Research methods assessing sodium hypochlorite cytotoxicity: A scoping review.

Sodium hypochlorite (NaOCl) cytotoxicity has been assessed using different methodologies, which has led to arbitrary interpretations. This scoping review aimed to discuss the different methodological protocols for assessing NaOCl cytotoxicity. A literature review performed in the PubMed and Embase databases up to July 2023 identified manuscripts reporting NaOCl cytotoxicity. The dataset identified 546 publications, further screened by 2 reviewers. Ninety studies were identified and mined methodologically to collect information on cell type, cytotoxicity assay, NaOCl dilution solutions, presence of fetal bovine serum (FBS), and NaOCl exposure time. The culture medium used in cytotoxicity assays contains buffering substances that neutralize the pH of NaOCl, thus reducing its cytotoxicity, an approach that may lead to bias when solutions with different pH are compared. For short exposure periods, as in simulations to evaluate the contact between irrigant and periapical tissue cells during chemo-mechanical preparation, NaOCl dilution should be performed with saline, which does not buffer the irrigant. For long exposure periods, as in simulations of irrigant extrusions, NaOCl should be diluted in the culture medium, to reproduce the expected buffering effect occurring in extrusions. The presence of FBS in culture medium can decrease NaOCl toxicity. There is no standardization of NaOCl cytotoxicity methodologies. This poses the risk of arriving at incorrect results and, therefore, pertinent tests must be refined.

Calcium hypochlorite cytotoxicity mechanism in fibroblasts and effect on osteoblast mineralization.

AIM: To determine the cytotoxicity mechanism of 2.5% calcium hypochlorite [Ca(OCl)2 ] in L929 fibroblasts and the effect of this solution on human osteoblast-like cells (Saos-2) mineralization, compared to that of 2.5% sodium hypochlorite (NaOCl). METHODOLOGY: L929 fibroblasts were exposed to Ca(OCl)2 and NaOCl at different dilutions for 10 min. Cell metabolism was assessed by methyl-thiazole-tetrazolium (MTT); lysosome integrity, by neutral red (NR) assay; type of cell death, by flow cytometry (apoptosis/necrosis); cytoskeleton, by actin and α-tubulin fluorescence and cell ultrastructure, by transmission electron microscopy (TEM). The alkaline phosphatase (ALP) activity and mineralized nodule formation were determined in Saos-2 by thymolphthalein release and alizarin red staining (ARS), respectively. The data were analysed by two-way anova and Bonferroni's post-test (α = .05). RESULTS: Ca(OCl)2 promoted higher cell viability and a lower percentage of apoptosis and necrosis than NaOCl (p < .05). Ca(OCl)2 and NaOCl decreased cell metabolism and lysosome integrity, induced the breakdown of microtubules and actin filaments, promoted alterations of rough endoplasmic reticulum and disruption of mitochondrial cristae. Additionally, Ca(OCl)2 did not induce ALP activity and had no effect on mineralized nodules formation. CONCLUSIONS: Although Ca(OCl)2 and NaOCl promoted the same cytotoxicity mechanism, Ca(OCl)2 was less cytotoxic than NaOCl. As for ALP activity, no differences were observed between NaOCl and Ca(OCl)2 . The production of mineralized nodules induced by Ca(OCl)2 was lower than those induced by NaOCl, but was not different from those induced by the control group.

Recombinant sugarcane cystatin CaneCPI-5 promotes osteogenic differentiation.

Cysteine proteases orchestrate bone remodeling, and are inhibited by cystatins. In reinforcing our hypothesis that exogenous and naturally obtained inhibitors of cysteine proteases (cystatins) act on bone remodeling, we decided to challenge osteoblasts with sugarcane-derived cystatin (CaneCPI-5) for up to 7 days. To this end, we investigated molecular issues related to the decisive, preliminary stages of osteoblast biology, such as adhesion, migration, proliferation, and differentiation. Our data showed that CaneCPI-5 negatively modulates both cofilin phosphorylation at Ser03, and the increase in cytoskeleton remodeling during the adhesion mechanism, possibly as a prerequisite to controlling cell proliferation and migration. This is mainly because CaneCPI-5 also caused the overexpression of the CDK2 gene, and greater migration of osteoblasts. Extracellular matrix remodeling was also evaluated in this study by investigating matrix metalloproteinase (MMP) activities. Our data showed that CaneCPI-5 overstimulates both MMP-2 and MMP-9 activities, and suggested that this cellular event could be related to osteoblast differentiation. Additionally, differentiation mechanisms were better evaluated by investigating Osterix and alkaline phosphatase (ALP) genes, and bone morphogenetic protein (BMP) signaling members. Altogether, our data showed that CaneCPI-5 can trigger biological mechanisms related to osteoblast differentiation, and broaden the perspectives for better exploring biotechnological approaches for bone disorders.

