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This study evaluated the antibiofilm efficacy of a calcium silicate-based intracanal medicament (Bio-C Temp) against Fusobacterium nucleatum (F. nucleatum). Dentin slices (n = 88) were inoculated with F. nucleatum biofilms and divided into experimental groups (n = 24 each) treated with Bio-C Temp, triple antibiotic paste (TAP), or Ca(OH) 2 for 1 or 2 weeks. Untreated dentin slices exposed to saline served as a positive control, while sterile dentin slices incubated anaerobically served as a negative control. Bacterial viability was assessed using confocal laser scanning microscopy after staining with LIVE/DEAD ® BacLight ™ dye. Samples treated with TAP demonstrated the highest percentage of dead bacteria (95%). The antibiofilm effect of TAP and Bio-C Temp significantly increased from Week 1 to Week 2. At Week 2, samples treated with Bio-C Temp showed a significantly higher percentage of dead bacteria (75.25%) than those treated with Ca(OH) 2 (58.65%). TAP showed superior efficacy against F. nucleatum biofilms. After an application time of 2 weeks, the antibiofilm efficacy of Bio-C Temp was superior to that of Ca(OH) 2 ., (© 2024. The Author(s).)

Different Streptococcal species including Streptococcus mutans, Streptococcus sobrinus and Enterococcus faecalis are commonly isolated in root canal infections including refractory, recurrent, and persistent cases. Calcium hydroxide (Ca (OH) 2 ) has been widely used in endodontics as an intracanal medicament. However, using new antimicrobial herbal alternatives offers promising potentials which can be additionally enhanced by using nanoparticles (NPs). In this study, we evaluate the antimicrobial efficacy and antibiofilm effect of Neem oil including its NPs preparations and we compare the effect of conventional Ca (OH) 2 to Ca (OH) 2 NPs using standard disc diffusion method and quantitative microtitre dish biofilm formation assay against common pathogens isolated from root canal samples. Molecular docking was used to test the binding of 10 Streptococcal macromolecules to 5 candidate neem active constituents. Neem NPs 0.125 mg/ml showed better antibacterial effect than both Neem 15 mg/ml and Neem 0.15 mg/ml. Ca (OH) 2 NPs 0.125 mg/ml also showed better antibacterial effect than each of Ca (OH) 2 10 mg/ml and Ca (OH) 2 0.1 mg/ml. Best biofilm mass inhibition was achieved by Neem oil 0.15 mg/ml at 74.55% ( IQ: 67.36-87.65) and Neem NPs 0.0125 mg/ml at 59.33% (IQ: 51--75.27). For Ca (OH) 2 , the best biofilm mass inhibition was observed with Ca (OH) 2 NPs 0.125 mg/ml at 54.7% (IQ: 42.37- 77.25). Both neem oil and neem NPs show promising antibacterial and antibiofilm potential against Mutans Streptococci group at low concentrations and hence are good candidates for use as endodontic medications. In silico analysis shows that both Sitosterol and Gedunin appear to be important active constituents of neem and possible drug candidates. Additionally, Ca (OH) 2 NPs showed significantly higher antimicrobial effect against Mutans streptococci group than conventional Ca (OH) 2 preparations., (© 2024. The Author(s).)

Background: The management of endodontic infections is a complex challenge, mainly due to the involvement of diverse microorganisms and their by-products. This study aimed to evaluate the efficacy of N-acetylcysteine (NAC), calcium hydroxide (Ca(OH) 2 ), and their combined application as intracanal medications in combating Enterococcus faecalis , Escherichia coli , and lipopolysaccharides (LPS) from E. coli ., Methods: A total of 60 single-rooted human teeth were carefully selected and divided into six groups. These tooth canals were deliberately exposed to E. faecalis (ATCC 29212) and E. coli (ATCC 25922) to induce biofilm formation. Subsequently, the specimens were treated with NAC, Ca(OH) 2 , or a combination of both substances. Three samples of the root canals were collected at three moments: the first sample (S1) was to confirm the initial contamination, the second sample (S2) was immediately post-instrumentation, and the third sample (S3) was collected after the use of the intracanal medication. The antimicrobial efficacy of these intracanal medications was assessed by enumerating colony-forming units per milliliter (CFU/mL). In addition to this, the kinetic chromogenic Limulus Amebocyte Lysate (LAL) assay by Lonza was used to quantify LPS from E. coli . Data tested for normality; then, Kruskal-Wallis and Friedman tests were used, and Dunn's for multiple comparisons., Results: The findings of this study showed significant reductions in the microbial load of E. faecalis and E. coli by S3. Notably, there were no statistically significant differences among the treatment groups concerning these microorganisms. However, it was observed that only the combination of NAC and Ca(OH) 2 led to a noteworthy decrease in the quantity of E. coli's LPS after 7-days, demonstrating a statistically significant difference from the other treatment groups. NAC + Ca(OH) 2 combination, applied for a duration of 7-days, proved to be more suitable in reducing the presence of E. faecalis , E. coli , and LPS from E. coli within the context of endodontic infections., Competing Interests: Amjad Abu Hasna is an Academic Editor for PeerJ., (© 2024 Khoury et al.)

Introduction: The aim of this study was to test the hypothesis that a combination of D-amino acids (DAAs) and trans-cinnamaldehyde (TC) demonstrates superior antibiofilm activity to calcium hydroxide (CH) and untreated controls., Methods: In this 3-part in vitro study, the concentration of DAAs (D-methionine, D-leucine, D-tyrosine, and D-tryptophan) that would significantly decrease Enterococcus faecalis and Actinomyces naeslundii biofilm biomass was first determined. Then, the effect of TC + selected DAAs on polymicrobial biofilms was characterized by quantifying the biomass and biofilm viability. Finally, the antibiofilm effects of TC + DAA was compared with CH and untreated controls by (i) determining bacterial viability and (ii) quantifying biofilm matrix composition using selective fluorescence-binding analysis. Statistical analysis was performed using one-way ANOVA and appropriate multiple comparisons test, with P < .05 considered as statistically significant., Results: TC (0.06%) + D-tyrosine (1 mM) + D-tryptophan (25 mM) significantly reduced the biomass and biofilm viability compared to the control (P < .05). While no significant difference was observed between TC + DAA and CH in the cultivable bacterial counts (P > .05), confocal microscopy demonstrated a significantly greater percentage of dead bacteria in TC + DAA-treated biofilms compared to CH and the control (P < .05). TC + DAA significantly decreased the biovolume and all the examined components of the biofilm matrix quantity compared to the control, while CH significantly reduced only the exopolysaccharide quantity (P < .05)., Conclusion: The combination of TC + D-tyrosine + D-tryptophan demonstrated superior antibiofilm activity (biofilm bacterial killing and reduction of matrix quantity) to CH and has potential to be developed as an intracanal medicament., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Objective: The primary goal is to evaluate the effects of two different intracanal medicaments, calcium hydroxide [Ca(OH)2] and double antibiotic paste (DAP), on radiographic outcomes during regenerative endodontic procedures (REP) of immature permanent mandibular first molars with symptomatic irreversible pulpitis and symptomatic apical periodontitis (SIP/SAP). Additionally, the secondary goal was to evaluate MMP-8 levels during REP using two different intracanal medicaments., Methodology: The study included 20 patients with immature mandibular first molars exhibiting SIP/SAP. Participants were randomly assigned into two groups based on the applied intracanal medicament. Ca(OH)2 (n=10) was prepared by mixing it with sterile distilled water, while the same amount of powdered metronidazole and ciprofloxacin were mixed and combined with sterile distilled water for DAP (n=10). MMP-8 in periapical samples were measured at baseline and on the 14th day using immunofluorometric assay. Image-J software with TurboReg plug-in was utilized to determine changes in root length, root width, radiographic root area (RRA) during the 12-month follow-up period. Data were analyzed by SPSS 25.0 (p<.05)., Results: Significant increase in MMP-8 on the 14th day compared to baseline in both groups (p<0.001). There was no significant difference between the two groups in terms of the increase in MMP-8 (p>0.05). Root length significantly increased in both groups (p=0.001), with Ca(OH)2 showing a greater increase (p=0.046). Root width and RRA increased similarly in both groups at 12th month., Conclusion: Both Ca(OH)2 and DAP applications resulted in a significant increase in periapical MMP-8 levels. Increase in radiographic root width and root area was similar between two groups, but Ca(OH)2 led to a significantly greater increase in root length. Further studies with larger sample sizes are necessary to validate our findings during REP of vital immature permanent mandibular molars. Clinical Trials database: NCT05581706.

Objectives: The mannose phosphotransferase system (Man-PTS) plays crucial roles in the adaptive metabolic activity of Enterococcus faecalis (E. faecalis) in adverse environments. The aim of this study was to evaluate the role of Man-PTS in the alkaline resistance of E. faecalis against calcium hydroxide (CH) and the effect of metformin (Met) on the alkaline resistance of E. faecalis to CH., Materials and Methods: The regulatory role of Man-PTS EII in the alkaline resistance of E. faecalis was firstly investigated using a wild-type highly alkaline-resistant E. faecalis XS 003, standard ATCC 29212 and Man-PTS EIID gene deficient (△mptD) and overexpressing (+mptD)  strains of E. faecalis. RNA sequencing of Met-treated E. faecalis was performed to further validate the effect of Met on Man-PTS. The effect of Met on CH resistance of E. faecalis was verified by evaluating the survival, membrane potential and permeability, intracellular pH and ATP, and the expression of Man-PTS EII and membrane transporter-related genes of E. faecalis. The effect of Met on the ability of CH to remove E. faecalis biofilm on the dentin surface was also tested. The in vivo therapeutic effect of Met plus CH (CHM) was further investigated in a rat apical periodontitis model induced by E. faecalis XS 003., Results: Man-PTS EII significantly promoted the survival ability of E. faecalis in CH and enhanced its resistance to CH. The inhibition of Man-PTS EII by Met resulted in reduced alkaline resistance of E. faecalis in the presence of CH, while also enhancing the antimicrobial properties of CH against E. faecalis biofilm on dentin. Additionally, Met plus CH showed the synergistically promoted intra-canal E. faecalis infection control and healing of periapical lesion in rats., Conclusions: Met could significantly reduce the alkaline resistance of E. faecalis against CH through the modulation of Man-PTS EII, and improved the antibacterial effect of CH against E. faecalis infection both in vitro and in vivo., Clinical Relevance: Met could significantly enhance the ability of CH to control E. faecalis infection through reducing the alkaline resistance of E. faecalis., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Calcium hydroxide is used as a reparative dentin inducer. The combination of cocoa pod husk and calcium hydroxide has also been studied as a reparative dentin inducer, whereas anchovy was identified as a good calcium source., Objectives: The aim of the study was to investigate the optimal ratio of a nano-combination of calcium hydroxide, cocoa pod husk extract and anchovy powder as a reparative dentin inducer by analyzing their material, antioxidant and cytotoxic properties., Material and Methods: The ratios of the 3 elements were established and their performance was compared in terms of viscosity, setting time, pH, and solubility rate. The optimal ratio was then examined for its antioxidant capacity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Additionally, its cytotoxicity against human dental pulp stem cells (hDPSCs) was evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay., Results: A ratio of 1:1:1 of calcium hydroxide, cocoa pod husk extract and anchovy powder was identified as the optimal ratio. It exhibited the greatest flowability, the shortest final setting time, had a pH range of 8.5-10.5, and the second highest solubility rate in comparison with other ratios. Moreover, it demonstrated active radical scavenging activity and stimulated the proliferation of hDPSCs., Conclusions: Based on the material, antioxidant and cytotoxic properties, a ratio of 1:1:1 of nano-calcium hydroxide, cocoa pod husk extract and anchovy powder was determined to be optimal for use as a reparative dentin inducer.

