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Unlike bones, teeth are remarkably resilient and can withstand severe trauma, making age assessment based on the dentition essential for forensic analysis. Modern techniques for age estimation focus on pulp-chamber volume measurements using radiographs and computerized tomography (CT); however, these are applicable only for complete teeth (i.e., with intact crown and root). In the current study, we developed a new approach using high-resolution micro-computerized tomography (μCT) to visualize the secondary dentin (SD), an inner layer surrounding the pulp which accumulates with age, thus facilitating age estimation of fragmented and broken teeth.The growth pattern of the SD with age was analyzed for 77 lower premolars from two anthropological collections. A comparison of SD virtual segmentation and histological measurement was highly correlative (ICC = 0.95). SD was measured per volume (mm 3 ) of a 1 mm thick slice directly below the cemento-enamel junction. Regression analysis using SD measurements increased the success rates of age estimation (82%) compared with the "gold-standard" pulp/dentin method (54%) in the range of ± 10 years. The accuracy of age estimation based on SD analysis was improved to a range of 7-8 years.The SD method thus allows age estimation with greater prediction rates and better accuracy based on only a small fragment of a tooth in a non-invasive manner. This novel methodology is easy to use, accessible, and bears implications in various fields such as forensic sciences and anthropological research.

Aim: To investigate whether static magnetic fields (SMFs) have a positive effect on the migration and dentinogenesis of dental pulp stem cells (DPSCs) to promote reparative dentine formation., Methodology: In vitro scratch assays and a traumatic pulp exposure model were performed to evaluate the effect of 0.4-Tesla (T) SMF on DPSC migration. The cytoskeletons of the DPSCs were identified by fluorescence immunostaining and compared with those of a sham-exposed group. Dentinogenic evaluation was performed by analysing the expressions of DMP-1 and DSPP marker genes using a quantitative real-time polymerase chain reaction (qRT-PCR) process. Furthermore, the formation of calcified deposits was examined by staining the dentinogenic DPSCs with Alizarin Red S dye. Finally, the role played by the p38 MAPK signalling pathway in the migration and dentinogenesis of DPSCs under 0.4-T SMF was investigated by incorporating p38 inhibitor (SB203580) into the in vitro DPSC experiments. The Student's t-test and the Kruskal-Wallis test followed by Dunn's post hoc test with a significance level of P < 0.05 were used for statistical analysis., Results: The scratch assay results revealed that the application of 0.4-T SMF enhanced DPSCs migration towards the scratch wound (P < 0.05). The cytoskeletons of the SMF-treated DPSCs were found to be aligned perpendicular to the scratch wound. After 20 days of culture, the SMF-treated group had a greater number of out-grown cells than the sham-exposed group (nonmagnetized control). For the SMF-treated group, the DMP-1 (P < 0.05) and DSPP genes (P < 0.05), analysed by qRT-PCR, exhibited a higher expression. The distribution of calcified nodules was also found to be denser in the SMF-treated group when stained with Alizarin Red S dye (P < 0.05). Given the incorporation of p38 inhibitor SB203580 into the DPSCs, cell migration and dentinogenesis were suppressed. No difference was found between the SMF-treated and sham-exposed cells (P > 0.05)., Conclusion: 0.4-T SMF enhanced DPSC migration and dentinogenesis through the activation of the p38 MAPK-related pathway., (© 2018 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

There is a need for dental age estimation methods after completion of the third molar mineralization. Degenerative dental characteristics appear to be suitable for forensic age diagnostics beyond the 18th year of life. In 2012, Olze et al. investigated the criteria studied by Gustafson using orthopantomograms. The objective of this study was to prove the applicability and reliability of this method with a large cohort and a wide age range, including older individuals. For this purpose, 2346 orthopantomograms of 1167 female and 1179 male Germans aged 15 to 70 years were reviewed. The characteristics of secondary dentin formation, cementum apposition, periodontal recession and attrition were evaluated in all the mandibular premolars. The correlation of the individual characteristics with the chronological age was examined by means of a stepwise multiple regression analysis, in which the chronological age formed the dependent variable. Following those results, R 2 values amounted to 0.73 to 0.8; the standard error of estimate was 6.8 to 8.2 years. Fundamentally, the recommendation for conducting age estimations in the living by these methods can be shared. The values for the quality of the regression are, however, not precise enough for a reliable age estimation around regular retirement date ages. More precise regression formulae for the age group of 15 to 40 years of life are separately presented in this study. Further research should investigate the influence of ethnicity, dietary habits and modern health care on the degenerative characteristics in question.

Objective: The present study aims to investigate whether reparative dentinogenesis could be guided at central pulpal sites or at a distance from the amputated pulp of miniature pig teeth, by using set calcium silicate-based carriers containing human recombinant bioactive molecules., Design: Pulp exposures were performed in 72 permanent teeth of 4 healthy miniature swine. The teeth were capped with pre-manufactured implants of set calcium silicate-based material containing BMP-7, TGFβ1 or WnT-1, for 3 weeks. Conical-shaped intrapulpal implants were exposed in the central pulp core, while disc-shaped extrapulpal implants were placed at a distance from the amputated pulp. Implants without bioactive molecules were used as controls. Thickness and forms of new matrix mineralized deposition were assessed histologically at post-operative periods of 3 weeks by light microscopy., Results: Intrapulpal applications: Calcified structures composed of osteodentine were found in contact with the BMP-7 implants. An inhomogeneous calcified tissue matrix was found around the WnT-1 carriers. A two-zone calcified structure composed of osteodentine and a thicker tubular matrix zone was seen at the TGFβ1 carrier-pulp interface. Extrapulpal applications: The space between WnT-1 implants and pulp periphery had been invaded by soft tissue with traces of calcified foci. Thick calcified structures composed of osteodentine were found surrounding pulp exposure sites in response to application of BMP-7. Spindle-shaped cells associated with atubular calcified matrix or elongated polarized cells associated with tubular dentine-like matrix were found along the cut dentinal walls of the TGFβ1 group., Conclusion: The present experiments indicated that set calcium silicate could be used as carrier for biologically active molecules. TGFβ1 was shown to be an effective bioactive molecule in guiding tertiary dentine formation., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Introduction: The purpose of this study was to compare the effectiveness of mineral trioxide aggregate (MTA) and calcium hydroxide (CH) as pulp capping materials in humans by means of a meta-analysis., Methods: The PubMed, Cochrane Library, Embase, and Web of Knowledge databases were used in the literature search from their establishment date until December 7, 2014. Studies that met the inclusion criteria were accepted, and necessary information was extracted by 2 authors independently using a standardized form. The success rate, inflammatory response, and dentin bridge formation were evaluated., Results: Thirteen studies met the inclusion criteria. There was no significant heterogeneity between studies, so a fixed-effects model was used. The MTA treatment groups showed a significantly higher success rate compared with CH-capped groups (randomized controlled trials: odds ratio [OR] = 2.26; 95% confidence interval [CI] = 1.33-3.85; P = .003; retrospective nonrandomized trials: OR = 2.88; 95% CI, 1.86-4.44; P < .00001). MTA was superior to CH in terms of the absence of an inflammatory response as well as dentin bridge formation, with the OR being 4.56 (95% CI, 2.65-7.83) and 3.56 (95% CI, 1.89-6.70), respectively., Conclusions: MTA has a higher success rate and results in less pulpal inflammatory response and more predictable hard dentin bridge formation than CH. MTA appears to be a suitable replacement of CH used for direct pulp capping., (Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Many dental pulp stem cells are neural crest derivatives essential for lifelong maintenance of tooth functions and homeostasis as well as tooth repair. These cells may be directly implicated in the healing process or indirectly involved in cell-to-cell diffusion of paracrine messages to resident (pulpoblasts) or nonresident cells (migrating mesenchymal cells). The identity of the pulp progenitors and the mechanisms sustaining their regenerative capacity remain largely unknown. Taking advantage of the A4 cell line, a multipotent stem cell derived from the molar pulp of mouse embryo, we investigated the capacity of these pulp-derived precursors to induce in vivo the formation of a reparative dentin-like structure upon implantation within the pulp of a rodent incisor or a first maxillary molar after surgical exposure. One month after the pulp injury alone, a nonmineralized fibrous matrix filled the mesial part of the coronal pulp chamber. Upon A4 cell implantation, a mineralized osteodentin was formed in the implantation site without affecting the structure and vitality of the residual pulp in the central and distal parts of the pulp chamber. These results show that dental pulp stem cells can induce the formation of reparative dentin and therefore constitute a useful tool for pulp therapies. Finally, reparative dentin was also built up when A4 progenitors were performed by alginate beads, suggesting that alginate is a suitable carrier for cell implantation in teeth., (Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

