To investigate the relationship between diabetes mellitus and local/systemic effects of both grey and white mineral trioxide aggregate (MTA) Angelus on bone marker expression.

Wistar rats were divided into two groups: healthy and diabetic (Alloxan induced), which were further divided into three subgroups (control, GMTA Angelus and WMTA Angelus). Polyethylene tubes filled with MTA materials or empty tubes were implanted in dorsal connective tissue. On days 7 and 30, blood samples were collected for calcium, phosphorus and ALP measurement. The animals were euthanized; implanted tubes were removed and processed for immunohistochemical analysis of osteocalcin (OCN) and osteopontin (OPN). Kruskal–Wallis followed by Dunn's multiple comparison test was performed for nonparametric data, and anova followed by Tukey's test for parametric data.

No difference in systemic serum calcium levels between both groups was observed. On day 7, serum phosphorus levels within the WMTA healthy group were higher than that of the diabetic group. On day 30, healthy rats exhibited lower phosphorus levels than diabetic ones. At both time points, the diabetic group was associated with more ALP activity than the healthy group. Immunohistochemical analyses of the healthy group revealed OCN- and OPN-positive cells in the presence of both MTA materials. However, under diabetic conditions, both OCN and OPN were absent.

Both MTA materials were associated with an increase in serum calcium, phosphorus and ALP, suggesting a potential systemic effect, along with triggered differentiation of OCN- and OPN-positive cells. Moreover, in diabetic conditions, an inhibitory effect on MTA-induced differentiation of OCN- and OPN-positive cells was detected.

To examine the efficacy of a novel supplementary irrigant agitating brush (Finisher GF Brush, MedicNRG, Kibbutz Afikim, Israel) on the debridement of root canals prepared with a novel stainless steel rotary instrumentation system (Gentlefile; MedicNRG), or nickel titanium rotary instruments in oval root canals.

Mandibular premolars (n = 72) were selected and divided randomly into three experimental groups (n = 24) after microCT scanning: group 1, canal preparation to rotary NiTi size 20, .04 taper (R20); group 2, rotary NiTi to size 25, .04 taper (R25) and group 3, Gentlefile size 23, .04 taper (GF). Specimens were subdivided into two subgroups: subgroup A, syringe-and-needle irrigation (SNI); subgroup B, Finisher GF Brush (GB). Ten untreated canals served as controls. Specimens were processed for histological evaluation, and the remaining pulp tissue (RPT) was measured. Data were analysed using Mann–Whitney and Kruskal–Wallis tests (P = 0.05).

All experimental groups had significantly less RPT than the control (P < 0.05). Group 3B (GF-GB) had significantly less RPT than groups 1B (R20-GB) and 2B (R25-GF; P < 0.05). When irrigated with SNI, there was no significant difference in the RPT between the three groups (P > 0.05). When instrumented with R20, there was no significant difference between SNI and GF (P < 0.05) whilst GB had significantly less RPT than SNI for R25 (P < 0.05).

Supplementary irrigant agitation with the Finisher GF Brush improved the debridement of canals prepared with Gentlefile and size 25, .04 taper rotary NiTi. Root canal debridement did not significantly differ between the instruments when syringe irrigation was used.

To investigate if crystallinity and ultrastructure is modified when cervical dentine is treated with four different nanogels-based solutions for remineralizing purposes.

Experimental nanogels based on polymeric nanoparticles (NPs) and zinc, calcium, or doxycycline-loaded NPs were applied on citric acid etched dentine to facilitate the occlusion of tubules and the mineralization of the dentine surface. Dentine surfaces were studied by X-ray diffraction and transmission electron microscopy through selected area diffraction and bright-field imaging.

Crystals at the dentine surface were identified as hydroxyapatite with the highest crystallographic maturity and crystallite size in dentine treated with Zn-NPs based-gel. Texture increased in all samples from 24 h to 7 d, except in dentine surfaces treated with Zn-NPs gel. Polyhedral, plate-like and drop-like shaped apatite crystals constituted the bulk of minerals in dentine treated with Zn-NPs gel, after 7 d. Polymorphic, cubic and needle-like shaped crystals distinguished minerals, with more amorphous characteristics in dentine treated with Ca-NPs gel after 7 d than that found when Zn-NPs were applied. Doxycycline-NPs produced the smallest crystallites with poor crystallinity, maturity and chemical stability.

Crystalline and amorphous phases of newly formed hydroxyapatite were described in both types of dentine treated with Zn-NPs as well as Ca-NPs gels with multiple shapes of crystallites. Crystal shapes ranged from rounded/drop-like or plate-like crystals to needle-like or polyhedral and cubic apatite appearance.

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To evaluate the association between the presence of selected bacterial species/groups in the apical root canal and expression of mediators of soft and bone tissue destruction in apical periodontitis lesions. Relationships between bacteria and some other features of apical periodontitis were also investigated.

Seventeen freshly extracted teeth with pulp necrosis and apical periodontitis were included. The apical root segment was sectioned and cryopulverized; DNA was extracted and evaluated for the presence of 9 bacterial species/groups using real-time polymerase chain reaction. Lesions were processed for histopathological and immunohistochemical analyses, which targeted matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9), receptor activator of NFκB (RANK), RANK ligand (RANKL) and osteoprotegerin (OPG). Associations of the target bacteria with expression of these mediators, presence of symptoms, lesion size and histopathological diagnosis were evaluated. Data were analysed using the chi-square, Fisher's exact, Mann–Whitney and Pearson tests. P values lower than 0.05 were considered significant.

All pulverized apical root samples were positive for bacteria. The most prevalent taxa were Actinobacteria (53%), Streptococcus species (35%), Fusobacterium species and Parvimonas micra (18%). The target mediators exhibited a high mean expression in the lesions (MMP-2: 82%; MMP-9: 73%; RANK: 78%; RANKL; 81%; OPG; 83%). Mean RANKL:OPG ratio was significantly higher in granulomas than cysts (P < 0.05, Mann–Whitney test). Actinobacteria were associated with granulomas, higher MMP-2 expression, lower OPG expression, and higher RANKL:OPG ratio (P < 0.05 for all, Fisher's exact test or Mann–Whitney test). No other significant associations were found.

Actinobacteria may play an important role in the active phase of soft and bone tissue destruction in apical periodontitis.

To evaluate the antimicrobial action of an irrigant containing silver nanoparticles in an aqueous vehicle (AgNp), sodium hypochlorite and chlorhexidine against Enterococcus faecalis biofilm and infected dentinal tubules.

Bovine dentine blocks were used for E. faecalis biofilm development for 21 days and irrigated with 94 ppm AgNp solution, 2.5% NaOCl and 2% chlorhexidine for 5, 15 and 30 min. For infection of dentinal tubules with E. faecalis, dentine specimens from bovine incisors were submitted to a contamination protocol over 5 days, with eight centrifugation cycles on every alternate day, and irrigated with the same solutions and time intervals used for the biofilm. The specimens were stained with the Live/Dead technique and evaluated using a confocal laser scanning microscope (CLSM). The bioImage_L software was used for measurement of the total biovolume of biofilm in μm³ and percentage of viable bacteria (green cells) in biofilm and in dentinal tubules found after the irrigation. Statistical analyses were performed using Kruskal–Wallis and Dunn's tests for quantification of viable cells in biofilm, the Friedman test for comparisons of viable bacteria in dentinal tubules in different areas of the root canal and the Mann–Whitney U-test to compare the action of the irrigants between the two methods (P < 0.05).

