Lara T. Friedlander

Dental stem cells and their potential role in apexogenesis and apexification

Injury to an immature permanent tooth may result in cessation of dentine deposition and root maturation leaving an open root apex and thin dentinal walls that are prone to fracture. Endodontic treatment is often complicated and protracted with an uncertain prognosis frequently resulting in premature tooth loss. Postnatal stem cells, which are capable of self-renewal, proliferation and differentiation into multiple specialized cell lineages have been isolated and identified within the dental pulp, apical papilla and periodontal ligament. The ability of these cells to produce pulp-dentine and cementum-periodontal ligament complexes in vivo suggest potential applications involving stem cells, growth factors and scaffolds for apexification or apexogenesis. Similar protein expression amongst dental stem cells possibly implicates a common origin; however, the dominant cells to repopulate an open apex will be directed by local environmental cues. A greater understanding of the structure and function of cells within their environment is necessary to regulate and facilitate cellular differentiation along a certain developmental path with subsequent tissue regeneration. This review focuses on development of the apical tissues, dental stem cells and their possible involvement clinically in closing the open root apex. MEDLINE and EMBASE computer databases were searched up to January 2009. Abstracts of all potentially relevant articles were scanned and their contents identified before retrieval of full articles. A manual search of article reference lists as well as a forward search on selected authors of these articles was undertaken. It appears that dental stem cells have the potential for continued cell division and regeneration to replace dental tissues lost through trauma or disease. Clinical applications using these cells for apexogenesis and apexification will be dependent on a greater understanding of the environment at the immature root end and what stimulates dental stem cells to begin dividing and then express a certain phenotype.

Expression of Toll-like receptors 2 and 4 and the OPG-RANKL-RANK system in inflammatory external root resorption and external cervical resorption

AIM To investigate and compare the expression of the pathogen recognition receptors Toll-like receptor (TLR) 2 and TLR4, and the hard tissue resorption triad osteoprotegerin (OPG)-receptor activator of nuclear factor kappa-B ligand (RANKL)-receptor activator of nuclear factor kappa-B (RANK) in external inflammatory root resorption of endodontic origin (ER) and external cervical root resorption (ECR) by immunohistochemistry. METHODOLOGY Formalin-fixed, paraffin-embedded archival specimens collected from teeth that were diagnosed clinically, radiographically and histopathologically with either ER (n = 9) or ECR (n = 9) were processed for immunohistochemistry to investigate and compare levels of TLR2, TLR4, OPG, RANKL, RANK, CD3, CD19 and CD83 expression. The histological features were evaluated via haematoxylin and eosin stain. Taylors modification of the Brown and Brenn Gram stain was used for examining the presence and distribution of bacteria. All stained slides were digitally photographed and qualitatively analysed, and F test and unpaired Students t-test were used for statistical analysis. RESULTS Both ER and ECR showed similar immuno-histopathology characteristics of a fibrovascular connective tissue with varying degrees of inflammatory infiltrate consisting of T and B lymphocytes, dendritic cells, polymorphonuclear lymphocytes and plasma cells. Colonies of bacteria were identified in the majority of lesions, and this correlated with the cellular expression of TLR2 and TLR4 in all lesions. Similarly, all lesions showed a significantly higher (P < 0.05) level of cells expressing RANKL than OPG, indicating hard tissue resorption processes where active in the lesions. CONCLUSION The immunohistopathology patterns of ECR samples were consistent with the bacteria-driven ER specimens, suggesting bacteria-induced inflammation may be involved in ECR.