The ultimate goal of vital pulp therapy is to regenerate rapidly dentin possessing an excellent quality using a biocompatible, bioactive agent. Dentin phosphophoryn (DPP), the most abundant noncollagenous polyanionic protein in dentin, cross-linked to atelocollagen fibrils was applied to direct pulp capping in rats. After 1, 2, and 3 weeks, the teeth applied were examined on the induction of reparative dentin formation and the response of pulp tissue, compared to calcium hydroxide-based agent conventionally used. The reparative dentin formation induced by DPP/collagen composite was more rapid than by calcium hydroxide. In the morphometrical analysis, the formation rate of reparative dentin by DPP/collagen composite was approximately the same as that by calcium hydroxide at 3 weeks. Nevertheless, the compactness of reparative dentin formed by DPP/collagen composite was much superior to what resulted from calcium hydroxide. Also, DPP/collagen composite showed high covering ability of exposed pulp. Moreover, DPP/collagen composite led only to slight pulp inflammation at the beginning whereas calcium hydroxide formed necrotic layer adjacent to the material and induced severe inflammation in pulp tissue at 1 week. The present study demonstrates a potential for DPP/collagen composite as a rapid biocompatible inducer for the formation of reparative dentin of excellent quality in rats.

Calcium hydroxide has been used extensively in dentistry for a century. Despite its widespread use as a pulp-capping agent, its mechanisms of action still remain ambiguous. Understanding its modes of action will lead to a broader understanding of the mechanisms associated with induced dentinogenesis and help in optimizing the currently available agents to target specific regenerative processes to obtain the best possible clinical outcomes. A literature search relating to mechanisms of dentinogenesis of calcium hydroxide up to December 2011 was carried out using pubmed and MEDLINE database searches as well as manual searching of cross-references from identified studies. Resulting suggestions regarding dentinogenic mechanisms of calcium hydroxide range from direct irritating action of the material to induction of release of biologically active molecules. The purpose of this article is to discuss various mechanisms through which calcium hydroxide may induce tertiary dentinogenesis in the light of observations made in included studies., (© 2012 International Endodontic Journal.)

We evaluated the effect of calcium hydroxide cement (CHC) and glass ionomer cement (GIC) on carious dentin and bacterial infections after partial caries removal and sealing. Sixty permanent teeth with deep lesions underwent partial caries removal, the application of CHC, GIC or wax, i.e. negative control (NC), and were then sealed for 3-4 months. After the partial caries removal and the sealing period, the dentin was clinically assessed (colour and consistency) and analysed by scanning electron microscopy to assess dentin organization and bacterial infections. The effect of the treatment in each group was assessed by the Wilcoxon and χ2 tests, differences among groups by the Kruskal-Wallis test and the correlations between variables by Spearman correlation. No clinical symptoms or radiographic signals of pulpits or pulp necrosis were observed during the study. Dentin darkening was observed after the sealing period in the CHC and NC groups (p < 0.05). However, there was no difference in the colour after treatment among the 3 groups (p > 0.05). Dentin hardening occurred in all groups after treatment (p < 0.05), also with no differences (p > 0.05). Dentin samples showed better organization after the sealing period than after partial caries removal, with total or partial obliteration of dentinal tubules (CHC p < 0.03, GIC p < 0.05, NC p < 0.01) and a reduction of bacterial infections (CHC p < 0.03, GIC p < 0.05, NC p < 0.03). No differences were observed. Correlations between the different criteria, except for colour and bacterial infection, were detected in all cases. Partial caries removal and sealing resulted in dentin hardening, decreased bacterial numbers and dentin reorganization, irrespective of the dentin protection used., (Copyright © 2013 S. Karger AG, Basel.)

Aim: To compare the long-term clinical and radiographic outcomes of pulpotomies in primary molars performed with white or gray Mineral Trioxide Aggregate (MTA) in combination with ferric sulfate (FS), when one package of MTA is used for multiple treatments., Design: Sixty eight children with 86 vital carious primary molars underwent pulpotomy with FS, and grey or white MTA. One package of MTA was used for 7-8 treatments. Clinical and radiographic evaluation was performed before and 6 to 47 months after treatment., Results: Success rates were similar for pulpotomies performed with white (60-teeth) and grey MTA (16 teeth) (p > 0.05), and for those performed with the addition of FS to white or gray MTA when one package of MTA was used for multiple pulpotomies compared to one package of MTA alone., Conclusion: Gray and white MTA in conjunction with FS induce comparable clinical and radiographic success rate. The use of one package of MTA for multiple pulpotomies, combined with FS, is a cost-effective treatment.

Aim: To examine the feasibility of using the pOBCol3.6GFPtpz [3.6-green fluorescent protein (GFP)] transgenic mice as an in vivo model for studying the biological sequence of events during pulp healing and reparative dentinogenesis., Methodology: Pulp exposures were created in the first maxillary molar of 12-16-week-old 3.6-GFP transgenic mice with CD1 and C57/Bl6 genetic background. Direct pulp capping on exposed teeth was performed using mineral trioxide aggregate followed by restoration with a light-cured adhesive system (AS) and composite resin. In control teeth, the AS was placed in direct contact with the pulp. Animals were euthanized at various time points after pulp exposure and capping. The maxillary arch was isolated, fixed and processed for histological and epifluorescence analysis to examine reparative dentinogenesis., Results: Analysis of teeth immediately after pulp exposure revealed absence of odontoblasts expressing 3.6-GFP at the injury site. Evidence of reparative dentinogenesis was apparent at 4 weeks in 3.6-GFP mice in CD1 background and at 8 weeks in 3.6-GFP mice with C57/Bl6 background. The reparative dentine with both groups contained newly formed atubular-mineralized tissue resembling a dentine bridge and/or osteodentine that was lined by cells expressing 3.6-GFP as well as 3.6-GFP expressing cells embedded within the atubular matrix., Conclusion: This study was conducted in a few animals and did not allow statistical analysis. The results revealed that the 3.6-GFP transgenic animals provide a unique model for direct analysis of cellular and molecular mechanisms of pulp repair and tertiary dentinogenesis in vivo. The study also shows the effects of the capping material and the genetic background of the mice in the sequence and timing of reparative dentinogenesis., (© 2012 International Endodontic Journal.)