Clinical Study of Antimicrobial Efficacy of Laser Ablation Therapy with Indocyanine Green in Root Canal Treatment.

INTRODUCTION: Laser ablation (LA) therapy is used as an adjunct to endodontic treatment to improve microbial reduction. However, studies evaluating the impact of LA with indocyanine green (ICG) are scarce. This study aimed to evaluate the antimicrobial efficacy of LA therapy with ICG in root canal treatment. METHODS: Sixty patients with periapical lesions in teeth with a single canal and absence of pain, edema, and previous treatment were selected. Patients were randomly allocated into 3 groups according to the apical sizes used (n = 20); 25/04, 30/04, and 35/04 were the final sizes used. In half the patients of each group, 2.5% sodium hypochlorite was used as an irrigating solution, and in the other half, saline solution was used. After instrumentation, all patients received LA therapy with ICG. Root canal sampling was performed before (S1) and after (S2) root canal instrumentation and immediately after LA therapy with ICG (S3). Colony-forming units were counted, and statistical tests were applied (P < .05). RESULTS: There was a significant reduction in colony-forming units from S1 to S2 in all treatment protocols (P < .05); 2.5% sodium hypochlorite as an irrigating solution showed a greater microbial reduction compared with saline solution (P < .05). LA therapy with ICG further reduced the microbial counts significantly (S2 to S3 and S1 to S3) whether sodium hypochlorite or saline was used (P < .05). CONCLUSIONS: LA therapy with ICG significantly increased microbial reduction in root canals regardless of instrumentation sizes or the irrigation solution used.

Removal of separated instruments from unfavourable locations: Case reports using the HBW ultrasonic ring or a surgical approach.

Two patients were admitted to the dental clinic to evaluate two right mandibular first molars (both FDI no. 46). In Case 1, there was an accidental instrument separation, which was thought to have been bypassed; however, the instrument was located beyond the apical foramen, in the alveolar bone. The 1-week follow-up revealed that the fragment was close to the inferior alveolar nerve, thus allowing surgical removal to be performed. In Case 2, two separated instruments located in the apical third of the mesiobuccal and mesiolingual canals were removed using the HBW ultrasonic ring coupled to ISO 10 and 15 hand instruments. The 3-month and 1-year follow-ups (Case 1), and the 2-year and 3-year follow-ups (Case 2) showed a favourable evolution in both cases. It can be concluded that both the surgical approach and the HBW ultrasonic ring are viable options for the removal of separated instruments in unfavourable locations.

Endodontic micro-resurgery and guided tissue regeneration of a periapical cyst associated to recurrent root perforation: a case report.

Although the success rates of microsurgery and micro-resurgery are very high, the influence of a recurrent perforation combined with radicular cyst remains unclear. A 21-year-old white female patient had a history of root perforation in a previously treated right maxillary lateral incisor. Analysis using cone-beam computed tomography (CBCT) revealed an extensive and well-defined periapical radiolucency, involving the buccal and palatal bone plate. The perforation was sealed with bioceramic material (Biodentine) in the pre-surgical phase. In the surgical phase, guided tissue regeneration (GTR) was performed by combining xenograft (lyophilized bovine bone) and autologous platelet-rich fibrin applied to the bone defect. The root-end preparation was done using an ultrasonic tip. The retrograde filling was performed using a bioceramic material (Biodentine). Histopathological analysis confirmed a radicular cyst. The patient returned to her referring practitioner to continue the restorative procedures. CBCT analysis after 1-year recall revealed another perforation in the same place as the first intervention, ultimately treated by micro-resurgery using the same protocol with GTR, and a bioceramic material (MTA Angelus). The 2-year recall showed healing and bone neoformation. In conclusion, endodontic micro-resurgery with GTR showed long-term favorable results when a radicular cyst and a recurrent perforation compromised the success.

