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Aim: To evaluate in vivo the bone tissue response of rats to varying amounts of infected and noninfected dentine debris., Methodology: Bone tissue reactions were evaluated histologically in 42 Wistar rats after 7, 30 and 60 days. For each animal, three surgical cavities were prepared on the femur and filled with varying amounts (5, 10 or 20 mg) of infected or noninfected dentine debris pellets. In the negative control group, the surgical cavities were not filled. At the end of each experimental period, the animals were euthanized. The samples were processed histologically and analysed using a light microscope. The presence and the severity of inflammatory reaction, as well as hard tissue deposition were evaluated. Data were subjected to statistical analysis and the effects of the dependent variables calculated using nonparametric tests Kruskal-Wallis and Mann-Whitney U with due Bonferroni corrections at P = 0.05., Results: At 7 days, the presence of infected debris significantly increased the histopathological scores for neutrophils (P < 0.05), and abscess formation (P < 0.05). Noninfected debris scored significantly higher for lymphocyte infiltrate compared with the control group and infected debris (P < 0.05). Both infected and noninfected debris equally triggered eosinophil cells compared with no-dentine (P < 0.05). As for giant cells and macrophages, no difference was detected amongst the dentine groups (P > 0.05). Hard tissue deposition was similar regardless of the presence or the bacteriological status of the dentine (P = 1.00). None of the above histopathological parameters was significantly influenced by the amount of debris (P > 0.05). For all parameters evaluated, at 7 days of analysis, the inflammatory response was significantly more intense compared with 30 and 60 days (P < 0.05). Inflammatory parameters were scored similarly for the evaluated groups after 30 and 60 days (P > 0.05). However, hard tissue deposition has significantly increased after 30 days (P < 0.05). No difference was seen between 30 and 60 days of analysis (P = 1.00) for all histological parameters evaluated., Conclusion: The assumption that the amount of extruded debris may negatively affect the inflammatory response of bone tissue was not validated in the present in vivo animal study. Infected dentine may trigger acute inflammatory parameters especially during the first 7 days of contact with the tissue; however, in the long term, these negative effects are mitigated., (© 2019 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

The aim of this scoping study was to evaluate the survival rate and nature of tissue formed inside root canals of human immature permanent teeth with necrotic pulps (NIPT) under root canal revascularization (RCR). The search was performed in SciVerse Scopus®, PubMed/MEDLINE, Web of Science®, BIREME and in the grey literature up to November 2015. The keywords were selected using MeSH terms and DECs. Two independent reviewers scrutinized the records obtained considering specific inclusion criteria. The included studies were evaluated in accordance with a modified Arksey and O' Malley's framework. From 375 studies that were evaluated, 75 were included. A total of 367 NIPT were submitted to RCR, from which only 21 needed further endodontic treatment. The weighted mean follow-up time was 17.6 months. The data were derived mainly from case reports (69%) or small case series (15%). NaOCl [0.5-6%] was applied as the disinfecting solution in almost all studies. Triple antibiotic paste was as effective as Ca(OH) 2 as on intracanal medicament. De novo tissue was cementum and poorly mineralized bone positive to bone sialoprotein (BSP) but negative to dentine sialoprotein (DSP). Failures were associated mainly with reinfection of the root canal. The majority of included studies reported a significant increase in both root length and width. However, as most of these data came from case reports, they must be interpreted with care, as most were focused on treatment successes (not failures). Therefore, well-designed randomized controlled trials comparing RCR with available apexification treatments are needed to address this gap in the literature., (© 2016 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Stem cell-based therapy (SC-BT) is emerging as an alternative for endodontic therapies. The interaction between stem cells and scaffolds plays a crucial role in the generation of a 'friendly cell' microenvironment. The aim of this systematic review was to explore techniques applied to regenerate the pulp-dentine complex tissue using SC-BT. An electronic search into the SciVerse Scopus (SS), ISI Web Science (IWS) and Entrez PubMed (EP) using specific keywords was performed. Specific inclusion and exclusion criteria were predetermined. The search yielded papers, out of which full-text papers were included in the final analyses. Data extraction pooled the results in four main topics: (a) influence of the chemical properties of the scaffolds over cell behaviour; (b) influence of the physical characteristics of scaffolds over cell behaviour; (c) strategies applied to improve the stem cell/scaffold interface; and (d) influence of cue microenvironment on stem cell differentiation towards odontoblast-like cells and pulp-like tissue formation. The relationship between the scaffolds, the environment and the growth factors released from dentine are critical for de novo pulp tissue regeneration. The preconditioning of dentine walls with ethylenediaminetetraacetic acid (EDTA) was imperative for successful pulp-dentine complex regeneration. An analyses of the grouped results revealed that pulp regeneration was an attainable goal., (© 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Knowledge about security and the potential damage originated by the gingival displacement techniques has not been described through randomised clinical studies. This crossover, double-blind, randomised clinical trial evaluated clinical and immunological factors related to conventional and cordless gingival displacement (GD) techniques, and patients' perceptions in 12 subjects with the employment of 2 GD techniques: conventional (gingival cord + 25% AlCl3 astringent gel) and cordless (15% AlCl3 astringent-based paste). In each subject, two anterior teeth were treated and a 10-day wash-out period separated the two treatments. Periodontal indices were evaluated before (baseline) and 1 and 10 days after GD. Interleukin 1β, interleukin 6 and tumour necrosis factor α concentrations in gingival crevicular fluid were measured before and 1 day after GD. Subjective parameters (pain, unpleasant taste and stress) were also evaluated. Data were analysed by one-way repeated-measures analysis of variance and Tukey's test (immunological factors), the Friedman test (periodontal parameters) and Fisher's exact or chi-squared test (subjective parameters), with a significance level of 95%. Gingival bleeding index, probing depth and plaque index values did not differ significantly between groups at any timepoint. Neither technique resulted in worse periodontal indices. Both techniques yielded similar results for pain and unpleasant taste, but conventional GD was significantly more stressful than cordless GD for volunteers. Both treatments significantly increased mean concentrations of the three cytokines, with the conventional technique producing the highest cytokine levels. Cordless GD is less stressful for patients and results in lower post-treatment levels of inflammatory cytokines compared with conventional GD., (© 2014 John Wiley & Sons Ltd.)