The main criteria used in dental age estimation in living young individuals are mineralisation and eruption of the third molars. In order to further diversify the spectrum of characteristics after completion of third molar development, tests were undertaken to determine whether the characteristics studied by Gustafson can be ascertained with the required forensic certainty with the aid of the orthopantomogram and whether the evaluation of these could render forensic odontological age diagnoses possible beyond the 18th year of life. For this purpose, 1,299 conventionally produced orthopantomograms of 650 female and 649 male Germans aged from 15 to 40 years were studied. The characteristics of secondary dentin formation, cementum apposition, periodontal recession and attrition were evaluated in all the mandibular premolars. The correlation of the individual characteristics with chronological age was studied with the aid of a multiple regression analysis in which chronological age formed the dependent variable. Depending on the tooth studied, the R values amounted to 0.65 to 0.73; the standard error of estimate was 5.3 to 5.7 years. Basically, the regression equations calculated can be recommended for age estimation in living individuals, although it should be borne in mind that the applicability of the new method presented is limited by the quality of the X-ray images.

Introduction: Vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP 2) are growth factors (GFs) identified within the dentine-pulp complex and involved into the cellular events connected to the pulp-healing response. It is well established that the expression of these GFs is increased in different tissues in diabetes mellitus. Because there are no data concerning the levels of VEGF and BMP 2 in human dental pulp, the aim of present study was to quantify VEGF and BMP 2 levels in intact dental pulp and dental pulp that underwent reactive dentinogenesis in healthy and diabetic human subjects., Methods: The study was conducted on 28 healthy and 28 subjects with controlled diabetes type II who underwent pulp extirpation as a part of prosthetic rehabilitation. Pulp were collected from intact teeth and teeth treated by indirect pulp capping. The levels of VEGF and BMP 2 were determined in the pulp tissue lysates with enzyme-linked immunosorbent assay., Results: The levels of VEGF and BMP 2 were significantly higher in intact teeth pulp of diabetic than in healthy subjects. The concentrations of these GFs were significantly lowered in teeth with indirect pulp capping both in healthy and diabetic persons. Furthermore, VEGF and BMP 2 levels were in strong positive correlation., Conclusions: Similar changes in the levels of VEGF and BMP 2 in intact and treated teeth of healthy and diabetic patients could be suggestive of associated roles of these GFs in responses of healthy and diabetic dental pulp., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: It is suggested that dental pulp stem cells are involved in tooth regeneration and play an important role in maintaining pulp homeostasis. Previously, normal dental pulps were more widely used for experimental models than carious dental pulps. The aim of this study was to isolate and culture the dental pulp stem cells from carious and normal teeth and to evaluate stem cell parameters., Methods: Pulp tissues were obtained and dissociated from normal and carious teeth. Single-cell suspensions were seeded into 6-well plates and purified by collecting multiple colonies. Normal dental pulp stem cells (DPSCs) and carious dental pulp stem cells (CDPSCs) were compared for morphologic appearance and for their capacity to differentiate into 3 lineages. Colony-forming and MTT assays, cell cycle analysis, gene expression, and alkaline phosphatase activity were also evaluated., Results: Stem cells were cultured successfully from normal and carious dental pulps. CDPSCs displayed increased proliferation ability compared with DPSCs. CDPSCs also showed enhanced ALP activity, mineralization ability, and expression of osteogenesis/dentinogenesis-related genes. All cultures differentiated into 3 cell types., Conclusions: Our data suggest that caries as a local microenvironment should be taken into account when DPSCs are intended to be used for investigations and application. Furthermore, the mechanism of the underlying changes in cell properties requires further study., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: The equilibrium of inhibitors and inducers plays an important role in the mineralization of dentin during dentinogenesis. High-temperature requirement protein A1 (HtrA1) is a novel bone mineralization inhibitor involved in physiological and pathological bone formation. However, the expression of HtrA1 in tooth mineralization is unknown. The purpose of the present study was to investigate the localization and quantity of HtrA1 and its possible related molecules during induced reparative dentin formation., Methods: Rats were randomly sacrificed after direct pulp capping on days 0, 7, 14, and 21. Maxillary segments were obtained and routinely prepared for histological analysis, immunohistochemistry, quantum dots-based double immunofluorescence, and CRi's Nuance imaging system (CRI, Woburn, MA)-based quantitative determination., Results: The square measure values of reparative dentin significantly increased on day 7 and continued to increase until day 21. HtrA1, matrix Gla protein (MGP), nestin, and bone sialoprotein were positively stained and colocalized in the odontoblasts and/or odontoblast-like cells zone and reparative dentin during induced reparative dentin formation. The expressions of HtrA1 and MGP were significantly enhanced after direct pulp capping on day 7 and did not significantly change between days 7, 14, and day 21. Both expressions of HtrA1 and MGP were positively correlated with the square measure values of reparative dentin; however, no correlation was found between the expressions of HtrA1 and MGP., Conclusions: HtrA1 could be observed and might possibly be involved in the process of reparative dentin formation associated with MGP., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This study was designed to compare the thickness of dentin formed associated with autogenous tooth transplantation in dogs, using either one- or two-stage surgical techniques. The study consisted of three Beagles, older than 5 months, in which six incisors and six premolars were transplanted to mechanically prepared recipient sockets. One group was transplanted using a one-stage method to recipient beds prepared immediately before transplantation. The second groups of teeth were transplanted using a two-stage method in which the recipient beds were prepared and left to heal for 7 days before transplantation. Dogs were injected with xylenol orange, calcein and oxytetracycline at 2 days before, 3 and 9 weeks after transplantation, respectively, for vital staining. Clinical examinations were carried out every week, and the animals were euthanized 9 weeks later. The jaws were resected, fixed in formaldehyde and embedded in resin. Undemineralized sections were cut and examined by fluorescent microscopy. The thickness of dentin formed in the third week after transplantation and 9 weeks was evaluated by undertaking histomorphometric analysis and analysed using the Mann-Whitney U test (P = 0.05). All the transplanted teeth in both groups survived, and the dentin was formed. No statistically significant difference was found in the thickness of dentin formed in the third week and formed in the third to ninth week between the treatment groups (P = 0.999 and P = 0.998, respectively). This study demonstrated that there was no difference between the two surgical techniques in terms of the thickness of dentin formed in transplanted teeth., (© 2011 John Wiley & Sons A/S.)

Connective tissue growth factor/CCN family 2 (CTGF/CCN2) has been considered to participate in tooth development. To date, the expression and role of CTGF/CCN2 in reparative dentinogenesis have been unclear. Our previous study revealed that matrix metalloproteinase-3 (MMP-3) stimulates cell migration via CTGF/CCN2 expression and secretion in human dental pulp cells, and that this is dependent on dynamin-related endocytosis and independent of protease activity. The objective of the present study was to determine the expression of CTGF/CCN2 in reparative dentin in human carious teeth and to examine the effect of CTGF/CCN2 on mineralization in cultured human dental pulp cells. Minimal expression of CTGF/CCN2 was evident in odontoblasts subjacent to the dentin-pulp junction in healthy teeth, whereas strong expression was detected in odontoblast-like cells lining the reparative dentin subjacent to dental caries. In human dental pulp cells, CTGF/CCN2 promoted mineralization but failed to induce proliferation, suggesting that this molecule has the ability to induce the differentiation of human dental pulp cells. Taken together, the data suggest that CTGF/CCN2 is likely involved in reparative dentinogenesis through formation of hard tissue in human carious teeth.