Strontium Carbonate and Strontium-Substituted Calcium Carbonate Nanoparticles Form Protective Deposits on Dentin Surface and Enhance Human Dental Pulp Stem Cells Mineralization.

Strontium acetate is applied for dental hypersensitivity treatment; however, the use of strontium carbonates for this purpose has not been described. The use of Sr-carbonate nanoparticles takes advantage of both the benefits of strontium on dentin mineralization and the abrasive properties of carbonates. Here in, we aimed to synthesize strontium carbonate and strontium-substituted calcium carbonate nanoparticles and test them as potential compounds in active dentifrices for treating dental hypersensitivity. For this, SrCO3, Sr0.5Ca0.5CO3, and CaCO3 nanoparticles were precipitated using Na2CO3, SrCl2, and/or CaCl2 as precursors. Their morphology and crystallinity were evaluated by electron microscopy (SEM) and X-ray diffraction, respectively. The nanoparticles were added to a poly (vinyl alcohol) gel and used to brush dentin surfaces isolated from human third molars. Dentin chemical composition before and after brushing was investigated by infrared spectroscopy (FTIR) and X-ray dispersive energy spectroscopy. Dentin tubule morphology, obliteration, and resistance of the coatings to acid attack were investigated by SEM and EDS. The cytotoxicity and ability of the particles to trigger the mineralization of hDPSCs in vitro were studied. Dentin brushed with the nanoparticles was coated by a mineral layer that was also able to penetrate the tubules, while CaCO3 remained as individual particles on the surface. FTIR bands related to carbonate groups were intensified after brushing with either SrCO3 or Sr0.5Ca0.5CO3. The shift of the phosphate-related FTIR band to a lower wavenumber indicated that strontium replaced calcium on the dentin structure after treatment. The coating promoted by SrCO3 or Sr0.5Ca0.5CO3 resisted the acid attack, while calcium and phosphorus were removed from the top of the dentin surface. The nanoparticles were not toxic to hDPSCs and elicited mineralization of the cells, as revealed by increased mineral nodule formation and enhanced expression of COL1, ALP, and RUNX2. Adding Sr0.5Ca0.5CO3 as an active ingredient in dentifrices formulations may be commercially advantageous since this compound combines the well-known abrasive properties of calcium carbonate with the mineralization ability of strontium, while the final cost remains between the cost of CaCO3 and SrCO3. The novel Sr0.5Ca0.5CO3 nanoparticles might emerge as an alternative for the treatment of dental hypersensitivity.

The Effect of Octenidine on Proliferation, Migration, and Osteogenic Differentiation of Human Dental Pulp and Apical Papilla Stem Cells.

INTRODUCTION: The research for alternative irrigating solutions is ongoing, since no "ideal" solution has yet been found. Octenidine dihydrochloride (OCT) has been indicated as an endodontic irrigant because it has adequate antimicrobial and biological properties. The present study aimed to assess the effects of OCT on proliferation, migration, and induction of the osteogenic phenotype of stem cells from human dental pulp and apical papilla. METHODS: Cells were collected from human third molars and exposed to different doses of OCT, chlorhexidine (CHX), sodium hypochlorite (NaOCl), and ethylenediaminetetraacetic acid (EDTA) to determine cell viability by alamarBlue assay; proliferation by bromodeoxyuridine incorporation; migration by the Transwell assay; alkaline phosphatase activity by thymolphthalein release; and production of mineralized nodules by alizarin red staining. The results were analyzed by 1- or 2-way analysis of variance and Tukey (α = .05). RESULTS: CHX promoted lower cell viability, followed by OCT, NaOCl, and EDTA, especially at intermediate doses (P < .05). Cells exposed to CHX had less proliferation than the other groups (P < .05). The Transwell assay revealed no differences among OCT, EDTA, and culture medium (control group) (P > .05). OCT and EDTA induced greater migration than CHX and NaOCl (P < .05). OCT and EDTA induced higher alkaline phosphatase activity than NaOCl and CHX (P < .05). No difference was detected among the groups using alizarin red staining (P > .05). CONCLUSIONS: OCT induced high migration, proliferation, and alkaline phosphatase activity of stem cells from human dental pulp and apical papilla, which could be advantageous for regenerative endodontic procedures.

