTIVES: Reattachment of the avulsed enamel-dentine coronal fragment to the remaining tooth structure has become an accepted clinical alternative to a resin composite build-up for the restoration of crown fractured teeth. Since little knowledge exists as to the pulpal response to this procedure, this study was designed to observe the condition of the pulp following experimentally induced crown fracture and restoration in monkeys. METHODS: Experiments were conducted in eight young green Vervet monkeys (Cercopithecus aethiops). In all, 64 fractured incisors were investigated. Light microscopic examination of pulp tissue specimens was carried out after 3 months of observation. RESULTS: The evaluation was restricted to specimens having a fracture plane within 2 mm of the pulp and no pulpal exposure. In general, pulp tissue was well preserved irrespective of the restorative procedure. Even if the restoration or the bonded tooth fragment had been lost during the follow-up period, the pulp generally remained in good condition. Inflammatory infiltrates where seen in only a few specimens and then as clusters of mononuclear leukocytes. Hard tissue repair was frequently observed and displayed various configurations from isolated hard tissue deposits to areas of extensive hard tissue repair in the coronal portion of the pulp. Pronounced hard tissue repair and occurrence of inflammatory cell infiltrates correlated with the presence of stainable bacteria on the fractured dentine surface. CONCLUSIONS: In the absence of direct exposure, reparative dentine is a frequent feature of the pulp's response to crown fracture and restoration with composite or reattachment of the crown fragment with dentine bonding. These restorative procedures appear to ensure continued function of the underlying pulp.     

Conventional endodontics and the operating microscope

Patients are increasingly wishing to undergo conventional endodontic treatment rather than to risk the loss of a tooth. Endodontic treatment in teeth that have been previously restored with extensive intracoronal or extracoronal restorations are often difficult to treat. The orientation of the root canals to the crown of the tooth may be lost, and this may often be compounded by the deposition of reparative dentin in the pulp chamber. The operating microscope allows better visualization of the working field, ensuring that the anatomy of the tooth is more readily inspected. This greatly enhances the clinician's ability to locate extra root canals and therefore increase the likelihood of a successful outcome. It should not be forgotten that the operating microscope also has a place in other fields of dentistry, especially restorative dentistry, and is an asset to both the specialist and the generalist.     

Injuries to the permanent teeth. Periodontal lesions

Tooth luxations are relatively common. In case of concussion or subluxation the tooth is not displaced. The treatment will consist of relief of the tooth. Most frequent complications are pulp necrosis and obliteration of pulpal tissues. In case of extrusive luxation pulpal tissues and the periodontal ligament are injured. When tooth mobility is increased flexible splinting should be considered. Endodontic treatment is necessary after extrusive luxation of a tooth with completed root formation. Teeth with open apex often show pulpal obliteration after extensive luxation. Lateral luxation is more complex than extrusive luxation since the alveolar bone is also damaged. Repositioning and splinting of the tooth are necessary. When the apical foramen in closed, endodontic treatment will be necessary. Teeth with incomplete root formation will develop pulp obliteration. Following lateral luxation, external root resorption and loss of marginal bone are not infrequent. Intrusive luxation is the type of trauma with most unfavorable prognosis. All intruded teeth will become necrotic and external root resorption and marginal bone loss are frequent. There is no consensus regarding the therapeutic approach. Orthodontic extrusion or surgical mobilisation are possible options. In case of avulsion, both the pulpal tissues and the periodontal ligament are disrupted. Preservation of the vitality of the periodontal ligament covering the root will determine the prognosis of the reimplanted tooth. Therefore the tooth will be repositioned as soon as possible. When this is not possible, milk or a specific solution are most appropriate for tooth conservation. When the reimplanted tooth has complete root formation, devitalization will be performed one week after after repositioning. In case of a tooth with open apex revascularisation can be awaited. Healing of the periodontal ligament will determine prognosis. When a normal ligament is obtained during healing or when surface resorption is obtained, the tooth can be preserved for a long period. When progressive replacement resorption (ankylosis) develops, most teeth can remain in position for about 10 years. When inflammatory resorption develops, the tooth will be lost within a short time.     

