Growth factor release from dentine matrix by pulp-capping agents promotes pulp tissue repair-associated events.

AIM: To characterize growth factor release from dentine by pulp-capping agents and to determine the effects of liberated dentine extracellular matrix (dECM) components on pulp cells in the key wound healing processes of migration and cell growth. METHODOLOGY: Powdered human dentine was exposed to solutions of calcium hydroxide, white and grey mineral trioxide aggregate (MTA) (ProRoot, (Dentsply Tulsa, Tulsa, OK, USA) over 14 days. The solubilized dECM components were dialysed and lyophilized and characterized using multiplex quantitative ELISA. Following dECM component extraction dentine was analysed using Fourier-transformed infrared spectroscopy (FTIR). Primary rat dental pulp cells (RDPCs) were exposed to dECM components (0.1-100 µg mL-1 ) released by calcium hydroxide, white and grey MTA, and cell growth and chemotactic responses were assessed. Statistical differences between the experimental and control groups were determined using one-way anova. RESULTS: A broad range of growth factors, many not previously reported in dentine, were liberated by these pulp-capping agents, including SCF, M-CSF, GM-CSF, IGFBP-1, NGF and GDNF. White and grey MTA liberated more growth factors than calcium hydroxide. FTIR analysis of dentine exposed to pulp-capping agents showed partial depletion of amide bands I, II and III, with little alteration in phosphate peaks compared to untreated dentine. dECM components released by white and grey MTA induced significantly more cell growth at low-to-moderate concentrations (P â‰¦ 0.05) examined in this study and significantly enhanced cell chemotaxis at all concentrations compared with controls (P â‰¦ 0.05). CONCLUSIONS: White and grey MTA solubilize a broad range of bioactive molecules from dentine, which can induce proliferation and chemotaxis in pulp cells.

Influence of root canal fillings on longevity of direct and indirect restorations placed within the General Dental Services in England and Wales.

AIM: To consider the survival of restorations in root filled and non-root filled teeth. METHODS: A data set was established consisting of patients, 18 years or older. For each patient on the database with a tooth restored with a direct or indirect restoration with or without a root filling, the subsequent history of intervention on that tooth was consulted, and the next date of intervention, if any could be found in the data set, was obtained. Thus a data set was created of restored teeth and whether they have also received root fillings, with the dates of restoration and root filling placement and the dates, if any, of re-intervention. Modified Kaplan-Meier statistical analysis was used to quantify the distribution of time to intervention. RESULTS: Data for over 80,000 different adult patients were analysed, of whom 46% were male and 54% female. A total of 538,967 restoration placements were obtained from the data over a period of 11 years, of which 30,073 were root fillings. CONCLUSIONS: Examination of the survival of restorations in teeth with and without root canal fillings indicated that those with root canal fillings have shorter intervals before re-intervention than teeth without root fillings. Restorations on root canal treated anterior teeth with post and cores had the lowest survival time.

Should pulp chamber pulpotomy be seen as a permanent treatment? Some preliminary thoughts.

AIM: To investigate the benefits of pulpotomy (to the level of the floor of the pulp chamber) as an endodontic treatment for teeth with vital pulps. METHODOLOGY: Seventeen patients, aged 7-54 years (mean of 37.2 year), were treated by pulpotomy and filling with ProRoot MTA(®) in premolar or molar teeth with vital pulps and without clinical evidence of irreversible pulpitis. The patients were then followed up for 12 to 24 months and the teeth then assessed by clinical and radiographic examination. Statistical analysis was performed with Kaplan-Meier survival probability statistics to estimate the survival of the treated teeth. RESULTS: At 24 months, the survival rate without any complementary treatment was estimated to be 82%. Two of the 17 treated teeth required root canal treatment for pain control and one for prosthetic reasons. CONCLUSIONS: Under the conditions of this study, pulpotomy offered a viable alternative to root canal treatment for teeth with vital pulps in the short term. However, there is insufficient clinical evidence to consider this technique for the treatment of every permanent tooth. Nevertheless, it should be considered as a potential alternative approach to be further developed for future applications.