Multispecies biofilm removal by a multisonic irrigation system in mandibular molars.

AIM: The aim of the study was to assess biofilm removal efficacy of GentleWave System and passive ultrasonic irrigation (PUI). METHODOLOGY: Twenty-two human mandibular molars with Vertucci's type II configuration in the mesial root were selected. Teeth were autoclaved, inoculated with dental plaque and incubated in a CDC biofilm reactor for two weeks. The mesial roots were instrumented up to 20.06 file (V-Taper) for the GentleWave group and up to 35.04 file (Vortex Blue) for PUI group. Irrigation was performed using GentleWave and PUI irrigation protocols (n = 11). Dentine debris on paper points samples were obtained for quantitative real-time polymerase chain reaction (qPCR) and 16S ribosomal RNA gene sequencing (next-generation aequencing-NGS). For qPCR, a non-parametric test (α = 0.05) was used. Next-generation sequencing data were analysed using mothur, with alpha diversity calculated as the Shannon and Chao1 indices and Bray-Curtis dissimilarities were used for beta diversity. Differences in alpha diversity and abundances of genera were evaluated using Kruskal-Wallis test. Differences in community composition were evaluated using analysis of similarity with Bonferroni correction for multiple comparisons. RESULTS: Quantitative real-time polymerase chain reaction results showed that the reduction estimated in percentages for both groups was equivalent (p > .05). NGS analysis showed that both techniques promoted a significant reduction in reads and OTUs number (p < .05). Shannon alpha diversity and Chao1 index showed no differences between pre- or post-treatment samples for both groups (p > .05). Additionally, pre-treatment communities differed from post-treatment samples in both groups regarding bacterial taxa reduction (ANOSIM R = 0.50 and 0.55, p < .001). CONCLUSIONS: Bacterial reduction in mesial roots of mandibular molars prepared to 35.04 with PUI was similar to those prepared to 20.06 with a multisonic irrigant activation system.

Physicochemical properties and penetration into dentinal tubules of calcium hypochlorite with surfactants.

The aim was to assess the physicochemical properties and the penetration into dentinal tubules of calcium hypochlorite solution [Ca(OCl)2], with or without surfactants. The surfactants benzalkonium chloride, cetrimide, Tween 80 and Triton X-100 were mixed at different concentrations with sodium hypochlorite solution (NaOCl), Ca(OCl)2 and distilled water (control). Once the critical micellar concentration (CMC) of the surfactants in Ca(OCl)2 and NaOCl was determined, pH, free chlorine, surface tension and free calcium ions were evaluated. The penetration into dentinal tubules of NaOCl and Ca(OCl)2, with or without benzalkonium chloride and Triton X-100 [surfactants that promoted the lowest surface tension of Ca(OCl)2], was assessed using human premolars stained with crystal violet. The statistical tests were one-way ANOVA and Tukey's post-test, Kruskal-Wallis and Dunn's post-test, two-way ANOVA and Bonferroni's post-test, and t-test; depending on the assay. The addition of surfactants reduced the surface tension of NaOCl and Ca(OCl)2, and did not alter the pH or the free available chlorine of either solution. The addition of all surfactants increased the availability of free calcium ions in Ca(OCl)2, especially benzalkonium chloride. Ca(OCl)2 exhibited lower penetration into dentinal tubules than NaOCl, and the addition of surfactants did not improve the penetration of Ca(OCl)2, but did increase the penetration of NaOCl. It can be concluded that the addition of surfactants to Ca(OCl)2 did not increase the penetration into dentinal tubules, but it did promote lower surface tension, without changing the pH or free available chlorine values, and higher availability of free calcium ions in Ca(OCl)2.

