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Abstract Background Intrauterine fetal death (IUFD) is a tragic event and, despite efforts to reduce rates, its incidence remains difficult to reduce. The objective of the present study was to examine the etiological factors that contribute to the main causes and conditions associated with IUFD, over an 11-year period in a region of North-East Italy (Friuli Venezia Giulia) for which reliable data in available. Methods Retrospective analysis of all 278 IUFD cases occurred between 2005 and 2015 in pregnancies with gestational age ≥ 23 weeks. Results The incidence of IUFD was 2.8‰ live births. Of these, 30% were small for gestational age (SGA), with immigrant women being significantly over-represented. The share of SGA reached 35% in cases in which a maternal of fetal pathological condition was present, and dropped to 28% in the absence of associated pathology. In 78 pregnancies (28%) no pathology was recorded that could justify IUFD. Of all IUFDs, 11% occurred during labor, and 72% occurred at a gestational age above 30 weeks. Conclusion The percentage of IUFD cases for which no possible cause can be identified is quite high. Only the adoption of evidence-based diagnostic protocols, with integrated immunologic, genetic and pathologic examinations, can help reduce this diagnostic gap, contributing to the prevention of future IUFDs.

Entender o comportamento mecânico de enrocamentos é, antes de tudo, garantir a segurança dos seres que residem à jusante de barragens. A Usina Hidrelétrica Machadinho teve sua construção finalizada no ano de 2002, tendo como principal característica a inserção de basalto vesicular em até 30% da constituição do corpo do aterro principal, cuja litologia predominante é o dacito. Os trabalhos mais recentes, disponíveis na literatura de enrocamento, têm alertado sobre a alterabilidade das propriedades mecânicas das rochas, uma vez que obras de barragens possuem tempo de vida útil muito longo. Este trabalho teve como objetivo avaliar as estruturas e mineralogia dos enrocamentos de Machadinho. Foram realizadas microscopias eletrônicas e análises petrográficas, tendo-se ainda avaliado os componentes químicos em áreas específicas por meio de espectrometria de dispersão. As constituições mineralógicas dos enrocamentos da UHE Machadinho indicaram elevadas porcentagens de vidro vulcânico nas estruturas, sobretudo o basalto vesicular, direcionando-se para a necessidade da avaliação da ação do intemperismo ao longo do tempo no comportamento mecânico deste material, como garantia de segurança da barragem.

Objectives: The objective of this research was to evaluate the volumetric and surface stability in vitro, to different pH levels, of calcium silicate-based repair cements ready-to-use and powder/liquid exposed., Materials and Methods: Sixty human teeth were retro-prepared to a depth of 3 mm and divided into two groups. The first group included powder/liquid cements Biodentine (Septodont, France) and MTA HP (Angelus, Brazil); the second group included ready-to-use cements Bio-C Repair (Angelus, Brazil) and ENDOCEM MTA (Maruchi, Republic of Korea). All samples were submerged in PBS solution with pH levels of 5, 7, and 12. Initially, the materials were evaluated for porosity, marginal adaptation, and volumetric change using micro-CT, and then re-evaluated after 7 and 30 days of exposure to PBS solution. The samples were characterized using SEM/EDS and RAMAN spectroscopy. Statistical evaluation was performed using ANOVA with a 5% significance level., Results: Bio-C Repair showed significant statistical differences (p < 0.05) at different pHs in 7 and 30 days compared to other materials, being greater in acidic medium. ENDOCEM, regardless of pH, at 30 days showed fewer voids when compared to other repair cements., Conclusion: This micro-CT study demonstrated that regardless pH, the clinic presentation of calcium silicate-based materials ready-to-use showed higher and loss volumetric than powder/liquid materials at 30 days., Clinical Relevance: Under neutral pH conditions, calcium silicate-based materials are expected to maintain properties such as bioactivity and low solubility. However, in clinical situations involving infection and inflammation, pH variations can impact the physicochemical properties of these materials regardless of their clinical presentation., Competing Interests: Declarations. Ethical approval: All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and national research committee. This research meets the requirements of the specific resolutions on ethics in research with human beings of the National Health Council– Ministry of Health and was approved by The Research Ethics Committee of the Faculdade de Odontologia de Piracicaba (FOP) of the Universidade Estadual de Campinas (UNICAMP) (53383921.5.0000.5418). Informed consent: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: To investigate volumetric changes, in vivo biocompatibility, and systemic migration from eight commercial endodontic sealer materials in paste/paste, powder/liquid, and pre-mixed forms., Materials and Methods: The sealers AH Plus Bioceramic, AH Plus Jet, BioRoot RCS, MTApex, Bio-C Sealer, Bio-C Sealer Ion+, EndoSequence BC Sealer and NeoSEALER Flo were studied. After characterisation by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), Raman spectroscopy and X-ray diffractometry (XRD), tubes were implanted in Wistar rats' alveolar bone and subcutaneous tissues. Micro-CT evaluated volumetric changes pre/post 30 days of implantation. Histological and immunohistochemistry analyses assessed biocompatibility. Kidney samples underwent spectrometry (ICP-MS) for tantalum, tungsten and zirconium. Statistical analysis determined normality and significance (udp < 0.05)., Results: Characterisation revealed calcium, silicon, and radiopacifiers in the materials. Volumetric changes showed greater alteration in subcutaneous tissues than alveolar bone; BioRoot RCS and MTApex (powder/liquid) were most stable. Histological analysis indicated intense inflammation for AH Plus Jet, moderate for others; IL-10 was marked positively for all materials. AH Plus Jet had an 18-fold higher tungsten and a 37-fold higher zirconium mass fraction in kidneys versus controls, while tantalum showed lower accumulation patterns., Conclusion: Root canal filling materials' responses varied by implantation site and form, demonstrating acceptable biocompatibility. Tantalum and zirconium oxide radiopacifiers appear systemically safe; tungsten-based radiopacifiers are unsuitable due to metal accumulation risks., Clinical Relevance: This study highlights the need for further in vivo studies on endodontic sealers' chemical, biological, and physical behaviors and their systemic migration., Competing Interests: Declarations. Ethical approval: This study was approved by the institutional ethics committee animal research of the Piracicaba Dental School (code CEUA 5888-1/2021 and 5889-1/2021). Consent to participate: This study did not involve humans. Competing interests: The authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Short tandem repeat analysis is a robust and reliable DNA analysis technique that aids in source identification of a biological sample. However, the interpretation, particularly when DNA mixtures are present at low levels, can be complicated by the presence of PCR artifacts most commonly referred to as stutter. The presence of stutter products can increase the difficulty of interpretation in DNA mixtures as well as low-level DNA samples down to a single cell. Stutter product formation is stochastic in nature and although methods exist that can estimate the magnitude of stutter product formation, it still is not well understood. With the increased sensitivity of forensic DNA analyses, it has become possible to obtain interpretable DNA profiles from as low as 6.6 pg of DNA, or a single human diploid cell. However, this presents an interpretational challenge because the stutter in these low-level DNA samples might stray from the expected patterns observed in high-level DNA samples. Therefore, this project focuses on characterizing stutter in single cell samples to help generate a deeper understanding of stutter and provide a guide for detecting and evaluating stutter in low-level samples. Stutter analysis was performed using data generated from 180 single cells isolated with the DEPArrayTM NxT, amplified using the PowerPlex Fusion 6 C amplification kit at 29 or 30 cycles. Stutter was successfully characterized in single cells and stutter percentages were highly elevated compared to high-level samples where the variance increased as the number of cells being analyzed decreased leading to potential high stutter at low DNA levels. Using empirical and simulated (resampled) data, this study also reinforces historically relevant patterns in stutter product formation and demonstrates the relative differences in stutter in n-1, n-2 and n + 1 stutter product formation in simple, complex and compound repeats., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Aim: Calcium silicate-based cements have been widely used in dentistry mainly due to their physicochemical and biological properties. Commercially available materials use radiopacifiers containing metals (bismuth, tantalum, tungsten and/or zirconium). To investigate volumetric changes, in vivo biocompatibility and systemic migration from eight commercially available materials, including powder/liquid and 'ready-to-use' presentations., Methodology: After characterization, tubes were implanted in healthy Wistar rats' alveolar bone and subcutaneous tissues. Micro-CT was used to evaluate volumetric change before/after 30 days of implantation. Histological and immunohistochemistry analysis were used to evaluate materials' biocompatibility. After euthanasia, kidney samples were retrieved, acidic digested and evaluated by inductively coupled plasma mass spectrometry (ICP-MS) for bismuth, tantalum, tungsten and zirconium mass fractions. Statistical analysis compared the results for normality and comparisons adopted a level of significance of 0.05., Results: Characterization photomicrographs and spectroscopy analysis revealed calcium, silicon and radiopacifiers for tested cements. Volumetric changes after implantation showed higher alteration in subcutaneous tissues than alveolar bone indicating that Biodentine, EndoSequence BC RRM Putty and ProRoot MTA were the most stable materials. Histological analysis found intense inflammation for NeoPUTTY and moderate for the other materials; osteocalcin and osteopontin were positively marked for all materials. Despite its volumetric stability, ProRoot MTA showed a 1000-fold higher mass fraction of bismuth accumulation and MTA Repair HP a 37-fold higher tungsten accumulation in kidney samples when compared with the nonexposed controls. All tantalum-analysed samples indicated a similar mass fraction with the nonexposed controls. Biodentine exhibited a significant lower kidney mass fraction of zirconium accumulation when compared with this control., Conclusions: Volumetric analysis revealed that Bio-C Repair, NeoPUTTY and MTA Repair HP presented greater volumetric loss when implanted in the subcutaneous tissue. NeoPUTTY presented more intense inflammatory infiltrate. Systemic migration analysis highlighted the predominance of bismuth in the ProRoot MTA group. These results suggest that endodontic repair cements are affected by their chemical composition, the type of implant tissue and different clinical settings., (© 2025 British Endodontic Society. Published by John Wiley & Sons Ltd.)