Dentin-pulp complex regeneration: from lab to clinic.

Dentistry is entering an exciting era in which many of the advances in biotechnology offer opportunities for exploitation in novel and more effective therapies. Pulp healing is complex and dependent on the extent of injury, among many other factors. Many of the molecular and cellular processes involved in these healing events recapitulate developmental processes. The regulation of odontoblast activity is clearly central to pulp healing, and an understanding of the mechanisms involved in these processes is necessary to enable laboratory studies to be translated to clinic application. Transcriptome analysis has identified changes in many odontoblast genes during the life-cycle of this cell and its responses to injurious challenge. The p38 MAPKinase pathway appears to be central to the transcriptional control of odontoblasts and may provide a key target for therapeutic intervention. The many recent advances in knowledge of pulpal stem cells and molecular signaling molecules within the tooth, now provide exciting opportunities for clinical translation to novel therapies. Such translation will require the partnership of researchers and skilled clinicians who can effectively apply advances in knowledge to appropriate clinical cases and develop novel therapies which can be realistically introduced into the clinic.

Molecular characterization of young and mature odontoblasts.

UNLABELLED: The odontoblast is the secretory cell responsible for primary, secondary and tertiary reactionary dentinogenesis. We provide evidence that the changes in secretory activity of odontoblasts reflect differential transcriptional control and that common regulatory processes may exist between dentine and bone. INTRODUCTION: Based on the hypothesis that differential dentine secretion (primary and secondary dentinogenesis) is associated with changes in the transcriptional control within the cell, we have investigated the transcriptome of odontoblasts at young and mature stages and subsequently used this information to identify key regulatory intracellular pathways involved in this process. MATERIALS AND METHODS: We used microarray analysis to compare the transcriptome of early stage (primary dentinogenesis) and late stage (secondary dentinogenesis) odontoblasts from 30 month old bovine teeth. Secondarily, we used post-array sqRT-PCR to confirm the differential expression of 23 genes in both populations of odontoblasts. Finally, immunohistochemistry was performed on bovine and murine tissues with antibodies to DMP1 and anti-phospho p38 proteins. RESULTS: DMP-1 and osteocalcin gene expression were up-regulated in the mature odontoblasts, whereas collagen I, DSPP, TGF-beta1 and TGF-beta1R gene expression were down-regulated. Microarray analysis highlighted 574 differentially regulated genes (fold change>2 - p<0.05). This study supports further existing similarities between pulp cells and bone cells. Using post-array Sq-RT-PCR we characterized transcript levels of genes involved in the p38 MAP kinase pathway (PTPRR, NTRKK2, MAPK13, MAP2K6, MKK3). Differential p38 gene activation was confirmed by immunohistochemistry for p38 protein in murine teeth. Finally, immunohistochemistry for DMP1 indicated that odontoblasts involved in primary and secondary dentinogenesis may coexist in the same tooth. CONCLUSION: As established in bone cells, the transcriptome of the odontoblast was shown here to evolve with their stage and functional maturity. Identification of the involved signalling pathways, as highlighted for p38, will enable the deciphering of physiology and pathology of mineralised tissue formation.

Root canal retreatment.

UNLABELLED: Root canal therapy is not always successful and an increasing number of patients are requesting retreatment to address intra-radicular infection. The armamentarium available to assist the dentist, some of which is described in this article, has never been greater. CLINICAL RELEVANCE: This paper discusses the causes of failure and reviews many of the techniques available to the practitioner to tackle the problem.

Ten-year outcome of root fillings in the General Dental Services in England and Wales.