Aim: Calcium hydroxide (CH) has been considered as a direct pulp capping materials (DPC) for the last decades despite having some limitations. Phosphorylate pullulan (PPL) incorporated with CH (CHPPL) is a novel biomaterial that was introduced as a promising DPC material. Thus, the aim of the study was to evaluate the inflammatory response and mineralized tissue formation (MTF) ability of PPL-based CH formulations on rat molars after DPC., Methodology: This study consisted of six groups: CH with 1% PPL (CHPPL-1); 3% PPL (CHPPL-3); 5% PPL (CHPPL-5); Dycal and NEX MTA Cement (N-MTA) as the positive control, and no capping materials (NC). One hundred twenty maxillary first molar cavities were prepared on Wistar rats. After capping, all the cavities were restored with 4-META/MMA-TBB resin and pulpal responses were evaluated at days 1, 7, and 28. Kruskal-Wallis followed by Mann-Whitney U-test was performed with a significance level of 0.05. Immunohistochemical expression of IL-6, Nestin, and DMP-1 was observed., Results: At day 1, CHPPL-1, N-MTA, and Dycal exhibited no to mild inflammation, whilst CHPPL-3, CHPPL-5, and NC showed mild to moderate inflammation, and the results were significantly different (p < .05). At day 7, mild to moderate inflammation was observed in CHPPL-1, N-MTA, and Dycal, whereas CHPPL-3, CHPPL-5, and NC exhibited moderate to severe inflammation. Significant differences were observed between CHPPL-1 and N-MTA with NC (p < .05), CHPPL-1 and CHPPL-3 with CHPPL-5 and Dycal (p < .05), and CHPPL-3 with N-MTA (p < .05). A thin layer of mineralized tissue formation (MTF) was observed in all groups. At day 28, CHPPL-1, Dycal, and N-MTA showed no to mild inflammation, whilst CHPPL-3, CHPPL-5, and NC exhibited mild to severe inflammation, and statistically significant difference was detected (p < .05). CHPPL-1, Dycal, and N-MTA exhibited continuous MTF, whilst CHPPL-3, CHPPL-5, and NC had thicker and interrupted MTF. Significant differences were observed between CHPPL-1, CHPPL-3, and N-MTA with NC group (p < .05). Variable expressions of IL-6, Nestin, and DMP-1 indicated differences in the materials' impact on odontoblast-like cell formation and tissue mineralization., Conclusions: These findings suggest that CHPPL-1 has the potential to minimize pulpal inflammation and promote MTF and had similar efficacy as MTA cement., (© 2024 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Objective: Evaluate the bacterial reduction effect of disinfection protocols used in lower molars infected by Enterococcus faecalis., Methods: Eighty extracted lower molars were prepared and inoculated with E. faecalis for 2 months. The teeth were then distributed into 2 control groups (n = 4) and 4 experimental groups (n = 18) in function of the disinfection protocol utilized: G1. WaveOne Gold (WOG), passive ultrasonic irrigation (PUI) and Ultracal; G2. WOG, PUI and calcium hydroxide (CH); G3. WOG, XP-Endo Finisher (XPF) and Ultracal; G4. WOG, XPF and CH. Bacteriological samples were collected previously (S1), after preparation (S2), final agitation (S3) and intracanal dressing (S4). Microbial growth was assessed according to culture turbidity and UV spectrophotometry. Statistical analyses used the Friedman test for paired samples and Kruskal-Wallis test for non-paired data (p<0.05)., Results: No protocol eliminated E. faecalis effectively. The S2, S3 and S4 samples were statistically different from the S1 samples in G1, G2, G3 and G4 (p<0.05). Statistical differences were observed in bacterial reduction between G1 and G2 and G1 and G3 after the intracanal medicament (S4) (p<0.05)., Conclusion: The WOG + PUI + CH disinfection protocol showed higher percentages of bacterial reduction.

Calcium Hydroxide-based endodontic sealer loaded with antimicrobial agents have been commonly employed in conventional root canal treatment. These sealers are not effective against E. faecalis due to the persistent nature of this bacterium and its ability to evade the antibacterial action of calcium hydroxide. Therefore, endodontic sealer containing Carbon nanodots stabilized silver nanoparticles (CD-AgNPs) was proposed to combat E. faecalis. The therapeutic effect of CD-AgNPs was investigated and a new cytocompatible Calcium Hydroxide-based endodontic sealer enriched with CD-AgNPs was synthesized that exhibited a steady release of Ag+ ions and lower water solubility at 24 hours, and enhanced antibacterial potential against E. faecalis. CD-AgNPs was synthesized and characterized morphologically and compositionally by Scanning Electron Microscopy, Fourier Transform Infrared Spectroscopy (FTIR), and UV-Vis Spectroscopy, followed by optimization via minimum inhibitory concentration (MIC) determination against E. faecalis by broth microdilution technique and Cytotoxicity analysis against NIH3T3 cell lines via Alamar Blue assay. Calcium hydroxide in distilled water was taken as control (C), Calcium hydroxide with to CD-AgNPs (5mg/ml and 10mg/ml) yielded novel endodontic sealers (E1 and E2). Morphological and chemical analysis of the novel sealers were done by SEM and FTIR; followed by in vitro assessment for antibacterial potential against E. faecalis via agar disc diffusion method, release of Ag+ ions for 21 days by Atomic Absorption Spectrophotometry and water solubility by weight change for 21 days. CD-AgNPs were 15-20 nm spherical-shaped particles in uniformly distributed clusters and revealed presence of constituent elements in nano-assembly. FTIR spectra revealed absorption peaks that correspond to various functional groups. UV-Vis absorption spectra showed prominent peaks that correspond to Carbon nanodots and Silver nanoparticles. CD-AgNPs exhibited MIC value of 5mg/ml and cytocompatibility of 84.47% with NIH3T3 cell lines. Novel endodontic sealer cut-discs revealed irregular, hexagonal particles (100-120 nm) with aggregation and rough structure with the presence of constituent elements. FTIR spectra of novel endodontic sealers revealed absorption peaks that correspond to various functional groups. Novel endodontic sealers exhibited enhanced antibacterial potential where E-2 showed greatest inhibition zone against E. faecalis (6.3±2 mm), a steady but highest release of Ag+ ions was exhibited by E-1 (0.043±0.0001 mg/mL) and showed water solubility of <3% at 24 hours where E-2 showed minimal weight loss at all time intervals. Novel endodontic sealers were cytocompatible and showed enhanced antibacterial potential against E. faecalis, however, E2 outperformed in this study in all aspects., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ali et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Bacteria and their byproducts are key contributors to the onset and perpetuation of pulpoperiapical pathosis. Intracanal medication is vital in achieving successful endodontic outcomes as it targets and eradicates remaining microorganisms following biomechanical preparation., Aim and Objective: The aim of the study was to compare and evaluate the antimicrobial efficacy of calcium hydroxide (CH) paste, triple antibiotic paste (TAP), and probiotics (PBs) as intracanal medicament in 12-17-year-old children undergoing root canal treatment for the management of infected pulpal tissues in young permanent teeth., Materials and Methods: A total of 30 patients aged 12-17 years indicated for endodontic therapy in maxillary incisors and with no systemic complications were selected. They were randomly divided into three groups, i.e., Group I - CH group, Group II - TAP, and Group III - PB allocating 10 teeth in each group. After access opening, the first sample (S1) was collected by inserting a paper point into the root canal, the second sample (S2) was collected immediately after biomechanical preparation, and the third sample (S3) was collected after 7 days, i.e., postintracanal medication. Samples were sent for microbiological analysis to assess the microbial count, and statistical analysis was done for the obtained data., Results: The three intracanal medicaments were successful in reducing the microbial counts of Enterococcus faecalis in the infected root canals. However, according to the results of the study, the PB group demonstrated greater effectiveness against E. faecalis compared to the CH group and displayed similar antimicrobial efficacy as the TAP group., Conclusion: PB exhibited antimicrobial efficacy comparable to TAP but greater than Ca (OH) 2 paste. Hence, PB can be utilized as an intracanal medicament in young permanent teeth., (Copyright © 2024 Copyright: © 2024 Journal of Indian Society of Pedodontics and Preventive Dentistry.)

Background: Azathioprine is one of the earliest immunosuppressants prescribed for several autoimmune diseases. Yet there is a lack of research on the impact of azathioprine on pulp healing following the pulp capping procedure., Aim: This study aimed to investigate the effect of azathioprine on the healing ability of mechanically exposed dogs' dental pulps following direct pulp capping with mineral trioxide aggregate (MTA), bio-aggregates (BA), and Calcium hydroxide (Ca(OH) 2 )., Methods: Four mongrel dogs were randomly assigned to two groups (two dogs/30 teeth in each group): immunosuppressed (group I) and control (group II). Group I received azathioprine for two months before surgical treatments and until the dogs were euthanized. Fifteen class V buccal cavities were performed in each dog. Each group was randomly divided into three subgroups (10 teeth each) based on the pulp capping substance. The pulps in subgroups A, B, and C were immediately capped with MTA, BA, and Ca(OH) 2 , respectively. Inflammation and dentine bridge development were histopathologically evaluated and scored at one and two months. The data were statistically analyzed., Results: The immunosuppressed group exhibited statistically greater inflammatory cell count and decreased dentine bridge thickness, compared to the control group in all subgroups ( p < 0.05)., Conclusion: Azathioprine has an adverse effect on the healing of exposed dogs' dental pulp following direct pulp capping with MTA, BA, and Ca(OH) 2 . Therefore, patients using azathioprine as an immunosuppressive medication may experience delayed healing of mechanically exposed pulps following capping with MTA, BA, or Ca(OH) 2 ., Competing Interests: The authors declare no conflicts of interest to report.