The AgNp solution eliminated fewer bacteria, but was able to dissolve more biofilm compared with chlorhexidine (P < 0.05). NaOCl had the greatest antimicrobial activity and biofilm dissolution capacity. AgNp solution had less antimicrobial action in infected dentinal tubules compared with NaOCl (P < 0.05). The AgNp solution after 5 min was more effective in eliminating planktonic bacteria in dentinal tubules than in biofilm, but at 30 min fewer viable bacteria were observed in the biofilm compared with intratubular dentine (P < 0.05).

AgNp irrigant was not as effective against E. faecalis compared to solutions commonly used in root canal treatment. NaOCl is appropriate as an irrigant because it was effective in disrupting biofilm and in eliminating bacteria in biofilms and in dentinal tubules.

To investigate the role played by silent information regulator 2 homologue 1 (SIRT1) during angiogenesis of periapical periodontitis.

Periapical granulomas were subjected to dual-colour immunofluorescence imaging and real-time polymerase chain reactions assaying the expression levels of SIRT1, vascular endothelial growth factor (VEGF) and VE-cadherin. The association between Ki-67 and SIRT1 expression was also examined. Human umbilical vein endothelial cells (HUVECs) were treated with a combination of lipopolysaccharide and resveratrol (a SIRT1 activator) or sirtinol (a SIRT1 inhibitor); and the levels of mRNAs encoding SIRT1, VEGF and VE-cadherin were determined. HUVEC tube formation was assayed in the presence of resveratrol or sirtinol. The Mann–Whitney U-test or the Tukey–Kramer test was used for statistical analysis.

Ki-67-expressing cells, including endothelial cells, lay adjacent to SIRT1-expressing cells in periapical granulomas. In addition, SIRT1-expressing cells were detected adjacent to VEGF-expressing cells and VEGF- or VE-cadherin-expressing endothelial cells. SIRT1, VEGF and VE-cadherin mRNA expression levels in periapical granulomas were significantly higher (P = 0.0054, 0.0090 and 0.0090, respectively) than those in healthy gingival tissues. HUVECs treated with resveratrol exhibited significantly higher expression of mRNAs encoding SIRT1, VEGF and VE-cadherin (P = 0.0019, 0.00005 and 0.0045, respectively) compared with controls, but sirtinol inhibited such expression. Resveratrol caused HUVECs to form tube-like structures, whilst sirtinol inhibited this process.

These findings suggest that SIRT1 may stimulate angiogenesis in periapical granulomas by triggering the proliferation of endothelial cells and inducing VEGF and VE-cadherin expression.

To study the demographics of Swedish adults who had received a root filling, followed by extraction during the following 5–6 years in comparison with subjects who had undergone a corresponding root filling with an uneventful outcome.

The root filled maxillary first molar was chosen as the comparison model. The Swedish Social Insurance Agency provided data on all teeth reported as root filled in Sweden during 2009. A comparison group, equally large as the study group, was constructed by randomly selecting subjects with root filled maxillary first molars, which had not subsequently been extracted, that is, an uneventful outcome. Demographic data on the subjects were obtained from Statistics Sweden: country of birth, disposable income, educational level, age, civil status and gender. Chi-square, t-tests and logistic regression were used for statistical analyses.

In the year 2009, 36 139 maxillary first molar teeth were reported to have been root filled, 4362 (12.1%) of which were then recorded as extracted during the following 5–6 year period. Only minor intergroup differences were noted: 86.5% of the study group were Swedish-born, compared with 84.4% of the comparison group (P = 0.007). Women comprised 53.2% of the study group and 50.5% (P = 0.01) of the comparison group. There was an association between extractions and gender as well as age; men had a lower odds ratio (OR) for extraction OR, 0.87; confidence interval (CI), 0.80–0.95. For every additional year, the chance for extraction was higher OR, 1.01; CI, 1.01–1.01. No other significant differences were detected.

There was only little or no demographic differences between the study group, comprising Swedish adults who had undergone root filling of one of their maxillary first molars in 2009 and subsequent extraction during the following 5–6 years, and the comparison group, with uneventful outcomes after a corresponding root filling.

To investigate whether a combination of mineral trioxide aggregate (MTA) and fluoride compounds affects bone cells.

Mineral trioxide aggregate (MTA) discs (ProRoot^®, Dentsply Sirona, Ballaigues, Switzerland) with and without the addition of 0.1%, 0.25% and 0.5% sodium fluoride were characterized for their surface roughness by laser scanning microscopy and for the adhesion of human alveolar osteoblasts by scanning electron microscopy. Using eluates from fluoride-enriched MTA discs, the cell proliferation was measured by monitoring the DNA incorporation of 5-bromo-2′-deoxyuridine. Further, gene expression was evaluated by qPCR arrays, extracellular matrix mineralization was quantified by absorption measurement of Alizarin red stains, and effects were calculated with repeated measures analysis and post hoc P-value adjustment.

Irrespective of fluoride addition, cell adhesion was similar on MTA discs, of which the surface roughness was comparable. Control osteoblasts had a curvilinear proliferation pattern peaking at d5, which was levelled out by incubation with MTA. The addition of fluoride partly restored the MTA-related reduction in the cellular proliferation rate in a dose-dependent manner. At the mRNA level, both fluoride and MTA modulated a number of genes involved in osteogenesis, bone mineral metabolism and extracellular matrix formation. Although MTA significantly impaired extracellular matrix mineralization, the addition of fluoride supported the formation of mineralized nodules in a dose-dependent manner.

The addition of fluoride modulated the biocompatibility of MTA in terms of supporting bone cell proliferation and hard tissue formation. Hence, fluoride enrichment is a trend-setting advancement for MTA-based endodontic therapies.

To assess the outcome of full pulpotomy using Biodentine in permanent teeth with carious exposures and symptoms indicative of irreversible pulpitis.

Sixty-four permanent molar teeth with symptomatic vital pulps in 52 patients aged 19–69 years were included. Preoperative pulpal and periapical diagnosis was established. After informed consent, the tooth was anaesthetized, isolated using rubber dam and disinfected with 5% NaOCl before caries excavation; subsequently, the pulp was amputated to the level of the canal orifices. Haemostasis was achieved, and a 3-mm layer of Biodentine (Septodont, Saint-Maur-des-Fosses, France) was placed as the pulpotomy agent. Resin-modified glass–ionomer liner was placed and the tooth restored with either resin composite or amalgam, and a postoperative periapical radiograph exposed. Clinical and radiographic evaluation was completed at 6 months and 1 year postoperatively. Pain levels were scored preoperatively and 2 days post-treatment.

Clinical signs and symptoms indicative of irreversible pulpitis were established in all teeth, and periapical rarefaction was present in nine teeth. After 2 days, 93.8% reported complete relief of pain. At 6 months, 63 of 64 attended recall with 98.4% clinical and radiographic success. At 1 year, 59 of 63 attended recall, with 100% clinical and 98.4 radiographic success. Seven of eight cases with periapical rarefaction who attended recall had improvement in the periapical index (PAI) score. A hard tissue barrier was detected radiographically in four cases.

Full pulpotomy using Biodentine was a successful treatment option for cariously exposed pulps in mature permanent molar teeth with clinical signs and symptoms indicative of irreversible pulpitis, up to 1 year.

To compare the cyclic fatigue resistance of the One G, ProGlider, HyFlex EDM and R-Pilot glide path NiTi files at body temperature.

Twenty One G (size 14, .03 taper), 20 ProGlider (size 16, .02 taper), 20 HyFlex EDM (size 10, .05 taper) and 20 R-Pilot (size 12.5, .04 taper) instruments were operated in rotation at 300 rpm (One G, ProGlider and HyFlex) or in reciprocation (R-Pilot) at 35 °C in artificial canals that were manufactured by reproducing the size and taper of the instrument until fracture occurred. The time to fracture was recorded in seconds using a digital chronometer, and the length of the fractured fragments was registered. Mean data were analysed statistically using the Kruskal–Wallis test and post hoc Tukey tests via SPSS 21.0 software. The statistical significance level was set at 5%.