AIM: To consider the survival of root canal treatment provided within the General Dental Services in England and Wales, with failure being defined as re-treating of a root canal, apical surgery or extraction. METHODS: A data set was established consisting of patients, 18 years or older, whose birthdays were included within a set of randomly selected dates and whose restoration records contained the placement of one or more direct restorations or crowns in courses of treatment between September 1990 and January 2002. The history of each root canal-treated tooth was consulted, and the next date for an intervention on the root canal of the tooth, defined as a re-treatment, apical surgery or extraction, was obtained. Thus, a data set was created of root canal-treated teeth, with the dates of root canal filling placement and the dates, if any, of re-intervention. RESULTS: Data for over 80,000 different adult patients were analysed, and a total of 30,843 root canal-treated teeth identified from the data over a period of 11 years. The proportion surviving without further treatment of the root canal was estimated at 74% within an observation time of up to 10 years, with survival being strongly correlated with the characteristics of the patient, including age and treatment history, with older patients having root canal treatment with earlier re-intervention than those of younger patients. CONCLUSION: Within the data set analysed, an estimated 74% of root canal-treated teeth pass through 10 years without re-treatment, apical surgery or extraction.

Balancing the role of the dental school in teaching, research and patient care; including care for underserved areas.

Inequalities within dentistry are common and are reflected in wide differences in the levels of oral health and the standard of care available both within and between countries and communities. Furthermore there are patients, particularly those with special treatment needs, who do not have the same access to dental services as the general public. The dental school should aim to recruit students from varied backgrounds into all areas covered by the oral healthcare team and to train students to treat the full spectrum of patients including those with special needs. It is essential, however, that the dental student achieves a high standard of clinical competence and this cannot be gained by treating only those patients with low expectations for care. Balancing these aspects of clinical education is difficult. Research is an important stimulus to better teaching and better clinical care. It is recognized that dental school staff should be active in research, teaching, clinical work and frequently administration. Maintaining a balance between the commitments to clinical care, teaching and research while also taking account of underserved areas in each of these categories is a difficult challenge but one that has to be met to a high degree in a successful, modern dental school.

Dental regeneration and materials: a partnership.

Considerable focus on the biocompatibility of dental materials over the last three decades has provided a platform for a wealth of studies on the cellular and molecular responses of the cells of the pulp to injury, both from the disease process and from subsequent restorative intervention. These studies have been fundamental to understanding not only how we can achieve a biocompatible response during restoration of dental disease but also how we can exploit the pulpal cellular responses to achieve wound healing and tissue regeneration in the dentine-pulp complex. This article examines the responses of the pulp to injury and the events leading to tissue regeneration. As new biologically based regenerative therapies emerge for the dental tissues, it is important that these develop in partnership with more traditional approaches using dental materials.

Comparison of operative procedure variables on pulpal viability in an ex vivo model.

AIM: To measure and compare the responses of pulp tissue to cavity preparation and restoration variables using a novel tooth slice culture model. METHODOLOGY: Experimental cavities (265) were continuously cut, under carefully controlled conditions, into the dentine of the labial aspect of 28-day-old Wistar rat incisors, and slices of these teeth maintained in organ culture for up to 2 weeks. The experimental variables examined were: the preparation method, remaining dentine thickness, coolant, drill speed, conditioning with EDTA and filling materials. The reactions of the dentine-pulp complex to the experimental variables were measured using pathohistometric analysis and the correlations between variables were determined using analysis of variance statistical tests. RESULTS: In rank order of surgically induced restorative pulpal injury, from the most to the least injurious were: remaining dentine thickness, absence of coolant during cavity preparation, bur speed, cavity conditioning treatments and the filling material. CONCLUSIONS: To reduce pulp injury and to promote pulpal repair activity, the correct use of appropriate materials are important. However, of relatively greater importance is the operative technique adopted, the need to avoid the excess removal of dentine and to minimize trauma during preparation.

Evaluation of an intranasal house dust mite provocation model as a tool in clinical research.

