Coronal Pulp Research Articles

Coronal pulp size in molars: a study of bitewing radiographs.

To: (i) study coronal pulp dimensions in human first molar teeth; (ii) investigate the effects of restorations on pulp size; (iii) determine differences in dimensions between teeth of Mongoloid patients and teeth of other ethnic groups; and (iv) record the presence of pulp stones. Bitewing radiographs of 121 subjects (mean age, 20.9 years) were taken under standardized conditions. The films were digitally scanned and nine measurements were made from the image of each first molar. The data were analysed in terms of presence or absence of restorations and in terms of racial group. Differences were examined using Student's t-test, Pearson correlations and Levene's test. A total of 445 teeth were analysed. Large crowns were correlated to large pulps. Teeth restored with occlusal and proximal restorations had significantly smaller pulps (P = 0.044 and 0.004, respectively), but no difference was found in pulp area in the clinical crown between the restoration types. Mongoloid crowns were shorter and more bulbous. Pulp areas of maxillary molars and pulp widths at the cervix of mandibular molars were significantly larger in Mongoloids. The pulp area in the clinical crown correlated to pulp horn height for Mongoloids and others and for maxillary and mandibular teeth. No differences were found between the heights of pulp horns in the Mongoloid and other teeth. Four of the pulp measurements demonstrated sexual dimorphism. Stones were present in almost 10% of the subjects, representing 4% of the tooth pulps examined. Even teeth with shallow occlusal restorations had reduced pulp spaces. There were significant differences in pulp and crown dimensions between the teeth of Mongoloid patients and those of other patients.

Porcine enamel matrix derivative enhances the formation of reparative dentine and dentine bridges during wound healing of amputated rat molars.

We examined the biological effects of porcine enamel matrix derivative (EMD; Emdogain) on the formation of reparative dentine and dentine bridges in rat molars after pulp amputation. The pulp chambers of upper molars of Wistar rats were perforated and the amputated pulp surfaces were directly capped with either EMD or its carrier propylene glycol alginate (PGA) as control. The cavities were then restored with glass-ionomer cement. On post-amputation days 4-30, the dissected maxillae were examined by light and electron microscopy. In PGA-capped pulp, reparative dentine had been formed over the dentine walls under the prepared cavity on day 7 post-amputation and its thickness extended until day 30. On day 30, as well as reparative dentine formation, diffuse calcification had occurred beneath the amputated wound surfaces. Dentine bridge formation under the amputated coronal pulp surface was observed in 18.2% of amputated pulp on day 30. In EMD-capped pulp, reparative dentine had already been formed by odontoblast-like cells over the dentine walls, already on day 4 post-amputation, and its thickness extended until day 30. The Ca and P weight % and Ca/P ratio of reparative dentine matrix were similar to those of pre-existing dentine matrix, and these values were not different between PGA and EMD-capped pulp. Dentine bridge formation was observed in 27.3% of EMD-capped pulp on day 30. Our results suggest that EMD enhances the formation of both reparative dentine and dentine bridges during wound healing of amputated rat molar pulp.

Immunohistochemical Localization of μ-Opioid Receptors in Human Dental Pulp

Studies in both animal and clinical models suggest that opioids exert their analgesic effects not only through activation of receptors in the CNS but also through interaction with peripheral opioid receptors. This study evaluated the presence and distribution of mu-opioid receptors in human dental pulp. Human third molars indicated for extraction were removed, fixed in 4% paraformaldehyde and 0.2% picric acid, and decalcified in 10% EDTA and 7.5% polyvinylpyrrolidone. The teeth were cut using a cryostat, and the avidin-biotin peroxidase immunohistochemistry technique was used. Immunostaining for mu-opioid receptors was detected along the nerve bundle of the radicular as well as coronal dental pulp. Positive immunostaining was also observed in the individual nerve fibers in the coronal region. This demonstration of opiate receptors on pulpal nerve fibers suggests a peripheral site in the dental pulp where endogenous or exogenous opioids can interact with mu-opioid receptors.

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.

An immunohistochemical study on hard tissue formation in a subcutaneously transplanted rat molar.