The cyclic fatigue resistance of the R-Pilot files was significantly greater than the other instruments, and the One G was significantly lower (P < 0.05). There was no difference between the HyFlex EDM and the ProGlider (P > 0.05). No significant difference (P > 0.05) was evident in the mean length of the fractured fragments of the various instruments.

The cyclic fatigue resistance of the R-Pilot reciprocating glide path file was significantly greater than that of the rotary HyFlex EDM, ProGlider and One G glide path files.

To compare the cyclic fatigue resistance of R-PILOT and WaveOne Gold Glider files in curved artificial canals.

A total of 60 new R-PILOT and WaveOne Gold Glider files were tested in artificial canals with 45° and 60° angles of curvature. Fifteen new files of each brand were tested in both canals. Cyclic fatigue resistance was determined by recording the time to file fracture in the artificial canals. The length of each fractured fragment was also recorded. An independent sample t test was used to analyze the data.

In the canal with a 45° angle of curvature, no significant differences were observed between the R-PILOT and WaveOne Gold Glider files (P > 0.05). In the canal with a 60° angle of curvature, WaveOne Gold Glider files had greater cyclic fatigue resistance than R-PILOT files (P < 0.05). There was no difference between the files in terms of the lengths of fractured fragments in canals with 45° and 60° angles of curvature (P > 0.05).

WaveOne Gold Glider files exhibited greater cyclic fatigue resistance than R-PILOT files in artificial canals with a 60° angle of curvature.

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To evaluate retrospectively the prevalence of vertical root fractures (VRF) in a cohort of patients during apical surgery and the factors possibly associated with VRF.

The sample consisted of 944 root filled teeth belonging to 768 patients (49.3% males and 50.7% females; mean age 43.5±11.2 years, range 22-68 years), consecutively referred for endodontic surgery over a six-year period. All patients underwent a clinical assessment of their signs and symptoms. Periapical radiographs of teeth that were candidates for endodontic surgery were taken. Sixty-eight teeth with VRF were identified. Vertical root fractures were identified in pre-surgical screenings in 32 cases (47.1%) and these did not undergo surgery. Another 36 cases of VRF were noted during the intervention for root-end resection. The influence of posts, post type, tooth type, periodontal probing defects, spontaneous pain, sinus tract, and follow-up duration was assessed using a logistic regression analysis.

Vertical root fractures occurred significantly more frequently (P<0.001) when a post was present (61 VRF out of 377 teeth with post, prevalence 16.2%) than in teeth without a post (1.2%). Threaded posts and cast posts were significantly more involved in VRF than fibre, silica or carbide posts (P<0.001). Most fractures (80.9%) occurred 1 to 5 years after root canal treatment. Sinus tracts, probing defects and spontaneous pain were significantly more associated with VRF cases than with non-fractured teeth.

In the present group of teeth, the major risk for VRF was represented by posts retained by actively engaging the canal via mechanical design (thread), or by frictional fit (cast).

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To compare the cyclic fatigue resistance and bending properties of R-Pilot and WaveOne Gold (WOG) Glider files, at intracanal temperature (35°C).

Forty R-Pilot and 40 WOG Glider files were subjected to a cyclic fatigue resistance test (n = 20), calculating the time to fracture (TTF) in an artificial stainless-steel canal. The length of the fractured file tips (FL) was also measured. The fracture surface of fragments was examined with a scanning electron microscope and the cross-sectional area of the fractured surfaces were measured. Flexibility of the tested files (n = 20) was determined by 45° bending test. Data were analyzed statistically using the Mann-Whitney U test at 5% significance level.

Time to fracture value was found to be significantly higher in the R-Pilot group compared to the WOG Glider (P < 0.05). There was no significant difference between groups according to the fracture length. The bending resistance of R-Pilot files was significantly higher than WOG Glider files (P < 0.05).

A significant greater cyclic fatigue resistance was observed for R-Pilot files compared to WOG Glider instruments, although the bending resistance of WOG Glider files was lower.

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To use micro-CT technology and metrology software to validate the use of contralateral premolars as samples in endodontic comparison studies by comparing them before and after canal instrumentation with one instrumentation system. Furthermore, to determine whether contralateral premolar roots (CPRs) will yield non-significantly different outcomes regarding shaping ability (volume), degree of twisting and three-dimensional shape changes. The null-hypothesis (H₀) is that there are no differences between the CPRs pre- or post-instrumentation.

Twenty-eight extracted human contralateral premolars (n = 44 contralateral roots) from 12 donor patients were scanned with microcomputed tomography before and after instrumentation. Root canal lengths (RCLs) were measured visually using a dental-operating microscope, electronic apex locator and micro-CT scans. Data were analysed statistically for differences between pre- and post-instrumentation.

Instrumentation increased the volume of the canals significantly (P < 0.05). Degree of twisting for a majority (83%) of the contralateral roots pairs did not change significantly (P > 0.05). There was no significant difference (P > 0.05) in the shape deviation analysis between contralateral pairs. There was no significant difference (P > 0.05) for RCL between the contralateral pairs for any of the three endometric methods.

Contralateral premolar root canals were associated with similar changes in terms of volume, three-dimensional shape and degree of twisting from pre- to post-instrumentation. There was no difference between the CPR pairs pre- and post-instrumentation, and the study validates contralateral premolars as samples for root canal comparison studies. The null-hypothesis (H₀) could not be rejected.

To evaluate lymphocyte-like cell activation (CD5-positive cells) and the expression of interleukin (IL)-6 and IL-17 in the pulp after tooth bleaching with two concentrations of hydrogen peroxide (H₂O₂).

The right and left maxillary molars from 40 rats were treated randomly with bleaching gel with 20% H₂O₂ (BLUE group, 1 application of 50 min), 35% H₂O₂ (MAXX group, three applications of 15 min), or placebo gel (control). After 2 and 30 days, the rats were killed (n = 10), and the jaws were processed for histological and immunohistochemistry analysis of the pulp tissue. The scores of inflammation and immunolabelling (IL-6/IL-17) were submitted to Mann–Whitney and Kruskal–Wallis followed Dunn tests, respectively; anova tests were used for comparisons of number of CD5-positive cells and pulp chamber area values (P < 0.05).

At 2 days, 60% of specimens of the BLUE group were associated with moderate inflammation in pulp horns, and in the MAXX group with necrosis (P < 0.05). At 30 days, the pulp was organized, and tertiary dentine was formed. The MAXX group had superior immunolabelling of IL-17 at 2 days differing significantly from other groups (P < 0.05). At 2 days, 90% of the specimens of the BLUE group had moderate immunolabelling of IL-6, and 50% of the MAXX group had severe immunolabelling, both significantly different from the control (P < 0.05). There was no significant difference between the groups at 30 days (P > 0.05). CD5-positive cells were present at 2 and 30 days, particularly in the bleached groups (P < 0.05), without significant difference between time periods (P > 0.05).

IL-6 and IL-17 participated in inflammation in the pulp tissue of rats after tooth bleaching, particularly at 2 days. The immunolabelling was greater with increasing H₂O₂ concentration. This process was accompanied by the prolonged activation of CD5-positive cells.

To analyse the differences in root canal system configuration in patients belonging to different age groups using cone beam computed tomography (CBCT) technology.

CBCT examinations from a pre-existing database were accessed. Patients were divided according to age groups: ‘≤20 years’, ‘21–40 years’, ‘41–60 years’ and ‘≥61 years’. Each group included tooth data regarding their root canal system configurations according to the Vertucci classification and its supplementary configurations. Cohen kappa coefficient of agreement was calculated to evaluate observer reliability.