This study evaluated the glass fibre post (GFP) removal using three different protocols with ultrasonic tips. Thirty-six extracted single-rooted teeth were endodontically treated and cemented with a 10-mm GFP. Three protocols were used for GFP removal: 20 s activated with cooling, 20 s activated without cooling and 40 s activated without cooling. The removal time and temperature increase on the external root surface were recorded. The amount of residual material and the volume of dentin loss were assessed using micro-computed tomography. Data were analysed using ANOVA with a 5% significance level. The removal time and the amount of residual material were not affected by the protocol. There was no difference in temperature increase between the 20-s groups. The 40s activated without cooling produced the highest temperatures, especially in the apical region, and greater dentin loss. It is not advisable to use the ultrasonic insert continuously for more than 20 s., (© 2024 Australian Society of Endodontology Inc.)

Aim: The present study examined the leaching and cytotoxicity of bismuth from ProRoot MTA and aimed to identify whether bismuth leaching was affected by the cement base and the immersion regime used., Methodology: The leaching profile of bismuth was examined from ProRoot MTA and compared with hydroxyapatite containing 20% bismuth oxide as well as hydroxyapatite and tricalcium silicate to investigate whether bismuth release changed depending on the cement base. Bismuth leaching was determined after 30 and 180 days of ageing immersed in Dulbecco's modified Eagle's medium (DMEM) using mass spectroscopy (ICP-MS). The media were either unchanged or regularly replenished. The pH, surface microstructure and phase changes of aged materials were assessed. Wistar rat femoral bone marrow stromal cells (BMSCs) and cutaneous fibroblasts were isolated, cultured and seeded for cell counting (trypan blue live/dead) after exposure to non-aged, 30- and 180-days-aged samples in regularly replenished DMEM. Aged DMEM in contact with materials was also used to culture BMSCs to investigate the effect of material leachates on the cells. Gene expression analysis was also carried out after direct exposure of cells to non-aged materials. Differences between groups were statistically tested at a significance level of 5%., Results: All materials exhibited alterations after immersion in DMEM and this increased with longer exposure times. The bismuth leached from ProRoot MTA as detected by ICP-MS. Aged ProRoot MTA samples exhibited a black discolouration and surface calcium carbonate deposition. ProRoot MTA influenced cell counts after direct exposure and its 180-days leachates reduced BMSC viability. After direct BMSC contact with non-aged ProRoot MTA an upregulation of metallothionein (MT1 and MT2A) expression and down-regulation of collagen-1a (Col-1a) and bone sialoprotein (BSP) expression was identified., Conclusions: Bismuth leaching was observed throughout 180-days observation period from all materials containing bismuth oxide. This negatively influenced cell viability and gene expression associated with bismuth exposure. This is the first study to report that metallothionein gene expression was influenced by exposure to ProRoot MTA., (© 2024 The Author(s). International Endodontic Journal published by John Wiley & Sons Ltd on behalf of British Endodontic Society.)

Microplastics (MPs) and nanoplastics are small synthetic organic polymer particles (<5 mm and <1 μm, respectively) that originate directly from plastic compounds or result from the degradation of plastic. These particles are a global concern because they are widely distributed in water, air, food, and soil, and recent scientific evidence has linked MPs to negative biological effects. Although these particles are difficult to detect in humans, MPs have been identified in different biological fluids and tissues, such as the placenta, lung, intestines, liver, blood, urine, and kidneys. Human exposure to MPs can occur by ingestion, inhalation, or dermal contact, potentially causing metabolic alterations. Data from experimental and clinical studies have revealed that the ability of MPs to promote inflammation, oxidative stress, and organ dysfunction and negatively affect clinical outcomes is associated with their accumulation in body fluids and tissues. Although evidence of the putative action of MPs in the human kidney is still scarce, there is growing interest in studying MPs in this organ. In addition, chronic kidney disease requires investigation because this condition is potentially prone to MP accumulation. The purpose of the present article is (i) to review the general aspects of MP generation, available analytic methods for identification, and the main known biological toxic effects; and (ii) to describe and critically analyze key experimental and clinical studies that support a role of MPs in kidney disease., (Copyright © 2024 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)

Background and Objective: Osteoporosis is associated with bone microarchitecture alterations, and the depletion of estrogen during menopause is a major contributing factor to its development. The literature highlights the noteworthy role of gut microbiota in bone metabolism, particularly in the progression of osteoporosis. Periodontal disease leads to alveolar bone loss, which may be influenced by estrogen deficiency, and this mechanism is intricately associated with an imbalance in systemic microbiota. The aim of this study was to evaluate the effects of Bifidobacterium animalis subsp. lactis HN019 (B. lactis HN019) and Lacticaseibacillus casei 01 (L. casei 01) administrations on an osteoporosis animal model., Materials and Methods: Thirty-three female rats were randomly divided into three groups: control (C-OVX), C-OVX-HN019 and C-OVX-LC01. All animals were ovariectomized. In groups C-OVX-HN019 and C-OVX-LC01, the probiotics were administered for 4 months. All animals were euthanized after 16 weeks from ovariectomy. Microtomographic, histopathological and immunohistochemical examinations were conducted on periodontal tissues, whereas histomorphometry, histopathological and immunohistochemical analyses were carried out on the intestine. The levels of estradiol were assessed in blood using an immunoenzymatic assay. The data were subjected to statistical analyses (p < .05)., Results: The C-OVX-LC01 group exhibited a significant reduction in alveolar bone porosity and an increase in connective tissue density compared to C-OVX (p < .05). The C-OVX-HN019 and C-OVX-LC01 groups presented reduced expression of TRAP and RANKL compared to the C-OVX (p < .05). The C-OVX group presented villi defects, mild neutrophil infiltration, decrease in both villous height and intestinal crypts and reduced expression of intestinal junctional epithelium markers e-cadherin and claudin 01 compared to C-OVX-HN019 and C-OVX-LC01 (p < .05). The C-OVX group had lower estradiol levels than C-OVX-HN019 and C-OVX-LC01 (p < .05)., Conclusion: The probiotic therapy promoted a reduction in alveolar bone destruction and intestinal permeability as well as an increase in estradiol levels in ovariectomized rats. Specifically, the probiotic strain Lacticaseibacillus casei 01 exhibited greater effectiveness compared to Bifidobacterium animalis subsp. lactis HN019, indicating strain-dependent outcomes., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

DNA mixture deconvolution in the forensic DNA community has been addressed in a variety of ways. "Front-end" methods that separate the cellular components of mixtures can provide a significant benefit over computational methods as there is no need to rely on models with inherent uncertainty to generate conclusions. Historically, cell separation methods have been investigated but have been largely ineffective due to high cost, unreliability, and the lack of proper instrumentation. However, the last decade has given rise to more innovative technology that can target and recover cells more effectively. This study focuses on the development and optimization of a method to selectively label and recover male cells in a mixture of male and female epithelial cells using a Y-chromosome labeling kit with DEPArray™ technology, whereby male cells are labeled and recovered into a single extraction-ready tube. Labeling efficiency was tested using freshly collected and aged buccal swabs where 70%-75% and 38% of male cells were labeled, respectively, with less than 1% false positives. DEPArray™ detection was assessed using single buccal epithelial cells where approximately 80% of labeled cells were identified as male. Mixtures (1:1, 1:10, male to female) yielded profiles that were predominantly single source male or those in which the male component was more easily interpreted. The male-specific labeling method was demonstrated to be both robust and reliable when used on freshly collected cells. While the DEPArray™ meditated detection and recovery had notable limitations, it still improved the interpretation of the male component in same-cell mixtures in more recently collected samples., (© 2024 The Authors. Journal of Forensic Sciences published by Wiley Periodicals LLC on behalf of American Academy of Forensic Sciences.)