This study explored the antimicrobial effects of ketoprofen, piroxicam, and celecoxib alone or combined with calcium hydroxide (CH) against two strains of Enterococcus faecalis (E. faecalis) and assessed the influence of such combinations on the pH of CH. Minimum inhibitory concentrations (MICs) of the three tested NSAIDs were determined. Tested pastes were placed into wells punched in seeded agar plates and the bacterial inhibition zones were measured. Antibiofilm activity was assessed against 3 weeks of biofilm induced in bovine dentine blocks. The pH of the pastes was measured at four-time intervals. MIC values were 3.12, 25, and 25 mg/ml for ketoprofen, piroxicam, and celecoxib, respectively, and were similar for both bacterial strains except for celecoxib, which showed 8% growth at the highest tested concentration against vancomycin-resistant E. faecalis. Ketoprofen had the largest mean inhibition zone that was comparable to CH. None of the six tested pastes exhibited antibiofilm activity of a significant level in comparison to CH. A noticeable increase in the antibiofilm activity was found when 20% NSAIDs were added to CH while maintaining an alkaline pH. Ketoprofen was found to be the most effective among the tested NSAIDs. Although its effect was comparable to CH, adding ketoprofen at a ratio of 20% resulted in 50% higher antimicrobial action than CH alone. Accordingly, incorporating NSAIDs in inter-appointment dressing has the potential to utilize their anti-inflammatory, local analgesic, and antibacterial actions, which overcome the limitations of CH and improve the outcome of root canal treatment., (© 2024. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

To evaluate and compare the effect of calcium hydroxide [Ca(OH) 2 ] and 2-hydroxyisocaproic acid (HICA) on the microhardness of root dentine. Fifty-one matured maxillary central incisors with straight root and type I canal configuration were decoronated to a standardized length of 16 mm. The root canals were cleaned and shaped using rotary instruments up to size F5. The tooth samples were then randomly assigned into three groups (n = 17) based on the intracanal medicament placed. Group A: control group with no intracanal medicament, Group B: root canals placed with Ca(OH) 2 , and Group C: root canals placed with HICA. After 1 week, the intracanal medicaments placed within the root canals were removed and the canals were dried. Subsequently, the specimens were split longitudinally into two halves and subjected to the Vickers microhardness test with indentations made at the coronal, middle, and apical-third root regions. The data were analyzed using one-way ANOVA and Tukey's post hoc tests. The control group showed significantly highest microhardness value (P < 0.001), followed by HICA and finally, Ca(OH) 2 at the coronal, middle, and apical-third root regions, respectively. Nonetheless, there were no statistically significant differences in microhardness values across different root regions within each group. HICA was found to have a significantly lesser reduction in root dentine microhardness which could be a potential alternative to Ca(OH) 2 . Nevertheless, future studies are warranted to verify the present outcomes in the clinical setting., (© 2023. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

Calcium hydroxide (Ca(OH) 2 NPs), calcium titanate (CaTiO 3 NPs) and yttrium oxide (Y 2 O 3 NPs) nanoparticles are prevalent in many industries, including food and medicine, but their small size raises concerns about potential cellular damage and genotoxic effects. However, there are very limited studies available on their genotoxic effects. Hence, this was done to investigate the effects of multiple administration of Ca(OH) 2 NPs, CaTiO 3 NPs or/and Y 2 O 3 NPs on genomic DNA stability, mitochondrial membrane potential integrity and inflammation induction in mouse brain tissues. Mice were orally administered Ca(OH) 2 NPs, CaTiO 3 NPs or/and Y 2 O 3 NPs at a dose level of 50 mg/kg b.w three times a week for 2 weeks. Genomic DNA integrity was studied using Comet assay and the level of reactive oxygen species (ROS) within brain cells was analyzed using 2,7 dichlorofluorescein diacetate dye. The expression level of Presenilin-1, tumor necrosis factor-alpha (TNF-α) and Interleukin-6 (IL-6) genes and the integrity of the mitochondrial membrane potential were also detected. Oral administration of Ca(OH) 2 NPs caused the highest damage to genomic DNA and mitochondrial membrane potential, less genomic DNA and mitochondrial damage was induced by CaTiO 3 NPs administration while administration of Y 2 O 3 NPs did not cause any remarkable change in the integrity of genomic DNA and mitochondrial membrane potential. Highest ROS generation and upregulation of presenilin-1, TNF-α and IL-6 genes were also observed within the brain cells of mice administrated Ca(OH) 2 NPs but Y 2 O 3 NPs administration almost caused no changes in ROS generation and genes expression compared to the negative control. Administration of CaTiO 3 NPs alone slightly increased ROS generation and the expression level of TNF-α and IL-6 genes. Moreover, no remarkable changes in the integrity of genomic DNA and mitochondrial DNA potential, ROS level and the expression level of presenilin-1, TNF-α and IL-6 genes were noticed after simultaneous coadministration of Y 2 O 3 NPs with Ca(OH) 2 NPs and CaTiO 3 NPs. Coadministration of Y 2 O 3 NPs with Ca(OH) 2 NPs and CaTiO 3 NPs mitigated Ca(OH) 2 NPs and CaTiO 3 NPs induced ROS generation, genomic DNA damage and inflammation along with restoring the integrity of mitochondrial membrane potential through Y 2 O 3 NPs scavenging free radicals ability. Therefore, further studies are recommended to study the possibility of using Y 2 O 3 NPs to alleviate Ca(OH) 2 NPs and CaTiO 3 NPs induced genotoxic effects., (© 2024. The Author(s).)

Aim: To investigate the effect of 38% SDF and its serial dilutions on the Stem cells from Human Exfoliated Deciduous teeth (SHED) and its ability to release growth factors from deciduous dentine., Methods: The viability of SHED post-exposure to 38%, 3.8%, 0.38%, 0.038%, and 0.0038% SDF were assessed at 2, 5, and 7 days using the CyQuant assay, and results were validated using the MTT assay. The osteogenic differentiation of the cells was also investigated post-exposure to 0.0038% SDF. The release of the growth factors; TGF-β1, FGF-b, BMP-2, and VEGF from deciduous dentin discs exposed to 38% SDF, 0.0038% SDF, Ca(OH)2, MTA, and 17% EDTA were examined using ELISA. Statistical analysis was performed using means and standard deviations (p < 0.05). Two-way ANOVA compared the means of more than two groups with Tukey's multiple comparison test. The unpaired t-test was also used to compare the differences between the two data sets., Conclusion: 38% SDF released dentinogenic growth factors from dentin discs, potentially explaining its role in reactionary dentinogenesis. Moreover, 0.0038% SDF resulted in a non-cytotoxic concentration that enhanced cellular proliferation and released bioactive molecules from dentin comparable to the 38% concentration. After further investigations, the 0.0038% dilution of SDF could present itself as a clinical concentration.

Aim: This study was done to clinically investigate the efficacy of glycyrrhizin-based intracanal medication (ICM) compared to calcium hydroxide Ca(OH) 2 and Ledermix on the bacterial counts inside the root canals of necrotic teeth with chronic periapical periodontitis., Methodology: Thirty-six patients having single-rooted and single-canaled necrotic teeth with chronic periapical periodontitis were enrolled in this research. Aseptic control measures were taken before clinical steps. Access cavity preparation was done. First bacteriological samples (S1) were collected immediately after access cavity preparation and before cleaning and shaping inside the root canals. Cleaning and shaping were performed on the root canals. Patients were randomly allocated into 3 groups according to the type of intracanal medicament used [Ca(OH) 2 , Ledermix, Glycyrrhizin]. Second bacteriological samples (S2) were collected after 1 week from placing the ICMs. S1 and S2 were transferred to the lab of microbiology for culturing on blood agar dishes in anaerobic conditions, and the bacteria on the plates were enumerated as colony-forming units (CFUs) by the manual counting method. The anti-bacterial efficacy of the ICM was estimated by the percentage reduction in the bacterial colonies from S1 to S2., Results: All tested ICM documented a significant reduction in the CFUs from S1 to S2 ( p < 0.05). Regarding S1, there was no significant difference between the three tested materials ( p > 0.05). Regarding S2, Glycyrrhizin and Ledermix showed significantly lower CFUs than Ca(OH) 2 . There was no significant difference between Glycyrrhizin and Ledermix., Conclusion: Under the conditions of the present study, Glycyrrhizin and Ledermix showed comparable antimicrobial effects that were better than the commonly used Ca(OH) 2 ICM. Glycyrrhizin may be a promising ICM., Clinical Significance: Herbal medicine can be considered as an alternative antimicrobial material for root canal disinfection because it has many benefits, like low toxicity, absence of microbial resistance, and favorable antimicrobial efficacy. This study highlights the clinical efficacy of Glycyrrhizin as a promising ICM. How to cite this article: Eltantawi AR, Abdel-Razik GM, Elhawary YM, et al. Efficacy of Glycyrrhizin as an Intracanal Medicament on Bacterial Load Reduction in Primary Infected Root Canals: A Randomized Clinical Trial. J Contemp Dent Pract 2024;25(6):540-546.