VEGF and VEGFR2 in dentigerous cysts associated with impacted third molars

Summary The aims of this study were to determine the presence and distribution of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR2) in dentigerous cysts compared with normal dental follicles as a control tissue and to evaluate endothelial cells and proliferating cells as indicators of angiogenic activity in these tissues. Twenty specimens histologically diagnosed as dentigerous cysts and 20 dental follicle specimens were included. Immunohistochemistry (IHC) using anti-VEGF and anti-VEGFR2 antibodies stained for the growth factor and its receptor, while anti-CD34 and anti-CD146 antibodies were used to identify endothelial cells. Anti-proliferating cell nuclear antigen (PCNA) antibody detected proliferating cells within the specimens. Slides were examined microscopically and results evaluated using kappa statistics, negative binomial regression and ordinal logistic regression. The mean age for patients with dentigerous cysts was 23 years and they were more common in males. Proteins for VEGF, VEGFR2, PCNA, CD34, and CD146 were expressed in all dentigerous cysts and dental follicles. VEGF and VEGFR2 were expressed on several cell types within the tissues, however there was a significantly greater percentage of positive staining in dentigerous cysts compared with dental follicles (odds ratio = 31.24, p < 0.001). CD34+, CD146+, and PCNA+ cells were observed in both dentigerous cysts and dental follicles but for all markers there were significantly more positive cells in dentigerous cysts (p < 0.001); this was especially evident in cases associated with inflammation. PCNA was seen in most endothelial cells lining small thin walled blood vessels suggesting endothelial proliferation. There was a high level of intra- and inter-examiner agreement (kappa 0.77 and 0.75, respectively). VEGF and VEGFR2 and angiogenic activity are present in dental follicles and dentigerous cysts and may contribute to local bone resorption for tooth eruption or the development and progression of dentigerous cysts.

Avulsion and replantation of a primary incisor tooth

Avulsion of a primary tooth is a serious dental trauma, and the guidelines of the International Association of Dental Traumatology and textbooks in paediatric dentistry do not recommend replantation. Such management can result in severe damage to the supporting structures, and together with avulsion itself is commonly associated with developmental disturbances of the permanent tooth. We report the case of replantation in a 9-month-old child with a successful outcome, in a unique situation where conditions were optimal and careful long-term follow up was possible.

The anatomy of the root apex: A review and clinical considerations in endodontics

Studies on the anatomy of the root apex are an area of interest to the endodontist; they have reported that the position of the apical constriction, apical foramen and the cemento-dentinal junction varies across the tooth types. These anatomical apical landmarks are considered extension limits for root canal instrumentation and filling. Achieving an optimum working length is thought essential for successful root canal treatment, so adopting any of these landmarks is associated with certain risks and benefits. The variability in the position of the apical constriction and apical foramen, for example, complicates their clinical detection, while the cemento-dentinal junction is a histological landmark that cannot be detected clinically. The radiographic apex does not always coincide with the anatomic apex of the tooth. The pre-operative status of the pulp must be considered while obtaining the working length. Most prognostic studies agree that extending the root filling to within 2-3 mm of the radiographic apex is associated with favorable treatment outcomes.

Influence of clinical experience on the radiographic determination of endodontic working length.

AIM To determine the influence of clinical experience on the accuracy and consistency of estimation of radiographic working length (WL) for the root canal treatment of single-rooted teeth. METHODOLOGY Forty conventional WL periapical radiographs that included variations in file length were selected. They were digitally scanned and arranged in PowerPoint presentations on CDs. These were distributed to three assessor groups; fourth-year undergraduates at two stages of training (Groups 1 and 2) and endodontic postgraduates (Group 3). Participants were asked to determine the adjustment needed in millimetres to position the file tip at the correct WL for each image. A gold standard file position was provided by three experienced endodontists. For inter-group comparison of scores, the Kruskal-Wallis, ANOVA and post hoc Bonferroni tests were used. Evaluation of intra-examiner consistency was with the Kappa test. To evaluate intra-group consistency, the Wilcoxon signed rank test was used to compare the frequency of weighted correct scorings. RESULTS File adjustments of Group 3 were significantly more accurate than those of Group 1 (P = 0.006). The scores of Group 3 were also better than those of Group 2, although the difference was not significant. When the scores of the undergraduate groups were compared, the difference was not statistically significant. The consistency of the groups was not affected by a 2-week pause between assessments, and no definite pattern could be detected across any of the groups with the Kappa test. CONCLUSION Clinical experience after graduation influenced the accuracy of estimating the adjustments needed for correct radiographic WL of single-rooted teeth. The most experienced group was significantly more accurate than the other groups.

Radiographic assessment of endodontic working length

The use of radiographs for working length determination is usual practice in endodontics. Exposing radiographs following the principles of the paralleling technique allows more accurate length determination compared to the bisecting-angle method. However, it has been reported that up to 28.5% of cases can have the file tip extending beyond the confines of the root canals despite an acceptable radiographic appearance. The accuracy of radiographic working length determination could be affected by the location of the apical foramen, tooth type, canal curvature and superimposition of surrounding structures. Variations among observers by virtue of training and experience may also influence the accuracy of the procedure. The interpretation of radiographs could be affected by film speed and viewing conditions, with the superiority of digital imaging over conventional radiography for working length determination remaining debatable. The combination of several methods is recommended for acquiring the most accurate working length.