Overall 12 325 teeth from 670 patients were included. Most of the root groups had higher or equal prevalence of Vertucci type I configurations in the younger groups whilst presenting a greater tendency for multiple root canal system configurations in older patients, mainly Vertucci type II in both maxillary and mandibular second premolars and in the distal root of the mandibular first molar. The Cohen kappa coefficient of agreement was 89.4 ± 1.8%.

Clinicians should be aware that the root canal system configuration changes over a lifetime. In this study, the most affected teeth were the second premolars and the distal root of mandibular first molars.

To investigate the combinatorial effects of lipopolysaccharide (LPS) and extracted dentine matrix proteins (eDMP) on regenerative and inflammatory responses in human dental pulp stem cells (DPSCs).

Culture media were supplemented with several concentrations of LPS, eDMP and combinations of both. Cell viability was assessed over 1 week by MTT assay; cell survival was evaluated after 24 h and 7 days by flow cytometry. The expression of mineralization-associated marker genes was determined by real-time quantitative polymerase chain reaction (RT-qPCR). To analyse the inflammatory response, secretion of interleukin 6 (IL-6) was quantified in the initial and the late phase of cell culture by enzyme-linked immunosorbent assay (ELISA). Data were treated nonparametrically and Mann–Whitney U-tests were performed to compare all experimental groups (α = 0.05).

Whereas LPS had no impact on viability, eDMP led to a concentration-dependent decrease, which was significant after 7 days (P ≤ 0.024). A moderate decline of cell survival induced by LPS was detected after 48 h (P ≤ 0.026), whereas eDMP was able to reverse this effect. eDMP alone caused increased expression of tested marker genes, LPS had no regulatory effect. Combined eDMP and LPS induced an upregulation of collagen type I and osteocalcin, whereas expression levels of dentine matrix acidic phosphoprotein and dentine sialophosphoprotein were similar to the control. IL-6-secretion was increased by LPS over time. eDMP markedly elevated initial production of IL-6 (P ≤ 0.002), but suppressed LPS-induced cytokine production in the later phase.

Lipopolysaccharide did not affect cell viability but interfered with odontoblast-like cell differentiation of DPSCs. Proteins from the dentine matrix may have a protective effect, attenuate the detrimental impact of LPS and thus play an important role during pulp repair.

To evaluate how additional information from Cone Beam CT (CBCT) impacts on periapical assessment and treatment planning based on clinical examination and periapical radiographs (PR) in cases followed up five to eleven years after surgical endodontic retreatment (SER).

Patients receiving SER during 2004–2010 were reinvited for follow-up examination including clinical examination, PR, and CBCT. In total, 108 patients (119 teeth) were reinvited, 74 patients (83 teeth) accepted to participate. Three observers initially assessed PR according to the four-scaled, increasing disease severity criteria by Rud et al. (International Journal of Oral Surgery, 1, 1972 and 195) and Molven et al. (International Journal of Oral and Maxillofacial Surgery, 16, and 432): ‘Radiographic assessment A’. By including clinical information ‘Treatment plan A’ was made as follows: 1) no treatment, 2) further observation, 3) SER reoperation (SER-R), or 4) extraction. Hereafter, the CBCT volume was assessed and the information incorporated for ‘Radiographic assessment B’ followed by ‘Treatment plan B’. Agreement between radiographic assessments and between treatment plans was recorded and assessed statistically by Stuart–Maxwell test for marginal homogeneity.

Nine teeth had been extracted; thus, the final analysis included 74 teeth (66 patients). The radiographic assessment was changed as a result of the CBCT evaluation in 38 cases (51.4%), of which 35 (47.3%) were to a higher Rud & Molven score, P < 0.001. The treatment plan was changed for 18 teeth (24.3%). For 14 teeth (18.9%), the change was from no treatment or further observation to a more invasive treatment plan (SER-R or extraction), P = 0.005.

The use of CBCT for long-term follow-up after SER led to more cases diagnosed with persisting or recurrent apical periodontitis and hence often to the recommendation of a more invasive treatment modality.

To evaluate ex vivo the efficacy of ProTaper Universal Retreatment files (Dentsply Sirona, Ballaigues, Switzerland) in removing Thermafil, GuttaCore (both Dentsply Sirona) or vertically compacted gutta-percha from curved root canals using micro-CT.

Sixty curved molar roots with the same mean canal curvatures and radii in two directions were prepared using ProFile instruments (Dentsply Sirona) to size 30 with .04 taper and obturated with either Thermafil, GuttaCore or vertically compacted gutta-percha and AH Plus (n = 20). Specimens were retreated using the ProTaper Universal Retreatment files D1, D2 and D3 to working length, and root canal preparation was completed with ProTaper Next (Dentsply Sirona) to size ×4. Percentages of residual filling material and dentine removal were assessed using micro-CT imaging. Working time and procedural errors were recorded. Statistical analysis was performed using Kruskal–Wallis and Wilcoxon tests.

No significant differences between carrier-based and warm vertical compaction regarding residual filling material (14.2–19.3%) and dentine removal (2.7–3.2 mm³) were detected (P > 0.05). Time to reach working length was significantly faster for canals filled with GuttaCore than that observed for Thermafil and warm vertical compaction (P < 0.05). Five lateral perforations with the D3 file occurred during retreatment, one in the Thermafil and four in the vertical compaction group.

Remaining filling material and dentine removal were similar for all canal filling techniques. Regaining working length was significantly faster for GuttaCore compared with Thermafil and vertically compacted gutta-percha. Procedural errors occurred during retreatment of severely curved root canals with the ProTaper Universal Retreatment files in 5 of 60 canals (8%).

To evaluate the frequency of dentinal microcracks after ultrasonic removal of fractured files from the middle third of root canals using micro-computed tomography (micro-CT).

Eighteen bilaterally matched pairs of human mandibular incisors extracted for periodontal reasons were included. The matched pairs of teeth were then divided into a control group and an experimental group, with one member of each pair assigned to each group. In the control group, the canals were instrumented using the ProTaper Next (PTN) system. In the experimental group, size 20 K-files were fractured in the middle third of the root canals, followed by their ultrasonic removal. Subsequently, the canals were instrumented with the PTN system. All teeth were scanned using high-resolution micro-CT before (preoperative) and after (intraoperative) file removal and after (postoperative) root canal preparation. Pre-, intra-, and postoperative cross-sectional images of the roots were screened to identify the presence of dentinal defects. Two experienced observers evaluated the images twice in a blinded manner. The incidence of dentinal microcracks was noted and statistically analyzed using Fisher's exact and McNemar's tests (P = 0.05), with the root cross-section and the tooth root as the units of analysis, respectively.

All fractured files in the experimental group were removed successfully. New microcracks were detected in 0.56% (93/16472) cross sections (8/18 specimens) generated after file removal in the experimental group. These microcracks were detected 4–6 mm below the root canal orifice and exhibited a width and length of 12–36 μm and 48–72 μm, respectively. They did not disappear or propagate after canal preparation. No new dentinal microcracks were observed in the control group. There was a significant difference in the incidence of new microcracks between the two groups (P < 0.05).

Ultrasonic removal of fractured files from root canals resulted in the formation of short microcracks in a small number of cross sections in approximately half the specimens. Further studies are necessary to determine the cause and consequences of this finding.

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To evaluate the number of healthy and functional root filled teeth of patients included in a recall program for at least 20 years.

Teeth were root filled by a single specialist following manual canal instrumentation, lateral/vertical compaction of gutta-percha and restored with glass-ionomer cements and bonding system/composite resin. In a large percentage of teeth a metal-ceramic crown was positioned during follow-up. Patients included in the recall program (n=130) were blindly assessed both clinically and radiographically (every 2 years) to evaluate clinical symptoms and periapical status (PAI). The following variables were analyzed: age, tooth location, tooth type, initial diagnosis, PAI, root filling length and coronal restoration type. Chi-square-test and Multilevel analysis were performed to detect variables associated with treatment functionality and disease/lesions (p<0.05). A Cumulative teeth survival curve was constructed by means of Kaplan-Meier using extractions as the endpoint.