Introduction: The aim of this study was to evaluate the anti-osteoclastic activity of calcium hydroxide-loaded poly(lactic-co-glycolic acid) nanoparticles [Ca(OH) 2 -loaded PLGA NPs] in comparison to calcium hydroxide nanoparticles [Ca(OH) 2 NPs]., Methods: RAW 264.7 cell lines (third-fifth passage) were cultured and incubated with soluble receptor activator of nuclear factor kappa B ligand in triplicate. Subsequently, Ca(OH) 2 -loaded PLGA NPs and Ca(OH) 2 NPs were added for 7 days to evaluate their effects on receptor activator of nuclear factor kappa B ligand-induced osteoclast differentiation of RAW 264.7 cells by tartrate-resistant acid phosphatase activity. Additionally, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was conducted to confirm the cytotoxicity of treatments to cells., Results: Tartrate-resistant acid phosphatase staining showed a significant reduction in the osteoclast number when treated with Ca(OH) 2 -loaded PLGA NPs compared with Ca(OH) 2 NPs (P < .01). In comparison to the control, the number of osteoclasts significantly reduced upon treatment with Ca(OH) 2 -loaded PLGA NPs (P < .05), but there was no significant difference in Ca(OH) 2 NPs. Furthermore, osteoclast morphology in both treatment groups exhibited smaller sizes than the control group. Neither Ca(OH) 2 -loaded PLGA NPs nor Ca(OH) 2 NPs demonstrated cytotoxic effects on RAW264.7 cells., Conclusions: Both Ca(OH) 2 NPs with and without poly(lactic-co-glycolic acid) have the ability to inhibit osteoclast differentiation. However, Ca(OH) 2 -loaded PLGA NPs exhibit greater potential than Ca(OH) 2 NPs, making them a promising intracanal medicament for cases of root resorption., (Copyright © 2024 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Purpose: To investigate the influence of methylene blue (MB)-mediated antimicrobial photodynamic therapy (aPDT) and calcium hydroxide (CH) medication on the mechanical characteristics, degree of conversion (DC), quantification, and volume of gaps at the adhesive interface of glass fiber posts (GFPs) luted to distinct thirds of root canal dentin. Additionally, the microhardness (MH), elastic modulus (Eit), morphology, and chemical structure of the intraradicular dentin were assessed., Materials and Methods: 6 experimental groups were formed by sorting 102 bovine incisors. Canals receiving deionized water irrigation as a negative control; canals receiving deionized water irrigation and filled with CH as a positive control; groups treated with CH + MB at 50 and 100 mg/L without irradiation; and groups treated with CH + MB at 50 and 100 mg/L irradiated by red laser for 60 s (660 nm; 100 mW; 6.5 J; 72 J/cm 2 ). MH, Eit, and DC properties were evaluated for both the resin cement layer and root dentin substrate (n = 8). Volume and quantification of gaps at the bonding interface (n = 6), and dentin morphology and chemical content were investigated (n = 3). Data were analyzed using a repeated-measures 2-way ANOVA followed by Tukey post hoc analysis (α = 0.05)., Results: The distinct intraradicular thirds and treatment with MB-mediated aPDT, whether activated or not, in combination with CH, had a significant impact on the mechanical characteristics of the root dentin. This effect was also observed in the MH, Eit, DC, quantification, and volume of gaps at the luting interface (P < .05). In general, a higher concentration of MB, whether activated by a red laser or not, led to lower values in the mechanical properties of the root dentin, as well as in MH, Eit, and DC at the adhesive interface (P < .05). Additionally, these groups exhibited higher values for quantification and volume of gaps at the luting substrate (P < .05). Scanning electron micrographs and energy dispersive X-ray spectra showed qualitative similarity among all groups, except for the negative experimental control group., Conclusions: MB-mediated aPDT at 50 mg/L, in combination with CH, demonstrated favorable physico-chemical and mechanical characteristics in intraradicular dentin, along with satisfactory mechanical features and the adhesive interface integrity for GFPs at all intraradicular depths., Clinical Significance: MB-mediated aPDT at a concentration of 50 mg/L combined to CH medication represents a suitable choice for photosensitization in the context of intracanal disinfection following the biomechanical procedure and prior to luting of intraradicular restorations., Competing Interests: Declaration of competing interest The authors have stated explicitly that there are no conflicts of interest in connection with this article., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the antimicrobial activity of calcium hydroxide-loaded poly(lactic-co-glycolic acid) nanoparticles (CH-loaded PLGA NPs) on multi-species biofilms. Human root blocks were prepared (n = 40), and multi-species suspensions of Candida albicans, Enterococcus faecalis and Streptococcus gordonii were incubated within the root canals for 21 days. Canals (n = 10/group) were then medicated with saline solution (negative control), chlorhexidine (positive control), calcium hydroxide and CH-loaded PLGA NPs for 7 days. Samples taken from the 0.1 mm root canal dentin were collected, and cell growth was detected by culture on BHI agar. The viable cell count of the Ca(OH) 2 , chlorhexidine gel and CH-loaded PLGA NPs group was significantly lower than the normal saline group (p < 0.001). CH-loaded PLGA NPs demonstrated a significant lower viable cell than Ca(OH) 2 (p < 0.001); it has potential as a medicament for endodontic therapy., (© 2023 Australian Society of Endodontology Inc.)

High expression of MMP-2 and MMP-9 in periapical lesions plays an important role in the degradation of the extracellular matrix. This study aimed to investigate the effect of epigallocatechin-3-gallate (EGCG)-based endodontic paste as an intracanal dressing on the expression of MMP-2 and MMP-9 in periapical lesions. Periapical lesions were experimentally induced in 35 mature beagle dog premolars randomly divided into healthy teeth, untreated periapical lesions, periapical lesions treated in a single session (control groups), and periapical lesions treated in two sessions with EGCG or calcium hydroxide-based pastes (experimental groups). After 120 days, specimens were obtained for histopathologic and immunofluorescence analyses to assess the expression of MMP-2 and MMP-9. The statistical analysis was performed using a p-value of 0.05. Endodontic treatment in two sessions using medication with EGCG and calcium hydroxide-based pastes provided similar repair of the apical and periapical tissues and neoformation of periodontal ligament fibers, cementum, and alveolar bone (p>0.05). The experimental groups treated in two sessions with both medications presented expression of MMP-2 and MMP-9 similar to that in healthy teeth (p>0.05), and significantly lower than teeth treated in a single session or untreated periapical lesions (p <0.001). Expression of MMP-2 and MMP-9 was observed in the cytoplasm of fibroblasts, osteoblasts, cementoblasts, cementocytes, and vascular endothelium. The use of EGCG-based endodontic paste reduced the expression of MMP-2 and MMP-9 and allowed repair of periapical lesions, similar to calcium hydroxide-based paste, and superior to treatment performed in a single session.

Aim: To study the effect of glycyrrhizin (GA) on the viability and proliferation of dental pulp stem cells (DPSCs) compared with intracanal medicaments., Materials and Methods: Third molars of an adult donor were used to obtain the DPSCs. Flow cytometry was utilized to conduct phenotypic analysis for DPSCs. The methyl-thiazol tetrazolium (MTT) test was used to detect the cell viability. Cell proliferation assay was conducted at distinct time intervals: 3, 5, and 7 days., Results: The flow cytometry analysis verified the positive expression of mesenchymal cell surface antigen molecules (CD73, CD90, and CD105) and the absence of hematological markers (CD14, CD34, and CD45) in the DPSCs. The cells that treated with concentrations more than 0.5 mg/mL of Ca(OH 2 ) and triple antibiotic paste (TAP) gave significant decrease in viability in comparison to the untreated cells ( p < 0.05). Also, the cells treated with concentrations 50 and 25 µM of GA showed no significant difference compared with the untreated cells ( p > 0.05), while concentrations 12.5 and 6.25 µM expressed a significant increase in viability compared with the untreated cells ( p < 0.05). At 7 days, cells treated with the three different concentrations of GA (12.5, 25, and 50 µM) demonstrated a significant increase in cell density compared with Ca(OH) 2 and TAP-treated cells ( p < 0.05)., Conclusion: Based upon the potential of GA on DPSCs proliferation compared with Ca(OH) 2 and TAP, It is conceivable to acknowledge that GA could be used as an intracanal medicaments for revascularization process of necrotic immature teeth., Clinical Significance: This study emphasizes the significance of assessing alternative root canal medicaments and their impact on the proliferation and viability of DPSCs. The results regarding GA, specifically its impact on the viability and growth of DPSCs, provide essential understanding for its potential application as an intracanal medicine. This study adds to the continuous endeavors in identifying safer and more efficient intracanal therapies, which are essential for improving patient outcomes in endodontic operations. How to cite this article: Alrashidi MA, Badawi MF, Elbeltagy MG, et al . The Effect of Glycyrrhizin on the Viability and Proliferation of Dental Pulp Stem Cells Compared to Intracanal Medicaments. J Contemp Dent Pract 2024;25(3):267-275.

Background: An innovative intracanal medication formulation was introduced in the current study to improve the calcium hydroxide (Ca(OH) 2 ) therapeutic capability against resistant Enterococcus faecalis (E. faecalis) biofilm. This in-vitro study aimed to prepare, characterize, and evaluate the antibacterial efficiency of Ca(OH) 2 loaded on Gum Arabic (GA) nanocarrier (Ca(OH) 2 -GA NPs) and to compare this efficiency with conventional Ca(OH) 2 , Ca(OH) 2 nanoparticles (NPs), GA, and GA NPs., Materials and Methods: The prepared nanoparticle formulations for the tested medications were characterized using Transmission Electron Microscope (TEM) and Fourier-Transform Infrared Spectroscopy (FTIR). 141 human mandibular premolars were selected, and their root canals were prepared. Twenty-one roots were then sectioned into 42 tooth slices. All prepared root canals (n = 120) and teeth slices (n = 42) were divided into six groups according to the intracanal medication used. E. faecalis was inoculated in the samples for 21 days to form biofilms, and then the corresponding medications were applied for 7 days. After medication application, the residual E. faecalis bacteria were assessed using CFU, Q-PCR, and SEM. Additionally, the effect of Ca(OH) 2 -GA NPs on E. faecalis biofilm genes (agg, ace, and efaA) was investigated using RT-PCR. Data were statistically analyzed at a 0.05 level of significance., Results: The synthesis of NPs was confirmed using TEM. The results of the FTIR proved that the Ca(OH) 2 was successfully encapsulated in the GA NPs. Ca(OH) 2 -GA NPs caused a significant reduction in the E. faecalis biofilm gene expression when compared to the control (p < 0.001). There were significant differences in the E. faecalis CFU mean count and CT mean values between the tested groups (p < 0.001) except between the Ca(OH) 2 and GA CFU mean count. Ca(OH) 2 -GA NPs showed the least statistical E. faecalis mean count among other groups. SEM observation showed that E. faecalis biofilm was diminished in all treatment groups, especially in the Ca(OH) 2 -GA NPS group when compared to the control group., Conclusions: Ca(OH) 2 and GA nanoparticles demonstrate superior anti-E. faecalis activity when compared to their conventional counterparts. Ca(OH) 2 -GA NPs showed the best antibacterial efficacy in treating E. faecalis biofilm. The tested NP formulations could be considered as promising intracanal medications., (© 2024. The Author(s).)