While dental pulp undergoes calcification following tooth replantation or transplantation, we actually know little about these mechanisms. We therefore conducted histological and immunohistochemical evaluations of mineralized tissue that formed in the pulp of rat maxillary molar transplanted into abdominal subcutaneous tissue. One, 2, 3, and 4 weeks post-transplantation, the teeth were investigated immunohistochemically using antibodies to osteocalcin (OCN), osteopontin (OPN), bone sialoprotein (BSP), dentin sialoprotein (DSP), and tissue non-specific alkaline phosphatase (TNAP). In the 1st week after transplantation, cell-rich hard tissue was formed at the root apex. At 2 weeks, formations of hard tissue, with few cells in the root canals and bone-like tissue in the coronal pulp chamber, were noted. After 3 and 4 weeks, the amounts of these hard tissues were increased. The immunolocalization of OCN, OPN, and BSP was seen strongly in coronal and apical hard tissues, but weakly in the root hard tissue. Conversely, DSP localized in the root hard tissue, but not in other newly formed hard tissues. At 1 week, TNAP localized along the periphery of the apical hard tissue and the lower surfaces of root predentin. These results demonstrate that the newly formed hard tissues in the pulp cavity of subcutaneously transplanted molars could be classified into three types, suggesting that these might be formed by type-specific cells.

Expression of adenosine triphosphate P2X3 receptors in rat molar pulp and trigeminal ganglia.

The objective of this study was to investigate the expression of adenosine triphosphate P2X3 receptor subunits in rat molar pulp and trigeminal ganglia and their relationship to substance P. Rat molar pulp and trigeminal ganglia were fixed and sectioned. Double immunostaining with anti-P2X3 and anti-substance P were used to localize P2X3 and substance P expression simultaneously with different stains. P2X3 immunoreactivity (IR) fibers were present in root and coronal pulp. P2X3-IR fibers formed subodontoblastic plexus and advanced into the predentin and dentin. P2X3-IR neurons were localized in small and medium-sized cells in trigeminal ganglia. Colocalization of P2X3 and substance P was not found in either molar pulp or trigeminal ganglia. The results of this study indicated that adenosine triphosphate in pulp tissues may stimulate a subpopulation of non-substance P trigeminal afferent fibers through activation of P2X3 receptors.

Comparative immunohistochemical analysis of the peptidergic innervation of human primary and permanent tooth pulp

This immunohistochemical study sought to determine whether there are any differences in the peptidergic innervation of these pulps and whether dental caries is associated with changes in neuropeptide expression. Mandibular first permanent molars and second primary molars ( n=120) were obtained from children requiring dental extractions under general anaesthesia. Extracted teeth were split longitudinally, placed in fixative, and categorized as intact, moderately carious or grossly carious. The coronal pulps were removed and 10-μm frozen sections were processed for indirect immunofluorescence. Double labelling employed combinations of the following antisera: (1) protein gene product 9.5, a general neuronal marker; (2) one of the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), galanin (GAL), enkephalin (ENK) and somatostatin (SOM). Image analysis was then used to determine the percentage area of immunostaining for each label within different anatomical regions of the coronal pulp. Sparse or absent immunoreactivity for GAL, ENK and SOM made analysis impossible. Analysis of CGRP, SP and VIP revealed significant interdentition differences, with their expression being significantly greater in permanent teeth, but this was not the case for NPY, with primary and permanent teeth demonstrating a similar amount of label for this peptide. Both dentitions showed significant increases in CGRP, SP, VIP and NPY expression with caries progression. These findings could have biological and clinical importance in connection with nociception, inflammation and healing.

