Shizuko Yamauchi

Tissue Engineering Strategies for Immature Teeth with Apical Periodontitis

INTRODUCTION Regenerative endodontic treatment on immature teeth with apical periodontitis is promising but still not well-established. The purpose of this study was to explore novel strategies to engineer a vital support structure within a root canal space by a combination of induced blood clot, exposure of dentin matrix, and a cross-linked collagen scaffold. METHODS Apical periodontitis was induced in 6 dogs with immature teeth (n = 64). After disinfection, the following groups were randomly assigned: blood clot (BC) alone, BC with a cross-linked collagen scaffold (CCS), BC with exposure of dentin matrix by ethylenediaminetetraacetic acid (EDTA), and BC with CCS and EDTA. Positive (infected only) and negative controls (untreated) were also included. The dogs were followed up for 3.5 months and killed. Periradicular healing and root wall thickening were radiographically analyzed and statistically evaluated. The jaws were then fixed, demineralized, and subjected to histologic analyses. Newly formed mineralized tissues were histomorphometrically analyzed, quantified, and statistically evaluated. RESULTS Radiographically there was significant difference in periradicular healing and root wall thickening (P < .05). Histomorphometric analysis showed significantly more mineralized tissue formation in the groups containing the scaffold (P < .05). Exposure of the dentin matrix by EDTA appeared to increase the adherence of the newly formed mineralized tissue to the root walls. CONCLUSIONS The use of cross-linked collagen scaffold and exposure of dentin matrix combined with blood clot might provide an efficient approach to generate a vital support structure for the treatment of immature teeth with apical periodontitis.

Immunohistological Characterization of Newly Formed Tissues after Regenerative Procedure in Immature Dog Teeth

INTRODUCTION In a previous report, we showed that 2 types of mineralized tissues were formed in the canal spaces of dogs after tissue engineering treatments of immature teeth with apical periodontitis: (1) dentin- associated mineralized tissue (DAMT) and (2) bony islands (BIs). The objective of this study was to characterize these mineralized tissues. METHODS The maturation and organization of collagen matrices in DAMT, BIs, and the interface between DAMT and the dentin wall were characterized using a histochemical method with picrosirius red staining under polarized light microscopy. In addition, the distribution of 2 noncollagenous proteins (ie, dentin sialoprotein and bone sialoprotein) in these tissues was investigated by immunohistochemical methods with specific antibodies. RESULTS The results showed that DAMT is distinct from dentin, bone, or BIs. Although it resembled cementum to an extent showing similar immunoreactivity to the noncollagenous proteins, the organization and maturation of collagen matrix was significantly different from cementum. BIs resembled a bone matrix in terms of morphology, collagen organization, and immunoreactivity. CONCLUSIONS The results indicate that DAMT and BIs formed in the canal space are distinct from each other, one exhibiting a unique mineralized tissue and the other a bone-like tissue.

Effects of gender and acute dental pain on thermal pain responses

OBJECTIVE Considerable research suggests that females exhibit greater sensitivity to laboratory pain procedures than do males; however, whether the presence of acute clinical pain influences this sex difference in pain sensitivity has not been investigated. The present experiment investigated the effects of sex and acute dental pain on laboratory pain responses. DESIGN Thermal pain onset and tolerance were determined in 46 dental patients (15 male, 31 female) experiencing pain due to acute irreversible pulpitis and in 33 healthy controls (13 male, 20 female). In addition, measures of mood and coping were obtained in all participants. All subjects participated in two experimental sessions. The first session took place immediately before the patients underwent endodontic treatment for relief of pulpal pain. The second session took place approximately 1-2 weeks later, when pulpitis patients were pain free after treatment. During each session, thermal pain onset and tolerance were assessed with a 1-cm2 contact thermode applied to the right volar forearm using an ascending method of limits. RESULTS During both sessions, thermal pain onset and tolerance were lower in control females than in control males; however, male and female pulpitis patients did not differ in their thermal pain responses during either session. Pulpitis patients also showed greater affective distress than controls. CONCLUSIONS These data suggest that the sex difference in thermal pain sensitivity frequently reported in pain-free subjects appears to be absent in patients presenting with acute dental pain. However, this effect cannot be explained solely based on the presence of clinical pain because the effect on pain threshold and tolerance persisted into session 2, when pulpitis patients were pain free. Potential explanations for these results are discussed.

Identification, Partial Characterization, and Distribution of Versican and Link Protein in Bovine Dental Pulp

The dynamics of changes in the cellularity and extracellular matrix composition of dental pulp varies considerably during tooth development and maturation. In this paper, we studied matrix proteoglycans where we hypothesized that they played important roles in structural, spatial, and transport aspects of pulpal development and maintenance. The pulpal tissue was collected from partially erupted bovine incisors, pulverized, and then extracted with 6 M guanidine-HCl. The extract was subjected to anion column chromatography (DEAE-8HR), and the fractions collected were screened by dot-blot immunoassay by means of monoclonal antibodies generated against 4- and 6-sulfated chondroitin sulfate isomers, and keratan sulfate, 2-B-6, 3-B-3, and 5-D-4, respectively. The chondroitin-6-sulfate was the major glycosaminoglycan species and occurred as a large-molecular-weight proteoglycan (> 500 kDa). After further purification, it was subjected to agarose/acrylamide composite gel electrophoresis, and it migrated as a single band stained with Stains-All. The band was immunopositive against antibody 3-B-3 by Western blot analysis. The partial amino acid sequence analyses of the core protein clearly indicated this molecule to be versican. The presence of link protein was also confirmed by Western blot analysis with an anti-link protein monoclonal antibody, 8-A-4. Furthermore, immunohistochemical study indicated that the distributions of versican and link protein coincide in the dental pulp and are enriched in the peripheral area of the tissue just beneath the odontoblast layer. Since the dental pulp contains hyaluronan, versican may bind to hyaluronan via its hyaluronan-binding domain, where this association is stabilized by link protein. This complex, then, could form large hydrated proteoglycan aggregates that fill the extracellular space, support odontoblasts, and/or facilitate the transport function of metabolites and nutrients within the tissue.