Aim: To assess the effectiveness of mineral trioxide aggregate (MTA) used as an indirect pulp-capping material in human molar and premolar teeth., Methodology: We conducted a clinical evaluation of 60 teeth, which underwent an indirect pulp-capping procedure with either MTA or calcium hydroxide cement (Dycal(®) ). Calcium hydroxide was compared with MTA and the thickness of the newly formed dentine was measured at regular time intervals. The follow-up was at 3 and 6 months, and dentine formation was monitored by radiological measurements on digitised images using Mesurim Pro(®) software., Results: At 3 months, the clinical success rates of MTA and calcium hydroxide were 93% and 73%, respectively (P = 0.02). At 6 months, the success rate was 89.6% with MTA, and remained steady at 73% with calcium hydroxide (P = 0.63). The mean initial residual dentine thickness was 0.23 mm, and increased by 0.121 mm with MTA and by 0.136 mm with calcium hydroxide at 3 months. At 6 months, there was an increase of 0.235 mm with MTA and of 0.221 mm with calcium hydroxide., Conclusions:   A higher success rate was observed in the MTA group relative to the Dycal(®) group after 3 months, which was statistically significant. After 6 months, no statistically significant difference was found in the dentine thickness between the two groups. Additional histological investigations are needed to support these findings., (© 2012 FDI World Dental Federation.)

GPC-1 (glypican-1) is a cell surface heparan sulfate proteoglycan that acts as a co-receptor for heparin-binding growth factors and members of the TGF-β (transforming growth factor beta-1) family. The function of cell-surface proteoglycans in the reparative dentine process has been under investigation. Gpc-1 was detected with similar frequency as tgf-β1 in the cDNA library using mRNA from the odontoblast-like cell-enriched pulp of rat incisors. The aim of this study was to test our hypothesis that gpc-1 may be related to reparative dentine formation. We examined the expression of this gene during the reparative dentine process, as well as the effect of gpc-1 on odontoblast-like cell differentiation using siRNA (small interfering RNA) to down-regulate gpc-1 expression. Immunohistological examination showed that GPC-1 was expressed in pulp cells entrapped by fibrodentine and odontoblast-like cells as well as TGF-β1. The mRNAs for gpc-1, -3 and -4, except for gpc-2, were expressed during odontoblast-like cell differentiation in pulp cells. The relative levels of gpc-1 mRNA were increased prior to the differentiation stages and were decreased during the secretory and maturation stages of pulp cells. Down-regulation of gpc-1 expression resulted in a 3.9-fold increase in tgf-β1 expression in pulp cells and a 0.3-fold decrease in dspp (dentine sialophosphoprotein) expression compared with control. These results suggested that gpc-1 and tgfβ-1 expression are necessary for the onset of differentiation, but should be down-regulated before other molecules are implicated in the formation of reparative dentine. In conclusion, gpc-1 expression in odontoblast-like cells is associated with the early differentiation but not with the formation of reparative dentine.

Aim: To examine the in vivo response of dental pulps in dogs to three pulp-capping agents: calcium hydroxide (CH), mineral trioxide aggregate (MTA) and a new endodontic calcium enriched mixture (CEM) cement., Methodology: Thirty-six second and third premolar teeth in six beagle dogs were randomly assigned to three experimental groups; CH, MTA or CEM cement. Following isolation and exposure, pulp tissues were removed with a fissure bur and haemostasis achieved. The pulps were dressed with appropriate materials, and the access cavity restored with amalgam. Histological analysis was performed 8 weeks after treatment; the samples were assessed by an independent observer for calcified bridge formation, pulp vitality and pulp inflammation. The data were analysed by Kruskal-Wallis, Mann-Whitney and one-way anova tests., Results: The number of root canals that showed calcified bridge formation, pulp vitality and lack of inflammation was significantly higher for teeth capped with either MTA or CEM cement in comparison with CH (P < 0.05). No significant difference was found between the CEM cement and MTA in terms of calcified bridge formation, pulp vitality and lack of inflammation (P > 0.05)., Conclusions: Mineral trioxide aggregate and CEM cement were associated with a similar favourable biological response to pulpotomy treatment and demonstrated a more effective induction of dentinal bridge formation compared to CH.

Introduction: Regenerative endodontic procedures use the differentiation potential of embryonic and adult pulp progenitor cell populations to reconstitute dental structures., Methods: An in-depth search of the literature was accomplished to review biologic knowledge from basic research on tooth morphogenesis and differentiation, root development, dentin-pulp regeneration, pulp revascularization and apexification, experimental and clinical studies on the dentinogenic differentiation potential of progenitor cells in the embryonic dental papilla, dental pulp, and associated mesenchymal tissues of the developing root., Results: Odontogenic potential is determined during early tooth morphogenesis in the odontogenic mesenchyme. Progenitor cells from the odontogenic mesenchyme give rise to primary dentin-forming cells (odontoblasts) in the presence of stage-specific enamel epithelium and/or basement membrane and tertiary dentin-forming cells (odontoblast-like cells) in experimental conditions. The specificity of odontogenic mesenchymal cells to form tertiary dentin might be related to the repertoire of signaling pathways operated by the temporospatial pattern of epithelial-mesenchymal interactions during tooth formation. Dental papilla cells isolated from tooth germs before the onset of odontoblast differentiation have not shown any competence to become odontoblasts in the absence of enamel epithelium. On the other hand, the specificity of progenitor cells in the mesenchymal cell populations of the developing root apex remains to be determined., Conclusions: It seems evident that the dental pulp might be only used as a source of progenitor cells with dentinogenic competence for the regeneration of the dentin-pulp complex. The nature of dental or apical papilla progenitor cells in terms of their specificity for dentin regeneration has to be first characterized., (Copyright (c) 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: To evaluate the effects of recombinant growth factors on tertiary dentine formation and intratubular mineralization after their application on deep dentinal cavities in dog's teeth., Methodology: Treatment included dentinal etching (37% phosphoric acid) and applications of bioactive molecules (1 microg mL(-1) TGF-beta1, 10 microg mL(-1) IGF-1, 10 microg mL(-1) bFGF, 10 microg mL(-1) OP-1 or 1 microg mL(-1) monoclonal anti-human TGF-beta1 in phosphate buffered saline, PBS) at the dentinal base of buccal Class V cavities. Control groups were treated with 0.1% dog serum albumin (DSA) in PBS omitting the growth factors. This was performed both with and without dentinal etching. The dentinal responses regarding tertiary dentine formation and intratubular mineralization were assessed after 3 and 8 weeks, respectively, using light and scanning electron microscopy. Some specimens were also subjected to dentine permeability testing., Results: The group treated with transforming growth factor-beta1 (TGF-beta1) and, to a lesser extent, the one treated with osteogenic protein-1 (OP-1) showed significantly greater (P < 0.05) tertiary dentine formation and intratubular mineralization over an 8-week period when compared with the control and the other experimental groups. There were no significant differences between groups in reduction in dentine permeability after treatment., Conclusion: Treatment of exposed dentinal tubules with biologically active molecules might induce intratubular mineralization and tertiary dentine formation. Further research is needed to substantiate any clinical benefits as opposed to traditional treatments of exposed dentine so as to provide a scientific base for the clinical regulation of dentine reactions.