Aim: This study aims to evaluate the impact of asiaticoside (AC) on the viability and proliferation of dental pulp stem cells (DPSCs), considering the known negative effects of routinely used intracanal medicaments. This evaluation will be compared with the outcomes from using traditional intracanal medicaments, specifically triple antibiotic paste (TAP) and calcium hydroxide [Ca(OH) 2 ]., Materials and Methods: The DPSCs were obtained from the third molars of an adult donor. The application of flow cytometry was employed to do a phenotypic analysis on DPSCs using CD90, CD73, CD105, CD34, CD14, and CD45 antibodies. The methylthiazol tetrazolium (MTT) assay was employed to assess cellular viability. The cells were treated with different concentrations of TAP and Ca(OH) 2 (5, 2.5, 1, 0.5, and 0.25 mg/mL), along with AC (100, 50, 25, 12.5, and 6.25 µM). A cell proliferation rate was performed at 3, 5, and 7 days., Results: The characterization of DPSCs was conducted by flow cytometry analysis, which verified the presence of mesenchymal cell surface antigen molecules (CD105, CD73, and CD90) and demonstrated the absence of hematopoietic markers (CD34, CD45, and CD14). Cells treated with concentrations over 0.5 mg/mL of TAP and Ca(OH) 2 showed a notable reduction in cell viability in comparison to the untreated cells ( p < 0.05). Additionally, the cells treated with different concentrations of AC 12.5, 6.25, 25, and 50 µM did not differ significantly from the untreated cells ( p > 0.05). Nevertheless, cells treated with concentrations of 100 µM showed a significant reduction in viability compared to the untreated cells ( p < 0.05). After a period of 7 days, it was noted that cells exposed to three different concentrations of AC (50, 25, and 12.5 µM) had a notable rise in cell density in comparison to TAP and Ca(OH) 2 (p < 0.05). Furthermore, cells that were exposed to a concentration of 12.5 µM exhibited the highest cell density., Conclusion: The cellular viability of the AC-treated cells was superior to that of the TAP and Ca(OH) 2 -treated cells. Moreover, the AC with a concentration of 12.5 µM had the highest degree of proliferation., Clinical Significance: This study underscores the importance of evaluating alternative root canal medicaments and their effects on DPSCs' growth and vitality. The findings on AC, particularly its influence on the survival and proliferation of DPSCs, offer valuable insights for its probable use as an intracanal medication. This research contributes to the ongoing efforts to identify safer and more effective intracanal treatments, which are crucial for enhancing patient outcomes in endodontic procedures. How to cite this article: Alazemi MJ, Badawi MF, Elbeltagy MG, et al . Examining the Effects of Asiaticoside on Dental Pulp Stem Cell Viability and Proliferation: A Promising Approach to Root Canal Treatment. J Contemp Dent Pract 2024;25(2):118-127.

Enterococcus faecalis was the main causative bacteria of refractory periapical periodontitis (PP), there is a pressing need to explore effective methods for eradicating E. faecalis in patients with refractory PP. This study aimed to assess the anti-infective effectiveness of phage PEf771 in treating periapical inflammation in rats. We developed a rat model of PP through E. faecalis YN771 induction. Micro-computed tomography and hematoxylin-eosin staining were utilized to evaluate bone destruction and inflammation in experimental teeth for seven consecutive weeks. Subsequently, rats with PP caused by E. faecalis YN771 were treated with phage PEf771, calcium hydroxide preparation, and 2% chlorhexidine gel. The healing progress of bone defects and inflammation in the apical region was monitored over three consecutive weeks using imaging and histopathology assessments. The PP rat model was successfully developed, and bone destruction and inflammatory cell infiltration in the apical region of the experimental tooth peaked at 4 weeks. The area of bone destruction in rats treated with phage PEf771, calcium hydroxide preparation, and 2% chlorhexidine gel was significantly smaller than that in the untreated group. Phage PEf771, calcium hydroxide preparation, and 2% chlorhexi-dine gel all have the effect of promoting the healing of apical lesions. Therapeutic effects of phage PEf771 on periapical inflammation infected by E. faecalis YN771 enhanced with time. Phage PEf771 promoted the healing of apical lesions, presenting a promising new approach for the treatment of refractory PP using bacteriophages.

Aim: We evaluated Diacetylcurcumin (DAC), a derivative of curcumin, for its antibacterial and antibiofilm properties against Enterococcus faecalis . Methods: Minimum inhibitory concentration (MIC) and minimum bactericidal concentration were determined, along with antibiofilm potential and toxicity in Galleria mellonella . Additionally, in silico computational analysis was performed to understand its mechanisms of action. Results & conclusion: DAC demonstrated significant antibacterial effects, with MIC and MBC values of 15.6 and 31.25 μg/ml, respectively, and reduced biofilm formation. A synergistic effect, reducing biofilm by 77%, was observed when combined with calcium hydroxide. G. mellonella toxicity tests confirmed DAC's safety at tested concentrations, suggesting its potential for use in root canal disinfection products.

Background: The placement of liners near the pulp area is essential for therapeutic effects and maintaining pulp health while stimulating the formation of tertiary dentin. This in vitro study aimed to evaluate the calcium release, pH, biocompatibility, solubility, and bioactivity of three resin-modified calcium hydroxide cavity liners., Methods: The disc specimens of each cavity liner were prepared using polyethylene molds of 7 mm in diameter and 2 mm in height (n = 10). Three light-cure liners evaluated include Ultra-Blend Plus (UB), Base-it (BI), and Master Dent (MD). The samples were then immersed in flasks containing 10 mL of distilled water. Calcium ion release, pH, and solubility were evaluated in two weeks of incubation. The cytotoxicity of extracts adjacent to the specimens was evaluated by MTT assay using NIH/3T3 cells after 1, 3, and 7 days of incubation. The ability to induce the nucleation of calcium phosphates (CaPs) after 28-day immersion in a simulated body fluid was investigated by SEM-EDX analysis. Statistical analysis was performed using ANOVA, Kruskal-Wallis, and repeated measures tests at the significant level of 0.05., Results: There was a significant difference in the release of calcium ions among the three liners investigated on days 1, 7, and 14 (p < 0.05). UB liners exhibited a significantly higher amount of calcium release than the other two liners, followed by BI, and MD. On day 1, there was no significant difference in the average pH among the three liners. However, after day 7, the MD liner showed a significant decrease in pH compared to the other two liners. BI liner demonstrated the highest level of biocompatibility, followed by the MD and UB liners. UB showed a high calcium release, solubility with no alkalizing activity, and the formation of more mature Ca-rich apatite deposits than the other two liners., Conclusion: Based on the results of this study, the cavity liner material's performance is material dependent. It can impact ion release, biocompatibility, and bioactivity which are important factors to consider in clinical practice. Further studies are needed to investigate the long-term effects of different liner materials on oral tissues., (© 2023. The Author(s).)

Due to the high failure rates associated to endodontic disinfection, this study aimed to investigate the antibacterial properties of poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) loaded with Ca(OH) 2 for endodontic disinfection procedures. Ca(OH) 2 NPs production and physicochemical characterization were carried out as well as multiple antibacterial tests using three bacterial strains and an ex vivo model of endodontic infection with extracted human teeth. Agar diffusion test and broth dilution determined the inhibition growth zones (n = 5) and the minimal inhibitory concentration (MIC, n = 5), respectively. Cell viability was assessed using Live/Dead staining with confocal microscopy (n = 5). Data was analysed using ANOVA followed by post-hoc analysis. After 24 h of incubation, Ca(OH)₂ NPs demonstrated a MIC of 10 µg/mL for Porphyromonas gingivalis (p < 0.001) and Enterococcus faecalis and 5 µg/mL for Fusobacterium nucleatum (p < 0.001). Although the agar diffusion test did not exhibit any inhibition area for Ca(OH) 2 nor for Ca(OH)₂ NPs, this was probably due to the buffering effect of the agar medium. However, the antibacterial capacity was confirmed in an ex vivo model, where instrumentalized teeth were infected with Enterococcus Faecalis and treated after 28 days of culture. A significant reduction in bacterial metabolic activity was confirmed for Ca(OH) 2 NPs (40 % reduction with a single dose) and confirmed by Live/Dead staining. In conclusion, Ca(OH)₂-loaded PLGA NPs present promising antibacterial efficacy for endodontic disinfection procedures., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Elena Sanchez-Lopez has patent #Composition comprising nanoparticles, method for the preparation of a composition comprising nanoparticles and uses of the composition for dental treatment. EP20382504.7 issued to Explotation rights: Universitat Internacional de Catalunya and Universitat de Barcelona. Regarding the patent mentioned other inventors are also included in this manuscript: García López, M. L.; Sánchez López, E.; Durán-Sindreu, F.; González Sánchez, J. A. and Elmasmari, F.]., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Aim: This study was aimed at evaluating the effect of acetyl-11-keto-β-boswellic acid (AKBA) on dental pulp stem cells (DPSCs) viability and proliferation to be used as a potential root canal medicament., Materials and Methods: Dental pulp stem cells were isolated from human third molars. The phenotypic characterization of DPSCs was verified by flow cytometry analysis. The viability assay was performed using the methyl-thiazoltetrazolium (MTT) assay. Cells were treated with different concentration of triple antibiotic paste (TAP) and calcium hydroxide Ca(OH 2 ) (5, 2.5, 1, 0.5, and 0.25 mg/mL), AKBA (10, 5, 1, 0.1, and 0.01 µM). All experiments were done in separate triplicate experiments . Results: Dental pulp stem cells were characterized by flow cytometry. Cells treated with Ca(OH) 2 (1, 2.5, and 5 mg/mL) showed significantly reduced viability compared with the control cells ( p < 0.05). Dental pulp stem cells treated with 1, 2.5, and 5 mg/mL TAP resulted in a significant decrease in viability ( p < 0.05). Cells treated with AKBA in concentrations (1, 0.1, and 0.01 µM) demonstrated higher viability than the control group ( p < 0.05), while AKBA in concentrations (5 and 10 µM) showed equal or decreased viability than the control group. ( p > 0.05). Regarding cell density assay, AKBA showed significant increase in cell density after 5 and 7 days compared with cells medicated with TAP and Ca(OH) 2 while TAP revealed marked reduction in cell density in all the tested intervals., Conclusion: Acetyl-11-keto-β-boswellic acid in lower concentrations (0.01, 0.1, and 1 µM) demonstrated superior cell viability than TAP and Ca(OH) 2 , and it may possess the potential to be an intracanal medicament in regenerative endodontics., Clinical Significance: Studying the effect of different potential root canal medicaments and their capability to induce DPSCs proliferation might be of value. The influence of AKBA on the viability and proliferation of DPSCs tested in this study sheds light on its use as a potential intracanal medication especially in regenerative endodontics. How to cite this article: Amer NA, Badawi MF, Elbeltagi MG, et al . Effect of Boswellic Acid on Viability of Dental Pulp Stem Cells Compared to the Commonly Used Intracanal Medications: An In Vitro Study. J Contemp Dent Pract 2023;24(12):957-966.