Age-related and site-specific changes in the pulpodentinal morphology of rat molars

The rat molars are frequently used as experimental models in endodontic research, but there is little systematic information available on the influence of age on the pulpodentinal organ in Wistar rat molars and it is often difficult to evaluate more subtle changes following experimental interventions. The aim here was to describe changes with age in first upper Wistar rat molars with specific reference to the pulpodentinal organ. Animals were perfused with glutaraldehyde at 19 days, 1, 3, 6, 8, 12, 16, or 24 months of age. First upper molars from 56 animals were demineralized in EDTA, embedded in Epon, and processed for light and transmission electron microscopy. Substantial variation in the structure of the dentine and odontoblasts was observed within the root canals and the coronal pulp chamber. In general, odontoblasts changed from a tall, columnar morphology in the coronal pulp chamber to a more cuboidal or flattened shape near the apex, particularly towards the interradicular space. Secondary dentine formation was more pronounced along the mesial aspect of the root chamber and corresponding to the bottom of fissures. Local tertiary dentine formation was layered in the upper pulp chamber, corresponding to occlusal attrition of the cusp. In several molars a local formation of irregular tertiary dentine was observed cervically in the mesial pulp chamber. After 1 year, a distinct protrusion of irregular dentine extended into the mesiocervical pulp, apparently corresponding to a denudation of cervical root dentine. Experimental pulp-capping studies frequently use first upper rat molars with perforations made through the mesial aspect of the crown; such perforations might be close to the irregular dentine in the mesiocervical region. In conclusion, this study identifies age-associated and regional changes of pulpodentinal morphology in first upper rat molars. Therefore, evaluation of morphological alterations following vital-pulp experiments should be done in specific age groups and at specific sites in the pulp.

The effect of pulpward pressure on the response to 50% lidocaine (lignocaine) applied to exposed dentine in cats

The responses of intradental nerves can be blocked by 50% lidocaine applied to exposed, etched dentine. The aim here was to analyse electrophysiologically the contribution of pulpward pressure to the anaesthetic effect of lidocaine applied to exposed dentine. In 17 adult cats with intact lower canines, approximately 1 mm of the coronal tip of the canine was removed and the exposed dentine acid-etched. Pulpward pressure was applied to the exposed dentine through a chamber fixed to the tip of the tooth. Cervical and coronal electrodes of silver wire and Ag/AgCl mixture were placed, the cervical 2 mm from the gingival margin of the canine and the coronal into the coronal pulp chamber. The inferior alveolar nerve was surgically exposed and single pulpal nerve fibres responding to electrical stimulation of the canine were identified. The chamber was filled with 50% lidocaine solution, and 0 (atmospheric pressure), 1.3, 5.0 or 10.0 kPa pressure was applied to the chamber for 20 min. Orthodromic and antidromic action potentials were compared before and after the lidocaine application. When lidocaine was applied at 0, 1.3 or 5.0 kPa ( n=5 each) for 20 min, all the orthodromic responses to cathodal electrical stimulation with the coronal electrode were blocked. Almost all orthodromic responses (12/15) remained unblocked when the cathodal electrical stimulation was applied with the cervical electrode. Applying 10.0 kPa pressure to the lidocaine solution blocked all the pulpal nerve responses ( n=5) to both stimulation modes. The amplitude of the antidromic compound action potential was significantly decreased when the applied pressure was increased. These results suggest that increased pulpward pressure may effectively force anaesthetic solution more rapidly through the dentinal tubules into the pulp.

Minéralisation de la pulpe dentaire : apports de l'ingénierie tissulaire aux thérapeutiques de demain en odontologie

When bioactive molecules such as bone sialoprotein (BSP), bone morphogenetic protein-7 (BMP-7, also termed OP-1) and chondrogenic Inducing Agents (CIA, A+4 and A-4) were implanted in the pulp of the first upper molars, mineralizations were induced. They were either limited to the formation of a reparative dentinal bridge closing the pulpal wound (CIA A+4), or filled the mesial part of the coronal pulp (BSP), or filled totally the pulp located in the root canal (BMP-7 and CIA A-4). Consequently, these molecules may change in the next future the every day practice in dentistry.

Expression of galanin receptor-1 (GALR1) in the rat trigeminal ganglia and molar teeth.