Introduction: This article describes an apexogenesis report of an 8-year old boy referred to our clinic 4 weeks after an impact trauma to the maxillary left central incisor that caused a complicated crown fracture and pulpal exposure., Methods: In the radiographic examination, the tooth was observed to be immature. After access cavity preparation, cervical pulpotomy was performed, and the remaining pulp was capped with calcium enriched mixture (CEM) cement. The crown was restored by using the fractured incisal segment on the next day., Results: The radiographic and clinical examinations on the 6-month and 12-month follow-ups showed that the tooth remained functional, root development was completed, and the apex was formed. A calcified bridge was produced underneath the capping material. No further endodontic intervention was necessary., Conclusions: Considering the healing potential of traumatized immature vital pulp, the use of CEM cement for apexogenesis might be an applicable choice; however, further clinical studies with longer follow-up periods are recommended., (Copyright (c) 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: Immunosuppressive drugs are used in clinical medicine for a variety of disorders, but their effects on the reparative capacity of the dental pulp are unknown. This study evaluated the influence of selected immunosuppressive drugs on pulpal tissue healing after direct pulp capping of mechanically exposed dog's teeth with mineral trioxide aggregate (MTA)., Methods: Ten healthy male dogs were assigned into 5 experimental groups: a control group in which no drug was received and 4 experimental groups in which the immunosuppressive drugs prednisone, mycophenolate mofetil, sirolimus, and cyclosporine A were administered 45 days before the operative procedures and until the dogs were killed. Class V cavities were prepared on the buccal surfaces of 12 teeth in each dog. In each cavity, the pulp was exposed and capped with MTA. The pulpal tissue responses to capping material were assessed 65 days postoperatively., Results: Compared with the control group, variable responses was recorded in the groups treated with mycophenolate mofetil, sirolimus, and cyclosporine A, which were characterized by moderate to severe inflammatory reactions, tissue necrosis, and total absence of hard tissue bridging. Pulpal tissue responses in the group treated with prednisone were characterized by inflammatory cell infiltration, limited tissue necrosis, as well as partial to complete hard tissue bridging., Conclusions: From these findings, it seemed evident that acceptable repair of the dentin-pulp complex, eg, wound healing with hard tissue formation after capping with MTA, is unlikely with mycophenolate mofetil, sirolimus, or cyclosporine A immunosuppressive drug therapy.

We investigated three-dimensional decrease in the volume of the pulp chamber caused by age-related secondary dentin formation using micro-CT and evaluated the applicability of the results to evaluation of age taking into account sex, age and tooth type. Decrease was slightly higher in females than in males, and a higher correlation between decrease and aging was observed in females. A comparison between age-groups revealed that decrease progressed between the fifties and sixties in males, and the forties and fifties in females. A stronger correlation between aging and decrease was observed in the mandibular central incisors than in the mandibular second premolars. This correlation was higher than other correlations between sexes and age-groups.

Introduction: The purpose of this study was to determine whether human recombinant bone morphogenetic protein-2 (rhBMP-2), calcitonin gene-related peptide (CGRP), calcitonin (CT), or parathyroid hormone-related protein (PTHrP) promoted reparative tertiary dentin or osteodentin formation in ferret canines., Methods: Ferrets had up to 4 pulpotomies performed under anesthesia. All pulps had sterile absorbable sponge of a standard size placed in contact with the pulp. The sponge contained sterile saline, rhBMP-2, CGRP, CT, or PTHrP. The opening was filled with an intermediate restorative material. After 6 weeks, the ferrets were anesthetized, and the pulpotomized teeth were extracted. The canines were fixed, decalcified, sectioned, and stained with hematoxylin-eosin. Sections were selected from the area of the opening, and the amount of reparative tertiary dentin and osteodentin was measured by using a digitizer., Results: Analysis of the photomicrographs showed that rhBMP-2 induced 0.58 +/- 0.19 mm(2) osteodentin and 0.56 +/- 0.18 mm(2) tertiary dentin. CGRP induced 0.46 +/- 0.05 mm(2) osteodentin and 0.38 +/- 0.04 mm(2) tertiary dentin. The amount of rhBMP-2-induced and CGRP-induced osteodentin and tertiary dentin was significantly (P < .001) more than that found in the sterile saline-treated teeth (0.29 +/- 0.03 mm(2) osteodentin and 0.14 +/- 0.03 mm(2) tertiary dentin) or CT (0.2 +/- 0.06 mm(2) osteodentin and 0.16 +/- 0.05 mm(2) tertiary dentin; P < .01). PTHrP significantly (P < .05) reduced the amount of osteodentin (0.17 +/- 0.02 mm(2)) observed in the saline-treated teeth but was not significantly different in the amount of tertiary dentin observed., Conclusions: RhBMP-2 and CGRP promoted more pulpal healing than either CT or PTHrP.

Introduction: Pulp regeneration therapy is important to overcome the limitations of conventional therapy to induce reparative dentinogenesis. In the present study, we examined the effects of controlled release of different dosages of fibroblast growth factor-2 (FGF-2) from gelatin hydrogels to regenerate the dentin-pulp complex., Methods: After the amputation of dental pulp of rat molars, gelatin hydrogels incorporating various dosages of FGF-2 were individually implanted into dentin defects above the sites of the amputated pulps. Histologic changes as well as the expression of dentin matrix protein-1 (DMP-1) and nestin in the dentin defect area above the amputated pulp were analyzed., Results: We found that controlled release of high doses of FGF-2 from gelatin hydrogels induced DMP-1-positive calcified particles in the proliferating pulp, whereas a moderate dose of FGF-2 induced DMP-1-positive dentinal bridge on the surface of the proliferating pulp. These findings indicate that the dosage of released FGF-2 has an influence on the structure of calcified tissue regenerated in dentin defects. In addition, pulp cells near calcified tissues regenerated in dentin defects were nestin-negative, suggesting that the calcified tissues might be osteodentin., Conclusions: Our results showed that the dentin regeneration on amputated pulp, not reparative dentin formation toward amputated pulp, can be regulated by adjusting the dosage of FGF-2 incorporated in biodegradable gelatin hydrogels.

Introduction: The current study investigated the effectiveness of enamel matrix derivative (EMD) and mineral trioxide aggregate (MTA) in inducing the differentiation of human dental pulp cells (HDPCs) into odontoblast-like cells in vitro., Methods: The effects of MTA and EMD on odontoblastic differentiation were indexed by alkaline phosphatase (ALP) activity and the expression of odontoblastic/osteoblastic markers, as determined by reverse-transcription polymerase chain reaction analysis. Mineralization was also evaluated by the staining of calcium deposits with Alizarin red. Cells were treated with a combination of MTA and EMD or with MTA alone., Results: Compared with MTA-treated cells on day 3, MTA/EMD-treated cells exhibited significantly greater increases in ALP activity and in dentin sialophosphoprotein and bone sialoprotein expression. Mineralization was significantly greater on day 7 in MTA/EMD-treated cells than in MTA-treated cells., Conclusions: These results suggest that a combination of MTA and EMD promotes more rapid differentiation in HDPCs than MTA alone and that this combination is thus a potential pulp-capping agent.

Objective: Conditioned media (CM) from human dental pulp cells (HDPC) was investigated for its effects on the proliferation and differentiation of HDPC and MG63 cells., Study Design: CM prepared from the primary culture of HDPC was used for the culture of HDPC and MG63. Cell growth, alkaline phosphatase (ALPase) activity, mRNA expression of differentiation-related genes, and mineralization of the HDPC and MG63 cells in the media containing CM were assessed., Results: HDPC CM increased the cell growth of HDPC but decreased MG63 cell growth. ALPase activity and the mineralization of both HDPC and MG63 were enhanced by HDPC CM. The CM also up-regulated the expressions of DSPP, DMP-1, and OCN mRNA in HDPC. Pretreatment of HDPC CM with a neutralizing antibody against TGF-beta completely eliminated the effect of CM on ALPase activity in HDPC., Conclusion: HDPC was able to secrete odontogenic differentiation-inducing factors, in which TGF-beta seems to a key element of the CM effects.

