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.

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.

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.

In vivo and in vitro anti-inflammatory and pro-osteogenic effects of citrus cystatin CsinCPI-2.

Cystatins are natural inhibitors of cysteine peptidases. Recently, cystatins derived from plants, named phytocystatins, have been extensively studied. Among them, CsinCPI-2 proteins from Citrus sinensis were identified and recombinantly produced by our group. Thus, this study described the recombinant expression, purification, and inhibitory activity of this new phytocystatin against human cathepsins K and B and assessed the anti-inflammatory effect of CsinCPI-2 in vitro in mouse and in vivo in rats. In addition, the pro-osteogenic effect of CsinCPI-2 was investigated in vitro. The inflammatory response of mouse macrophage cells stimulated with P. gingivalis was modulated by CsinCPI-2. The in vitro results showed an inhibitory effect (p < 0.05) on cathepsin K, cathepsin B, IL-1ß, and TNF-α gene expression. In addition, CsinCPI-2 significantly inhibited in vivo the activity of TNF-α (p < 0.05) in the blood of rats, previously stimulated by E. coli lipopolysaccharide (LPS). CsinCPI-2 had a pro-osteogenic effect in human dental pulp cells, demonstrated by the increase in alkaline phosphatase (ALP) activity, deposition of mineralized nodules, and the gene expression of the osteogenic markers as bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (Runx-2), ALP, osteocalcin, and bone sialoprotein (BSP). These preliminary studies suggested that CsinCPI-2 has a potential anti-inflammatory, and at the same time, a pro-osteogenic effect. This may lead to new therapies for the control of diseases where inflammation plays a key role, such as periodontal disease and apical periodontitis.

Antimicrobial Activity and pH of Calcium Hydroxide and Zinc Oxide Nanoparticles Intracanal Medication and Association with Chlorhexidine.

AIM: To evaluate pH and antibacterial activity of pastes with calcium hydroxide [Ca(OH)2] and zinc oxide (ZnO) microparticles (micro) or nanoparticles (nano) and association with 0.4% chlorhexidine against Enterococcus faecalis. MATERIALS AND METHODS: The following pastes were analyzed: Ca(OH)2/ZnO micro, (2) Ca(OH)2/ZnO nano, (3) Ca(OH)2/ ZnO micro + 0.4% chlorhexidine, (4) Ca(OH)2/ZnO nano + 0.4% chlorhexidine. Antibacterial activity against E. faecalis was evaluated by agar diffusion test. The direct contact test on planktonic cells of E. faecalis was performed for 30 and 60 seconds. Root canals from bovine teeth were filled with the pastes and pH was evaluated after 1, 7, 14, 21, 30 and 60 days. The data obtained were submitted to the statistical tests analysis of variance (ANOVA) and Tukey or Kruskal-Wallis and Dunn test, with a 5% significance level. RESULTS: Calcium hydroxide and zinc oxide nano, and the pastes with 0.4% chlorhexidine were more effective in agar diffusion test. In the direct contact test, the pastes with chlorhexidine showed the highest effect after 30 seconds. All pastes eliminated E. faecalis after 60 seconds. All pastes promoted an increase in pH. The highest increase in pH was observed with nanoparticle medications after 1 and 7 days (p < 0.05). After this period, the pastes presented similar pH increase. CONCLUSION: It was concluded that calcium hydroxide and zinc oxide nanoparticles promoted greater initial alkalinization. The antimicrobial activity of the pastes against E. faecalis is favored by the association with chlorhexidine. CLINICAL SIGNIFICANCE: Although nanoparticles of calcium hydroxide and zinc oxide promoted antibacterial effect, the activity against E. faecalis is favored by association with chlorhexidine.

Antimicrobial activity of root canal irrigants associated with cetrimide against biofilm and planktonic Enterococcus faecalis.