The expression of galanin receptor-1 (GALR1) was investigated in the rat trigeminal ganglion by using immunocytochemistry and in situ hybridization. In addition, the regional distribution of GALR1-immunoreactive pulpal nerves and their ultrastructure were examined in the molar teeth. In the trigeminal ganglion, the immunoreactivity for GALR1 was recognizable in about 30% of the total number of neurons. Most of the cell bodies were small to medium in size. Analysis of serially cut sections alternately stained with GALR1 and galanin antisera demonstrated that some GALR1-positive cells displayed immunoreactivity for galanin. In situ hybridization analysis, expression of GALR1 mRNA was detected in trigeminal ganglion cells. The cell size distribution was similar to that of GALR1-immunoreactive cells. In the dental pulp, a small number of nerve fibers displayed immunoreactivity for GALR1. The labeled fibers formed terminal arbors in the coronal pulp around and within the odontoblast cell layer, but never penetrated into the predentin and dentin. Ultrastructurally, GALR1 immunoreactivity in the dental pulp was confined to the axoplasm of unmyelinated nerve fibers. The present study provided new evidence that unmyelinated primary afferents innervating dental pulp possessed galanin receptor, and suggests the existence of nociceptive primary afferents functioning as autocrine cells.

Osteogenic proteins (bone sialoprotein and bone morphogenetic protein-7) and dental pulp mineralization.

Bone sialoprotein (BSP) cross-linked to collagen/gelatin was implanted in the pulp of rat's upper molars. Comparison was carried out with a sham group (non implanted), with a group of rats receiving the carrier alone, and a group of molars where the perforated pulps were capped with calcium hydroxide. The cavities were occluded with a glass-ionomer cement (GIC). After 8, 14 and 30 days respectively the rats were killed by intracardiac perfusion of the fixative and processed for light microscopy. Dentin and predentin debris pushed into the pulp during the preparation enhanced self-repair processes, with large pulp remnants. The carrier alone induced slight inflammation, and calcium hydroxide the formation of a reparative dentin bridge. BSP stimulated the recruitment of cells which produced an homogeneous atubular dentin-like structure, filling after one month the mesial third of the crown pulp. Osteogenic protein (OP-1) used in the same experimental conditions induced the formation of osteodentin in the coronal pulp and the radicular part of the pulp was totally filled by a mineralized material. The differences reported here suggest two possible different therapeutic approaches with the two osteogenic proteins, BSP inducing pulp mineralization in the crown part, and OP-1 occluding the root part of the pulp.

Differential repair responses in the coronal and radicular areas of the exposed rat molar pulp induced by recombinant human bone morphogenetic protein 7 (osteogenic protein 1)

Bone morphogenetic protein 7 (BMP 7), also termed osteogenic protein 1, a member of the transforming growth-factor superfamily, was examined for its efficacy in inducing reparative dentinogenesis in the exposed pulps of rat molars. To determine if the reaction was dose-dependent, collagen pellets containing 1, 3 or 10 μg of recombinant BMP 7 were inserted in intentionally perforated pulps (10–12 pulps per group) in the deepest part of half-moon class V-like cavities cut in the mesial aspect of upper first molars. As controls, the collagen carrier (CC group) alone and calcium hydroxide (Ca group) were used as capping agents. All cavities were then restored with a glass-ionomer cement. Half of the animals were killed after 8 days and the other half after 28 days, by intracardiac perfusion of fixative. The molars were processed for histological evaluation by light microscopy. No difference in effect could be detected between the three concentrations of BMP 7 groups at either time interval. After 8 days, all groups showed varying inflammation, from mild of severe, and the Ca group demonstrated early formation of a reparative dentine bridge. At 28 days the CC group displayed irregular osteodentine formation, leaving some unmineralized areas at the exposure site and interglobular unmineralized areas containing pulp remnants. In the Ca-treated pulps, the initial formation of thick reparative osteodentine bridges that sealed more or less completely the pulp perforation was followed, in the deeper part, by irregular tubular dentine. In most BMP 7-treated specimens, the initial inflammation has resolved at 8 days and at 28 days heterogeneous mineralization or osteodentine filled the mesial coronal pulp. They also had complete filling of the radicular pulp by homogenous mineralization in the mesial root; this reaction was found in 11 teeth in the BMP 7 group, one tooth in the CC group an none of the Ca group. These results emphasize the biological differences the coronal and radicular parts of the pulp, and the potential of bioactive molecules such as BMP 7 to provide an a alternative conventional endodontic treatments.