Recent reports have suggested that the stromal cell-derived factor (SDF)-1alpha-CXCR4 axis has a direct effect on stem and progenitor cell recruitment in muscle and neural tissue repair after injury. No information is available about SDF-1alpha or CXC chemokine receptor 4 (CXCR4) in dental tissues. The aim of this study was to assess the expression of SDF-1alpha and its receptor, CXCR4, in healthy or inflamed human dental pulp and to evaluate the effects of SDF-1alpha on dental pulp cells (DPCs) in both proliferation and migration in vitro. Immunohistochemical staining and reverse-transcription polymerase chain reaction detected weak expression of SDF-1alpha and CXCR4 in healthy dental pulp and strong expression of SDF-1alpha and CXCR4 in inflamed dental pulp. A methylthiazol tetrazolium assay showed that SDF-1alpha could not promote DPCs proliferation. A transmigration assay, however, indicated that SDF-1alpha enhanced DPCs migration, which could be abolished by anti-CXCR4 antibodies. Taken together, these results imply that the SDF-1alpha-CXCR4 axis may play a role in the recruitment of CXCR4-positive DPCs toward the damaged sites.

Endodontic treatment options for immature, nonvital teeth conventionally include surgical endodontics, apexification with calcium hydroxide, or single visit mineral trioxide aggregate plug. A new treatment option of revascularization has recently been introduced. It involves disinfecting the root canal system, providing a matrix of blood clot into which cells could grow, and sealing of the coronal access. The present pilot clinical study was undertaken to evaluate the efficacy of revascularization in 14 cases of infected, immature teeth. Endodontic treatment was initiated, and after infection control, revascularization was performed. The access cavity was sealed with glass ionomer cement. The cases were followed up at regular intervals of 3 months; the range in follow-up was 0.5-3.5 years. The outcomes were as follows. Radiographic resolution of periradicular radiolucencies was judged to be good to excellent in 93% (13 of 14) of the cases. In the majority of cases, a narrowing of the wide apical opening was evident. In 3 cases, thickening of apical dentinal walls and increased root length were observed. The striking finding was complete resolution of clinical signs and symptoms and appreciable healing of periapical lesions in 78% (11 of 14) of cases. Thickening of lateral dentinal walls was evident in 57% (8/14) of cases, and increased root length was observed in 71% (10/14) of cases. None of the cases presented with pain, reinfection, or radiographic enlargement of preexisting apical pathology. This pilot study documented a favorable outcome of revascularization procedures conducted in immature nonvital, infected permanent teeth.

This study investigated the reparative process of mechanically exposed pulps capped with mineral trioxide aggregate (MTA). Maxillary first molars of 8-week-old rats were MTA-capped for 1-14 days, and 5-bromo-2'-deoxyuridine-labeled proliferating cells and immunoreactivity for nestin and osteopontin were analyzed. MTA capping caused mild necrotic changes followed by progressive new matrix formation and calcified bridging. Proliferating cells peaked at 3 days when matrix formation was inconspicuous. Nestin-expressing cells appeared at 3 days, were arranged beneath the newly formed matrix at 5 days, and showed odontoblast-like morphology by 14 days. Osteopontin immunoreactivity was detected just beneath the necrotic area after 1 day. These findings suggest that pulpal responses to MTA capping involve proliferation and migration of progenitors followed by their differentiation into odontoblast-like cells, a mechanism basically similar to those to calcium hydroxide. Osteopontin might play a triggering role in initiation of the pulpal reparative process.

This study was conducted to evaluate the pulpal response to direct capping with either mineral trioxide aggregate (MTA) or calcium hydroxide (CH) cement in humans, with a focus on dentin bridge formation and dentin sialoprotein (DSP) and heme oxygenase-1 (HO-1) expression. Direct pulp capping was performed in 20 cases of caries-free human third molars. The pulps were exposed and capped with either MTA or hard-setting CH. After 2 months, the teeth were extracted, and the specimens were prepared for histologic and immunohistochemical evaluations. Histologically, 100% of the MTA group and 60% of the CH group developed dentin bridges. The mean thickness of the dentin bridges observed in the MTA group was statistically greater than that of CH group. In addition, DSP and HO-1 were expressed in the odontoblast-like cells and pulp fibroblasts beneath the dentin bridge; furthermore, significantly greater immunostaining was observed in the MTA group than in the CH group. Collectively, these results indicate that MTA is superior to CH in terms of inducing the dentinogenic process in human pulp capping.

Some clinical case reports have shown that immature permanent teeth with periradicular periodontitis or abscess can undergo apexogenesis after conservative endodontic treatment. A call for a paradigm shift and new protocol for the clinical management of these cases has been brought to attention. Concomitantly, a new population of mesenchymal stem cells residing in the apical papilla of permanent immature teeth recently has been discovered and was termed stem cells from the apical papilla (SCAP). These stem cells appear to be the source of odontoblasts that are responsible for the formation of root dentin. Conservation of these stem cells when treating immature teeth may allow continuous formation of the root to completion. This article reviews current findings on the isolation and characterization of these stem cells. The potential role of these stem cells in the following respects will be discussed: (1) their contribution in continued root maturation in endodontically treated immature teeth with periradicular periodontitis or abscess and (2) their potential utilization for pulp/dentin regeneration and bioroot engineering.

To evaluate the pulp healing to bacterial contamination beneath a hard-setting calcium hydroxide (DY: Dycal, L.D. Caulk Co.) and a self-etching adhesive resin (2V: Clearfil Liner Bond 2V, Kuraray Medical Inc.) following dentin bridge formation. Class V cavities were prepared on 30 monkey teeth, and the pulps were exposed with a carbide bur through the cavity floor. Each exposed pulp was capped with either DY or 2V. The cavities were restored with a hybrid resin composite. The resin composite was removed at 180 days after capping, and then cavities were left open to the oral environment for 2 weeks to obtain bacteria contamination DY (BDY) and 2V (B2V; n = 10). A non-bacterial-contaminated group capped with DY was used as control. After bacterial challenges, inflammatory cell infiltration, incidence and differentiation of dentin bridges were evaluated histologically. There were significant differences in the presence of inflammatory cell infiltration among all groups (P < 0.05). No moderate or severe inflammatory reaction was found in Group DY. Group BDY showed moderate or severe inflammatory cell infiltration in 50%, and showed four necrotic specimens. Although no statistically significant difference was found in the formation and differentiation of dentin bridges among all groups, tunnel defects in dentin bridges were detected in 70% (DY), 80% (BDY), and 50% (B2V). Group B2V showed a significantly lower presence of inflammatory cell infiltration than Group BDY (P < 0.05). Bonding agent is supposed to seal the exposure site, and the remaining bonding agent on the cavities was effective as the barrier in the dentin bridges after bacterial challenges.