At the 20-year recall, 72 patients (31 M, 41 F; mean age 57.7±8.29 years; 196 teeth) completed the follow-up. Thirty-six patients were excluded for medical complications or died before the end of the study. Drop-outs consisted of 22 patients (17%) who did not complete the follow-up. Single metal-ceramic crowns were positioned after 4-6 months in 40% of teeth. Composite restorations were replaced with single metal-ceramic crowns during the follow-up in 53% of teeth after 8-19 years. Out of 196 teeth, 155 were classified as Survived (79%), 128 of which (65%) were Healthy (PAI≤2). Thirty-nine teeth (20%) were extracted for non-endodontic reasons. Twenty-nine teeth (15%) were classified as: re-exacerbation (11 teeth; 5.6%) or persistent asymptomatic lesions (18 teeth; 9%). Only 2 re-exacerbated teeth were extracted. Multilevel analysis confirmed the clinical relevance of tooth type (p=0.001) on Survived and Healthy teeth (p=0.007). Tooth location (p=0.0045) and initial diagnosis (p=0.019) significantly affected only Healthy teeth.

Root filled teeth were more frequently extracted for non-endodontic reasons rather than for endodontic disease. The majority of teeth with adequate root fillings, adequate restorations and included in a recall program remained functional and healthy for more than 20 years.

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To evaluate the views of final year dental surgery students (BDS; G1) at Cardiff University and general dental practitioners (GDPs; G2) within the geographic area of Cardiff, Wales, on antibiotic prescribing for endodontic conditions, and investigate the potential differences between the two groups.

A cross-sectional online questionnaire-based survey of 12 qualitative and quantitative questions was distributed to 76 final year BDS Cardiff University students and 55 dental practices within Cardiff, UK. Six questions recorded general information, and the remaining questions included a series of hypothetical clinical scenarios, where the participants were asked to state whether they would or would not prescribe antibiotics. The data were analysed using spss version 23 to produce descriptive statistics, contingency tables and to run chi-square (χ²) tests, Fisher's exact tests and relative risk calculations.

The response rate was 60% (n = 79). All G1 participants were aware of the consequences of antibiotic overuse. Approximately 60% of responders were aware of guidelines for antibiotic use in endodontic therapies, and 83% would only use antibiotics for a limited selection of patients (e.g. patients with systemic complications). G1 responses to clinical scenarios indicated overall that they were comparable to the ideal answers except for acute apical abscess (64% believed that antibiotics were indicated). The majority of G2 were aware of the consequences of antibiotic overuse. Only 28% of G2 were aware of guidelines for antibiotic use in endodontic therapies. Overall responses revealed that antibiotics would be prescribed for: systemic complications (78%), acute apical abscess (72%) and symptomatic apical periodontitis (28%). The clinical scenarios revealed G1 were more likely to prescribe antibiotics compared to G2 for cases of necrotic pulp with symptomatic apical periodontitis without systemic complications (incorrect answer) and less likely to other clinical scenarios such as necrotic pulp and asymptomatic apical periodontitis for patients with a history of rheumatic fever (ideal answers), symptomatic irreversible/reversible pulpitis, failure to achieve anaesthesia, chronic apical abscess for patients with diabetes. The recognition of antibiotic prescription for cases with signs of spreading infection was more evident in G2.

Final year undergraduate students were aware of the antibiotic resistance crisis, although a third was not aware of guidelines for use of antibiotics in endodontic conditions; their responses to clinical scenario were generally compatible with the guidelines. General dentists were less aware of the implications of overuse of antibiotics and the existence of guidelines, and their responses were occasionally incompatible with antibiotic guidelines for endodontic therapies.

To evaluate the cytocompatibility of Biodentine and iRoot FS with human periodontal ligament cells (hPDLCs).

Human periodontal ligament cells were characterized by flow cytometry and immunocytochemical analysis. Human periodontal ligament cell adhesion was assessed by scanning electron microscopy at day 3; proliferation by live/dead assay at days 1, 3 and 7; and osteogenic differentiation by alkaline phosphatase (ALP) activity staining, ALP quantification analysis and qRT-PCR at days 7 and 14. Data were analysed with anova and independent sample t-tests with SPSS 21.0.

Both iRoot FS and Biodentine increased the adhesion of hPDLCs at day 3. Compared to Biodentine, iRoot FS positively increased hPDLC proliferation on days 3 (P = 0.03) and 7 (P = 0.00). Osteogenic marker ALP was observed consistently in all samples, with iRoot FS having significantly higher ALP activity at day 14 (P = 0.00). Compared with Biodentine, iRoot FS significantly increased the mRNA level of ALP, COL1 and Runx2, and OCN increased only on day 14 (P < 0.05).

iRoot FS had a positive effect on the adhesion, proliferation and biomineralization of hPDLCs compared with Biodentine.

To investigate whether apical periodontitis lesions infected by Epstein–Barr virus (EBV) exhibit higher levels of oxidative stress biomarkers [8-hydroxydeoxyguanosine (8-OHdG) and oxidized glutathione (GSSG)] and bone resorption regulators [receptor activator of nuclear factor (NF-κB) ligand (RANKL) and osteoprotegerin (OPG)] compared to EBV-negative periapical lesions and healthy pulp tissues.

The experimental group consisted of 30 EBV-positive and 30 EBV-negative periapical lesions collected in conjunction with apicoectomy. The pulp tissues of 20 impacted third molars were used as healthy controls. The qualitative and quantitative analysis of EBV was performed by nested and real-time polymerase chain reaction (PCR), respectively. The levels of RANKL and OPG were analysed by reverse transcriptase real-time PCR. The levels of 8-OHdG and GSSG were determined by enzyme-linked immunosorbent assay (ELISA). Mann–Whitney U-test and Spearman's correlation were used for statistical analysis.

The levels of RANKL, OPG, 8-OHdG and GSSG were significantly higher in apical periodontitis lesions compared to healthy pulp controls (P = 0.001, P < 0.001, P < 0.001 and P < 0.05, respectively). RANKL and OPG mRNA expression was significantly higher in EBV-positive compared to EBV-negative periapical lesions (P < 0.05). There was no significant correlation between EBV copy numbers and levels of RANKL, OPG, 8OH-dG and GSSG in apical periodontitis.

Levels of bone resorption regulators and oxidative stress biomarkers were increased in apical periodontitis compared to healthy pulp tissues. EBV-positive periapical lesions exhibited higher levels of RANKL and OPG compared to EBV-negative periapical lesions. EBV may contribute to progression of apical periodontitis via enhanced production of bone resorption regulators.

To evaluate the effects of 2.8% or 10% calcium chloride (CaCl₂) in calcium aluminate cement (CAC) with either bismuth oxide (Bi₂O₃) or zinc oxide (ZnO) as radiopacifiers on the progression of osteogenic cell cultures.

Rat calvaria-derived cells were grown on Thermanox^® coverslips for 24 h and exposed to samples of (i) CACb: with 2.8% CaCl₂ and 25% Bi₂O₃; (ii) CACb+: with 10% CaCl₂ and 25% Bi₂O₃; (iii) CACz: with 2.8% CaCl₂ and 25% ZnO; or (iv) CACz+: with 10% CaCl₂ and 25% ZnO, placed on inserts. Nonexposed cultures served as the control. Calcium and phosphorus contents in culture media were quantified. The effects of the cements on cell apoptosis, cell viability and acquisition of the osteogenic cell phenotype were evaluated. Data were compared by Kruskal–Wallis test (α = 5%).