Physicochemical properties and penetration into dentinal tubules of calcium hypochlorite with surfactants

Abstract The aim was to assess the physicochemical properties and the penetration into dentinal tubules of calcium hypochlorite solution [Ca(OCl)2], with or without surfactants. The surfactants benzalkonium chloride, cetrimide, Tween 80 and Triton X-100 were mixed at different concentrations with sodium hypochlorite solution (NaOCl), Ca(OCl)2 and distilled water (control). Once the critical micellar concentration (CMC) of the surfactants in Ca(OCl)2 and NaOCl was determined, pH, free chlorine, surface tension and free calcium ions were evaluated. The penetration into dentinal tubules of NaOCl and Ca(OCl)2, with or without benzalkonium chloride and Triton X-100 [surfactants that promoted the lowest surface tension of Ca(OCl)2], was assessed using human premolars stained with crystal violet. The statistical tests were one-way ANOVA and Tukey's post-test, Kruskal-Wallis and Dunn's post-test, two-way ANOVA and Bonferroni's post-test, and t-test; depending on the assay. The addition of surfactants reduced the surface tension of NaOCl and Ca(OCl)2, and did not alter the pH or the free available chlorine of either solution. The addition of all surfactants increased the availability of free calcium ions in Ca(OCl)2, especially benzalkonium chloride. Ca(OCl)2 exhibited lower penetration into dentinal tubules than NaOCl, and the addition of surfactants did not improve the penetration of Ca(OCl)2, but did increase the penetration of NaOCl. It can be concluded that the addition of surfactants to Ca(OCl)2 did not increase the penetration into dentinal tubules, but it did promote lower surface tension, without changing the pH or free available chlorine values, and higher availability of free calcium ions in Ca(OCl)2.

Resumo O objetivo foi avaliar as propriedades físico-químicas e a penetrabilidade nos túbulos dentinários da solução de hipoclorito de cálcio [Ca(OCl)2], com ou sem surfactantes. Os surfactantes cloreto de benzalcônio, cetrimida, Tween 80 e Triton X-100 foram misturados em diferentes concentrações com a solução de hipoclorito de sódio (NaOCl), Ca(OCl)2 e água destilada (controle). Uma vez determinada a concentração micelar crítica (CMC) dos surfactantes em Ca(OCl)2 e NaOCl, foram avaliados o pH, cloro livre, tensão superficial e íons de cálcio livre. A penetrabilidade nos túbulos dentinários de NaOCl e Ca(OCl)2, com ou sem cloreto de benzalcônio e Triton X-100 [surfactantes que promoveram a menor tensão superficial de [Ca(OCl)2] foi avaliada utilizando pré-molares humanos corados com cristal violeta. Os testes estatísticos foram ANOVA de uma via e pós-teste de Tukey, Kruskal-Wallis e pós-teste de Dunn, ANOVA de duas vias e pós-teste de Bonferroni, e teste t; dependendo do ensaio. A adição de surfactantes reduziu a tensão superficial do NaOCl e Ca(OCl)2, e não alterou o pH ou cloro livre das soluções. A adição de todos os surfactantes aumentou a disponibilidade de íons de cálcio livre de Ca(OCl)2, principalmente o cloreto de benzalcônio. Ca(OCl)2 apresentou menor penetrabilidade nos túbulos dentinários do que NaOCl, e a adição de surfactantes não aumentou a penetrabilidade de Ca(OCl)2, mas aumentou a penetrabilidade de NaOCl. Pode-se concluir que a adição de surfactantes no Ca(OCl)2 não aumentou a penetrabilidade nos túbulos dentinários, mas promoveu menor tensão superficial, sem alterar os valores de pH ou cloro livre, e maior disponibilidade de íons de cálcio livre em Ca(OCl)2.

Outcomes of the GentleWave system on root canal treatment: a narrative review.

This study aimed to describe the outcomes of the GentleWave system (GW) (Sonendo) on root canal treatment. Published articles were collected from scientific databases (MEDLINE/PubMed platform, Web of Science, Scopus, Science Direct and Embase). A total of 24 studies were collected from August/2014 to July/2021, 20 in vitro and 4 clinical. GW System was not associated with extrusion of the irrigant, promoted faster organic dissolution than conventional syringe irrigation (CSI), passive ultrasonic irrigation (PUI) continuous ultrasonic irrigation (CUI) and EndoVac, reduced more bacterial DNA and biofilm than PUI and CUI, promoted higher penetration of sodium hypochlorite into dentinal tubules than PUI and CUI in vitro, and removed more intracanal medication than CSI and PUI. GW was able to remove pulp tissue and calcifications. Moreover, its ability to remove hard-tissue debris and smear layer was better than that of CSI, and its ability to remove root canal obturation residues was lower or similar to that of PUI, and similar to that of CSI and EndoVac. Regarding root canal obturation of minimally instrumented molar canals, GW was associated with high-quality obturation. Clinically, the success rate of endodontic treatment using GW was 97.3%, and the short-term postoperative pain in the GW group was not different from CSI. Further research, mainly clinical, is needed to establish whether GW has any advantages over other available irrigation methods.