Introduction: EDTA plays a crucial role in regenerative endodontic therapy (RET) because of its significant biological effects. However, EDTA is also recognized as the preferred anticoagulant for hematologic tests. Thus, this study aimed to assess the influence of different EDTA activation techniques on the morphology of blood clots after conditioning the root canal dentin., Methods: Forty extracted human teeth were prepared to simulate immature teeth and divided into the following 5 groups: (1) saline solution (negative control), (2) EDTA 17% + saline solution (CNI), (3) CNI + ultrasonic activation, (4) CNI + Easy clean activation, and (5) CNI + XP-endo Finisher activation. After irrigation, the roots were cleaved, and the root canals were filled with human blood to clot formation. The morphology and density of erythrocytes, platelets, and the fibrin network were observed using a scanning electron microscope. The fibrin network density was classified using a 4-point scale. Data were analyzed using the Friedman test and the Kruskal-Wallis test with Bonferroni adjustment (α = 5%)., Results: All groups exhibited consistent blood clot morphology characterized by a high density of erythrocytes, platelets, and white blood cells throughout the entire length of the root canal. The negative control group showed statistically significant high scores of fibrin density compared with the CNI group in all root thirds (P < .05). However, there was no statistical difference in the scores for the fibrin network density between the groups irrigated with EDTA with and without activation (P > .05)., Conclusions: EDTA may impair the fibrin network formation compared with the saline group. However, EDTA activation did not significantly change the effects on the blood clot in contact with the conditioned intraradicular dentin., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Objectives: The aim of this work is to test experimental cements, doped with a silicate based bioactive nanoparticle (NanoBiosilicate). Methods, we synthesized a glass nanoparticle by Sol-Gel Stöber method, used to be incorporated in a dental material for endodontic uses., Materials and Methods: We assess the mineralizing properties and biocompatibility. Besides the crystallography characterization of the resultant new crystals. Results, After analysis, and comparison with commercial materials, the material tested was similar in mechanical properties required by ISO, The ion release was effective after 2 hr. of setting and the novel material was cell compatible accepted by ISO., Results: We found new formed Calcium Phosphate peaks in the spectroscopic analysis (FTIR), remarkably the crystals formed were comparable to hydroxyapatite when analyzed with a Selected Area Electron Diffractometer, with rings of 2.84 Å for 002, and the 2.77 Å is also visible for 210. The 6.83 Å and 6.88 Å, for respective 222 and 004. The incorporation of Chlorhexidine was not detrimental for this property, Significance, the features mentioned represented a progress in biomineralization field that was associated to an improved mineral structure formation with increased crystallographic similarity to natural hydroxyapatite. When chlorhexidine was added a favorable biomodification of the remaining collagen in dentinal walls and antimicrobial activity potential were also observed., Competing Interests: Declaration of Competing Interest We wish to confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome.We confirm that the manuscript has been read and approved by the authors and that there are no other persons who satisfied the criteria for authorship but are not listed.We further confirm that we have given due consideration to the protection of intellectual property associated with this work and that there are no impediments to publication. In so doing we confirm that we have followed the regulations of our institutions concerning intellectual property., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Dental referrals to the Endodontics Specialty Clinic (ESC) are routine owing to the complexity of endodontic treatments. To obtain a better prognosis for treatment, students/dentists must perceive their technical limits. This study sought to investigate the referrals of patients to the ESC from different clinics of the Piracicaba Dental School, State University of Campinas - SP, Brazil, and check: a) the demographic profile of patients and the most commonly affected tooth; b) the clinic with the largest number of referrals; c) the reasons for referrals; d) the complexity of the cases; e) and the difficulty in assessing the referred cases based on the classification provided by the American Association of Endodontists (AAE) and Souza-Filho. The study sample consisted of patients' electronic dental referral records from February 2015 to June 2019. A total of 1,707 patients were referred to the ESC during the study period, and 62.4% were female. Lower molars were the most frequently involved teeth (34.8%), and 60.7% of the cases were referred due to the presence of root curvature. The AAE classification showed prevalence of highly difficult cases (71.3%), whereas Souza-Filho classification demonstrated a high rate of class III cases (85.8%). This study highlights the difficulties encountered by undergraduate students before or during endodontic treatments, reinforcing the need for clear criteria for selecting cases appropriate for each education level, thus improving endodontic treatment prognosis.

The objective of this study was to investigate the presence of culturable bacteria, endotoxins (LPS) and lipoteichoic acid (LTA) levels in teeth with normal vital pulp (NVP) with intact crowns (IC) and those with coronal restoration (CR) limited to the enamel level. A total of 20 teeth indicated for endodontic treatment due to prosthetic reasons were selected. Samples were collected from the root canals. The levels of cultivable bacteria, LPS and LTA were assessed. Statistical analyses were performed at significance level set at 5%. None of the teeth presented microbial growth. In the IC group, the LPS levels were limited to the lowest concentration of LPS. On the contrary, higher LPS and LTA levels were detected in teeth with CR. It was concluded that teeth with NVP and IC were negative for bacteria, LPS and LTA; while teeth with CR were positive for bacterial virulence factors., (© 2023 Australian Society of Endodontology Inc.)

Most commercially available STR amplification kits have never been fully validated for low template DNA analysis, highlighting the need for testing different PCR kits and conditions for improving single-cell profiling. Here, current strategies rely mainly on adjusting PCR cycle number and analytical threshold settings, with a strong preference for using 30 amplification cycles and thresholds at 30-150 RFU for allele detection. This study aimed to (1) determine appropriate conditions for obtaining informative profiles utilizing a dilution series, and (2) test the outcome on single cells using the DEPArray™ technology. Four routinely applied forensic STR kits were compared by using three different amplification volumes and DNA dilutions down to 3.0 pg, while two well-performing kits were used for single/pooled leucocyte and sperm cell genotyping. Besides reduced costs, the results demonstrate that a 50%-75% PCR volume reduction was beneficial for peak height evaluation. However, this was counteracted by an increased artifact generation in diluted DNA volumes. Regarding profile completeness, the advantage of volume reduction was only prominent in samples processed with Fusion 6C. For single and pooled cells, ESIFast and NGMDetect provided a solid basis for consensus profiling regarding locus failure, although locus dropouts were generally observed as stochastic events. Amplification volume of 12.5 μL was confirmed as appropriate in terms of peak heights and stutter frequencies, with increased stutter peaks being the main artifact in single-cell profiles. Limitations associated with these analyses are discussed, providing a solid foundation for further studies on low template DNA., (© 2023 The Authors. Journal of Forensic Sciences published by Wiley Periodicals LLC on behalf of American Academy of Forensic Sciences.)

Introduction: Regenerative endodontic procedures provide conditions for the continuity of apical formation and increase of the dentin wall thickness, being a useful technique to resume root development of necrotic teeth with incomplete apex. This study aimed to evaluate the coronal discoloration promoted by a new intracanal calcium silicate-based dressing and compare it with other intracanal dressings used in regenerative endodontic procedures., Methods: Sixty bovine incisors were prepared to simulate teeth with incomplete apexes and divided according to the intracanal dressing used: triple antibiotic paste-double antibiotic paste-DAP, calcium hydroxide associated with 2% chlorhexidine gel, calcium hydroxide associated with saline solution, and Bio-C Temp. As control, no dressing was performed. The intracanal dressings remained in the simulated root canal for 21 days. Luminosity (ΔL) and color difference (ΔE ab and ΔE 00 ) were evaluated using long-term (1 year) spectrophotometric analysis. The results were analyzed using the ANOVA test with Tukey post hoc with a significance of 5%., Results: Triple antibiotic paste showed the highest values of ΔE ab and ΔE 00 and lowest values of ΔL (P < .05). Bio-C Temp showed higher ΔE ab than the other groups from 6 months (P < .05), higher ΔE 00 and lower ΔL after 1 year (P < .05)., Conclusions: Bio-C Temp induced noticeable coronary discoloration after the 1-year analysis., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: To comparatively analyse the levels of culturable bacteria, endotoxins (LPS), tumour necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and substance P in teeth with symptomatic irreversible pulpitis (SIP) and vital normal pulp (VNP) tissues., Methodology: Thirty-two patients were included (20 teeth with SIP and 12 teeth with VNP tissues) in this cross-sectional study. Samples were collected from the full length of the root canals (microbial analysis) and periapical tissues (2 mm beyond the apex for immunological analysis), using sterile absorbent paper points. The levels of culturable bacteria (culture method), endotoxins (LAL Pyrogent 5000), TNF-α, IL-1β and substance P (ELISA) were assessed. The Mann-Whitney test was used for comparisons between the levels of CFU/mL, LPS, TNF-α, IL-1β and substance P in the SIP and VNP groups. The statistical analysis was performed with the significance level set at 5%., Results: Culturable bacteria were recovered from all teeth with SIP. On the other hand, no positive cultures were observed in the VNP tissues group (p > .05). The levels of LPS were approximately four times higher in teeth with SIP than in teeth with VNP tissues (p < .05). Higher levels of TNF-α and substance P were detected in teeth with SIP (p < .05). On the other hand, no difference in the levels of IL-1β was detected between the two groups (p > .05)., Conclusion: Teeth with symptomatic irreversible pulpitis present higher levels of culturable bacteria, endotoxins, TNF-α and substance P than those with vital normal pulp tissues. On the other hand, the levels of IL-1β were similar in teeth from both groups suggesting reduced implications of this inflammatory mediator in the early stages of infection., (© 2023 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Calcium silicate-based materials are used to block the communication between the root canal and the periodontal ligament space. This brings the materials into contact with tissues and the potential for local and systemic elemental release and movement. The aim of the study was to evaluate the elemental release of bismuth from ProRoot MTA in contact with connective tissues after 30 and 180 days as well as any accumulation in peripheral organs using an animal model. Tricalcium silicate and hydroxyapatite containing 20% bismuth oxide (HAp-Bi) were used as controls. The null hypothesis was that bismuth migrates from tricalcium silicate-based materials when associated with silicon. The materials were examined using scanning electron microscopy, energy dispersive spectroscopy (SEM/EDS) and X-ray diffraction prior to implantation as well as using SEM/EDS, micro X-ray fluorescence and Raman spectroscopy after implantation to assess elemental presence in surrounding tissues. Histological analysis was used to evaluate the changes in tissue architecture and inductively coupled plasma mass spectrometry (ICP-MS) was used to investigate the elemental deposition. For the systemic investigation, routine blood analysis was performed and organs were obtained to evaluate the presence of bismuth and silicon using ICP-MS after acid digestion. In the histological analysis of the implantation sites, macrophages and multinucleated giant cells could be observed after 30 days which after 180 days became a chronic infiltrate; although, no major differences were identified in red and white blood cell analyses and biochemical tests. Implantation altered the materials as observed in the Raman analysis and bismuth was detected both locally and within kidney samples after both periods of analysis, indicating the potential for accumulation of bismuth in this organ. Smaller amounts of bismuth than observed in the kidney were also detected in blood, liver and brain for the ProRoot MTA and HAp-Bi after 180 days. Bismuth was released from the ProRoot MTA locally and was detected systemically and in samples without silicon; thus, the null hypothesis was rejected. The bismuth release demonstrated that this element accumulated both locally and systemically, mainly in the kidneys in comparison with brain and liver regardless of the material base., (© 2023. The Author(s).)