Introduction: This study aimed to compare the postoperative pain level changes resulting from using calcium silicate- (EndoSeal MTA) and calcium hydroxide-based (Sealapex) root canal sealers in mandibular first and second molar teeth with symptomatic apical periodontitis., Methods: A total of 60 patients with symptomatic apical periodontitis in their lower molar teeth were randomly allocated into 2 groups according to sealer type (n = 30). Demographic data, including gender, age, and smoking habit, and preoperative pain measures were recorded. Root canal treatments were performed in a single visit. Postoperative pain measurements and analgesic intake were measured at 6, 12, 24, and 48 hours and after 3, 5, and 7 days using the visual analog scale. The data were statistically analyzed using a chi-squared test (to compare gender, age, smoking habit, analgesic intake, and sealer extrusion), the Mann-Whitney U test (to compare pain levels), Friedman tests (for the evaluations of the reduction in pain levels over time), and Spearman's correlation test (to analyze the relationships of age, gender, smoking habit factors with postoperative pain) (P = .05)., Results: The statistical analysis showed no significant differences between the groups in postoperative pain and analgesic intake at any of the time intervals evaluated (P > .05)., Conclusions: Patients treated with calcium silicate- and calcium hydroxide-based root canal sealers experienced similar postoperative pain and no statistically significant differences were observed in analgesic intake., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This study evaluated the antimicrobial efficacy of a new intracanal drug based on Pentaclethra macroloba extract, a plant of Amazonian origin, against Enterococcus faecalis using macrodilution test and intratubular evaluation with Confocal Laser Scanning Microscopy (CLSM). The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of the pure extract of Pentaclethra macroloba andits association with calcium hydroxide and ultracall were determined. Then, thirty-three dentin cylinders were prepared and inoculated with E. faecalis, to evaluate the antimicrobial effect of the medications on the dentinal tubules with CLSM. The data was analyzed using the Kruskal-Wallis and Dunn tests. The extract in association with calcium hydroxide showed a lower CBM, and in the intratubular test all tested medications were effective against E. faecalis (P >0.05). The new intracanal drug based on P. macroloba extract has an antimicrobial effect against E. faecalis and further studies are needed for its clinical use.

Background: The study aims to compare the synergistic antibacterial efficacy of different combinations of calcium hydroxide as an intracanal medicament against E. faecalis., Material and Methods: The current study included four hundred extracted human permanent mandibular premolar teeth. After complete chemo-mechanical preparation, the middle third of the root was sectioned using a rotary diamond disc and a total of 400 samples were obtained. The specimens were inoculated with E. faecalis for 21 days. After that, specimens were divided into five groups (n = 80) based on materials used for the disinfection of samples: Group I, calcium hydroxide alone; Group II, calcium hydroxide + 2% chlorhexidine gel; Group III, calcium hydroxide + 2% chitosan gel; Group IV, calcium hydroxide + 0.02% silver nanoparticle gel; Group V, calcium hydroxide + Bioactive glass S53P4. Dentin shavings from the apical third were obtained from the inner third of dentin were obtained using gates glidden no.1 to the apical depth, followed by no.2, 3, 4 and 5 analyzed for E. faecalis using the culture method. One-way analysis of variance (ANOVA) was used for data analysis, followed by post-hoc Tukey's test for multiple comparisons of means to check the difference in bacterial inhibition between the groups., Results: ANOVA results revealed a significant reduction of bacterial counts in all the groups compared (p < 0.001). Intergroup comparison showed maximum bacterial reduction (p < 0.001) with calcium hydroxide + bioactive glass S53P4 compared with other groups., Conclusion: Synergistic effect of calcium hydroxide showed better bacterial reduction compared to calcium hydroxide alone. Among the combinations evaluated, calcium hydroxide with bioactive glass, found to be most effective compared to other groups., (© 2023. The Author(s).)

Background: Vital pulp therapy, based on the use of stem cells, has promising research and therapeutic applications in dentistry. It is essential to understand the direct effect of capping materials on the dental pulp stem cells of primary teeth, which contribute to the healing powers of the tooth. The aim of this study is to evaluate the effect of different capping materials (Calcium Hydroxide (DyCal®) - Glass Ionomer (Fuji IX®) and light-cured resin modified calcium silicate (TheraCal LC®)) on the viability, proliferation, and differentiation of stem cells from human exfoliated deciduous teeth (SHEDs)., Methods: SHEDs were isolated from extracted primary teeth, then divided into four groups and each of the capping materials were applied to the stem cells as follows: group I the controls, group II with Ca(OH)2, group III with the GIC, and group IV with the Theracal LC. For all groups assessment of viability and proliferation rate was done using the MTT cell proliferation assay. Also, Differentiation was evaluated by measuring the gene expression of Alkaline phosphatase enzyme activity (ALP) and Dentin matrix protein-1 (DMP1) through quantitative real-time PCR. Morphological assessment was conducted using Alizarin Red S staining. All evaluations were performed after 7 and 14 days of culture., Results: TheraCal LC showed the highest values of proliferation, which was significant only compared to the control group after 2 weeks (p = 0.012). After one week, TheraCal LC showed the highest significant values of ALP and DMP1 compared to all other groups (p < 0.001)., Conclusion: The three materials under study are biocompatible, maintain viability, and stimulate the proliferation and differentiation of SHEDs. However, TheraCal LC allows better proliferation of SHEDs than Dycal Ca(OH)2 and Fuji IX GIC., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Different calcium hydroxide [Ca(OH) 2 ] formulations are available. The aim was to assess differences in hydroxyl ion release with different formulations. Sixty-six teeth were divided one control (n = 6) and four experimental (n = 15) groups: (i) Pulpdent Paste; (ii): DT Temporary Dressing; (iii): Ca(OH) 2 powder/saline; (iv): Ca(OH) 2 points; (v): no medicament. pH was measured in inner dentine and outer dentine cavities over 12 weeks. Inner dentine pH rose rapidly for all groups except the points and controls. Peak pH was reached by day 2 before dropping and stabilising (8.0-9.2). Outer dentine pH rose steadily until day 21 for aqueous solutions and then stabilised (8.0-8.5). The points had minimal pH increase for the entire period. There were no significant differences in hydroxyl ion release between the aqueous solutions. Type of paste base did not affect release and diffusion of hydroxyl ions which continued for 84 days., (© 2018 Australian Society of Endodontology Inc.)

The health of dental pulp tissue is critical to maintaining normal tooth function from the eruption of permanent teeth to the formation of the apex. The study evaluated the inflamed pulp response to the mineral trioxide aggregate (MTA) after direct pulp capping with the mechanical pulp exposure in rats' incisor. Forty-eight mandibular central incisors of twenty-four Sprague-Dawley rats which were prepared with the cavities of one mm diameter, and the pulp exposures were randomly assigned into two groups: MTA group and calcium hydroxide (Ca(OH) 2 ) group. The direct pulp capping was performed after three days and samples histological observations conduction within eight weeks. In both MTA and Ca(OH) 2 groups, dentin -like structures were observed in the pulp tissues of some teeth. The number of teeth with reparative tissue in MTA group was statistically significantly higher than that in Ca(OH) 2 group ( p = 0.041). Inflammatory cell infiltration was found in the crown pulp tissues in two groups, and no statistical difference was observed between the two groups ( p = 0.243). Pulp necrosis occurred in both groups, and there was no statistical difference between the two groups ( p = 0.622). The results in this paper suggest that MTA promotes direct pulp capping and hence has certain potential clinical applications value in the treatments for the preservation of inflamed pulp., Competing Interests: The authors declare no conflict of interest., (©2023 The Author(s). Published by MRE Press.)

This study is aimed to assess the efficacy of different final irrigation agitation techniques on dentin tubule penetration of nanoparticle calcium hydroxide (NCH) dressing. Ninety-six extracted upper incisors were shaped up to #40file. Then, Four experimental groups were formed according to the final irrigation procedure: conventional needle irrigation (CNI), manual dynamic agitation (MDA), sonic agitation (SA) and ultrasonic irrigant agitation (UIA). According to the intracanal drug used, these groups were divided into two subgroups: calcium hydroxide (CH) and NCH. Prepared CH preparations were labelled with Rhodamine B, and CH or NCH were placed in root canals. In the UIA group, both CH and NCH had the highest penetration depth and percentage compared to the other groups (p < 0.05). The penetration depth and percentage of NCH in the UIA and SA groups were significantly higher than in the CH groups (p < 0.05). UIA is more effective than other groups in increasing dentinal tubule penetration of CH and NCH., (© 2023 Australian Society of Endodontology Inc.)

Aim: Pulp vitality is essential for tooth integrity. Following pulp exposure, choosing a suitable pulp-capping material is crucial to maintain pulp vitality. However, the reparative dentine bridge created by calcium hydroxide (Ca(OH) 2 ) is generally porous and incomplete. The aim of the current study is to assess the in vitro and in vivo bioactivities of nano eggshell-based slurry (NES), using NES as a direct pulp-capping material, compared with Ca(OH) 2 in rabbit animal model., Methodology: Nano eggshell powder (NE) was characterized for particle morphology, chemical composition and ion release. In vitro bioactivity was tested by immersion in simulated body fluid (SBF) for 7 days. For histopathological evaluation, 36 adult New Zealand rabbits (72 pulp exposures) were divided into nine groups (n = 8) according to the pulp-capping material (NES, Ca(OH) 2 and no capping as negative control group) and the animals were sacrificed after 7, 14 or 28 days. The pulps of the two lower central incisors were exposed and then directly capped by Ca(OH) 2 or NES or left untreated. The cavities were then sealed with glass ionomer cement. Teeth were collected for histopathological evaluation using an optical microscope. Pulp haemorrhage, inflammation, fibrosis and calcific bridge formation were assessed. Results were statistically analysed using anova and Tukey's tests., Results: Nano eggshell particles were spherical with a 20 nm diameter and were composed mainly of calcite. Statistical analysis showed that there was a significant increase in the release of all investigated ions between days 1 and 28, except for copper. NES group showed a significantly higher release of all elements as compared to Ca(OH) 2 . Environmental scanning electron microscope micrographs of NES incubated for 7 days in SBF showed the formation of HAp with a Ca/P ratio (1.686). For histopathological evaluation, the difference between groups was statistically significant. At day 28, 75% of the pulps of the Ca(OH) 2 group showed mild calcific bridge in comparison with 100% moderate calcific bridge in the NES group. The NES group showed significantly less inflammation at days 7 and 28, and higher fibrosis at day 7 compared with Ca(OH) 2 ., Conclusions: Nano eggshell-based slurry represents a promising novel direct pulp-capping material with favourable pulp tissue response., (© 2023 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Background: Thorough disinfection of root canals in primary molars may be complicated by the complex root canal morphology. This in-vitro study aimed to compare direct and residual antimicrobial effect of 2% chlorhexidine (CHX) gel, 500 mg/ml double antibiotic paste (DAP) and chitosan-chlorhexidine nanoparticles (CS-CHX NPs) as intracanal medicaments against Enterococcus faecalis (E. faecalis) and Candida albicans (C. albicans) in primary molars., Methods: Mesial roots of 63 mandibular second primary molars were infected with E. faecalis and C. albicans. Teeth were divided into 9 groups: Ia: (CS-CHX NPs), IIa: (CHX), IIIa: (DAP), IVa: chitosan nanoparticles (CSNPs) in which medicaments were placed for 3 days, groups Ib: (CS-CHX NPs), IIb: CHX, IIIb: (DAP), IVb: (CSNPs) in which medicaments were placed for 7 days, and Group V (control): teeth were infected, irrigated with saline, and sampled 3- and 7-days post-infection. Microbiological samples were obtained after infection, 3, and 7 days after medicament placement and 7 days after medicament removal for both time points. One-way ANOVA, Tukey's post hoc test and paired t-test were used at p < 0.05., Results: CS-CHX NPs had the highest anticandidal effect which was comparable to CHX and significantly higher than other medicaments (p < 0.001) at both time points. CS-CHX NPs had the highest effect against E. faecalis which was comparable to DAP and significantly higher than other medicaments (p < 0.001) at 3 days. All medicaments showed similar effect against E. faecalis after 7 days. The 7-days placement significantly increased the antimicrobial effect against both micro-organisms in all groups, except CS-CHX NPs which showed an insignificant increase. CS-CHX NPs showed the highest residual effect against both micro-organisms that increased with 7-days placement., Conclusion: CSNPs and CHX combination showed a synergistic effect against both micro-organisms. CS-CHX NPs displayed a higher effect at a shorter period compared to other medicaments, yet its residual effect was higher with 7-days placement., (© 2023. The Author(s).)