Different formulations of peracetic acid: effects on smear layer removal, dentine erosion, cytotoxicity and antibiofilm activity.

OBJECTIVE: To assess the effects of different peracetic acid (PAA) formulations on smear layer (SL) removal, dentine erosion, cytotoxicity, and antibiofilm activity. METHODOLOGY: SL removal and dentine erosion were assessed using 90 premolars, distributed into six groups, according to final irrigation: PAA formulations (1% Sigma, 1% Bacterend OX, 1% Arposept, and 0.09-0.15% Anioxyde), 17% ethylenediaminetetraacetic acid (EDTA), and water (control). Cytotoxicity was assessed by methyl-thiazol-tetrazolium (MTT) and neutral red assays. Antibacterial and antibiofilm effectiveness was evaluated against Enterococcus faecalis. For cytotoxicity and antibiofilm activity assessment, the 2.5% NaOCl was also included. RESULTS: EDTA, Sigma, and Bacterend OX removed more SL than Arposept, Anioxyde, and water (p<0.05). EDTA caused more severe dentine erosion than Sigma and Bacterend OX (p<0.05). Sigma and Bacterend OX had higher cytotoxicity than the other solutions (p<0.05). NaOCl, Bacterend OX, Sigma, and Anioxyde significantly reduced E. faecalis colony-forming units (CFU) (p<0.05). The 2.5% NaOCl solution promoted greater biofilm biomass reduction (p<0.05) than the other solutions. All PAA formulations promoted greater biomass reduction than 17% EDTA (p<0.05). CONCLUSIONS: Although Sigma and Bacterend OX had higher cytotoxicity, they had a SL removal capability similar to that of EDTA, were as effective as NaOCl against E. faecalis biofilm, and promoted less dentine erosion than EDTA. Arposept and Anioxyde failed to remove the SL, had lower cytotoxicity, and showed less bacterial activity than NaOCl.

How can biophotonics help dentistry to avoid or minimize cross infection by SARS-CoV-2?

Biophotonics is defined as the combination of biology and photonics (the physical science of the light). It is a general term for all techniques that deal with the interaction between biological tissues/cells and photons (light). Biophotonics offers a great variety of techniques that can facilitate the early detection of diseases and promote innovative theragnostic approaches. As the COVID-19 infection can be transmitted due to the face-to-face communication, droplets and aerosol inhalation and the exposure to saliva, blood, and other body fluids, as well as the handling of sharp instruments, dental practices are at increased risk of infection. In this paper, a literature review was performed to explore the application of Biophotonics approaches in Dentistry focusing on the COVID-19 pandemic and how they can contribute to avoid or minimize the risks of infection in a dental setting. For this, search-related papers were retrieved from PubMED, Scielo, Google Schoolar, and American Dental Association and Centers for Disease Control and Prevention databases. The body of evidence currently available showed that Biophotonics approaches can reduce microorganism load, decontaminate surfaces, air, tissues, and minimize the generation of aerosol and virus spreading by minimally invasive, time-saving, and alternative techniques in general. However, each clinical situation must be individually evaluated regarding the benefits and drawbacks of these approaches, but always pursuing less-invasive and less aerosol-generating procedures, especially during the COVID-19 pandemic.

Multispecies biofilm removal by XP-endo Finisher and passive ultrasonic irrigation: A scanning electron microscopy study.