Hypercholesterolemia and aortic collagen synthesis in rabbit aortas

Male adult New Zealand rabbits were fed a 2% cholesterol diet for 30 of 60 days in order to determine the effect of hypercholesterolemia on aortic collagen synthesis. Collagen synthetic activity was estimated by measuring tissue prolyl hydroxylase activity and the amount of tissue collagen was estimated by measuring tissue hydroxyproline levels. Following 30 or 60 days of feeding there was a significant increase in both tissue and serum cholesterol indicating the onset of hypercholesterolemia. Measurement of collagen synthetic activity and tissue collagen levels demonstrated no increase over control tissues. These data therefore indicate that hypercholesterolemia is not a direct stimulus of tissue collagen synthetic activity.     

Dental innervation: functions and plasticity after peripheral injury

Oral tissues including the periodontal ligament, gingiva, and tooth pulp have a relatively dense sensory innervation and a rich vascular supply. Teeth and supporting tissues are susceptible to tissue injury and inflammation, partly due to lack of collateral blood and nerve supply and to their low compliance. This review focuses on dental nerve functions and adaptive changes in the trigeminal ganglion and tooth pulp after peripheral injuries. An overview of the peptidergic innervation of oral tissues is presented, followed by a discussion of plasticity in neuropeptide expression in trigeminal peripheral neurons after local insults to teeth and peripheral nerve injuries. The functional implications of these adaptive changes are considered, with special reference to nerve regeneration, inflammation, and hemodynamic regulation.     

Inputs to trigeminal brain stem neurones from facial, oral, tooth pulp and pharyngolaryngeal tissues: I. Responses to innocuous and noxious stimuli

Responses evoked in anaesthetized or decerebrate cats by stimulation of afferents supplying the face, mouth, pharynx, larynx, tooth pulp and jaw muscles were recorded from single neurones located in the trigeminal (V) main sensory nucleus, V nucleus oralis, and adjacent regions. Many cells (both V-thalamic relay and non-relay with localized V mechanoreceptive cutaneous fields could be activated by stimulation of a number of these afferents. A particularly prominent short-latency (often monosynaptic) input was noted from the canine tooth pulp, stimulation of which is generally considered to elicit only responses of pain in man. Control experiments showed that pulp-evoked responses were not the result of stimulus spread to tissues outside the pulp. The interaction of these various inputs to neurones at this level of the V brain stem complex typically resulted in a prolonged period of inhibition that was sometimes preceded by a short-lasting facilitatory phase. This inhibitory effect was also apparent in neurones located outside the complex, although a late facilitatory phase was frequently also noted. Our findings indicate a significant nociceptive input to V main sensory-oralis neurones, a proportion of which relay directly to the ventrobasal thalamus. The interactions described may be involved in perceptual and reflex aspects of responses to noxious and innocuous V stimuli.     

Effects of CO2 laser irradiation in vivo on rat alveolar bone and incisor enamel, dentin, and pulp

Previous studies have shown that a surgical 'window' can be drilled in alveolar bone for experimental manipulation of the underlying enamel organ and enamel. To determine whether similar and/or improved access could be obtained by use of the surgical capabilities of laser optics, and to note the effects of laser irradiation in vivo on the extracellular matrices and cells of bone, enamel, and dentin, tissue responses to laser-created lesions were examined histologically. Briefly, samples were prepared in which the alveolar bone along the inferior mandibular border of Wistar rats was exposed, and a continuous-wave CO2 laser equipped with a custom-made micromanipulator was used to penetrate the bone and to create lesions within the lower incisor. Animals were perfusion-fixed at either 10 min or 10 days post-treatment, and affected tissues were processed for light and transmission electron microscopy. At 10 min, all lesions consisted of a void of ablated tissue containing some organic debris. Tissues immediately surrounding the lesion were generally intact, but showed some damage, presumably resulting from elevated temperature effects. At 10 days, lesions in the bone, dentin, odontoblast layer, or pulp showed morphological evidence of tissue repair represented by the presence of cell infiltrates, new bone, or reparative dentin. In lesions that were created during the secretory stage of amelogenesis that had moved into the maturation stage, there was evidence of delayed or incomplete maturation of enamel (i.e., retention of organic matrix normally lost during maturation) related to the enamel organ affected by the laser treatment. In the bone lesion at 10 days, new bone formation was observed, while bone fragments originally created at the time of lasing were surrounded by mononuclear and large multinucleated giant cells. It is thus concluded that the application of this laser system is an alternative method for exposing unerupted dental tissues for experimental manipulation, and that laser irradiation may also be useful for the study of mineralized tissue repair.     