Successful endodontic treatment requires advanced materials to eliminate biofilm This study aims to assess the penetration depth and the effectiveness of Boswellia sacra as a novel intracanal medicament compared with calcium hydroxide against Enterococcus faecalis biofilm. 60 single-rooted teeth were decoronated, prepared and sterilised. Fifty teeth were contaminated with a culture of E. faecalis (ATCC 19433) for 21 days. Two teeth were used to confirm the biofilm using scanning electron microscope. For colony-forming unit (CFU), 40 samples were divided into one control group (calcium hydroxide) and the other experimental group (B. sacra). Each group was divided into two subgroups to be tested at 3 and 7 days. The minimum inhibitory concentration (MIC) of B. sacra was determined, and the B. sacra's ethanolic extract medicament was prepared. Eight discs divided into groups similar to CFU were used to evaluate live/dead bacteria using confocal laser scanning microscopy (CLSM). Ten teeth were selected for penetration depth using CLSM. The intracanal medicaments were mixed with 0.1% rhodamine B. were inserted into the root canals 0.2 slices were dissected and viewed under CLSM. The MIC of B. sacra was 1.25 mg/ml. The CFU evaluation proved that B. sacra are more effective than calcium hydroxide in the 3 days groups. However, it was statistically insignificant compared with calcium hydroxide after 7 days. The depth of penetration of B. sacra exceeds that of calcium hydroxide. B. sacra is an effective intracanal medicament., (© 2022 Australian Society of Endodontology Inc.)

The aim of this integrative review was to identify whether the disinfection procedures performed prior to regenerative endodontic treatment were effective on biofilm removal from the root canals. The research was based on PubMed, Latin American and Caribbean Health Sciences Literature (Lilacs) and Scientific Electronic Library Online (SciELO) databases. Four articles were selected; one of the studies was in vivo and the others ex vivo. Different disinfection procedures were studied, characterised mainly by the use of intracanal medication, highlighting the double antibiotic paste, triple antibiotic paste and calcium hydroxide paste. Disinfection ability was evaluated against Enterococcus faecalis and multispecies biofilms by using the fluorescence technique and colony forming unit counting, for 7 to 21 days. Double antibiotic paste and triple antibiotic paste demonstrated excellent antibiofilm activity, unlike CH paste that showed limited disinfection, even when associated with different antimicrobial agents. Triple antibiotic paste was the most effective medication against biofilm., (© 2022 Australian Society of Endodontology Inc.)

The purpose of this study was to evaluate, in vivo, the biocompatibility, biomineralization, collagen maturation and the in vitro antibacterial and cytotoxicity of resinous endodontic sealers containing calcium hydroxide. Forty rats were implanted with polyethylene tubes containing Sealer 26, Sealer Plus, Dia-ProSeal and an empty tube, examined after 7, 15, 30 and 60 days. Antimicrobial activity was evaluated against Enterococcus faecalis by Agar Diffusion Test (ADT) through inhibition zones. For cytotoxicity, undifferentiated pulp cells (OD-21) were cultured and assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, exposed to dilution of serial extracts at 6, 24, 48h. Cytotoxicity was analyzed by two-way ANOVA and Bonferroni correction. Kruskal-Wallis test followed by Dunn test was performed for nonparametric data (p<0.05). MTT assay revealed cell proliferation affected by sealers extract in all periods (p<0.0001), except for Dia-Proseal and Sealer Plus ⅛ dilution. Subcutaneous analysis showed at day 7th moderate inflammatory infiltration. After 30 days, Sealer 26 still showed moderate inflammatory infiltrate compared to mild inflammation from control and Dia-ProSeal (p = 0.006). At day 60th, all groups showed similar mild inflammatory infiltrate (p>0.05). Sealer 26 induced more biomineralization than other sealers in all periods. At 7 and 15 days, all sealers had significant percentage of immature collagen fibers. After 60 days Sealer 26 showed more mature fibers compared to other sealers (p<0.001). All sealers had a smaller zone of inhibition than chlorhexidine, but with no significant difference among any group (p>0.05). All sealers showed satisfactory biological responses with in vitro/in vivo biocompatibility and antimicrobial activity against planktonic bacteria. Sealer 26 induced more biomineralization than Sealer Plus and Dia-ProSeal., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Bueno et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objectives: To evaluate the incorporation of halloysite nanotubes (HNTs) loaded with one of two calcium sources (i.e., calcium hydroxide/CaOH 2 or beta-tricalcium phosphate/β-TCP) on the physicochemical and biological properties of an experimental resin-based dual-cured endodontic sealer., Materials and Methods: HNTs were encapsulated with CaOH 2 or β-TCP at 10 wt.%. HNTs containing CaOH 2 or β-TCP were added into the experimental sealers at 50 wt.%. The control sealers were the calcium-free HNT-modified resin-based experimental sealer and AH Plus™, a commercially available endodontic sealer. Degree of conversion, setting time, flow, film thickness, radiopacity, dimensional stability, and calcium ions release were determined. Antibiofilm properties and cytocompatibility of the formulated sealers and commercial control were also evaluated. One and two-way ANOVA analysis followed by Tukey's post hoc test was conducted to evaluate the effect of the independent variable on the evaluated properties., Results: FTIR confirmed the encapsulation of calcium sources into HNTs. Regarding flow and film thickness, the values obtained from these sealers were in accordance with the specifications provided by ISO 6876. For radiopacity, AH Plus™ achieved the highest radiopacity (p<0.05). Among the experimental formulations, all experimental HNT-containing compositions exhibited values below 3 mm Al. The experimental sealers showed greater dimensional changes when compared to the commercial (AH Plus™) control. The release of calcium ions was observed for the HNT&#95;CaOH 2 and HNT&#95;β-TCP sealers without statistical differences. Experimental sealers containing HNT&#95;CaOH 2 and HNT&#95;β-TCP significantly reduced the CFU/mL count and showed cell compatibility., Conclusions: The findings of this study demonstrate that the incorporation of HNT&#95;CaOH 2 or HNT&#95;β-TCP into resin-based experimental sealers promoted antimicrobial effects and gradual calcium release without impairing cytocompatibility or physicochemical properties of the sealers. Still, an adjustment to reach the minimal radiopacity established by ISO 6876 is needed., Clinical Relevance: The experimental resin-based sealers seemed to be an alternative for endodontics. The incorporation of calcium sources exerts promising antimicrobial effects while displaying low cell toxicity., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Context: Antimicrobial intracanal medicaments play a vital role in successful outcome of any endodontic procedure. One such plant extract Cuminium cyminium, as intracanal medicaments needs to be researched., Aims: The purpose of this study was in vitro assessment of the antibacterial activity of ethanol extract of C. Cyminium in comparison to Calcium hydroxide (Ca[OH] 2 ) as intracanal medicament against the pathogens of endodontic infection, at an interval 1 h, 24 h, 48 h, and 72 h., Settings and Design: The study was conducted in the central research laboratory of our institute. Freshly prepared C. cyminium extract was procured from AYUSH approved laboratory and direct contact test (DCT) was utilized, which is based on turbidometric determination of microbial growth in a 96-well microplate, carrying 6 times for each bacteria., Methodology: Three groups were assigned for each material in a 96 microwell plate for DCT. Bacterial growth kinetics was monitored at intervals of 1 h, 24 h, 48 h, and 72 h using spectrophotometer at 595 nm. The optical density of T2 (Test group), P2 (Positive control), and N2 (Negative control) was considered., Statistical Analysis Used: After compiling the data, based on the normality of data, further statistical analysis was conducted using Kolmogorov-Smirnov test, Paired t-test, and pairwise comparisons by Turkey's multiple post hoc procedures. The level of statistical significance was set at P = 0.05., Results: The comparison of mean optical density values of C. cyminium in comparison with Ca(OH) 2 against the microorganisms of endodontic origin showed a statically significant decrease in bacterial viability at the end of 24 h, 48 h, and 72 h., Conclusion: Based on the results of the study, it can be concluded that C. cyminium has significant antibacterial action against endodontic origin, at interval of 24 h, 48 h, and 72 h.

Background: To assess the antibacterial activity of multi-strain probiotics supernatants (MSP); Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus acidophilus as an intracanal medication on Enterococcus faecalis (E. faecalis) biofilm in a tooth model., Methods: Sixty extracted human single-rooted teeth with single canals were instrumented, sterilized, and inoculated with E. faecalis. After 21 days of incubation, four specimens were randomly selected to validate the biofilm formation by scanning electron microscope (SEM). The remaining specimens were randomly divided (n = 14), according to the intracanal medication (ICM) received into: Ca(OH) 2 : calcium hydroxide paste (35% Ultra Cal XS Ca(OH) 2 ), Probiotics supernatants: MSP in poloxamer gel vehicle Poloxamer: poloxamer gel vehicle and, Control: E. faecalis biofilm only. The tested groups were further subdivided into two equal subgroups (n = 7) according to the incubation period (24 h and 7 days). Shaved dentin chips were obtained and collected by H-files and paper points, respectively for bacterial culture. The antibacterial activity was assessed after each incubation period quantitatively and qualitatively using bacterial colony-forming units per milliliter (CFUs/ml) and SEM, respectively., Results: The lowest CFUs/ml was found in Ca (OH) 2 with a significant difference compared to other groups after 24 h. After 7 days, a similar outcome was found with a further significant reduction of CFUs/ml in all groups with no statistical difference between Ca(OH) 2 and probiotics supernatants groups. Ca (OH) 2 and Probiotics supernatants groups showed a significant (p < 0.05) percentage of overall bacterial reduction (100.00 ± 0.00% and 70.30 ± 12.95%, respectively) compared to poloxamer and control groups (27.80 ± 14.45 and 28.29 ± 19.79). SEM images showed a bacteria-free state in the Ca(OH) 2 group after 7 days while few bacteria were found in the probiotics supernatants group. An extensive invasion of bacteria was found in poloxamer and controls groups., Conclusion: MSP has a potential antibacterial effect on E. faecalis growth closely similar to the routinely used Ca (OH) 2 ., (© 2023. The Author(s).)