BACKGROUND: As a result of the all-year-round exposure to house dust mite (HDM), perennial rhinitis patients never have a clear symptom-free period. In this study, we investigated whether, despite these symptoms, we can still use nasal HDM provocations to study perennial allergic rhinitis and the effects of treatment. METHODS: In a parallel-group study, after 1 week treatment with either fluticasone propionate aqueous nasal spray (FPANS) or placebo, 20 patients, allergic to HDM, registered symptoms (nasal obstruction, rhinorrhoea, sneezing, pruritus and eye symptoms) using three different scoring methods [Lebel, categorical and visual analogue scale (VAS)] and peak nasal inspiratory flow (PNIF) after HDM provocations. Provocations were performed with 1000 biological units/ml and 24 h later with 10,000 biological units/ml of HDM. Before and after the provocations, nasal mucosa biopsies were taken for immunohistochemical staining to determine the number of eosinophils. RESULTS: House dust mite provocations resulted in an increase in symptoms and a decrease in PNIF. Even at high-dose provocation, the FPANS group registered significantly lower symptoms than the placebo group for nasal blockage, sneezing, eye symptoms, and PNIF in both early and late phases. FPANS also suppressed rhinorrhoea during the late phase and the influx of eosinophils in the lamina propria. CONCLUSION: Despite the high background of symptoms, allergic responses can be induced in this perennial rhinitis model. The VAS score seems most suited to detect these changes and the suppression of symptoms by 7 days of FPANS treatment. Epithelial eosinophilia at baseline was correlated positively with the severity of the reaction after the first provocation.

Thermal changes and cure depths associated with a high intensity light activation unit.

OBJECTIVES: The objectives of this study were to evaluate the thermal emission and curing characteristics of a high intensity halogen light (Astralis 10-Ivoclar Vivadent, Schaan. Leichtenstein) alone and on curing a conventional and a fast-curing micro-hybrid composite. METHODS: A bead thermistor was placed in the base of a standard model cavity. The cavity was irradiated using the light unit whilst empty, and when filled with either composite. Temperature rises were recorded using the light in four different output modes. Further samples were prepared to assess depth of cure via a digital penetrometer, light transmission using a computer-based radiometer, and microhardness with a Wallace hardness tester. RESULTS: Mean peak temperature rises recorded during polymerisation of the composites ranged from 6.9 degrees C for the product InTen-S cured with the Adhesive programme (Adh) to 11.0 degrees C for the product Tetric Ceram HB cured with the High Power (HI P) programme. A significantly greater depth of cure was obtained for InTen-S in line with the greater light transmission obtained for this material. DISCUSSION: The very high thermal emission characteristics reported in a recently published investigation for this light unit were not confirmed. CONCLUSIONS: As the extent of thermal trauma that can be tolerated by the dental pulp is unknown consideration should be given to the choice of light activation unit and curing programme when polymerising light activated resin based restorations in deep cavities close to the pulp.

A new insight into the oscillation characteristics of endosonic files used in dentistry.

The aim of this study was to assess the oscillation characteristics of unconstrained endosonic files using a scanning laser vibrometer (SLV). Factors investigated included file vibration frequency and node/antinode location as well as the variation in file displacement amplitude due to increasing generator power setting. A 30 kHz Mini Piezon generator (Electro-Medical Systems, Switzerland) was used in conjunction with a #15 and #35 K-file. Each file was fixed in position with the long axis of the file perpendicular to the SLV camera head. The laser from the SLV was scanned over the length of the oscillating file for generator power settings 1 to 5 (minimum to half power). Measurements were repeated ten times. The fundamental vibration frequency for both files was 27.50 kHz. Scans of each file showed the positions of nodes/anti-nodes along the file length. The #15 file demonstrated no significant variation in its mean maximum displacement amplitude with increasing generator power, except at power setting 5, where a decrease in displacement amplitude was observed. The #35 file showed a general increase in mean maximum displacement amplitude with increasing power setting, except at power setting 4 where a 65% decrease in displacement amplitude occurred. In conclusion, scanning laser vibrometry is an effective method for assessing endosonic file vibration characteristics. The SLV was able to demonstrate that (unloaded) file vibration displacement amplitude does not increase linearly with increasing generator power. Further work is being performed on a greater variety of files and generators. Vibration characteristics of files under various loads and varying degrees of constraint should also be investigated.

Odontoblast morphology and dental repair.