To compare the pulpal responses to light-cured and self-cured resin cements, cervical cavities were prepared in monkey's teeth, followed by application of etching gel and adhesive (Single Bond). A dual-cure resin cement (RelyX ARC) was applied, and hybrid composite inlays (Estenia) were bonded to the cavities. In one group, the cavities were photoirradiated for 20 seconds and the resin cement light-cured. In the other group, the resin cement was self-cured for six minutes without any photoirradiation. After experimental periods of seven, 28, and 70 days, histological features of pulp tissue were evaluated and compared. Results showed no significant differences in the histological features of the pulp tissues between the two curing methods. Both light-cured and self-cured resin cements showed acceptable biological compatibility with the monkey pulp. No bacterial penetration along the cavity walls was detected with either curing method.

Objective: To evaluate the pulpal response of beagles following direct pulp capping with Clearfil SE Bond (SB), a self-etching adhesive., Study Design: One hundred thirty sound teeth from 6 male beagles were used. One hundred twenty teeth had their pulp mechanically exposed at the bottom of the class V cavities and were divided into 2 groups. In group 1, teeth were capped with SB; group 2 teeth were capped with calcium hydroxide cement (CH). Ten teeth were kept intact as an untreated control group. After 7, 30, and 90 days, 40 teeth from 2 beagles were extracted and processed for light microscopic examination., Results: After the 7-day observation period, inflammatory reaction in the SB group was similar to that of the CH group (P > .05). After the 30-day and 90-day observation periods, inflammatory reaction was slight in both groups, but less dentin bridge formation was observed in specimens from the SB group than in those from the CH group (P < .05)., Conclusion: SB showed good biocompatibility with pulp, but its ability to induce reparative dentin was lower than that of CH.

The aim of this study was to investigate the expression of 2 extracellullar matrix glycoproteins, fibronectin (FNC) and tenascin (TNC), following direct pulp capping with calcium hydroxide (CH). Third molars scheduled for extraction were used. Standardized class I cavities with pulp exposures were prepared. After control of bleeding, CH powder was applied in the exposure sites, which were covered with CH cement (Dycal; Dentsply) and the cavities were filled with zinc oxide-eugenol cement. Three teeth were extracted at each post-treatment period (1, 7, 14, and 30 days). Demineralized and paraffin-embedded specimens were stained for histologic technique (hematoxylin-eosin) and for immunohistochemical analysis. Anti-TNC and anti-FNC monoclonal antibodies were used with the streptavidin-biotin complex method. Generally, similar patterns of immunohistochemical expression were observed for TNC and FNC in the pulp tissue as a whole. In the exposure site, TNC immunostaining increased over time, exhibiting a thicker immunostaining pattern within 30 days. The imunohistochemical technique showed expression of both glycoproteins during pulp healing process.

Aim: To report the multidisciplinary care of an unaesthetic geminated maxillary lateral incisor tooth, which allowed its preservation in the mouth., Summary: Preoperative examination of an unsightly geminated maxillary lateral incisor (tooth 22) demonstrated two pulp chambers and open apices, with normal pulp sensitivity responses. At surgery, a periodontal mucoperiosteal flap was reflected and the distal part of the geminated tooth was removed. The exposed root canal of the preserved tooth was sealed with mineral trioxide aggregate (MTA). The extraction socket and osseous defect was grafted with decalcified freeze-dried bone allograft (DFDBA) before flap closure. During follow-up, distal caries in tooth 22 and a diastema between tooth 22 and 23 were managed with composite resin restorations. Forty-two months postoperatively, normal thermal and electrical pulp sensitivity tests confirmed pulp health. Convincing apexogenesis and dentinogenesis of the developing root was confirmed by radiographic examination. Acceptable periodontal health including 3-4 mm clinical probing depths was achieved. Optimizing aesthetics and occlusion was accomplished without orthodontic treatment.

Purpose: To (1) study the injury and healing activity of the pulp tissue to calcium hydroxide [Ca(OH)2], resin composite (RC) and resin-modified glass-ionomer (RMGI) materials when used as direct pulp capping agents, and (2) compare the incidence of healing defects between these materials., Methods: 135 Class V pulp exposed cavities were prepared in non-human primate teeth. Direct pulp capping was conducted over 6 to 730 days with hard set Ca(OH)2, RMGI and CR materials. Healing defects recorded were: (1) bacterial leakage with McKays stain; (2) operative debris including dentin fragments and particles of capping material; (3) pulpal inflammatory activity according to FDI standards; (4) area and absence of dentin bridge formation; and (5) presence of tunnel defects in bridge. Statistical analysis was evaluated using ANOVA., Results: The capping materials were associated with varying levels of pulp healing defects, including tunnel defects (P= 0.0001); operative debris (P= 0.0001); pulpal inflammatory cell activity (P= 0.0073) and bacterial leakage (P= 0.0260). Other healing defects, and the area of dentin bridge were not influenced by capping materials (P> 0.05).

Purpose: To evaluate, clinically and radiographically, the dentin bridge formation potential of white mineral trioxide aggregate (white MTA) when used in pulpotomy treatments in primary molars., Methods: A total of 23 primary molars received a pulpotomy treatment using white MTA followed by stainless steel crown restoration and controlled 6 months after treatment., Results: Clinical and radiographic success was 100% as none of the molars showed any clinical or radiographic pathological signs. Reparative dentin deposition was found in some of the cases: stenosis was present in 69.2% of the pulp canals from mandibular molars and formation of dentin bridges was seen in 11.5% of the pulp canals from mandibular molars 6 months after treatment with white MTA.

Aim: To investigate the hypothesis that different haemostatic agents could impair the histological response of human pulps capped with calcium hydroxide., Methodology: Forty-five third molars scheduled for extraction were selected. Class I cavities with pulp exposures were prepared. Three agents were used to control bleeding: 0.9% saline solution (control, n = 14), 5.25% sodium hypochlorite (n = 16) and 2% chlorhexidine digluconate (n = 15). The pulps were dressed with hard-setting calcium hydroxide cement. After 7, 30 or 90 days, teeth were extracted, formalin-fixed and prepared for histochemical techniques. The biological response was categorized using the following criteria: inflammatory response, soft tissue organization, reactionary dentine and reparative dentine. Data were submitted to statistical analysis, using nonparametric Kruskal-Wallis one-way analysis of variance on ranks. Differences amongst groups were detected using Dunn's method., Results: The statistical analysis disclosed that whilst inflammatory response decreased over time, reactionary dentine deposition and reparative dentine formation increased in the latter periods of evaluation (P < 0.05). The three agents had similar performances for all criteria evaluated. The conventional pulp response to calcium hydroxide was observed over time, and complete pulp healing was observed in 88% of the specimens after 90 days., Conclusion: The three haemostatic agents did not impair the healing process following pulp exposure and capping with calcium hydroxide at different time intervals investigated.