Objective: The objective of this study was to evaluate the microbiological profile of root canals indicated for endodontic retreatment due to secondary endodontic infections evidenced by the presence of chronic apical periodontitis (G1) or for prosthetic reasons, without clinical and radiographic signs of endodontic reinfection (G2)., Methods: Microbiological samples were collected from thirty teeth (N=30) out of which 15 were indicated for retreatment due to the presence of chronic apical periodontitis (G1) and 15 were indicated exclusively for prosthetic reasons (G2). Samples were collected from root canals before (S1), after chemomechanical preparation (S2), and after 30 days of intracanal medication composed of calcium hydroxide and 2% chlorhexidine gel (S3). The molecular analysis was performed using Nested-PCR for the detection of 17 bacterial species. The efficacy of each stage of the retreatment in reducing the microbial load was verified by counting colony-forming units (CFU). The statistical analysis considered a significance level of 5%., Results: The results showed that bacteria were detected in 100% of the cases in S1, in both groups, by Nested PCR. The most frequently found species in S1 in both groups were Enterococcus faecalis, Fusobacterium nucleatum, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Parvimonas micra. The microbial load of G1 was higher than G2 in the initial samples (S1). Endodontic retreatment was effective in reducing the microbial load in G1 and G2. Statistically significant associations were found between specific bacteria and clinical signs and symptoms., Conclusion: It was concluded that the microbial community present in the root canal of teeth indicated for endodontic retreatment is mixed and heterogeneous. G1 and G2 differ in the number of species per case and microbial load., Clinical Relevance: Endodontic retreatment was effective in reducing the microbial load. Statistically significant associations were found between specific bacteria and clinical signs and symptoms., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: To evaluate the biocompatibility, physical and chemical properties of three pre-mixed calcium silicate-based sealers and an epoxy resin-based material were assessed. Pre-mixed sealers supposedly obtain water from the root canal moist to hydrate and set., Materials and Methods: Polyethylene tubes were filled with the materials Bio-C Sealer Ion+, Bio-C Sealer, EndoSequence BC Sealer and AH Plus Jet, or left empty and surgically implanted in the subcutaneous tissue of Wistar rats. The animals were euthanised and the tubes and tissue were removed for histological analysis and scanning electron microscopy (SEM) coupled with energy-dispersive spectrometry (EDS). Materials' surface chemical characterisation was assessed using Raman spectroscopy and SEM/EDS. Flow, setting time (in two conditions), solubility, radiopacity and pH were also analysed. ANOVA and Bonferroni correction were performed for comparisons (P < 0.05)., Results: Inflammatory response observed in the tissues subsided from 7 to 30 days. Tungsten migration could be detected in the surrounding tissue following AH Plus Jet implantation. All calcium silicate-based sealers exhibited zirconium oxide (radiopacifier) and tricalcium silicate peaks before and after implantation. All materials exhibited flow values above 17 mm. An approximately tenfold difference was observed between the plaster- and metal-mould setting times of the calcium silicate cements indicating its sensitivity to moist variations and solubility above 8% was also observed for these materials., Conclusions: Pre-mixed materials exhibited variable setting time and solubility with a decreasing inflammatory response., Clinical Relevance: The variable moist-dependant setting time with high solubility poses a concern for the clinical use of these pre-mixed sealers., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objective: To evaluate the effectiveness of supplemental photodynamic therapy for improving the bacterial removal and the levels of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) by conducting a clinical trial., Methodology: Twenty-four root canals with pulp necrosis and periapical lesion were selected and randomly divided into conventional group using endodontic treatment with chemo-mechanical preparation (CMP) alone (n = 12) and a group using antimicrobial photodynamic therapy (aPDT) after CMP (n = 12). The samples were collected before and after CMP (conventional group) and after photodynamic therapy (aPDT group). A photosensitizer (0.005% methylene blue) was applied to the root canal for 3 minutes after CMP, whereas aPDT was performed by using a red laser with a power of 30Mw and energy density of 9J/cm 2 for 90 s per root canal. Culture technique was performed to determine the bacterial colony forming units. LPS and LTA levels were quantified by using limulus amoebocyte lysate (LAL) assay and enzyme-linked immunosorbent assay (ELISA), respectively., Results: All samples showed growth of viable bacteria on Fastidious Anaerobe Agar (FAA), with an average of 5.19 × 10 5 CFU/ mL. CMP was effective in decreasing viable bacteria (p < 0.05), whereas there was a significant decrease (p < 0.05) in the samples treated with aPDT compared to those submitted to CMP. LPS and LTA were detected in all initial samples, with mean values of 20.561 EU/mL and 430.91 pg/mL, respectively. Both CMP and aPDT groups significantly decreased the levels of LPS and LTA (p < 0.05), with a statistical difference between the groups regarding aPDT (p < 0.05)., Conclusion: Photodynamic therapy as an adjunct to CMP proved to be effective in improving root canal disinfection and reducing the LPS and LTA levels in teeth with primary endodontic infection., Competing Interests: Declaration of Competing Interest The authors have declared that no conflict of interest exists., (Copyright © 2023. Published by Elsevier B.V.)

The effects of bismuth toxicity on the kidney-the main organ responsible for blood filtration-were systematically reviewed. This review was motivated by availability of several sources of bismuth in contact with humans including environmental, medications, dental materials, and cosmetics, potentially leading to kidney filtration of this chemical. No previous studies have systematically reviewed the literature considering this association. A total of 22 studies with a total of 46 individuals met the inclusion criteria, 19 being case reports with only one patient enrolled. The included studies publication dates ranged from 1961 to 2021 and the countries of publication were the United States of America, United Kingdom, Germany, Turkey, Switzerland, and Canada. Bismuth sources affecting the kidneys were uniquely reported as from medical purposes and mostly associated to overdoses with several symptoms, apparently with dose-dependent consequences. Patient history of renal impairment seemed to affect the outcome of the case. Several therapies were conducted following bismuth intoxication, and few studies performed renal biopsies describing its histological findings. It is crucial to reconsider the nephrotoxicity of bismuth compounds, mainly in patients with previous history of renal impairment.

The substitution of calcium with strontium in bioactive materials has been promising but there has been some concern over the material instability and possible toxicity. The aim of this research was the synthesis and characterization of calcium and strontium substituted bioactive materials and assessment of interactions with local tissues and peripheral elemental migration in an animal model. A bioactive glass, hydroxyapatite and hydraulic calcium silicate with 50% or 100% calcium substitution with strontium were developed and the set materials were characterized immediately after setting and after 30 and 180-days in solution. Following subcutaneous implantation, the local (tissue histology, elemental migration) and systemic effects (elemental deposition after organ digestion) were assessed. The strontium-replaced silicate cements resulted in the synthesis of partially substituted phases and strontium leaching at all-time points. The strontium silicate implanted in the animal model could not be retrieved in over half of the specimens showing the high rate of material digestion. Tissue histology showed that all materials caused inflammation after 30 days of implantation however this subsided and angiogenesis occurred after 180 days. Strontium was not detected in the local tissues or the peripheral organs while all calcium containing materials caused calcium deposition in the kidneys. The tricalcium silicate caused elemental migration of calcium and silicon in the local tissues shown by the elemental mapping but no deposition of calcium was identified in the peripheral organs verified by the assessment of the digested tissues. Strontium can substitute calcium in bioactive materials without adverse local or systemic effects., (© 2022 The Authors. Journal of Biomedical Materials Research Part A published by Wiley Periodicals LLC.)

Calcium silicate-based root canal sealers have been developed in powder/liquid or premixed ready-to-use forms. The evaluation of the physico-chemical properties of a prototype powder/liquid MTApex Sealer (Ultradent) in comparison to a ready-to-use material EndoSequence BC Sealer (Brasseler) was performed. The paste/paste epoxy resin-based AH Plus (Dentsply) served as control for comparisons. The sealers were evaluated (n = 6) regarding setting time (in dry and moist environments), flow and radiopacity, following the ISO-6876/2012 standard. Also, the pH was assessed. Material's surface and chemical characterization was evaluated using scanning-electron-microscopy (SEM) and energy-dispersive-spectrometry (EDS). Mixed ANOVA, Shapiro-Wilk, Levene, and post-hoc analysis with Bonferroni correction were performed at a significance level of 5%. MTApex Sealer exhibited the highest flow and EndoSequence BC Sealer had a significantly longer setting time in dry compared to the moist environment; however, for MTApex Sealer and AH Plus no significant changes occurred when additional moisture was provided. All materials exceeded 7 mm Al of radiopacity and showed a decreasing alkalinity over the 21 day-analysis. SEM/EDS evaluation resulted in peaks of calcium, silicon, and the respective radiopacifier. The prototype powder/liquid MTApex Sealer had the highest flow and similar setting time in both dry and moist environments; opposingly, EndoSequence BC Sealer was crucially influenced by external moisture. This suggests that the powder/liquid materials' setting seems to be more predictable.