OBJECTIVES: To investigate the changes in morphology and activity of pulp odontoblasts in response to cavity restoration variables and patient factors. METHODS: Class V non exposed cavities were prepared in the intact 1st or 2nd premolar teeth of 27 patients, aged between 9 and 17 years-old. Following tooth extraction, the area of reactionary dentine and the area of the odontoblasts were measured using computerised histomorphometry. RESULTS: The cytoplasm to nucleus ratio of the odontoblasts was found to increase beneath cut dentinal tubules, following the secretion of reactionary dentine. However, none of the patient or preparation variables were found to be correlated with changes in the odontoblast cytoplasm to nucleus ratio. CONCLUSIONS: Morphological changes in human odontoblasts is directly related to their capacity to repair dentine injuries and provide pulp protection. Changes in odontoblast morphology reflect secretory activity.

A pilot investigation of operator variability during intra-oral light curing.

OBJECTIVE: To test the hypothesis that operator experience influences the efficacy of light curing in a typical posterior intra-oral location. To investigate whether short cure cycles affect performance. DESIGN: A cross-sectional single-centre study designed to assess the efficacy of experienced and inexperienced operators when undertaking simulated intra-oral curing. SETTING: An in vitro laboratory based investigation conducted in a dental school during 2001. MATERIALS AND METHODS: A computer-based technique was used to monitor light intensity in a clinical simulation. Dentists and student operators were tested for their ability to cure a posterior restoration effectively. Relative light intensity was assessed against time for each operator and test run. RESULTS: Experienced (qualified) operators produced more effective and consistent cure results than less experienced undergraduate students. Operator performance was not affected by variations in irradiation time. CONCLUSIONS: This cross-sectional pilot investigation demonstrates that operator experience is a factor in successful clinical photo-curing of posterior restorations. Stable and accurate light guide positioning are required throughout the entire irradiation cycle to optimise intra-oral cure of light-activated restorations. Further investigations are planned to assess the potential of this novel method of assessment for use as a routine teaching aid in clinical practice.

Evaluation of smear layer removal by EDTAC and sodium hypochlorite with ultrasonic agitation.

AIM: To evaluate smear layer removal by different irrigating solutions under ultrasonic agitation. METHODOLOGY: Twenty recently extracted mandibular incisors with a single root canal were divided into four equal groups. Three groups were instrumented using the modified double-flared technique, the fourth remained unprepared. Each group was irrigated with either distilled water, 1.0% sodium hypochlorite alone or associated with 15% EDTAC between each file size. The final group was not instrumented but irrigated with 1.0% sodium hypochlorite and 15% EDTAC. A size 15 file energised by ultrasound was used with small amplitude filing movements against the canal walls in all groups. The teeth were split longitudinally and the roots measured to provide three sections of the same size (cervical, middle and apical). Samples were examined under the scanning electron microscope and assessed for the amount of smear layer by three independent and calibrated examiners. The scoring system ranged from 1 (no smear layer) to 4 (all areas covered by smear layer). Due to the non-parametric nature of the data, Friedman's test was used for statistical analysis. RESULTS: Canal walls were covered with smear layer in the group irrigated with 1% sodium hypochlorite alone and the group irrigated with distilled water. Canals irrigated with 1.0% sodium hypochlorite associated with 15% EDTAC had less smear layer throughout the canal (P < 0.001). There were no statistical differences for the amount of smear layer found on the cervical, middle and apical thirds when each group was analysed separately. CONCLUSIONS: Under ultrasonic agitation, sodium hypochlorite associated with EDTAC removed the smear layer from root canal walls, whereas irrigation with distilled water or 1.0% sodium hypochlorite alone did not remove smear layer.

Effect of exposure intensity and post-cure temperature storage on hardness of contemporary photo-activated composites.