Objectives: Successful endodontic therapy is based on the reduction of infecting bacteria by cleaning, shaping, and disinfecting of the root canal system, thus the use of intracanal dressing is necessary for optimal success of root canal treatment. This study was designed to evaluate the effect of chitosan and propolis as intracanal medicaments against Enterococcus faecalis compared to calcium hydroxide in primary root canals., Material and Methods: Ninety-six extracted primary second molars were collected. Teeth preparation was completed to size 30 K-file. They were randomly divided into four groups; (A): chitosan, (B): propolis, (C): calcium hydroxide, and (D): control group (saline). The tooth specimens were inoculated with E. faecalis. Then, tested materials were applied for all groups in accordance to the groups each tooth belonged to. Following this, the bacterial colonies were counted after 24 h, 72 h, and 1 week of applying dressing materials and incubation. Finally, one-way analysis of variance and Fisher's least significant difference tests were used for statistical comparisons between the groups at a significance level of .05., Results: No statistically significant difference was found between groups A, B, and C for both 24 h and a week (p ≥ .05). Yet, a statistical difference between groups A, B, C, and D after 72 h and 1 week were seen (p ≤ .05)., Conclusions: Chitosan and propolis medicaments were as effective as calcium hydroxide against E. faecalis in primary root canal treatment and might be considered as an alternative dressing material between treatment sessions., (© 2023 The Authors. Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

The study aims to investigate the quality of dentin barriers and pulp reaction to EndoSequence Root Repair Material (ERRM) combined with low-level laser application. In eight dogs, pulps were exposed via class V, half of the samples received low-level diode laser at 870 nm. Thereafter, cavities were capped with fast-set or regular-set ERRM. The specimens were processed for histomorphological and immunohistochemical examination after 2 weeks and 2 months. Dentin bridges were observed in all samples, and 87.5% were complete. The low-level laser group had significantly more reparative dentin area than the non-lased group (p < 0.05). The dentin bridges were found to have an unprecedented tubularity of 43%-89%. Tiny dentin island formation was observed within the material particles. Initial mild-to-moderate inflammatory reactions were observed, which subsided after 2 months. RUNX2 and osteocalcin staining were evident for all samples at both time intervals. Low-level laser combined with bioactive ERRM is effective in inducing reparative dentin formation., (© 2022 Australian Society of Endodontology Inc.)

Aim: This study was performed to evaluate the antibacterial efficacy of two commercially available probiotics (BIFILAC and VSL 3) as intracanal medicament against Enterococcus faecalis in endodontic therapy., Materials and Methods: Microorganisms from commercially available probiotics (BIFILAC and VSL 3) were extracted via the manufacturer's recommendations and mixed by weight. About 30 microliters were then placed on sterile discs. The pathogenic test organism was E. faecalis set to a 1 McFarland standard challenge. A two-probiotic disc template on blood agar plates was inoculated with E. faecalis and incubated at 37°C for 48 hours and 1 week respectively. Phase-1 of the study was conducted by a disc diffusion assay test to evaluate zones of inhibition (ZOI) in millimeters (mm). Phase-2 was conducted by mixing 9 mL of 30% poloxamer 407 and MRS broth in a test tube, together with the two probiotic mixtures and E. faecalis , set at a 2 McFarland standard. Serial dilutions up to 108 were done and the mixture was placed inside root canals and incubated at 37ºC for 36 hours and evaluated for colony-forming unit (CFU)/mL counts., Results: The results of phase-1 showed that probiotics Lactobacillus rhamnosus and Bifidobacterium species are effective in fighting against E. faecalis with the acceptable zone of inhibition. The results of phase-2 showed that both the probiotics are effective against E. faecalis with a reduction in the number of CFU after probiotic usage., Conclusion: Commercially available probiotics can be used effectively as an intracanal medicament to fight against E. faecalis , Poloxamer 407 is a promising vehicle for delivering probiotics inside the root canal system. Further in vitro and in vivo studies are needed to determine the full potential of "Bacteriotherapy" with an application of probiotics., Clinical Significance: If probiotics are proved to be an effective intracanal medicament against E.faecalis they can be used as an alternative to calcium hydroxide as intracanal medicament with no side effects to the host.

Objective: To construct a model of Enterococcus faecalis ( E. faecalis ) infection in dentinal tubules by gradient centrifugation and to evaluate the antibacterial effect of low-temperature plasma on E. faecalis in dentinal tubules., Methods: Standard dentin blocks of 4 mm×4 mm×2 mm size were prepared from single root canal isolated teeth without caries, placed in the E. faecalis bacterial solution, centrifuged in gradient and incubated for 24 h to establish the model of dentinal tubule infection with E. faecalis . The twenty dentin blocks of were divided into five groups, low-temperature plasma jet treatment for 0, 5 and 10 min, calcium hydroxide paste sealing for 7 d and 2% chlorhexidine gel sealing for 7 d. Scanning electron microscopy and confocal laser scanning microscope were used to assess the infection in the dentinal tubules and the antibacterial effect of low-temperature plasma., Results: The results of scanning electron microscopy and confocal laser scanning microscopy showed that after 24 h of incubation by gradient centrifugation, E. faecalis could fully enter the dentinal tubules to a depth of more than 600μm indicating that this method was time-saving and efficient and could successfully construct a model of E. faecalis infection in dentinal tubules. Low-temperature plasma could enter the dentinal tubules and play a role, the structure of E. faecalis was still intact after 5 min of low-temperature plasma treatment, with no obvious damage, and after 10 min of low-temperature plasma treatment, the surface morphology of E. faecalis was crumpled and deformed, the cell wall was seriously collapsed, and the normal physiological morphology was damaged indicating that the majority of E. faecalis was killed in the dentinal tubules. The antibacterial effect of low-temperature plasma treatment for 10 min exceeded that of the calcium hydroxide paste sealing for 7 d and the 2% chlorhexidine gel sealing for 7 d. These two chemicals had difficulty entering deep into the dentinal tubules, and therefore only had a few of antibacterial effect on the bacterial biofilm on the root canal wall, and there was also no significant damage to the E. faecalis bacterial structure., Conclusion: Gradient centrifugation could establish the model of E. faecalis dentin infection successfully. Low-temperature plasma treatment for 10 min could kill E. faecalis in dentinal tubules effectively, which is superior to the calcium hydroxide paste sealing for 7 d and the 2% chlorhexidine gel sealing for 7 d.

This study evaluated the bioactive potential of a macro-porous chitosan scaffold incorporated with calcium hydroxide (CH-Ca) and functionalized with bioactive doses of simvastatin (SV) for bone tissue regeneration. Initially, the bioactive dose of SV in osteoblastic cells (SAOS-2) was determined. For the direct contact experiment, SAOS-2 cells were plated on scaffolds to assess cell viability and osteogenic differentiation. The second assay was performed at a distance using extracts from scaffolds incubated in culture medium to assess the effect of conditioned medium on viability and osteogenic differentiation. The initial screening showed that 1 μM SV presented the best biostimulating effects, and this dose was selected for incorporation into the CH-Ca and pure chitosan (CH) scaffolds. The cells remained viable throughout the direct contact experiment, with the greatest cell density in the CH-Ca and CH-Ca-SV scaffolds because of their higher porosity. The CH-Ca-SV scaffold showed the most intense bio-stimulating effect in assays in the presence and absence of osteogenic medium, leading to an increased deposition of mineralized matrix. There was an increase in the viability of cells exposed to the extracts for CH-Ca, CH-SV, and CH-Ca-SV during the one-day period. There was an increase in ALP activity in the CH-Ca and CH-Ca-SV; however, the CH-Ca-SV scaffold resulted in an intense increase in the deposition of mineralized nodules, approximately 56.4% at 7 days and 117% at 14 days, compared with CH (control). In conclusion, functionalization of the CH-Ca scaffold with SV promoted an increase in bioactivity, presenting a promising option for bone tissue regeneration.

Objectives: This RCT investigated the impact of N-acetylcysteine (NAC) and calcium hydroxide [Ca(OH) 2 ] intracanal medications (ICMs) in primary endodontic infection with apical periodontitis (PEIAP)., Materials and Methods: Thirty-six teeth with PEIAP were randomly divided into groups according to the ICM: NAC, Ca(OH) 2  + saline solution (SSL), and Ca(OH) 2  + 2% chlorhexidine-gel (2% CHX-gel) (all, n = 12). Root canal samples (RCSs) were collected before (s1) and after instrumentation (s2) and after 14 days of ICM (s3). Chemomechanical preparation (CMP) was performed with a Reciproc file and 2.5% NaOCl. Checkerboard DNA-DNA hybridization was used to assess 40 target bacteria species., Results: At s1, bacterial DNA was detected in 100% of RCSs (36/36). All 40 bacterial species were found in PEIAP. The mean number of species per RCS was 17.92 ± 13.18. The most frequent bacteria were S. mitis (65%), E. nodatum (63%), E. faecalis (63%), F. nucl sp vicentii (58%), T. forsythia (58%), and F. periodonticum (56%). CMP reduced the mean number of species per RCS to 6.8 ± 2.36 (p < 0.05). At s3, the intragroup analysis revealed a broader antimicrobial activity for Ca (OH) 2  + 2% CHX-gel and NAC than Ca(OH) 2  + SSL (p < 0.05). NAC eliminated 8/12 bacteria species resistant to both Ca (OH) 2 ICMs, including P. micra, P. nigrescens, T. denticola, A. israelii, P. endodontalis, P. acnes, C. ochracea, and E. corrodens., Conclusions: Ca (OH)2 + 2% chlorhexidine gel (2% CHX gel) showed a greater bacterial elimination over the number of bacterial species; however, NAC eliminated 8/12 bacteria species resistant to both Ca (OH)2 ICMs (RBR-3xbnnn)., Clinical Relevance: The use of intracanal medication with a broad antimicrobial activity can optimize root canal disinfection. Ca(OH) 2  + 2% CHX gel and NAC showed a broader antimicrobial activity than Ca(OH) 2  + SSL against endodontic pathogens in primary root canal infection., Trial Registration: Brazilian Registry of Clinical Trials (REBEC), No. RBR-3xbnnn., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)