Background and Objectives: Discrimination due to race and/or ethnicity can be a pervasive stressor for Black college students in the United States beyond general negative life events and has demonstrated associations with adverse health and alcohol outcomes. Genetics may confer individual differences in the risk of drinking to cope with discrimination-related stress. This study tested whether associations of racial/ethnic discrimination with coping drinking motives and alcohol use differ as a function of a well-documented variant in the alcohol dehydrogenase 1B gene (ADH1B*3)., Methods: Cross-sectional data were obtained from 241 Black students (M age  = 20.04 [range = 18-53]; 66% female) attending a predominantly White university in the northeastern United States. Participants provided a saliva sample for genotyping and self-reported on their racial/ethnic discrimination experiences, coping drinking motives, and past-month total alcohol quantity., Results: Path models demonstrated that associations of discrimination with alcohol quantity directly or indirectly through coping drinking motives did not differ as a function of ADH1B*3, after controlling for gender, age, negative life events, and potential confounding interactions of covariates with model predictors. Regardless of ADH1B*3, greater experience of negative life events was associated with higher coping drinking motives, which in turn were associated with greater alcohol quantity. CONCLUSION AND SCIENTIFIC SIGNIFICANCE: Findings represent a novel investigation into gene-environment interplay in associations of alcohol use with racial/ethnic discrimination. Findings demonstrate coping-motivated drinking associated with negative life events within Black college drinkers regardless of ADH1B*3. Future research should leverage longitudinal designs to characterize associations of genetics, stressful experiences, and coping-motivated drinking over time., (© 2022 American Academy of Addiction Psychiatry.)

Forensic DNA analysis is among the most well-recognized and well-developed forensic disciplines. The field's use of DNA markers known as short tandem repeats (STRs) offer a robust means of discriminating individuals while also introducing challenges to the analysis. One of these challenges, stutter, is the result of a non-biological artifact introduced during PCR. The formation and amplification of these stutter products can occur at rates as high as 15-20% of the parent allele. The challenge inherent in this process is differentiating stutter artifacts from true alleles, particularly in the presence of a minor contributor. Traditionally, DNA profiles are obtained using capillary electrophoresis (CE), where amplified DNA fragments are separated by size, not sequence, and the identification of stutter is performed on a locus-specific level. The use of CE-based fragment data rather than sequence-based data, has limited the community's understanding of the precise behavior of stutter. Massively parallel sequencing (MPS) data provides an opportunity to better characterize stutter, permitting a more accurate means of detecting both size- or longest uninterrupted stretch (LUS)-based stutter but also allele and motif-specific stutter characteristics. This study sheds light on the value of characterizing motif- and allele-specific stutter, including non-LUS stutter, when using MPS methods. Analysis and characterization of stutter sequences was performed using data generated from 539 samples amplified with the ForenSeq and PowerSeq 46GY library preparation kit and sequenced on the Illumina MiSeq FGx. Assessment of non-LUS stutter begins with calculating stutter rates for all potential stutter products at a given locus (and allele), additionally, the occurrence of these discrete stutter products were quantified. Results show that although the LUS sequence stutters at a higher rate than non-LUS motifs, the non-LUS stutter products do occur at detectable levels and potentially influence sequence-based mixture analysis. The data indicate that the stutter from one motif or allele can be distinguished from another motif or allele based on their unique stutter rates; however, the number of stutter products from each motif or allele may similarly make up the overall pool of stutter products. Motif- and allele-specific stutter models provide the most comprehensive analysis of sequence stutter rates and provide the ability to differentiate stutter sequences more accurately from true allele stutter. This information provides a foundation for including the characterization of non-LUS stutter products when analyzing DNA profiles, specifically mixtures with potential low-level contributors., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Aim: To evaluate the physicochemical properties of five root canal sealers and assess their effect on an ex vivo dental plaque-derived polymicrobial community., Methodology: Dental plaque-derived microbial communities were exposed to the sealers (AH Plus [AHP], GuttaFlow Bioseal [GFB], Endoseal MTA [ESM], Bio-C sealer [BCS] and BioRoot RCS [BRR]) for 3, 6 and 18 h. The sealers' effect on the biofilm biomass and metabolic activity was quantified using crystal violet (CV) staining and MTT assay, respectively. Biofilm community composition and morphology were assessed by denaturing gradient gel electrophoresis (DGGE), 16S rRNA sequencing and scanning electron microscopy. The ISO6876:2012 specifications were followed to determine the setting time, radiopacity, flowability and solubility. Obturated acrylic teeth were used to assess the sealers' effect on pH. Surface chemical characterization was performed using SEM with coupled energy-dispersive spectroscopy. Data normality was assessed using the Shapiro-Wilk test. One-way anova and Tukey's tests were used to analyze data from setting time, radiopacity, flowability and solubility. Two-way anova and Dunnett's tests were used for the data analysis from CV, MTT and pH. 16S rRNA sequencing data were analyzed for alpha (Shannon index and Chao analysis) and beta diversity (Bray-Curtis dissimilarities). Differences in community composition were evaluated by analysis of similarity (p < .05)., Results: The sealers significantly influenced microbial community composition and morphology. All sealers complied with ISO6876:2012 requirements for setting time, radiopacity and flowability. Although only AHP effectively reduced the biofilm biomass, all sealers, except BRR, reduced biofilm metabolic activity., Conclusion: Despite adequate physical properties, none of the sealers tested prevented biofilm growth. Significant changes in community composition were observed. If observed in vivo, these changes could affect intracanal microbial survival, pathogenicity and treatment outcomes., (© 2022 The Authors. International Endodontic Journal published by John Wiley & Sons Ltd on behalf of British Endodontic Society.)

The present study aimed to compare the filled volume by gutta-percha and sealer in the apical region of the main canal and ramifications, after instrumentation at two different working lengths using nano-computed tomography (nano-CT). Twenty-two premolars with apical ramifications were selected after micro-computed tomography evaluation and were randomly divided into groups for further endodontic instrumentation at two different working lengths: G1 - Root canals shaped 1 mm short of the apical foramen (n=11), and G2 - Root canals shaped at the apical foramen (n=11). After completing root treatment, nano-CT images were acquired, and the filled volume by gutta-percha and sealer in the main canal apical 0-4 mm and 0-1 mm ranges, and apical ramifications were objectively measured by an operator specialized in both radiology and endodontics, blinded for both groups. The Mann-Whitney test was applied to compare both groups regarding the filling of the main canal apical ranges and apical ramifications with a significance level of 5% (α ≤ 0.05). It was observed that root canals shaped at the apical foramen had a larger volume of the main canal filled than root canals shaped 1 mm short of the apical foramen, at both apical ranges (0-4 and 0-1 mm) (p<0.05). Regarding the filling of the apical ramifications, there was no significant difference between groups (p>0.05). In conclusion, the root canals shaped at apical foramen exhibited increased filling volume of the main canal in the apical region. However, neither of both working lengths influenced filling of the apical ramifications.

Introduction: The objective of this study was to investigate the action of photodynamic therapy on pain control after endodontic treatment in asymptomatic teeth with a primary infection, within a single visit., Methods: Sixty (60) single-rooted teeth with pulp necrosis and periapical lesions were selected and randomly divided into two (2) groups (n = 30), according to the protocol; a control group (CG) and a group using photodynamic therapy (aPDT). The canals were instrumented with Reciproc files # 25 up to 40 along the entire length of the canal, using 2% chlorhexidine gel as the auxiliary chemical substance, followed by irrigation with sterile saline. aPDT consited of 0.005% methylene blue as photosensitizer, using AsGaAl diode laser, 660 nm wavelength, 100 mW of power and 9 J of energy, using optical fibers with 365 μm in diameter. The canals were filled with Endomethasone N cement., Results: Pain intensity was assessed at 8, 12, 24, 48, 72 h and 1 week after endodontic treatment using a visual analogue scale. The level of pain was classified as none (0), mild (1-3), moderate (4-7) or severe (8-10). The data were at a significance level of 5%. There was a statistically significant difference (p<0.05) in the periods of 8, 12, 24, 48 and 72 h between the control group and the aPDT group. After 1 week, there was no statistically significant difference., Conclusions: It is concluded that photodynamic therapy had a significant effect on decreasing post-endodontic treatment pain in teeth with necrotic pulp and asymptomatic periapical lesions., (Copyright © 2021. Published by Elsevier B.V.)