An Immunohistochemical Study of Osteoprotegerin in the Human Dental Pulp

This study investigated the expression of osteoprotegerin (OPG) in healthy and inflamed dental pulps. Histological sections 7 microm thick of 47 teeth, either caries-free or affected by gross caries, were used. Sections were stained with hematoxylin-eosin, and other sections of the same specimen were subjected to the avidin-biotin peroxidase complex immunohistochemical procedure for detection of OPG. The study focused on the coronal pulp that was divided into peripheral and central regions. In the peripheral pulp healthy and inflamed specimens showed high OPG immunoreactivity of the odontoblastic layer. When no inflammation was present in the central pulp OPG immunoreactivity was light. Fibroblasts and endothelial cells showed immunoreactivity ranging from none to intense. When inflammation was present in the central pulp the chronic inflammatory cells showed intense immunoreactivity.

Pattern of transmission of laser light through carious molar teeth.

The aims of this study were to investigate the pattern of transmission of laser light through carious human molar teeth, and to examine the effect on light transmission of reactions to caries in the dentine. Sectioned carious human molars were examined whilst illuminated from their external surfaces by a helium-neon laser probe of 0.5 mm diameter. The pattern of light transmission was observed and selected specimens were photographed. The passage of light through the surrounding dentine toward the pulp space was investigated by confocal microscopy. Further carious and restored teeth were illuminated from three buccal and three lingual sites to establish the probability of light being able to reach the coronal pulp space. For all 31 teeth examined, it was possible to illuminate the pulp space from a position on the buccal side. The presence of intracoronal restorations did not necessarily prevent light transmission to the pulp. Confocal microscopy demonstrated conduction of light within dystrophic deposits of irregular secondary dentine. The presence of carious lesions or restorations in molar teeth does not necessarily interfere with transmission of laser light to the pulp space. Light is conducted within irregular secondary dentine. With appropriate probe placement, assessments of pulp health in diseased posterior teeth may be possible using laser Doppler flowmetry.

Enamel matrix derivative promotes reparative processes in the dental pulp.

During odontogenesis, amelogenins from the preameloblasts are translocated to differentiating odontoblasts in the dental papilla, suggesting that amelogenins may be associated with odontoblast changes during development. In the present study, we have explored the effects of enamel matrix derivative (EMD) on the healing of a pulpal wound. Coronal pulp tissue of permanent maxillary premolars of miniature swine were exposed through buccal class V cavities. The exposed pulp was capped with EMD. The contralateral teeth served as controls and were capped with a calcium hydroxide paste (Dycal). The cavities were sealed with glass-ionomer cement. After 2 and 4 weeks, the histology of the teeth was analyzed. In the EMD-treated teeth, large amounts of newly formed dentin-like hard tissue with associated formative cells outlined the pulpal wound separating the cavity area from the remaining pulp tissue. Inflammatory cells were present in the wound area but not subjacent to the newly formed hard tissue. Morphometric analysis showed that the amount of hard tissue formed in EMD-treated teeth was more than twice that of the calcium-hydroxide-treated control teeth (p < 0.001), suggesting that EMD is capable of promoting reparative processes in the wounded pulp more strongly than is calcium hydroxide.

Localized intrinsic staining of teeth due to pulpitis and pulp necrosis in dogs.

Dental charts and records of 71 dogs were received and 84 intact tooth crowns were noted to have been entirely discolored in shades of pink, purple, grey, or tan. Endodontic therapy was performed on 49 of these teeth allowing gross examination of the entire pulp. In exploratory pulpotomy was performed in 15 teeth prior to exodontic therapy in order to assess the integrity of the coronal pulp. Gross signs of partial or total pulp necrosis were present in 59 (92.2%) of these teeth. The pulp was not observed grossly in 20 teeth which received exodontic therapy. Overall, of the 84 teeth evaluated, 36 (42.9%) had no radiographic signs of endodontic disease. Gross or radiographic signs of endodontic disease were absent in only 2 (4.1%) discolored teeth receiving endodontic therapy. Based on the low incidence of vital, affected teeth in the dogs of this study, endodontic on exodontic therapy is recommended for discolored teeth.