AIM: To evaluate the antibacterial activity of sodium hypochlorite (NaOCl) and chlorhexidine (CHX) alone or associated with cetrimide (CTR), and QMiX against biofilm and planktonic Enterococcus faecalis (E. faecalis) [American type culture collection (ATCC) 29212]. MATERIALS AND METHODS: The solutions 2.5% NaOCl, 2.5% NaOCl + 0.2% CTR, 2% CHX, 2% CHX + 0.2% CTR, 0.2% CTR, and QMiX were evaluated. E. faecalis biofilms were induced for 14 days on bovine dentin blocks. The irrigants were evaluated after contact with E. faecalis suspension and biofilm for 1 and 3 minutes. After that, serial decimal dilutions were made and plated on tryptic soy agar (TSA) medium. Plates were incubated for 24 hours at 37°C and the colony-forming unit (CFU) 1 ml was determined. Data were subjected to ANOVA and Tukey's tests at 5% significance. RESULTS: All microorganisms were eliminated by direct contact of the irrigants with planktonic cells. Only NaOCl and NaOCl + CTR were able to completely eliminate the microorganisms by direct contact with E. faecalis biofilm. CHX presented effectiveness similar to CHX + CTR CTR, and QMiX after 1 minute of contact and similar to NaOCl and NaOCl + CTR after 3 minutes (p > 0.05), but was unable to completely eliminate the microorganisms. CTR and QMiX did not differ from each other. CONCLUSION: CTR addition to CHX and NaOCl solutions did not improve the antimicrobial activity against biofilm. All evaluated irrigants and associations presented activity against planktonic E. faecalis. Only NaOCl and NaOCl + CTR eliminated biofilm after 1 and 3 minutes of direct contact.

A method for retrieving endodontic or atypical nonendodontic separated instruments from the root canal: a report of two cases.

AIM: This clinical report presents a new method for retrieving separated instruments from the root canal with minimally invasive procedures. BACKGROUND: The presence of separated instrument in root canal may interfere in the endodontic treatment prognosis. There are several recommended methods to retrieve separated instruments, but some are difficult in clinically practice. CASE REPORT: This study describes two cases of separated instrument removal from the root canal using a stainless-steel prepared needle associated with a K-file. Case 1 presented a fractured gutta-percha condenser within the mandibular second premolar, it was separated during incorrect intracanal medication calcium hydroxide placement. Case 2 had a fractured sewing needle within the upper central incisor that the patient used to remove food debris from the root canal. After cervical preparation, the fractured instruments were fitted inside a prepared needle and then an endodontic instrument (#25 K-file) was adapted with clockwise turning motion between the needle inner wall and the fragment. CONCLUSION: The endodontic or atypical nonendodontic separated instrument may be easily pull on of the root canal using a single and low cost device. CLINICAL SIGNIFICANCE: The methods for retrieving separated instruments from root canal are difficult and destructive procedures. The present case describes a simple method to solve this problem.

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.

Evaluation of Solubility, and Volumetric and Morphological Alterations of Bioceramic Filling Material for Primary Teeth: A New Methodological Approach.

Objective: To evaluate the solubility and the volumetric and morphological alterations of bioceramic filling material (Bio-CP) for primary teeth. Materials and Methods: Bio-CP, Calen thickened with zinc oxide (Calen-ZO), and with zinc oxide eugenol (ZOE) were placed in 1- or 2-mm-diameter polyethylene tubes and immersed in water or phosphate-buffered saline (PBS) for 30 days. The solubility (mass loss) was assessed using methodology modified from ISO 6876. Filling capacity, volumetric changes, and presence of voids were assessed by microcomputed tomography (micro-CT). The surface distribution of the chemical elements and the crystalline phases was evaluated by energy scattering X-ray scanning electron microscopy (SEM-EDX) and X-ray diffraction (XRD) to detect hydroxyapatite precipitate and components. The Shapiro-Wilk, Kruskal-Wallis, and Dunn's or two-way ANOVA and Tukey post hoc test were used (α = 0.05). Results: The solubility was ZOE > Calen-ZO = Bio-CP. Calen-ZO and Bio-CP were more soluble in water than in PBS. All the materials showed greater solubility in 2-mm tube diameter in both PBS and distilled water, except for Bio-CP in distilled water, which showed no difference between both tube diameters (1 and 2 mm). Only Calen-ZO and ZOE were analyzed by micro-CT, because Bio-CP separated into two phases during scanning. Calen-ZO had greater volumetric loss and presence of voids than ZOE in water, but there was no difference in PBS. The hydroxyapatite precipitate on the surface of Bio-CP and Calen-ZO was detected after immersion in PBS. Conclusion: Although Bio-CP had acceptable solubility and filling capacity, its composition did not allow a proper volumetric and void assessment. From a clinical perspective, Bio-CP has the potential to become a suitable material for root canal filling in primary teeth. Nonetheless, its composition must first be revised to achieve better chemical stability prior to its recommendation.