Introduction: The objective of this study was to monitor the effects of chemomechanical preparation (CMP) performed with 6% sodium hypochlorite and calcium hydroxide-based intracanal medication (ICM) on the levels and diversity of bacteria, endotoxins (lipopolysaccharides [LPS]), and lipoteichoic acid (LTA) in root canals of teeth with symptomatic irreversible pulpitis., Methods: Samples were collected from 10 teeth with symptomatic irreversible pulpitis before CMP (S1), after CMP (S2), and after ICM (S3). The levels of bacteria, LPS, and LTA were assessed by using checkerboard DNA-DNA hybridization, LAL Pyrogent 5000, and enzyme-linked immunosorbent assay, respectively. Wilcoxon test, repeated-measures analysis of variance, and Tukey post hoc test were used for statistical analysis at a significance level of 5%., Results: Forty species were detected at S1. Two species were eliminated after CMP and 5 after ICM. Resistant and pain-related species were detected in the root canals. Higher levels of culturable bacteria were detected at S1. However, CMP and ICM effectively reduced the microbial load in the root canals. Higher levels of LPS and LTA were detected at S1. CMP was effective in reducing both LPS and LTA (P < .05). ICM produced additional reduction in the levels of LPS (P > .05) and LTA (P < .05)., Conclusions: Chemomechanical preparation using 6% sodium hypochlorite and calcium hydroxide-based intracanal medication were effective in reducing the levels of bacteria, LPS, and LTA in teeth with vital pulp and irreversibly inflamed pulp., (Copyright © 2021 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Objectives: This study aimed to detect the presence of specific bacteria and to evaluate the levels of lipopolysaccharides (LPS) and lipoteichoic acid (LTA) in symptomatic necrotic root canals associated with acute apical abscess (symptomatic group - GI). It also aimed to compare the findings with those presented by asymptomatic necrotic root canals (asymptomatic group - GII) in the different stages of the endodontic treatment., Materials and Methods: Microbiological samples were collected from 20 root canals, including purulent collection from acute apical abscesses, before and after chemo-mechanical preparation (CMP) preparation (CMP) with chlorhexidine gel 2% and after 30 days of intracanal medication (ICM) with (Ca[OH] 2 ) + chlorhexidine. The culture method was used to evaluate the efficacy of CMP and ICM. Nested PCR was used to detect the presence of 17 specific bacteria. Levels of LPS were measured by using limulus amebocyte lysate, whereas enzyme-linked immunosorbent assay was used for the quantification of LTA., Results: CMP was effective in reducing the microbial load in both groups (P < 0.05). LPS levels were higher in GI than in GII (P < 0.05). There was a significant reduction in the LPS levels after CMP and ICM (P < 0.05) in GI and GII. LTA levels were significantly reduced in GI after ICM and in GII after CMP and ICM (both P < 0.05). Fusobacterium nucleatum and Enterococcus faecalis were frequently identified in both groups, alone or in combination with each other., Conclusion: Different species were detected in all stages of the endodontic treatment. CMP was able to reduce bacterial content and the levels of LPS, but not of LTA in the symptomatic group. High levels of LPS were correlated with spontaneous pain and pain to percussion in the symptomatic group., Clinical Relevance: This clinical study showed that chemo-mechanical preparation was able to reduce bacterial load and levels of LPS, but not of LTA in the symptomatic group. Elevated levels of LPS were correlated with the presence of symptomatology., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Aim: This study evaluated the effect of four intracanal medications commonly used in regenerative endodontic procedures on the bond strength of four calcium silicate-based materials, in which two are powder-to-liquid products (MTA and MTA-HP) and the other are ready-to-use materials (EndoSequence Root Repair Material Fast Set Putty [ERRM] and Bioceramic Repair [BIO-C Repair])., Methods: Ten bovine central incisors were selected and 4 slices (1.0 ± 0.1 mm) were prepared from each root. Next, four 0.8-mm wide holes were drilled in each slice and specimens were filled with one of the following intracanal medications: triple antibiotic paste, double antibiotic paste, calcium hydroxide with distilled water, and calcium hydroxide with 2% chlorhexidine gel. After 21 days, holes were filled with one of the materials: MTA, MTA-HP, ERRM, or BIO-C Repair. After storage, push-out test and failure analysis were performed. Data were submitted to analysis of variance in a 4 × 4 factorial scheme. Tukey's test was used for multiple comparisons., Results: The use of different interappointment dressings did not influence the results ( P > 0.05). ERRM and BIO-C Repair presented significantly higher values than MTA and MTA-HP ( P < 0.0001). Specimens showed a 100% occurrence of adhesive failures., Conclusion: The use of different intracanal medications presented similar impact on bond strength of calcium silicate-based materials. Ready-to-use ERRM and BIO-C Repair materials presented the best push-out values to dentine, whereas powder-to-liquid MTA and MTA-HP cements showed the lowest results., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Conservative Dentistry.)

Endodontic biomaterials have significantly improved dental treatment techniques in several aspects now that they can be used for vital pulp treatments, as temporary intracanal medication, in definitive fillings, in apical surgeries, and for regenerative procedures. Calcium silicate-based cement is a class of dental material that is used in endodontics in direct contact with the dental structures, connective tissue, and bone. Because the material interacts with biological tissues and stimulates biomineralization processes, its properties are of major importance. The main challenge in endodontic treatments is the elimination of biofilms that are present in the root canal system anatomical complexities, as it remains even after chemical-mechanical preparation and disinfection procedures. Thus, an additional challenge for these biomaterials is to exert antimicrobial activity while maintaining their biological properties in parallel. This article reviews the literature for studies considering the antimicrobial properties of calcium silicate-based dental biomaterials used in endodontic practice. Considering the reviewed studies, it can be affirmed that the reduced antimicrobial effect exhibited by calcium silicate-based endodontic materials clearly emphasizes that all clinical procedures prior to their use must be carefully performed. Future studies for the evaluation of these materials, and especially newly proposed materials, under poly-microbial biofilms associated with endodontic diseases will be necessary.

Aim: To evaluate possible modifications in root canal sealers subjected to a variety of heating conditions using vibrational spectroscopy and analysis of physical and chemical properties., Methodology: EndoSequence BC Sealer HiFlow, Bio-C Sealer, BioRoot RCS and AH Plus were analysed chemically using Raman spectroscopy (25-220 °C) and Fourier-transform infrared spectroscopy (FT-IR) (37-100 °C ). For FT-IR, the materials were tested individually and mixed with root dentine powder. Scanning electron microscopy (SEM) and coupled energy dispersive spectroscopy (EDS) were used to evaluate surface and chemical elements. ISO 6876-2012 and ASTM-C266-07 specifications were followed to evaluate flow, setting time (moist and dry), solubility and radiopacity. Also, pH analysis at 37 and 100 °C was performed. Shapiro-Wilk and Mixed ANOVA (within and between the effects of the subjects), Levene, and a post hoc analyses with Bonferroni correction were performed (P < 0.05)., Results: Vibrational spectroscopy revealed peaks of tricalcium silicate, dicalcium silicate and zirconium dioxide. Chemical changes in the Raman spectra during heating were discrete, as the inorganic content predominated the signalling for all root canal sealers. FT-IR analysis exhibited spectral changes in water absorption for EndoSequence BC Sealer HiFlow and Bio-C Sealer, probably related to dehydration. For BioRoot RCS and AH Plus, no significant chemical changes were observed. Bio-C Sealer exhibited a band of polyethylene glycol only after heating to 100 °C, probably related to its thermal decomposition. SEM/EDS analysis corroborated the composition results observed in vibrational spectroscopy for all materials. Heating to 100 °C significantly changed the flowability of all calcium silicate-based sealers with a wide variation in setting times at both temperatures, along with solubility levels above ISO standards. For all tested sealers, radiopacity fulfilled the requirements, and pH exhibited alkaline values., Conclusions: The tested calcium silicate-based sealers were affected by heating. Calcium silicate-based root canal sealers had high solubility which is a concern for their clinical use. AH Plus was the only root canal sealer that was stable after heating., (© 2021 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

This study evaluated the physicochemical, biological, and antimicrobial properties of a new hydraulic calcium silicate-based modified material, and compared it with MTA Repair HP and MTA Angelus. The materials were assessed regarding color luminosity (L), color change, radiopacity, setting time, and ISO 6876:2012 linear flow. Volumetric filling and volume change were evaluated using microcomputed-tomography (µCT). Chemical characterization after 28 days in Hank's Balanced Salt Solution (HBSS) and pH analysis were also assessed. Biological characterization of cytotoxicity and microbiological assessment were also undertaken. Shapiro-Wilk, ANOVA, Levene and post hoc analyses with Bonferroni correction were performed, adopting a 5% significance level (p <0.05). Bio-C Pulpo exhibited the highest L values after 90 days. All tested materials demonstrated color change during the analyses, and had radiopacity above 5 mm Al. MTA Repair HP set faster than Bio-C Pulpo, whereas the latter had the highest linear flow. MTA Repair HP had the highest volumetric filling in µCT analysis. Bio-C Pulpo showed the highest alkalinity during all tested periods, and the highest volumetric loss (above 9%), in comparison with MTA Repair HP and MTA Angelus. Bio-C Pulpo did not form calcium hydroxide after hydration. MTA Repair HP demonstrated the highest cytocompatibility, and Bio-C Pulpo, the highest cytotoxicity. No inhibition halos were observed for any material, and similar higher turbidity values were seen after direct contact. Composition additives used in Bio-C Pulpo modified its properties, and both the absence of calcium hydroxide deposition after hydration, and the related cytotoxicity of this material are of particular concern.