Characterization of α-smooth muscle actin positive cells in mineralized human dental pulp cultures

In response to injury, pulp precursor cells can differentiate into odontoblast-like cells that produce reparative dentine. In culture, pulp cells form mineralizing nodules, but the characteristics of the cells involved in this process are still not fully known. Human pulp cells for culture were obtained from coronal pulp isolated from non-erupted molars, and were maintained in RPMI 1640 medium supplemented with fetal calf serum. Nodules were forming in all human pulp primary cultures (HPPc) and human pulp subcultures observed until their fifth passage (HPSc<5). Mineralization of the nodules was confirmed by the presence of calcium and phosphate that were quantified by X-ray microanalysis. Specific immunolabeling revealed α-smooth muscle actin and vimentin in both HPPc and HPSc<5 cells. Cells positive for α-smooth muscle actin were either isolated or gathered together in the nodules. Under transmission electron microscopy, some cells in primary pulp cultures exhibited features typical of myofibroblasts or pericytes, such as stress fibers, fibronexus, indented nuclei and gap-junctions. These cells were frequently in close contact with mineral deposits. This work demonstrates for the first time the presence of pericytes or myofibroblasts in mineralized human pulp cultures, but further investigation is required to determine their origin, role and degree of differentiation.

Tablet fluoridation influences the calcification of primary tooth pulp.

This study was conducted to determine the influence of long-term tablet fluoridation on primary pulp calcification by light microscopy. Twenty-four caries-free primary molars (after continuous postpartally initiated 1- to 10-year tablet fluoridation) were compared to 17 primary molars of children without fluoride prophylaxis. Pulp calcification in children with tablet fluoridation was significantly more frequent and more pronounced than in untreated children (p = 0.001). Besides the known pulp stones, the prophylaxis group evidenced a special form of calcification consisting of fibrodentin-like hard tissue not observed in the untreated children. These hard tissue bodies developed "intramurally" on the pulp floor and the inside of the dental roots with an irregular extramural spread into the coronal and radicular pulp by displacement and fibrotization of the pulp tissue. Moreover, some of the teeth had more or less extensive areas of interglobular dentin. The affected teeth were ankylosed in the area of the bi- and trifurcation and on the inside of the roots and were thus infra-occluded. Although the duration of tablet fluoridation has no statistically significant influence on pulp calcification, there is a correlation between extensive pulp calcification, postnatally initiated fluoride prophylaxis and the infraocclusion of primary molars.

Alteration in the expression of heat shock protein (Hsp) 25-immunoreactivity in the dental pulp of rat molars following tooth replantation.

The regeneration process of dental pulp following tooth replantation in rat molars was investigated by immunocytochemistry for heat shock protein (Hsp) 25 and protein gene product 9.5 (PGP 9.5). In control teeth at postnatal 4 weeks, the odontoblasts showed intense Hsp 25-immunoreactivity in the coronal dental pulp, but little or no immunoreactivity in the root and floor pulp. In contrast, the Hsp 25-negative odontoblasts in the latter areas displayed immunoreactivity for PGP 9.5. Tooth replantation caused loss of Hsp 25- and PGP 9.5-immunoreactions in the dental pulp during postoperative days 1-3. At postoperative day 5, plump cells with clear nucleoli and several fine processes--presumably newly differentiated odontoblasts--at the pulp-dentin border became immunopositive for Hsp 25. These data suggest that the expression of Hsp 25- and PGP 9.5-immunoreactivity reflects the status of differentiation of the odontoblasts. Furthermore, some pulpal nerve fibers as well as the Schwann cells in the dental pulp, ordinarily negative in Hsp 25-immunoreaction, acquired their immunoreactivity by postoperative day 5, but lost it thereafter, suggesting the involvement of Hsp 25 in the regeneration of pulpal nerve fibers. In the case of bone-like tissue formation in the pulp space, on the other hand, no Hsp 25-immunoreactive odontoblasts were recognized in the pulp-dentin border. Thus, the alignment of Hsp 25-immunopositive odontoblasts along the pulp-dentin border indicates a decisive factor for inducing the reparative dentin formation after tooth replantation.