Responses of macrophage-associated antigen-expressing cells in the dental pulp of rat molars to experimental tooth replantation

Bacterial infection of the dental pulp is a major hindrance to successful pulp regeneration after tooth replantation. This study examined how macrophages and class II molecule-expressing cells of the pulp respond to tooth replantation, on the hypothesis that they contribute to the defence and repair of the traumatized pulp. Upper right first molars of 5-week-old male Wistar rats were replanted immediately after extraction; contralateral untreated teeth served as controls. Pulpal cells expressing macrophage-associated antigens were immunohistochemically demonstrated at 0 h (immediately after the replantation) to 84 days postoperatively using antirat monoclonal antibodies OX6 (anti-class II molecules), ED1 (pan-macrophage antibody, reactive also with dendritic cells) and ED2 (anti-resident macrophages). Between 3 and 7 days postoperatively, ED1+ and OX6+ cells, but not ED2+ cells, were concentrated in areas of degeneration formed in the coronal pulp, and frequently showed a marked accumulation along the pulp-dentine border of the cuspal area. Confocal laser scanning microscopy revealed that some of the OX6+ cells with a dendritic profile extended several cytoplasmic processes into the dentinal tubules communicating with the enamel-free area at the tip of the cusp. From 14-84 days, approx. two-thirds of specimens exhibited pulp-tissue regeneration with increasing formation of reparative dentine. Following the formation of sound reparative dentine, cells positive to each antibody were distributed more centrally in the pulp than in the controls, and thus did not show any accumulation along the pulp-dentine border. However, in the other specimens where a bone-like hard tissue had formed in the pulp chamber, many ED1+ and OX6+ cells were still concentrated in the remaining pulp tissue and showed a marked accumulation along the pulp dentine border. Few ED2+ cells were observed in these specimens. These findings suggest that, following tooth replantation, exudative macrophages are actively engaged in eliminating dentinal tubule-derived infectious stimuli and that class II molecule-expressing cells, most probably containing dendritic cells, are positioned strategically at the outermost portion of the injured pulp to monitor incoming antigens. The intensity of the pulpal defence reaction may be dependent on the status of hard-tissue formation, which influences the amount of incoming antigens.     

Migration of newly generated neurons upon ependymally derived radial guide cells in explant cultures of the adult songbird forebrain

The adult songbird forebrain undergoes neuronal production throughout adulthood, with the production of new neurons in discrete regions of the neostriatal ventricular zone. Upon mitogenesis, these new neurons migrate into the subjacent brain parenchyma along radially directed guide fibers. In long-term ventricular zone explant cultures, derived from the higher vocal center of the adult canary, newly migratory neurons were found to associate preferentially with a characteristic substrate cell type. These small, parvonuclear substrate cells formed tightly packed epithelioid sheets, in which ciliated ependymal cells were common, as recognized by both live observation and electron microscopy. A subpopulation of these cells was immunostained by monoclonal antibody 3A7, which preferentially stains the guide fiber network of the adult avian brain. These 3A7+ cells included ependymal cells and bipolar radial cells, as well as morphologically defined astrocytes. As they matured in vitro, the 3A7+ bipolar radial cells extended long, unbranching fibers, which ultimately traversed the culture substrate. Like ependymal cells, they supported neuronal migration. These cells were likely homologous to radial guide cells in vivo. Thus, neuronal migration in adult avian forebrain culture occurred upon guide cells of ependymal derivation.     

The influence of endodontic treatment upon periodontal wound healing

The interrelationship between periodontal and endodontic disease has aroused much speculation, confusion, and controversy. Pulpal and periodontal problems are responsible for more than 50% of tooth mortality today. Diagnosis is often difficult since these diseases have been studied primarily as separate entities. The toxic substances of the pulp may initiate periodontal defects through canal ramifications and patent dentinal tubules, thus impairing wound healing in regenerative procedures. Although no studies exist addressing the direct effect of pulpal infection on the outcome of guided tissue regeneration (GTR) procedures, several studies do indicate that pulpal status may play a significant role toward the end results of GTR. This review article discusses the potential influence of endodontic treatment on the long-term outcomes of GTR. Potential pathways between the pulp and periodontal ligament, which may be responsible for the failure of the regeneration of new periodontal attachment apparatus, are explored. Examination and review of the clinical and research findings in the literature relating to perio-endo lesions are made to demonstrate that a negative influence may exist between GTR outcomes and the status of the pulp.     