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.

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.

Fracture strength of incisor crowns after intracoronal bleaching with sodium percarbonate.

OBJECTIVES: To compare the fracture resistance of bovine teeth after intracoronal bleaching with sodium percarbonate (SPC) or sodium perborate (SP) mixed with water or 20% hydrogen peroxide (HP). MATERIALS AND METHODS: Fifty extracted bovine teeth were divided into four experimental groups (G1-G4) and one control (n = 10) after endodontic treatment. Following root canal obturation, a glass ionomer barrier was placed at the cemento-enamel junction. After that, the pulp chambers were filled with: G1 - SP with water; G2 - SP with 20% HP; G3 - SPC with water; and G4 - SPC with 20% HP. No bleaching agent was used in the control group. Coronal access cavities were sealed with glass ionomer and specimens were immersed in artificial saliva. The bleaching agents were replaced after 7 days, and teeth were kept in artificial saliva for an additional 7 days, after which the pastes were removed and the coronal access cavities were restored with glass ionomer. Crowns were subjected to compressive load at a cross head speed of 0.5 mm min(-1) applied at 135° to the long axis of the root by an EMIC DL2000 testing machine, until coronal fracture. Data were statistically analysed by anova and Tukey test. RESULTS: No differences in fracture resistance were observed between the experimental groups (P > 0.05). However, all experimental groups presented lower fracture resistance than the control group (P < 0.05). CONCLUSION: SPC and SP led to equal reduction on fracture resistance of dental crowns, regardless of being mixed with water or 20% HP.

Soft tissue profile changes after rapid maxillary expansion with a bonded expander.

The aim of this study was to evaluate the short- and long-term treatment effects of rapid maxillary expansion (RME) on the soft tissue facial profile of subjects treated with a modified acrylic-hyrax device. The sample comprised 10 males and 10 females in the mixed dentition. Their average age was 9.3 years ± 10 months pre-treatment (T1), with a narrow maxilla and posterior crossbite, treated with a modified fixed maxillary expander with an occlusal splint. Lateral cephalometric radiographs obtained at T1, immediately post-expansion (T2), and after retention (T3) were used to determine possible changes in the soft tissue facial profile. The means and standard deviations for linear and angular cephalometric measurements were analysed statistically using analysis of variance and Tukey's test (α = 0.05). The measurements at T2 differed significantly from those at T1 and T3. However, RME did not produce any statistically significant alteration (P > 0.05) in the soft tissue profile for any of the cephalometric landmarks evaluated when compared at T1 and T3. The use of a fixed expander associated with an occlusal splint did not cause significant alterations in the soft tissue facial profile at T3. This modified device is effective for preventing the adverse vertical effects of RME such as an increase anterior face height in patients with a crossbite.

Tooth embedding medium influences the accuracy of electronic apex locator.

This study evaluated the influence of tooth embedding media on the accuracy ofan electronic apex locator. The root canal length of 20 human mandibular canines was measured by inserting a 15 K-file into the root canal up to the apical foramen. The distance was measured with a digital caliper. The embedding media evaluated were alginate, saline, floral foam or gauze soaked in saline. Electronic root canal length measurement was performed with Root ZX II. Data were analysed using ANOVA for repeated measurements and Tukey test, at a significance level of 5%. There was no difference between the actual root canal length measurement and the electronic reading recorded with alginate medium. The readings obtained with the other media differed from the actual root canal length measurements. Alginate provided greater accuracy in electronic root canal length determination by Root ZX II than saline, floral foam and gauze.

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.

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.

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.

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 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.