Purpose: To evaluate the clinical, radiographical and histological findings in human third molars in which mechanical pulp exposures were capped with white ProRoot mineral trioxide aggregate (WMTA)., Methods: Forty-eight human third molars, caries-free or with incipient caries, scheduled to be extracted, were used and randomly divided into two groups: Group A: (n= 24) received WMTA and control Group B: (n= 24) received chemical set calcium hydroxide (Dycal). The teeth were isolated with rubber dam and Class I cavities prepared. Pulp exposure was performed using a sterile diamond bur and confirmed by frank bleeding. A sterile cotton pellet dipped in saline solution was placed over the exposure for 60 seconds. The preparation was then lightly rinsed with water and gently air-dried. WMTA or CH was placed over the exposure site followed by a small amount of a light-cured compomer. After etching with 35% phosphoric acid gel for 15 seconds, rinsing and blot drying, Prime and Bond NT adhesive was applied and light-cured. The cavity was then restored with a resin composite and light-cured. Evaluations were performed by phone after 7 days and clinically at 30 +/- 5 and 136 +/- 24 days, using standardized tests and radiographs. The teeth were extracted after 136 +/- 24 days; the roots were cut +/- 4-5 mm from the apex to allow for rapid fixation in 10% neutral buffered formalin. They were then processed for routine histological evaluation, embedded in paraffin, sectioned and stained with hematoxylin and eosin and Brown and Brenn for recognition of bacteria. Statistical analyses were performed using a Mann-Whitney U-test, a Chi-square test, a Fisher's exact test and an ANOVA., Results: No significant differences in post-operative sensitivity were reported after 7 days between the two materials (P> 0.05). Clinical examination demonstrated no significant differences at 30 +/- 5 days (P> 0.05) and at 136 +/- 24 days (P> 0.05). Histological findings: 45 of 48 teeth were suitable for microscopic evaluation (22 with WMTA and 23 with CH). Twenty from the WMTA and 18 from the CH group had developed a bridge. No statistically significant differences were found for superficial and deep inflammatory cell response (P> 0.05), presence of a dentin bridge (P> 0.01), and pulp vitality (P> 0.01), between WMTA and calcium hydroxide. A statistically significant difference was found for the diameter of exposure (P< or = 0.05) between WMTA (x= 0.35 +/- 0.19 mm) and CH (x= 0.25 +/- 0.09 mm). Only a minimal association between clinical and histological findings could be established for either material.

Aim: This was the radiographic evaluation of 15 maxillary incisors apexification treatment carried out on 7-9 years old patients with periapical pathology due to pulpal necrosis after dental trauma., Materials and Methods: The treatment consisted of two phases. The aim of first phase was the formation of a calcified apical barrier through calcium hydroxide medications repeated twice with a three months interval. The aim of the second phase was the obturation of the root canal system once achieved a radiographic evidence of the formation of an apical barrier. The treated teeth were radiographed at six months and then periodically once a year for a period of 7 and 13 years., Results: The radiographic analysis allowed to underline three different kinds of apical formation: a physiological development of the apical portion with a final root length equal to the contralateral tooth; the formation of a cap tissue and an apical development with the final root length slightly shorter than the contralateral tooth; or the formation of different layers of mineralised tissue that aggregate together trough the years., Conclusion: Apexification is not a static phenomenon and the apexified area undergoes through the years to a conspicuous readjustment involving bone, apical root tissues and root filling material.

Pulpal responses to gallium-aluminum-arsenide (GaAlAs) laser irradiation applied to the tooth remains to be elucidated. This study aimed to evaluate the effect of the GaAlAs laser on odontoblasts using immunohistochemistry for heat-shock protein (HSP)-25, which labels mature and newly differentiated odontoblasts. The mesial surface of the upper right first molar of 8-wk-old Wistar rats was lased at an output power of 0.5-1.5 W for 180 s. The animals were perfusion-fixed at intervals of 6 h to 30 d after irradiation. At 6 h to 7 d, the intensity of HSP-25-immunoreactivity was found to be disturbed in the coronal odontoblast-layer in an energy-dependent manner. At 30 d, tertiary dentin with/without bone-like tissue was formed abundantly in the dental pulp. Statistical analysis revealed that the area occupied by the new hard tissues was significantly wider in 1.5 W-lased specimens than in 0.5 W-lased specimens. An intense HSP-25 immunoreactivity was seen in the odontoblasts underlying the tertiary dentin, whereas immunoreactivity was weak around the bone-like tissue. It was concluded that the GaAlAs laser may induce the formation of tertiary dentin by influencing the secretory activity of odontoblasts. However, higher energies may cause irreversible changes to the pulp, often leading to the formation of an intrapulpal bone-like tissue.

Aim: To examine the success of the one-appointment pulpotomy technique with three different medicaments on primary molar teeth., Methods: The study was conducted on 104 primary molars in 104 children with an indication for pulpotomy treatment on at least one primary molar. Primary teeth were treated with either formocresol (FC) (34 teeth), calcium hydroxide (CA) (33 teeth) or ferric sulphate (FS) (37 teeth) using standardised criteria for the pulpotomy procedures. Teeth were clinically and radiographically evaluated during the examination period of 18 months., Results: The clinical success rate at 18 months for the FC and FS groups was 90.9% and 89.2%, respectively. The CA showed an overall clinical success rate of 82.3%, with no statistical difference compared with either the FC or FS groups. The overall radiographic success for each technique was: FC 84.8%, CA 76.5%, and FS 81.1%. The presence of a dentine bridge above the pulp amputation site was observed radiographically for CA (47%), and FS (40.5%) pulpotomies without any statistical difference. Radiographic examination did not reveal the presence of a dentine bridge for any of the teeth treated with FC pulpotomy., Conclusion: Favourable clinical and radiographic success rates of ferric sulphate pulpotomy, comparable to formocresol were obtained. Therefore, ferric sulphate can be recommended as a pulpotomy medicament.

Aim: To investigate the effect of Emdogain Gel (Biora AB, Malmo, Sweden), consisting of a enamel matrix derivative (EMD) in a propylene glycol alginate (PGA) vehicle, on experimentally exposed human pulps and to register postoperative symptoms., Methodology: Nine pairs of contralateral premolars scheduled for extraction on orthodontic indications were included. Following a superficial pulp amputation performed with a small (016) diamond bur, either EMDgel or a mix of calcium hydroxide and sterile saline was placed at random in contact with the pulp wound. The subjects made records of symptoms and were also interviewed about pain/discomfort by a blinded examiner. After 12 weeks the teeth were extracted, prepared and subjected to light microscopic examination in which the inflammation and newly formed hard tissue in the pulp were analysed. Immunohistochemistry was performed using affinity-purified rabbit anti-EMD polyclonal antibodies., Results: Postoperative symptoms were less frequent in the EMDgel-treated than in the calcium hydroxide-treated teeth, especially during the first six weeks. In the EMDgel-treated teeth, new tissue partly filled the space initially occupied by the gel and hard tissue was formed alongside the exposed dentine surfaces and in patches in the adjacent pulp tissue. EMD was detected in the areas where new hard tissue had been formed. The wound area of the EMDgel-treated teeth exhibited inflammation in the majority of the teeth whereas less inflammation was seen in the calcium hydroxide-treated teeth where the hard tissue was formed as a bridge., Conclusions: In the EMDgel-treated teeth, postoperative symptoms were less frequent and the amount and pattern of hard tissue formation were markedly different than in the teeth treated with calcium hydroxide. However, the operative procedure and the formulation with EMD in a PGA vehicle do not seem to be effective for the formation of a hard tissue barrier.

The ultimate goal of a regenerative pulp treatment strategy is to reconstitute normal tissue continuum at the pulp-dentin border, regulating tissue-specific processes of tertiary dentinogenesis. Experimental investigations in mature teeth have shown that a network of extracellular matrix molecules and growth factors signal tertiary dentinogenesis. Application of dentin matrix components or growth factors in deep dentinal cavities stimulated up-regulation of biosynthetic activity of primary odontoblasts (reactionary dentin formation). Pulp-capping studies with a broad spectrum of biological agents, including growth factors and extracellular matrix molecules, showed formation of osteodentin and/or tertiary dentinogenesis (reparative dentin formation). Promising biologically active substances should be subjected to careful evaluation in well-designed preclinical investigations as well as in long-term clinical trials before their introduction in clinical practice., (Copyright 2004 S. Karger AG, Basel)