CACb+ promoted the highest levels of calcium in the culture media; CACz+, the lowest levels of phosphorus (P < 0.05). CACz+ and CACb increased cell apoptosis (P < 0.05). CACb reduced cell viability (P < 0.05) and the expression of the osteoblastic phenotype. CACz+ and CACb+ promoted greater cell differentiation and matrix mineralization compared to CACz and CACb (P < 0.05).

For CAC with the lower CaCl₂ content, the use of Bi₂O₃ was detrimental for osteoblastic cell survival and differentiation compared to ZnO, while CAC with the higher CaCl₂ content supported the acquisition of the osteogenic cell phenotype in vitro regardless of the radiopacifier used. Thus, CAC with 10% CaCl₂ would potentially promote bone repair in the context of endodontic therapies.

To establish whether irrigant activation techniques (IATs) result in greater intracanal smear layer and debris removal than conventional needle irrigation (CNI).

Six electronic databases were searched to identify scanning electron microscopy studies evaluating smear layer and/or debris removal following the use of manual dynamic activation (MDA), passive ultrasonic irrigation (PUI), sonic irrigation (SI) or apical negative pressure (ANP) IATs in mature permanent teeth. Meta-analyses were performed for each canal segment (coronal, middle, apical and apical 1 mm) in addition to subgroup analyses for individual IATs with respect to CNI. Outcomes were presented as standardized mean differences (SMD) alongside 95% confidence intervals (95% CI) and chi-squared analysis.

From 252 citations, 16 studies were identified. The meta-analyses demonstrated significant improvements in coronal (SMD: 1.15, 95% CI: 0.72–1.57 / SMD: 0.54, 95% CI: 0.29–0.80), middle (SMD: 1.30, 95% CI: 0.59–2.53 / SMD: 0.8, 95% CI: 0.58–1.13) and apical thirds (SMD: 1.22, 95% CI: 0.83–1.62 / SMD: 1.86, 95% CI: 0.76–2.96) for smear layer and debris removal, respectively. In the apical 1 mm IATs improved cleanliness; however, differences were insignificant (SMD: 1.15, 95% CI: -0.47–2.77). Chi-squared analysis revealed heterogeneity scores of 79.3–92.8% and 0.0–93.5% for smear layer and debris removal, respectively.

IATs improve intracanal cleanliness across a substantial portion of the canal, and therefore, their use is recommended throughout root canal preparation. However, current data is too heterogeneous to compare and identify superiority of an individual technique highlighting the need to standardize experimental protocols and develop a more representative research model to investigate the in vivo impact of IATs on clinical outcomes and periapical healing following root canal treatment.

To evaluate the torsional properties of pathfinding nickel-titanium (NiTi) rotary instruments manufactured from several NiTi alloys, ProGlider (M-wire), Hyflex GPF (conventional NiTi Wire and controlled memory wire), Logic (conventional NiTi wire and controlled memory wire) and Mtwo (conventional NiTi wire).

A total of 56 NiTi instruments from Glidepath rotary systems (n = 8) were used: Logic (size 25, .01 taper), Logic CM (size 25, .01 taper), ProGlider (size 16, .02 taper), Hyflex GPF (size 15, .01 taper), Hyflex GPF CM (size 15, .02 taper; size 20, .02 taper) and Mtwo (size 10, .04 taper). The torsion tests were performed based on ISO 3630-1 (1992). Three millimetres of each instrument tip was clamped to a small load cell by a lever arm linked to the torsion axis. Data were analysed using a one-way analysis of variance (anova) and Tukey test with a significance level at a = 5%.

The Logic size 25, .01 taper had significantly higher torsional strength values (P < 0.05). The ProGlider was significantly different when compared with Hyflex GPF size 15, .01 taper and size 15, .02 taper (P < 0.05). The Logic CM size 25, .01 taper had significantly higher torsional strength than Hyflex GPF size 15, .01 taper and size 15, .02 taper (P < 0.05). No difference was found amongst Mtwo size 10, .04 taper and Hyflex GPF groups (size 15, .01 taper; size 15, .02 taper; size 20, .02 taper). In relation to the angle of rotation, Logic CM size 25, .01 taper and Hyflex GPF size 15, .01 taper had the highest angle values (P < 0.05). The ProGlider had the lowest angle values in comparison with all the groups (P < 0.05) followed by Mtwo size 10, .04 taper. The Logic size 25, .01 taper had significantly higher angle of rotation values than ProGlider and Mtwo size 10, .04 taper (P < 0.05).

The Logic size 25, .01 taper instrument made of conventional NiTi alloy had the highest torsional strength of all instruments tested. In addition, the ProGlider instrument manufactured from M-Wire alloy had the lowest angle of rotation to fracture in comparison with the other instruments.

To investigate the feasibility of decellularizing the entire dental pulp using a mild treatment protocol to develop a decellularized biological extracellular matrix scaffold for use in regenerative endodontic procedures.

Decellularized human dental pulps were assessed using histological and immunohistochemical methods, scanning electron microscope and DNA quantification assay. Cytotoxicity assays to determine decellularized scaffold biocompatibility were also performed. Decellularized scaffolds were seeded with human dental pulp stem cells and cell viability assessed using Live/Dead^® stain. Quantitative data were analysed statistically using Student's t-test and one-way analysis of variance to compare mean values between groups depending on group numbers.

Assessment of decellularized tissues revealed an acellular matrix with preservation of native tissue histoarchitecture and composition. Decellularized tissues showed no evidence of cytotoxicity, with cell growth in direct contact with the scaffold and no reduction in cellular activity following extract incubation. Furthermore, the scaffold was able to support human dental pulp stem cell viability and attachment following recellularization.

Promising results were observed in developing a decellularized biological scaffold derived from the dental pulp with the perseveration of extracellular structural components which are required for tissue-specific regeneration.

To investigate in situ Enterococcus faecalis biofilm removal from the lateral canal of a simulated root canal system using passive or active irrigation protocols.

Root canal models (n = 43) were manufactured from transparent resin materials using 3D printing. Each canal was created with an 18 mm length, apical size 30, a .06 taper and a lateral canal of 3 mm length, with 0.3 mm diameter. Biofilms were grown in the lateral canal and apical 3 mm of the main canal for 10 days. Three models from each group were examined for residual biofilm using SEM. The other forty models were divided into four groups (n = 10). The models were observed under a fluorescence microscope. Following 60 s of 9 mL of 2.5% NaOCl irrigation using syringe and needle, the irrigant was either left stagnant in the canal or activated using gutta-percha, sonic or ultrasonic methods for 30 s. Images were then captured every second using an external camera. The residual biofilm percentages were measured using image analysis software. The data were analysed using generalized linear mixed models. A significance level of 0.05 was used throughout.

The greatest level of biofilm removal was obtained with ultrasonic agitation (66.76%) followed by sonic (45.49%), manual agitation (43.97%) and passive irrigation groups (38.67%), respectively. The differences were significant between the residual biofilm in the passive irrigation and both sonic and ultrasonic groups (P = 0.001).

Agitation resulted in better penetration of 2.5% NaOCl into the lateral canal of an artificial root canal model. Ultrasonic agitation of NaOCl improved the removal of biofilm.

To evaluate apical transportation and centring ability during root canal preparation in mesial root canals of mandibular molars associated with ProTaper Gold (PTG), ProDesign S (PDS), Hyflex CM (HCM), Hyflex EDM and ProDesign Logic (PDL).