The aim was to assess the effect of XP-endo Finisher (XPF) on multispecies biofilm removal, in comparison with passive ultrasonic irrigation (PUI) and conventional syringe irrigation (CSI), by scanning electron microscope (SEM). Fifty mandibular first premolars were instrumented, longitudinally sectioned. The split halves were incubated for 4 days with a broth obtained from three bacteria strains: Enterococcus faecalis, Eikenella corrodens and Streptococcus anginosus. Subsequently, the re-approximated split halves were irrigated with 4% sodium hypochlorite (NaOCl) or water using CSI, and the final irrigation protocols were CSI with 4% NaOCl (CSI+4%NaOCl), PUI+4%NaOCl, XPF+4%NaOCl and CSI+water. The analysis of biofilm removal was performed using SEM images. There were no differences between PUI and XPF (P > 0.05), and both groups promoted higher biofilm removal than CSI+4%NaOCl and CSI+water groups (P < 0.05). It can be concluded the multispecies biofilm removal was significantly improved using XPF and PUI when compared to CSI.

Different formulations of peracetic acid: effects on smear layer removal, dentine erosion, cytotoxicity and antibiofilm activity

Abstract Objective: To assess the effects of different peracetic acid (PAA) formulations on smear layer (SL) removal, dentine erosion, cytotoxicity, and antibiofilm activity. Methodology: SL removal and dentine erosion were assessed using 90 premolars, distributed into six groups, according to final irrigation: PAA formulations (1% Sigma, 1% Bacterend OX, 1% Arposept, and 0.09-0.15% Anioxyde), 17% ethylenediaminetetraacetic acid (EDTA), and water (control). Cytotoxicity was assessed by methyl-thiazol-tetrazolium (MTT) and neutral red assays. Antibacterial and antibiofilm effectiveness was evaluated against Enterococcus faecalis. For cytotoxicity and antibiofilm activity assessment, the 2.5% NaOCl was also included. Results: EDTA, Sigma, and Bacterend OX removed more SL than Arposept, Anioxyde, and water (p<0.05). EDTA caused more severe dentine erosion than Sigma and Bacterend OX (p<0.05). Sigma and Bacterend OX had higher cytotoxicity than the other solutions (p<0.05). NaOCl, Bacterend OX, Sigma, and Anioxyde significantly reduced E. faecalis colony-forming units (CFU) (p<0.05). The 2.5% NaOCl solution promoted greater biofilm biomass reduction (p<0.05) than the other solutions. All PAA formulations promoted greater biomass reduction than 17% EDTA (p<0.05). Conclusions: Although Sigma and Bacterend OX had higher cytotoxicity, they had a SL removal capability similar to that of EDTA, were as effective as NaOCl against E. faecalis biofilm, and promoted less dentine erosion than EDTA. Arposept and Anioxyde failed to remove the SL, had lower cytotoxicity, and showed less bacterial activity than NaOCl.

Accidental Extrusion of Sodium Hypochlorite in a Patient Taking Alendronate: A Case Report With an 8-Year Follow-up.

Extrusion of sodium hypochlorite (NaOCl) is a very rare accident with the most common complications including pain, swelling, and hematoma. It can occur even if procedural guidelines, predisposing conditions, and risk factors are taken into consideration. A 59-year-old female patient was admitted to the endodontics department to treat a left maxillary first premolar (World Dental Federation tooth 24). The medical history included osteoporosis and systemic medication with alendronate. Initially, the diagnosis was "symptomatic irreversible pulpitis" with "normal apical tissues." During instrumentation of the buccal canal, accidental extrusion of 2.5% NaOCl occurred into the periapical tissues, resulting from misinterpretation of the working length, and caused severe pain and bleeding. The canal was immediately flushed using copious irrigation with saline solution. An analgesic and corticosteroid were prescribed. At 3 and 7 days, ecchymosis and slight hematoma were observed extraorally in the area of the affected tooth and an intraoral ulceration at the apex of the affected tooth. At 15 days, minimal signs of ecchymosis were observed, and the treatment was resumed. At 30 days after the accident, there was complete remission of the sequelae. The clinical, radiographic, and tomographic assessment after 1 month, 1 year, and 8 years showed favorable case evolution. During the 8-year follow-up, the patient maintained the systemic use of alendronate. It can be concluded that NaOCl extrusion is an accident that causes patients and clinicians to experience an unpleasant consequence. The radiographic working length should always be determined carefully. A successful outcome can be achieved in patients who take alendronate.

Physicochemical properties and effect of bioceramic root canal filling for primary teeth on osteoblast biology.