The regenerative potential of oxytalan fibers. An experimental study in the monkey

Although a number of studies have described the oxytalan fibers as being a natural component of the periodontal ligament, little information exists about the regenerative potential of these connective tissue fibers. The aim of the present study was to examine whether oxytalan fibers have the capacity to reform after regenerative periodontal therapy. Intrabony defects were produced surgically at the mesial aspects of teeth 37, 35, 45, 47 and at the distal aspects of teeth 11, 21, 31, 41 in one monkey (Macaca fascicularis). After 3 months, the defects were exposed using a full-thickness flap procedure. The root surfaces were debrided and subsequently PDGF-growth factors were placed in the defects. 4 of the 8 sites were covered with a bioresorbable membrane before closure of the wound. Post-surgically, antibiotics were given systemically for 1 week, and tooth cleaning was carried out 1x a week during the entire experimental period. After 5 months, the animal was sacrificed and the oral tissues were fixed by perfusion with 10% buffered formalin. Specimens containing the defects and surrounding tissues were dissected free and histological sections were cut in the mesio-distal direction, parallel to the long axes of the teeth. The sections were stained with hematoxylin and eosin or with the oxone-aldehyde-fuchsin-Halmi staining method and subsequently examined in the light and in the electron microscope. The results revealed that new oxytalan fibers oriented mainly in an apico-occlusal direction had developed in the regenerated periodontal ligament. Many of the newly-formed fibers were inserted into the new cementum, thus suggesting a strong relationship between this tissue and the oxytalan fibers. It is concluded that the regenerated periodontal ligament connective tissue formed after surgery contains oxytalan fibers similar to those present in the original tissue. These results demonstrate that oxytalan fibers develop de novo in the newly-formed periodontal ligament.     

Dental innervation and CGRP in adult p75-deficient mice

Adult dental tissues have unusual neurotrophin biology. Pulpal fibroblasts express nerve growth factor (NGF) and the low-affinity p75 neurotrophin receptor, their sensory nerve fibers express p75 and trk A, and pulpal sympathetic fibers lack p75. Following tooth injury, there is increased pulpal NGF, sprouting of sensory nerve endings, and increased immunoreactivity for the sensory neuropeptide calcitonin gene-related peptide (CGRP). In the present study, we have analyzed tooth structure and innervation of pulp and periodontal ligament in young (6-8 weeks, 3 months) and older (5-12 months) adult mice carrying a null mutation in the p75 gene and compared the results with those of age-matched wild-type controls. Our hypotheses were that tooth structure would be abnormal and that pulpal innervation would be greatly reduced because it consists primarily of nociceptive fibers that have been found to be severely depleted in skin of p75(-/-) mice. Tissues were fixed, X-rayed for gross dental morphology, decalcified, and analyzed for immunoreactivity for CGRP and for a general nerve marker, protein gene product 9.5. Radiographs showed worn-down molar crowns in p75-deficient mice. Light microscopy confirmed the accelerated molar wear and showed intense CGRP immunoreactivity in pulp nerve endings of mutant mice, compared with a gradual decrease in CGRP intensity in controls during normal aging. The CGRP intensity in 5-12-month-old pairs of mice was threefold greater in the mutants (P < 0.03), and in younger mice the mutant always had more CGRP than its matched control. The innervation of molar ligament in all p75-deficient mice was similar to that of controls except there was nerve sprouting near bone loss in mutants. The incisors of mutant mice did not have unusual wear and their pulpal CGRP immunoreactivity remained normal, but their periodontal ligament had fewer thin branched nerve endings at all ages. Thus, most innervation of teeth and their supporting tissues developed normally, and the only neural changes in p75(-/-) mutant mice were the reduction of incisor ligament sensory receptors and increased molar CGRP. Sensory nerves in teeth gradually lose neuropeptide intensity during aging, but that did not happen in the mutant mice, suggesting that the accelerated molar wear stimulated persistent high levels of CGRP.     