Sixty mandibular first molars with two separate canals in the mesial root were selected after root anatomy pairing by microcomputed tomography (microCT). The teeth were randomly divided into five groups (n = 24); the root canal volume was calculated to ensure sample homogeneity. All the root canals were prepared up to size 25 in accordance with the instructions of each rotary system manufacturer. After root canal preparation, the teeth were scanned by microCT to analyse apical transportation, root canal centralization and the pre- and post-preparation root canal volume at the apical and cervical levels. Kruskal–Wallis and Dunn tests were used for comparisons amongst groups for transportation values. For volume changes, the parametric ANOVA and Tukey’s tests were used

There were no significant differences in apical transportation amongst the rotary systems (P > 0.05). All the systems created apical transportation; values ranging from 0.031 mm (PDL) to 0.072 mm (PTG), and enlargements between 39% (HCM) and 91.1% (PDS) were observed. In relative to cervical transportation, significant differences were observed amongst the systems (P < 0.05). Mean transportation values between 0.07 mm (HCM) and 0.172 mm (PTG) were found, with enlargements between 35.4% (HCM) and 51.5% (PDS).

All the thermally treated systems resulted in similar apical transportation. In the cervical region, the Hyflex CM and Prodesign Logic systems were associated with more centred preparations.

To evaluate the efficacy of sonic irrigation (EndoActivator^®) using various polymer tips and power settings in a stained collagen ex vivo model.

The root canals of fifty human, straight single-rooted extracted teeth were prepared to size 40, .08 taper. The roots were split longitudinally; stained collagen applied to the canal surfaces, photographed and re-assembled. The canals were subjected to syringe without supplementary (group 1, n = 10) or with supplementary sonic (groups 2–5, n = 10) irrigation. EndoActivator^® tip sizes (size 15, .02 taper for groups 2 and 3; size 35, .04 taper for groups 4 and 5) and power settings (low for groups 2 and 4; high for groups 3 and 5) were tested. After irrigation, the canals were re-photographed and the area of residual stained collagen was quantified using the UTHSCA Image Tool program (Version 3.0). The data were analysed using Wilcoxon signed rank test and general linear mixed models.

Supplementary sonic irrigation using EndoActivator^® resulted in significantly (P < 0.0001) less residual collagen compared with syringe irrigation only. Agitation of irrigant using the large EndoActivator^® tip with high power resulted in significantly less (22.4% – 29.5%) residual collagen compared to other combinations (large tip/low power P = 0.001; small tip/low power P = 0.01; small tip/high power P = 0.04). There was no significant difference amongst the latter three groups (P > 0.5).

Supplementary sonic irrigation using the EndoActivator^® system was significantly more effective in removing stained collagen from the canal surface than syringe irrigation alone. EndoActivator^® used with large tip (size 35, .04 taper) and high power setting in size 40, .08 taper canals was more effective than other combinations.

To investigate whether hypertension affects mineralization associated with white and grey mineral trioxide aggregate (MTA Angelus^®) implanted subcutaneously into rats by assaying osteoblastic biomarkers.

Polyethylene tubes containing grey MTA Angelus^®, white MTA Angelus^®, intermediate restorative material (IRM; positive control) or an empty tube (negative control) were implanted into the dorsal connective tissue of spontaneous hypertensive (n = 12) and Wistar (normotensive; n = 10) rats. Half of the rats in each group were killed after 7 days, and the remaining after 30 days. Tubes with surrounding tissue were removed, and immunostaining was performed to detect RUNX-2, OPN and OCN proteins. The normality of data was analysed using the Shapiro–Wilk test. Comparison of two independent groups was performed using the Mann–Whitney U-test, to detect a significant difference. A post hoc test accounting for multiple comparisons was performed following Tukey's test (P < 0.05).

Under hypertensive conditions after 30 days, both MTA materials were associated with immunolabelling for RUNX-2 from low to moderate, which was less than that observed at normal blood pressure and the 7-day groups (P < 0.05). The expression of OPN and OCN proteins under both MTA conditions was considered low after both 7 and 30 days for the hypertensive condition, and was less than that in animals with normal blood pressure after 30 days (P < 0.05). No immunostaining for any biomarkers in the control and IRM groups was observed (P < 0.05).

Hypertension decreased the immunostaining of RUNX-2, OPN and OCN biomarkers in response to MTA. Thus, hypertension can jeopardize the mineralization ability of MTA and may have a negative impact on endodontic treatment outcomes.

To investigate the effects of the pro-inflammatory and Th17-polarizing mediator IL-17 on HDPFs-mediated IL-23 production and the molecular mechanism involved.

Interleukin (IL)-17R expression was determined by semi-quantitative reverse transcriptase-polymerase chain reaction and Western blot in cultured human dental pulp fibroblasts (HDPFs). Quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay were used to determine IL-23 mRNA and protein levels in IL-17-stimulated HDPFs, respectively. The nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) signalling pathways that mediate the IL-17-stimulated production of IL-23 was investigated using Western blot and specific signalling inhibitor analyses. Statistical analyses were performed using Kruskal–Wallis tests followed by the Mann–Whitney U-test. Statistical significance was considered when the P value < 0.05.

Primary HDPFs steadily expressed IL-17R mRNA and surface-bound protein. IL-17 stimulated the expression of IL-23 mRNA and protein in cultured human dental pulp fibroblasts, which was attenuated by IL-17 or IL-17R neutralizing antibodies. In accordance with the enhanced expression of IL-23, IL-17 stimulation resulted in rapid activation of p38 MAPK, extracellular signal-regulated kinase (ERK) 1/2, c-Jun-N-terminal kinase (JNK) and NF-κB in HDPFs. Inhibitors of p38 MAPK, ERK 1/2 or NF-κB significantly suppressed, whereas blocking JNK substantially augmented IL-23 production from IL-17-stimulated HDPFs.

HDPFs expressed IL-17R and responded to IL-17 to produce IL-23 via the activation of the NF-κB and MAPK signalling pathways. The findings provide insights into the cellular mechanisms of the participation of IL-17 in the activation of HDPFs in inflamed pulp tissue.

To evaluate the effects of progressive apical enlargement on the amount of unprepared root canal surface area and remaining dentine thickness.

The root canals of 30 extracted mandibular incisors with Vertucci′s type I configuration were instrumented with rotary HyFlex CM instruments (Coltene-Whaledent, Altstätten, Switzerland) up to 4 instruments larger than the first one that bound at the working length (WL). Teeth were scanned in a micro-computed tomography (micro-CT) device before canal preparation and after instrumentation with the 2nd, 3rd and 4th larger instruments. The amount of unprepared surface area in the full canal or in the apical 4 mm as well as the remaining dentine thickness at 10 mm from the WL were calculated and compared. The general linear model for repeated measures adjusted by Bonferroni's post hoc test was used for statistic analysis.

There was a significant reduction in the amount of unprepared areas after each increase in preparation size (P < 0.01). This was observed for both the full canal length and the 4-mm apical segment. The amount of remaining dentine was also significantly reduced after each file size (P < 0.01). However, dentine thickness always remained greater than 1 mm, even after using the largest instrument.

Apical preparations up to 4 instruments larger than the first one to bind at the WL caused a significant progressive reduction in the unprepared canal area.

To assess the stability of NaOCl solutions when combined with a novel product for clinical use, Dual Rinse HEDP, which contains etidronate (1-hydroxyethane 1,1-diphosphonate).

Mixtures of NaOCl solutions with Dual Rinse HEDP were prepared so that they initially contained 5.0%, 2.5% or 1.0% NaOCl and always 9.0% of dissolved Dual Rinse HEDP powder per total weight. NaOCl solutions alone were used as controls. The stability of these solutions over 8 h was assessed in transparent borosilicate glass bottles at ambient temperature (23 °C). Subsequently, the effects of heating (60 °C) or storing the solutions at 5 °C were studied in polypropylene syringes. NaOCl concentrations were measured by iodometric titration, that is free available chlorine contents. Experiments were performed in triplicate.