Aim: To evaluate the effect of four tooth storage temperature-based methods on quality of RNA obtained from cells retrieved from human dental pulps and human pre-dentine., Methodology: RNA was isolated from dental pulp tissue and from cells retrieved by scraping the pre-dentine of freshly extracted human third molars (n = 15) using TRIzol(®) reagent. Teeth were randomly assigned to the following temperature conditions: immediate RNA isolation after tooth extraction, liquid nitrogen (24 h), -80 °C (24 h), 20 °C (24 h) and 4 °C (6 h). RNA integrity was checked by the density of 28S and 18S ribosomal RNA. RT-PCR was used to analyse the expression of odontoblast makers (DSPP, DMP1 and MEPE) and the housekeeping gene GAPDH., Results: All experimental conditions evaluated preserved RNA integrity. The three odontoblastic markers were amplified from the pulp tissue and from the cells associated with pre-dentine., Conclusion: The four storage options allowed RNA isolation for RT-PCR analysis. These findings may facilitate the use of clinically derived human dental pulp and odontoblasts for endodontic research., (© 2011 International Endodontic Journal.)

Aim: To evaluate the influence of acid viscosity, endodontic sealer and root canal region on the push-out bond strength of a glass fibre post., Methodology: Seventy-eight single-rooted human teeth were selected (60 for push-out and 18 for scanning electron microscopy characterization, SEM, n = 13 per group). The root canals were prepared with a step-back technique and then filled with Endofill or AH Plus sealer. Teeth without root fillings were used as controls. The preparation of the post-space was to a length of 11 mm using standardized rotary instruments. The root dentine was treated with 37% phosphoric acid (gel or liquid). The fibre posts (Reforpost) were silanized, and resin cement (Enforce) was used for luting procedures. Each root was cross-sectioned, and samples from the cervical and apical regions were subjected to a push-out bond strength test. Specimens from each group were sectioned longitudinally and subjected to SEM characterization for the dentine/cement/post interface. Statistical analysis for push-out tests was carried out using factorial anova followed by Tukey's test (P < 0.05)., Results: The three factors under evaluation (acid, endodontic sealer and region) and their interaction significantly influenced bond strength values (P < 0.05). In general, liquid phosphoric acid had significantly higher bond strength values in the apical region (P < 0.05), with hybrid layer formation, while endodontic sealers reduced bond strength values compared to the control (P < 0.05)., Conclusion: The use of a liquid acid etchant created higher bond strength values in the apical region., (© 2011 International Endodontic Journal.)