Aim: This cross-sectional study aimed to investigate the microbial profile and to quantify the levels of endotoxins (LPS) and lipoteichoic acid (LTA) present in periapical lesions associated with root filled teeth and those that had received root canal retreatment. It also aimed to investigate the association between microorganisms and their virulence factors with clinical and radiographic features., Methodology: Patients with periapical lesions in teeth with post-treatment endodontic disease following primary root canal treatment (n = 19) and unsuccessful root canal retreatment (n = 13) were treatment planned for endodontic microsurgery, where the periapical lesions were collected. Clinical and radiographic data were also collected. For microbiological analysis, nested polymerase chain reaction was used to detect 17 bacterial species. Levels of LPS and LTA were determined using limulus amebocyte lysate and enzyme-linked immunosorbent assays, respectively. The Student t-test or Wilcoxon-Mann-Whitney tests were applied to compare the data on LPS and LTA with clinical and radiographic features. The associations between the clinical and radiographic features and the bacterial species were analysed using the Fisher's exact test. A significance level of 5% was adopted., Results: Bacterial DNA, LPS and LTA were detected in all samples. Parvimonas micra was the most commonly detected species in all groups, followed by Enterococcus faecalis, Fusobacterium nucleatum and Porphyromonas endodontalis. The type of endodontic treatment, whether a primary root canal treatment or retreatment, was not associated with the presence of any bacterial species in periapical lesions. The levels of LPS and LTA in periapical lesions of root filled teeth were not significantly different from those that had been retreated. Associations between the levels of LPS and LTA with clinical signs and symptoms were found. No association was found between specific bacteria and clinical features., Conclusion: Periapical lesions associated with teeth after primary root canal treatment and retreatment had similar polymicrobial composition. The levels of LPS and LTA in periapical lesions associated with teeth after primary root canal treatment and retreatment were similar, and both were associated with the same symptomatology., (© 2020 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

This review aims to evaluate whether root canal obturation with calcium silicate-based (CSB) sealers reduces the risk and intensity of endodontic postoperative pain when compared to epoxy resin-based (ERB) sealers. The review was registered at PROSPERO (CRD42020169255). Two independent reviewers conducted an electronic search in PubMed, Scopus, EMBASE, Web of Science, Cochrane Library and LILACS until November 2020 and included only randomized clinical trials with adult health participants undergoing root canal treatment. After selection, the JBI Critical Appraisal tool was used to assess the risk of bias. A fixed-effect meta-analysis was performed to summarize the results of pain risk and pain intensity at time intervals of 24 and 48 hours. Finally, the certainty of evidence was assessed using the GRADE approach. The search resulted in 1,206 records, of which five studies ( n = 421 patients) met the eligibility criteria and presented moderate to low risk of bias. There was no significant difference between groups in the risk of pain in the first 24 hours (relative risk or RR = 0.83, 95% confidence interval or CI: 0.60, 1.16, I 2 =) or 48 hours (RR = 0.56, 95% CI: 0.26, 1.21, I 2 =). Silicate-based sealers led to lower pain intensity only at 48 hours (mean and standard deviation = 0.37, 95% CI: 0.69, 0.05). All analyses revealed low heterogeneity ( I 2 < 25%). The evidence presented moderate level of certainty. Currently available evidence has shown that there is no difference between CSB and ERB sealers in the risk or intensity of postoperative pain., Competing Interests: None declared., (European Journal of Dentistry. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial-License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).)

Forensic science has yet to take full advantage of single cell analysis. Its greatest benefit is the ability to alleviate the challenges associated with DNA mixture analysis, which remains a significant hurdle in forensic science. Many of the factors that cause complexity in mixture interpretation are absent in single cell analyses-multiple contributors, varied levels of contribution, and allele masking. This study revisits single cell analyses in the context of forensic identification, introducing previously unseen depth to the characterization of data generated from single cells using a novel pipeline that includes recovery of single cells using the DEPArray NxT and amplification using the PowerPlex Fusion 6c kit with varied PCR cycles (29, 30, and 31). The resulting allelic signal was assessed using analytical thresholds of 10, 100, and 150RFU. The mean peak heights across the sample sets generally increased as cycle number increased, 75.0 ± 85.3, 147.1 ± 172.6, and 226.1 ± 298.2 RFU, for 29, 30, and 31 cycles, respectively. The average proportion of allele/locus dropout was most significantly impacted by changes in the detection threshold, whereas increases in PCR cycle number had less impact. Overall data quality improved notably when increasing PCR from 29 to 30 cycles, less improvement and more volatility was introduced at 31 cycles. The average random match probabilities for the 29, 30, and 31 cycle sets at 150RFU are 1 in 2.4 × 10 18  ± 1.46 × 10 19 , 1 in 1.49 × 10 25  ± 5.8 × 10 25 , and 1 in 1.83 × 10 24  ± 8.09 × 10 24 , respectively. This demonstrates the current power of single cell analysis in removing the need for complex mixture analysis.

Objectives: To evaluate a new material containing tantalum oxide as an alternative radiopacifier, and a water-based gel for hydration, in comparison with two calcium silicate-based cement: ProRoot MTA and Biodentine., Materials and Methods: ProRoot MTA (Dentsply), Biodentine (Septodont), and a new hydraulic calcium silicate cement White-MTAFlow (Ultradent) (in 'thin' consistency) were characterized using X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive spectroscopy (EDS). The interaction with dentin was also assessed using SEM and EDS. Physical and chemical properties radiopacity, setting time, linear flow, volumetric central filling, and lateral flow, pH, and volume change were investigated together with the color luminosity (L) and color change (ΔE). The agar diffusion and direct contact antimicrobial activity, and methylthiazolyldiphenyl-tetrazolium-bromide (MTT) cytotoxicity using human fibroblast cells were also evaluated. Data were statistically analyzed at a 5% significance level., Results: All materials were composed of tricalcium and dicalcium silicate but had different radiopacifiers, and calcium hydroxide (portlandite) deposition was detected in XRD analysis. White-MTAFlow exhibited radiopacity values in accordance with ISO standard, and the longest setting time. The water-based gel provided the highest linear flow, a comparable cavity central filling, and the highest groove-lateral flow in the volumetric flow analysis. White-MTAFlow exhibited an alkalinity reduction, and Biodentine, a progressive increase of pH values after 28 days. However, similar volume loss for White-MTAFlow was assessed in comparison to Biodentine after the 28-day immersion. White-MTAFlow showed the highest L value (91.5), and ProRoot MTA the lowest (78.1) due to dentin staining caused by bismuth migration. None of the materials exhibited inhibition halos against the tested bacteria, and similar turbidity values were obtained after 48 h in direct contact with E. faecalis, indicating an upregulation to bacterial growth. White-MTAFlow showed MTT cytocompatibility similarly to the control group., Conclusions: White-MTAFlow in 'thin' consistency presents comparable physicochemical, biological, and antimicrobial properties to ProRoot MTA and Biodentine, and does not cause color alteration in dentin., Clinical Relevance: White-MTAFlow is a suitable material for use as reparative endodontic cement. Further studies considering its biocompatibility are necessary.

Objective: Pulp revascularization is an effective treatment for immature necrotic teeth. Calcium hydroxide has been used in pulp revascularization as an intracanal medication due to its antimicrobial action and the non-exhibition of crown discoloration and cytotoxicity for stem cells from the apical papilla. Our study aimed to investigate the clinical success and quantitative radiographic changes of root development in immature traumatized teeth using calcium hydroxide plus 2% chlorhexidine gel as intracanal medication., Methodology: In this retrospective study, 16 patients were treated with a standardized pulp revascularization protocol. Calcium hydroxide and 2% chlorhexidine gel were manipulated in a 1:1 (v/v) ratio and inserted into root canals with Lentulo spirals (Dentsply Maillefer, Baillaigues, Switzerland). Patients were followed up for a period from 9 to 36 months for the evaluation of clinical and radiological data. Radiographic measurements of root length, root width, apical diameter, and MTA placement from the apex were quantified using software ImageJ. Wilcoxon test and t-test were used, according to nonparametric or parametric data, respectively, for changes over time in root length, root width, and apical diameter., Results: Fifteen teeth survived during the follow-up period (93.75%) and met the criteria for clinical success. Although the changes seem to be very small in many cases, significant increases in the average root length (14.28%, p<0.0001), root width (8.12%, p=0.0196), and decrease in apical diameter (48.37%, p=0.0007) were observed. MTA placement from the apex and age at the time of treatment was not significantly associated with the quantitative radiographic outcomes., Conclusions: Pulp revascularization in traumatized immature teeth treated with calcium hydroxide plus 2% chlorhexidine gel as intracanal medication had high success and survival rates, showing periodontal healing and resolution of signs and symptoms. However, concerning the continued root development, the outcomes can still be considered unpredictable.

Aim: The objective of this study was to evaluate the presence of microorganisms in the root canals (RC) of teeth submitted to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis., Material and Methods: Twenty teeth referred to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis were included in this study. Gutta-percha (GP) was collected from each third of the RC. After GP removal, microbial samples were also collected from the full extension of RC using paper points and files. The samples were cultivated in Fastidious Anaerobe Agar and the colony-forming units (CFU/mL) were counted. They were also semi-quantitatively analyzed by checkerboard DNA-DNA hybridization using 40 DNA bacterial probes., Results: Microorganisms were found in all samples. The coronal third of GP was more contaminated than the apical third (p ≤ 0.05). There was prevalence of Enterococcus hirae and Enterococcus faecalis in all RC thirds and also in the samples collected from the full extension of RCs., Conclusion: Microorganisms were found in all cases referred to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis. Enterococcus was the genus most frequently detected., Clinical Significance: An endodontic retreatment should be considered before replacing a prosthesis.