In the glass bottles at 23 °C, the 5.0% NaOCl/9.0% Dual Rinse HEDP solution lost 20% of the available chlorine after 1 h, whilst the corresponding 2.5% NaOCl and 1.0% NaOCl solutions retained this relative amount of available chlorine for 2 and 4 h, respectively. Results obtained in the glass bottles were similar to those achieved in the syringes. Heating of the NaOCl/Dual Rinse HEDP mixtures had a detrimental effect on available chlorine, with a complete loss after 1 h. In contrast, storing the NaOCl/Dual Rinse HEDP mixtures in a refrigerator at 5 °C kept the available chlorine high for 7 h, with the expected loss after a further hour of storage at 23 °C.

Initial NaOCl concentration and temperature both affected short-term storage stability of combined solutions containing Dual Rinse HEDP.

To characterize the potential of human periodontal ligament fibroblasts (HPLF) to synthesize CRP and Th-related cytokines in response to IL-6 in periodontal health and apical inflammation.

Primary HPLF stimulated with IL-6, soluble(s) IL-6 receptor (R) and controls were assayed for CRP, Th1, Th2, Th17 and Treg-related cytokines by quantitative real-time PCR and ELISA, respectively. IL-6R mRNA expression and its soluble protein levels were screened in HPLF cultures, and ex vivo samples of healthy periodontal ligaments (n = 5) and apical lesions (n = 13). Data were analysed with ANOVA or unpaired t-test.

0.5 ng mL⁻¹ IL-6 plus 1 ng mL⁻¹ of its soluble receptor (sIL-6R) for 24 h was effective in inducing CRP production. IL-6 alone had a mild dose-dependent effect; co-stimulation with sIL-6R significantly enhanced this effect, whereas it was completely abolished by the addition of IL-6R blocking antibody (P < 0.05). Similarly, higher mRNA expression and protein levels of Th1, Th17 and partially Treg-related cytokines were found for IL-6 combined with its soluble receptor versus the nonstimulated group and IL-6R antibody (P < 0.05). IL-6R mRNA expression was slightly induced by IL-6 compared to THP-1 cells, but sILR-6 protein could not be detected in HPLF. High sIL-6R levels were detected in apical lesions and were immunolocalized to mononuclear inflammatory cells and proliferating epithelium.

IL-6 trans-signalling induced Th1 and Th17-related cytokines and represents an extra-hepatic mechanism for PCR synthesis in human periodontal ligament fibroblasts, contributing to explain the bone-destructive phenotype of apical lesions and eventually its systemic complications.

To investigate the proliferation and differentiation potential of human dental pulp stem cells (DPSCs) in a three-dimensional culture model (TDM) by incorporation of VEGF and BMP-2.

TDM was established using fibrin gel (fg) as a soft tissue matrix and demineralized dentine disc (dd) as a hard tissue matrix. DPSCs and vascular endothelial growth factor (VEGF) were encapsulated in fibrin gel (fg-VEGF) and then inserted into bone morphogenetic protein (BMP-2)-coated demineralized dentine discs (dd-BMP-2). DPSCs were incubated for 28 days in various fg/dd combinations in the absence or presence of VEGF and BMP-2. Proliferation and morphology of DPSCs in fibrin gel were analysed using MTT and Live&Dead assays. Release profiles of VEGF and BMP-2 from fibrin gel and dentine discs were quantified using ELISA, and the expressions of angiogenic and odontogenic differentiation markers were determined with RT-qPCR analysis. Data were analysed statistically using Wilcoxon signed rank tests, Kruskal–Wallis tests with Mann–Whitney U tests and Bonferroni adjustment. The level of significance was set at P < 0.05.

DPSCs were able to proliferate and showed interconnected cellular elongations in fibrin gel depending on fibrinogen concentration whilst monolayer control group showed typical fibroblast-like cell morphology. Encapsulating of VEGF in fibrin gel and BMP-2 in gelatin that was used to coat dentine discs allowed the controlled releases of growth factors, which induced angiogenic and odontogenic gene expressions by DPSCs. Higher expressions of PECAM as an angiogenic factor, and BSP, DMP-1, OCN and CBFA as odontogenic factors, were observed in TDM as compared to the other fg/dd combinations and the monolayer control group (P < 0.05).

TDM consisting of fibrin gel and dentine matrix allowed cell–cell interactions. TDM was highly effective in delivering both VEGF and BMP-2 that enhanced the angiogenic and odontogenic potential of DPSCs.

To characterize chemical degradation of the principal constituents of dentine after exposure to NaOCl and EDTA using Infrared Spectroscopy (ATR–FTIR).

Ground dentine particles, from extracted permanent human molars, were passed through sieves of 38 to 1 000 μm to provide six size ranges. Portions (250 mg) of each size range were reacted with 5 mL of 2.5% NaOCl for 2-10 min; or 17% EDTA for 5-1440 min. Powders larger than 75 μm were also sequentially exposed to NaOCl/EDTA/NaOCl each for 10 min. All experiments were repeated five times. Reacted and unreacted powders were washed and dried. Particles larger than 75 μm were then reground. FTIR spectra of unground and reground reacted particles enabled assessment of particle surface versus bulk chemistry, respectively, plus estimation of reaction depth. Changes in the ratio of the 1 640 cm⁻¹ collagen: 1 010 cm⁻¹ phosphate peak height or its inverse were obtained. These were used to estimate surface and bulk fraction reacted and thus depth to which collagen or phosphate was reduced following immersion in NaOCl or EDTA, respectively. The data were analysed descriptively.

Surface collagen fraction declined by ~40% within 2 min of NaOCl exposure, and plateaued at ~60% between 6-10 min. Bulk spectra showed average depth of collagen loss at 10 min was 16 ± 13 μm. Ten minute EDTA exposure caused ~60% loss of surface phosphate. Average depth of phosphate loss was 19 ± 12 μm and 89 ± 43 μm after 10 and 1 440 min EDTA immersion, respectively. Sequential NaOCl/EDTA immersion yielded a 62 ± 28 μm thick phosphate-depleted surface. Sequential NaOCl/EDTA/NaOCl treatment resulted in approximately 85 μm of collagen loss.

Data revealed the sequential depletion of collagen by NaOCl and apatite by EDTA in dentine, simultaneously exposing the other moieties. Alternate exposure to NaOCl and EDTA therefore enhances the depth of erosion.

To compare the pre-sterilization cleaning of rotary Ni-Ti files of different sizes previously used a. ex vivo and b. clinically by a washer–disinfector, a regular ultrasonic bath, and the same ultrasonic bath in combination with a recently developed cavitation intensifying method.

Two sets of two hundred rotary Ni-Ti files, one previously used ex vivo and another one used clinically, were collected from the undergraduate and postgraduate clinics of the Academic Centre for Dentistry Amsterdam (ACTA). The instruments were immersed in an enzymatic solution and subsequently cleaned either by a washer–disinfector, a regular ultrasonic bath combined with a glass beaker, the same bath combined with a beaker lined with two cavitation intensifying sheets or with two standard plastic sheets. The positive control consisted of used files that did not undergo any cleaning and the negative control included new unused files. The instruments were then stained to reveal remaining protein material and scored under a stereoscopic microscope. The results were analysed by nonparametric statistical tests (α = 0.05).

No significant difference was found between the combination of the ultrasonic bath and the regular glass beaker and the same ultrasonic bath with the beaker lined with the cavitation intensifying sheets. The washer–disinfector left significantly more debris compared to the latter group when clinically used files were evaluated (P ≤ 0.001). The effect of instrument size on cleaning was not consistent.

None of the tested methods was able to remove all residual protein material from the files; however, it could be noted that this study did not follow the reprocessing protocol provided by the manufacturer.