Aim: To compare and contrast two colorimetric assays used for the measurement of proliferation using two dental pulp cell types: dental pulp stem cells (DPSC) and human dental pulp fibroblasts (HDPF)., Methodology: Dental pulp stem cells or HDPF were seeded at 0.25×10(4) cells per well in 96-well plates. Cell proliferation was evaluated after 24-72h. At the end of the experimental period, the sulforhodamine B (SRB) assay or a water-soluble tetrazolium salt (WST-1) assay was performed. Optical densities were determined in a microplate reader (Genius; TECAN). Data were analysed by Student's t-test (comparison between cell types) and one-way anova followed by Tukey test (time-point intervals). Pearson' correlation tests were performed to compare the two assays for each cell line., Results: Both assays showed that DPSC had higher proliferation rates than HDPF. A positive significant correlation between the two colorimetric assays tested for both cell types DPSC (Pearson's correlation coefficient=0.847; P<0.05) and HDPF (Pearson's correlation coefficient=0.775; P<0.05)., Conclusion: Both tests demonstrated similar trends of cell proliferation, and thus are both appropriate for the evaluation of DPSC and HDPF. The choice of assay is therefore one of the practical applications. SRB stained plates can be dried and stored so may have utility in laboratories where data may require review or when access to analytical equipment is limited. WST-1 assays have the benefit of both ease and speed and may have utility in laboratories requiring either high throughput or rapid analyses., (© 2010 International Endodontic Journal.)

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.

Aim: The purpose of the present study was to evaluate microleakage of a fourth generation dentine-bonding agent and composite restoration during a walking bleach treatment., Methodology: Thirty extracted non-carious incisors were selected and conventional root canal treatment was performed. Teeth were randomly divided into 3 groups (n = 10): group A (control), access cavities were restored with a fourth generation dentine-bonding agent, and incrementally restored with composite resin; group B, a paste composed of sodium perborate and hydrogen peroxide was placed into the pulp chamber and sealed with glass ionomer cement for 7 days, teeth were then restored in the same manner as group A; group C, a paste of sodium perborate and hydrogen peroxide was placed in the pulp chamber for 7 days and then after removal of the bleaching mixture, pulp chambers were filled with a calcium hydroxide paste and cavities sealed with glass ionomer for 1 week. The cavities were then restored with bonded restorations as in groups A and B. Teeth were subjected to thermal cycling and immersed in methylene blue for 8 h. Teeth were sectioned from buccal to lingual, through the centre of the restoration, using a diamond disk. Leakage was assessed using a standard scheme, under magnification (x20). Data were submitted to statistical analysis using nonparametric Kruskal-Wallis test., Results: Groups A and C exhibited similar leakage patterns, and both demonstrated less leakage values than group B (P < 0.05)., Conclusions: It was concluded that bleaching with sodium perborate and hydrogen peroxide increases microleakage; short-term use of a calcium hydroxide medicament did not increase microleakage.

Aim: The purpose of this study was to evaluate clinically and radiographically pulpotomies carried out under intrapulpal injection of anaesthetic solution., Methodology: Forty-one permanent mandibular molar teeth presenting with deep carious lesions and/or exposed pulps, with or without periapical changes on radiographic examination, were treated with pulpotomy and dressed with calcium hydroxide. The teeth were divided into three groups. Group A consisted of 15 teeth, where intrapulpal anaesthesia was administered by a slow injection of lidocaine hydrochloride 2%. Group B, with 14 teeth, where intrapulpal anaesthesia was obtained with lidocaine hydrochloride 2% with adrenaline 1:100,000. Group C consisted of 12 teeth in which anaesthesia was performed with a mandibular block using prilocaine hydrochloride 3% with felypressin 1: 100,000. Healing was evaluated using clinical and radiographic criteria: dentine barrier formation, absence of clinical symptoms and resolution of periapical involvement., Results: After an observation time of 6-8 weeks (postoperative control) and 24-32 weeks (intermediate control), healing occurred in 13 teeth from group A (87%), in 11 teeth from group B (79%) and in 10 teeth from group C (83%). No statistical difference was demonstrated between the three groups (Fisher's exact test)., Conclusions: Based on the methodology adopted, intrapulpal injection of anaesthetic solution did not impair healing in pulpotomized teeth.