Aim: To evaluate in a clinical trial the efficacy of reciprocating and ultrasonic activation of 6% sodium hypochlorite (NaOCl) in the microbial composition and reduction in microbial load as well as in levels of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) in teeth with primary endodontic infections., Methodology: Samples were collected from 24 root canals with pulp necrosis and periapical lesions, before and after chemo-mechanical canal preparation. The teeth were randomly divided according to the activation protocol as follows: control group without activation (WA, n = 8), reciprocating activation group using Easy Clean tip (EC, n = 8) and ultrasonic activation group using Irrisonic insert (US, n = 8). Microbiological specimens were processed using a culture technique and microbiota composition was analysed using the checkerboard technique. The levels of LPS and LTA were quantified using limulus amebocyte lysate (LAL) and enzyme-linked immunosorbent assay (ELISA), respectively. The Fisher's exact test, Kruskal-Wallis, Dunn's and Wilcoxon's test with a significance level of P < 0.05 were used for statistical analysis., Results: All initial specimens had growth of viable bacteria in fastidious anaerobe agar (FAA), with an average of 10 5  CFU mL -1 , whereas only one case had such growth after chemo-mechanical canal preparation. LPS and LTA were recovered in 100% of the cases. Chemo-mechanical canal preparation significantly decreased the levels of LPS and LTA (P < 0.05), but no significant differences were found between the groups (P > 0.05). Through the checkerboard technique, bacteria were found in 100% of the initial specimens with concentrations between <10 5 and 10 6 . The most frequently identified microorganisms were Prevotella nigrescens and Enterococcus hirae. After chemo-mechanical canal preparation, many species were not detected in any of the three groups tested. A significant reduction occurred in Group US, followed by Groups EC and WA., Conclusions: Activation of 6% NaOCl reduced the levels of LPS and LTA with no differences between the groups. However, ultrasonic activation was associated with a greater reduction in microbial load within root canals., (© 2019 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Tricalcium silicate-based cement are materials used in reparative and regenerative procedures in endodontics. A recently proposed formulation aimed to enhance handling during clinical use with a versatile material applicable by syringe. Although, the use of bismuth oxide as radiopacifier and grey raw powder are drawbacks considering aesthetics. Objectives Evaluate physicochemical, biological, and antimicrobial properties of Grey-MTAFlow (Ultradent) and assess whether the addition of zinc oxide (ZnO) prevents dentinal discoloration caused by bismuth oxide. Methodology Grey-MTAFlow was manipulated in 'thin' consistency for all tests. Luminosity, color change, ion migration to dentine, radiopacity, setting time, ISO 6876:2012 linear flow, volumetric lateral flow and central filling of simulated grooves scanned using micro-computed tomography (μCT), pH, calcium release, volumetric change using μCT, chemical characterisation, cytotoxicity, and antimicrobial activity were assessed. Addition of 5% ZnO to Grey-MTAFlow and a bismuth-containing experimental composition were comparatively tested. Statistical analyses used Shapiro-Wilk, T-test, ANOVA, and Kolmogorov-Smirnov (p<0.05). Results The addition of ZnO to Grey-MTAFlow prevented dentine darkening after 90 days due to bismuth migration reduction, although no statistical difference was found (p=0.863). ZnO addition significantly enhanced Grey-MTAFlow radiopacity without differences in initial setting time. Grey-MTAFlow presented an ISO linear flow of 10.9 mm and a balanced volumetric lateral flow with central filling in μCT evaluation. All compositions presented an alkaline pH after immersion. Grey-MTAFlow had a significantly higher calcium ion release after 28 days in comparison to 24 hours (p=0.011) and volumetric expansion of 0.4±1.8% after immersion. ZnO addition altered the hydrated cement matrix once calcium hydroxide (portlandite) could not be detected in characterisation. Neither of the materials produced inhibition halos nor reduced bacterial turbidity, but all presented cytocompatibility above 100%. Conclusion Grey-MTAFlow expanded after immersion and exhibited higher luminosity values after the evaluation period when ZnO was added, but chemical modifications after this addition occurred.

DNA mixture interpretation remains one of the major challenges in forensic DNA analysis. DNA mixture samples are inherently complex due to several factors including the variations in the quantity of DNA, the presence of non-allelic artifactual peaks and the presence of multiple contributors with variable levels of allele sharing. The Probabilistic Assessment for Contributor Estimation (PACE) is a fully continuous probabilistic machine learning-based method to predict the number of contributors (n) in a sample, and was previously developed for use with the Identifiler amplification kit. This system required manual preprocessing of data and was limited, exclusively, to samples amplified using said kit. This study introduces PACE™ v1.3.7 for use with both the GlobalFiler and PowerPlex Fusion 6c amplification kits. An automated artifact identification and management system has been added to accompany the rapid estimation of the number of donors in a given mixture. The artifact management module, when evaluated using previously unseen data, identified true allelic peaks and removed artifacts such as elevated baseline noise, stutter, and pull-up with accuracy over 93.5%. The systems yield the correct n classifications in over 90% of the samples, and demonstrate consistent accuracies as the number of donors and the overall mixture complexity increase. Misclassified samples generally exhibited high levels of allele sharing among donors, low DNA template amounts and high incidence of allelic dropout. This system offers a means for both artifact management and n estimation as well as a quantitative and reproducible method of assessing the quality of a profile., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

While DNA detection using capillary electrophoresis has enabled improvements in both resolution and throughput, the use of CE - particularly with multiple dye channels - can introduce artifacts that can complicate analyses. Undetected pull-up artifacts can pose a challenge to investigators, especially in low-level samples, while partial pull-up peaks can distort peak height balance within a locus and impact the downstream likelihood ratio. Current methods for addressing pull-up are typically manually implemented. This study presents an effective alternative: a series of mathematical models, created using symbolic regression achieved through genetic programming. The models estimate the amount of pull-up expected in a peak from a true allele for a given dye-dye relationship and instrument type. This leads to the removal of artifactual pull-up peaks and peak height corrections when pull-up is present within true alleles. When models are used in conjunction with a dynamic threshold, pull-up peaks were automatically detected and removed with an accuracy rate of 96.1%. The removal of partial pull-up from true allele peaks led to a more accurate heterozygote balance for the affected locus. These models have been optimized for use with any analytical threshold and can be implemented by any lab using a 3100 or 3500 instrument series., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objectives: The purpose of the present study was to evaluate the effect of calcium hydroxide with 2% chlorhexidine gel (HCX) or distilled water (HCA) compared to triple antibiotic paste (TAP) on push-out bond strength and the cement/dentin interface in canals sealed with White MTA Angelus (WMTA) or Biodentine (BD)., Materials and Methods: A total of 70 extracted human lower premolars were endodontically prepared and randomly divided into 4 groups according to the intracanal medication, as follows: group 1, HCX; group 2, TAP; group 3, HCA; and group 4, control (without intracanal medication). After 7 days, the medications were removed and the cervical third of the specimens was sectioned into five 1-mm sections. The sections were then sealed with WMTA or BD as a reparative material. After 7 days in 100% humidity, a push-out bond strength test was performed. Elemental analysis was performed at the interface, using energy-dispersive spectroscopy. The data were statistically analyzed using analysis of variance and the Tukey test ( p < 0.05)., Results: BD presented a higher bond strength than WMTA ( p < 0.05). BD or WMTA in canals treated with calcium hydroxide intracanal medications had the highest bond strength values, with a statistically significant difference compared to TAP in the WMTA group ( p < 0.05). There were small amounts of phosphorus in samples exposed to triple antibiotic paste, regardless of the coronal sealing., Conclusions: The use of intracanal medications did not affect the bond strength of WMTA and BD, except when TAP was used with WMTA., Competing Interests: Conflict of Interest: No potential conflict of interest relevant to this article was reported.

Objectives: Addition of aluminum fluoride (AlF 3 ) to MTA was tested to inhibit dental discoloration., Materials and Methods: MTA Angelus with 0, 5, 15, and 45% AlF 3 were tested. The set cements were characterized using scanning electron microscopy, energy-dispersive spectroscopy, and X-ray diffraction. Radiopacity and setting time were analyzed according to ANSI/ADA 57 and ASTM C266-08. Volume change was evaluated using volumetric micro-CT analysis. The pH and calcium ion release were assessed after 3 and 24 h and 28 days. Dental discoloration in contact with the cements was assessed after 24 h and 28 and 90 days of contact with bovine and human dentine. Tissue reaction to subcutaneous implantation in rats was examined after 30 and 60 days., Results: AlF 3 altered the microstructure of MTA. The addition of 5% AlF 3 did not significantly alter the radiopacity, setting time, and volume change (p > 0.05). pH and calcium ion release significantly increased with addition of AlF 3 (p > 0.05). All the tested proportions of AlF 3 prevented the dental darkening verified for MTA Angelus in bovine and human teeth. AlF 3 did not interfere in inflammatory response of MTA in all periods of analysis; otherwise, lower amounts showed less intense inflammatory infiltrate., Clinical Relevance: AlF 3 prevents destabilization of bismuth oxide and consequent tooth darkening, frequently verified in clinical practice when using white MTA., Conclusions: The use of 5% of AlF 3 in combination to MTA resulted in a cement that did not result in dental discoloration and did not affect significantly physical, chemical, and biological properties.