OBJECTIVES: The effect of variation in post-exposure storage temperature (18 vs. 37 degrees C) and light intensity (200 vs. 500mW/cm(2)) on micro-hardness of seven light-activated resin composite materials, cured with a Prismetics Mk II (Dentsply) light activation unit, were studied. METHODS: Hardness values at the upper and lower surfaces of 2mm thick disc shaped specimens of seven light-cured resin composite materials (Herculite XRV and Prodigy/Kerr, Z100 and Silux Plus/3M, TPH/Dentsply, Pertac-Hybrid/Espe, and Charisma/Kulzer), which had been stored dry, were determined 24h after irradiation with a Prismetics Mk II (Dentsply) light activation unit. RESULTS: Hardness values varied with product, surface, storage temperature, and curing light intensity. In no case did the hardness at the lower surface equal that of the upper surface, and the combination of 500mW/cm(2) intensity and 37 degrees C storage produced the best hardness results at the lower surface. CONCLUSIONS: Material composition had a significant influence on surface hardness. Only one of the seven products (TPH) produced a mean hardness values at the lower surface >80% of the maximum mean upper surface hardness obtained for the corresponding product at 500mW/cm(2) intensity/37 degrees C storage temperature when subjected to all four test regimes. Despite optimum post-cure storage conditions, 200mW/cm(2) intensity curing for 40s will not produce acceptable hardness at the lower surface of 2mm increments of the majority of products tested.

Ultrastructural localisation of TGF-beta exposure in dentine by chemical treatment.

Transforming growth factor-beta (TGF-beta) sequestered in dentine matrix has an important role in dental tissue repair after injury and its exposure at sites of injury may stimulate tertiary dentinogenesis. This study aimed to investigate the expression of TGF-beta isoforms in mature human dentine matrix and the ability of chemical treatments to expose TGF-beta on the cut surface of dentine using gold immunolabelling and subsequent scanning electron microscopy examination. TGF-beta1 was the only isoform that could be detected in human dentine and the nature of the chemical treatment of the tissue influenced its detection. EDTA treatment provided good exposure of TGF-beta1 on the dentine surface, whilst citric acid and sodium hypochlorite treatments revealed lesser amounts of this isoform. Only minimal staining for TGF-beta1 was observed in samples treated with phosphate-buffered saline. TGF-beta2 and -beta3 could not be detected in the specimens with any of the treatments. This study suggests that TGF-beta1 is the only TGF-beta isoform expressed by human odontoblasts to be sequestered in dentine implying that differences in isoform-extracellular matrix interactions may exist. Information on chemical treatment of tissue specimens for immunostaining may provide a useful basis for selection of tissue preparation techniques for clinical restorative treatment procedures to facilitate TGF-beta mediated reparative processes at sites of dental injury.

Tooth slice organ culture for cytotoxicity assessment of dental materials.

The purpose of this work was to develop a tooth slice organ culture method to assess the response of the cells of the dental pulp to commonly used dental materials and products. Wistar rat tooth slices were grown in culture for two and ten days in the presence of dental materials. After culture, the tooth tissues were processed and the responses of the pulpal cells were analysed histomorphometrically. Cytotoxic cell destruction was observed following the direct application of test materials to tooth slices (n = 298) after 10 days in culture (MANOVA, P = 0.0001), whilst the restoration of prepared deep dentine cavities (n = 30), with test products, did not result in a significant amount of pulpal injury (MANOVA, P = 0.287). In rank order of causing pulpal injury, the test materials from the most to the least cell destructive, was; Salicylic acid. Calcium hydroxide, Kalzinol zinc oxide eugenol, high-mercury Amalgam, Prime & Bond, Dycal, Barium sulphate, Hypocal, Scotchbond, Calasept, Life and One-step. Tooth slice organ culture, provided a cytotoxicity screening method for dental materials, bearing a closer physiological resemblance to the clinical situation than cell culture screening methods. Tooth slice culturing may have the potential to replace some types of in vivo animal experimentation, as there is a clear need to reduce this form of testing.

The metalloproteinase inhibitor GI5402 inhibits endotoxin-induced soluble CD27 and CD16 release in healthy humans.

Metalloproteinases have been implicated in the cleavage of a number of cell surface immune receptors. Oral administration of the metalloproteinase inhibitor GI5402 attenuated the release of soluble CD27 and CD16 into the circulation after intravenous endotoxin injection in healthy humans.