Microglial ramification requires nondiffusible factors derived from astrocytes

It is generally accepted that process-bearing microglial cells origina te from ameboid macrophage-like mesodermal cells. This transformation, often called ramification, accompanies down regulation of macrophage-like properties, but the mechanisms involved in ramification have not been clarified. We investigated factors to promote ramification in culture. Isolated ameboid microglial cells were seeded on living or paraformaldehyde-fixed astrocyte monolayers. About 80% of the cells ramified on the fixed astrocytes in astrocyte-conditioned medium as well as on the living astrocytes. In fresh culture medium, 50% of the cells on the fixed astrocytes ramified. On the other hand, ameboid cells rarely ramified on noncoated glass coverslips even in the conditioned medium. Ameboid cells cultured on extracellular matrices dervied from astrocytes ramified more than on those coated with plasma fibronectin or collagen type I. A synthetic peptide containing Arg-Gly-Asp sequence or a tyrosine kinase inhibitor genistein partially reversed the ramification induced on the fixed astrocyte monolayers. These results show that some nondiffusible factors derived from astrocytes are essential for microglial ramification. A part of the nondiffusible factors are present in the extracellular matrices, and the effects might be mediated by integrins. Some diffusible factors secreted by astrocytes seem to promote ramification, if the nondiffusible factors are present. The experiments using the fixed astrocyte monolayers may be useful to identify the diffusible factors responsibile for ramification.     

Tooth replantation in germ-free and conventional rats

The purpose of the present study was to evaluate the influence of bacterial infection on the pulpal and periodontal tissues in replanted teeth using germ-free and conventional rats. Forty maxillary and mandibular first molars from ten 6-week-old germ-free male Wistar rats were used. The animals and all materials were maintained in a germ-free environment inside vinyl isolators throughout the experimental periods. Twenty conventional male Wistar rats served as controls. The first molars were intentionally replanted immediately after extraction. At 3 days, and 1, 2, 4 and 8 weeks after replantation, animals were sacrificed and the replanted teeth were histopathologically evaluated. Diversity of pulp tissue response was notable in conventional rats, which initially showed various degrees of neutrophil infiltration and then displayed different types of response, including revascularization with reparative dentin formation and complete necrosis. Pulpal responses of germ-free rats were less variable, being characterized by an almost complete lack of neutrophil infiltration and a high frequency of bone-like tissue ingrowth. Typical inflammatory resorption was detected only in conventional rats, whereas a higher incidence of ankylosis was notable in germ-free rats. The present results may corroborate the concept that bacterial infection is a major cause of serious healing complications following tooth replantation, such as pulp necrosis and inflammatory root resorption. The difficulty in optimally controlling bacterial infection seems to be highly relevant to the complexity and unpredictability of the outcome of this procedure. It should also be emphasized that extensive mechanical damage to the periodontal tissues may trigger the development of unfavorable healing complications as ankylosis, even under strictly aseptic conditions.     

Clinical and histological evaluation of thermal injury thresholds in human teeth: a preliminary study

The effect on healthy dental pulp of thermal increases ranging from 8.9 to 14.7 degrees C was evaluated. These temperature increases correspond approximately to those caused by certain restorative procedures, such as tooth preparation with high-speed instruments and the fabrication of direct provisional crowns. Two criteria of evaluation have been used in conjunction, a clinical (symptomatic) and a histological one, to assert with greater precision potential damage to the pulp. The results suggest a low susceptibility of cells to heat, which does not appear to be a major factor of injury, at least in the short term. The main cause of postoperative inflammation or necrosis of the pulp is probably the injury of the dentine, a tissue in direct functional and physiological connection with the pulp.     

Expression and localization of membrane type 1 matrix metalloproteinase in tooth tissues

Matrix metalloproteinases (MMPs) have been detected in forming dental enamel and are thought to play an important role during enamel biomineralization. Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a membrane bound member of the MMP gene family that has previously been shown to be expressed by cells associated with bone and cartilage formation (osteoclasts, osteoblasts and chondrocytes). Thus, we asked if MT1-MMP was also expressed by the cells responsible for the formation of enamel and dentin. A porcine MT1-MMP cDNA composed of 3284 bp was isolated from an enamel organ-specific cDNA library. Multiple tissue Northern blot analysis revealed that the MT1-MMP message was expressed highly in the enamel organ and pulp organ when compared to the expression levels observed in other non-mineralizing tissues. Northern blot analysis of stage-specific enamel organs (early secretory, late secretory, or maturation stage) and their corresponding pulp organs revealed that MT1-MMP expression increased as the dentin matured. In the enamel organs, however, the MT1-MMP message level became reduced only during the late secretory stage. Immunohistochemical analysis showed that MT1-MMP was present on the surface of the cells (ameloblasts and odontoblasts) responsible for dentin and enamel formation. Thus, MT1-MMP is highly expressed in developing tooth tissues and may play a role in the biomineralization of enamel and dentin.