OBJECTIVE: Bio-C Pulpecto (Bio-CP) was recently developed as the first bioceramic root filling material for primary teeth. To evaluate the physicochemical properties of radiopacity, setting time, pH, cytocompatibility and potential of Bio-CP to induce mineralisation, compared with (1) Calen thickened with zinc oxide (Calen-ZO), and (2) zinc oxide and eugenol (ZOE). METHODOLOGY: Physicochemical properties were evaluated according to ISO 6876. Saos-2 (human osteoblast-like cell line) exposed to extracts of the materials were subjected to assays of methyl thiazolyl tetrazolium, neutral red, alkaline phosphatase (ALP) activity and mineralised nodule production. The results were analysed using one-way or two-way ANOVA and Tukey's or Bonferroni's post-tests (α=0.05). RESULTS: All the materials showed radiopacity higher than 3 mm Al. Bio-CP had lower pH than Calen-ZO, but higher pH than ZOE. Calen-ZO and Bio-CP did not set. The setting time for ZOE was 110 min. The cytocompatibility order was Calen-ZO > Bio-CP > ZOE (1:2, 1:4 dilutions) and Calen-ZO > Bio-CP = ZOE (1:12, 1:24 dilutions) and Calen-ZO = Bio-CP > ZOE (1:32 dilution). Bio-CP induced greater ALP activity at 7 days, and greater mineralised nodule production, compared to Calen-ZO (p<0.05). Conclusions Bio-CP showed adequate physicochemical properties, cytocompatibility and potential to induce mineralisation.

Cystatin-like protein of sweet orange (CsinCPI-2) modulates pre-osteoblast differentiation via ß-Catenin involvement.

Phytocystatins are endogenous cysteine-protease inhibitors present in plants. They are involved in initial germination rates and in plant defense mechanisms against phytopathogens. Recently, a new phytocystatin derived from sweet orange, CsinCPI-2, has been shown to inhibit the enzymatic activity of human cathepsins, presenting anti-inflammatory potential and pro-osteogenic effect in human dental pulp cells. The osteogenic potential of the CsinCPI-2 protein represents a new insight into plants cysteine proteases inhibitors and this effect needs to be better addressed. The aim of this study was to investigate the performance of pre-osteoblasts in response to CsinCPI-2, mainly focusing on cell adhesion, proliferation and differentiation mechanisms. Together our data show that in the first hours of treatment, protein in CsinCPI-2 promotes an increase in the expression of adhesion markers, which decrease after 24 h, leading to the activation of Kinase-dependent cyclines (CDKs) modulating the transition from G1 to S phases cell cycle. In addition, we saw that the increase in ERK may be associated with activation of the differentiation profile, also observed with an increase in the B-Catenin pathway and an increase in the expression of Runx2 in the group that received the treatment with CsinCPI-2.

Physicochemical properties and effect of bioceramic root canal filling for primary teeth on osteoblast biology

Abstract Bio-C Pulpecto (Bio-CP) was recently developed as the first bioceramic root filling material for primary teeth. Objective To evaluate the physicochemical properties of radiopacity, setting time, pH, cytocompatibility and potential of Bio-CP to induce mineralisation, compared with (1) Calen thickened with zinc oxide (Calen-ZO), and (2) zinc oxide and eugenol (ZOE). Methodology Physicochemical properties were evaluated according to ISO 6876. Saos-2 (human osteoblast-like cell line) exposed to extracts of the materials were subjected to assays of methyl thiazolyl tetrazolium, neutral red, alkaline phosphatase (ALP) activity and mineralised nodule production. The results were analysed using one-way or two-way ANOVA and Tukey's or Bonferroni's post-tests (α=0.05). Results All the materials showed radiopacity higher than 3 mm Al. Bio-CP had lower pH than Calen-ZO, but higher pH than ZOE. Calen-ZO and Bio-CP did not set. The setting time for ZOE was 110 min. The cytocompatibility order was Calen-ZO > Bio-CP > ZOE (1:2, 1:4 dilutions) and Calen-ZO > Bio-CP = ZOE (1:12, 1:24 dilutions) and Calen-ZO = Bio-CP > ZOE (1:32 dilution). Bio-CP induced greater ALP activity at 7 days, and greater mineralised nodule production, compared to Calen-ZO (p<0.05). Conclusions Bio-CP showed adequate physicochemical properties, cytocompatibility and potential to induce mineralisation.