Comparative bioactivity and immunomodulatory potential of the new Bioroot Flow and AH Plus Bioceramic sealer: An in vitro study on hPDLSCs.

OBJECTIVES: To evaluate the cytocompatibility, bioactivity, and anti-inflammatory potential of the new pre-mixed calcium silicate cement-based sealers Bioroot Flow (BrF) and AH Plus Bioceramic Sealer (AHPbcs) on human periodontal ligament stem cells (hPDLSCs) compared to the epoxy resin-based sealer AH Plus (AHP). MATERIALS AND METHODS: Standardized discs and 1:1, 1:2, and 1:4 eluates of BrF, AHPbcs and AHP after setting were prepared. The following assays were performed: cell attachment and morphology via SEM, cell viability via a MTT assay, cell migration/proliferation via a wound-healing assay, cytoskeleton organization via immunofluorescence staining; cytokine release via ELISA; osteo/cemento/odontogenic marker expression via RT-qPCR, and cell mineralized nodule formation via Alizarin Red S staining. HPDLSCs were isolated from extracted third molars from healthy patients. Comparisons were made with hPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media. Statistical significance was established at p < 0.05. RESULTS: Both BrF and AHPbcs showed significantly positive results in the cytocompatibility assays (cell metabolic activity, migration, attachment, morphology, and cytoskeleton organization) compared with a negative control group, while AHP showed significant negative results. BrF exhibited an upregulation of at least one osteo/cementogenic marker compared to the negative and positive control groups. BrF showed a significantly higher calcified nodule formation than AHPbcs, the negative and positive control groups, while AHPbcs was higher than the negative control group. Both were also significantly higher than AHP group. CONCLUSION: BrF and AHPbcs exhibit adequate and comparable cytocompatibility on hPDLSCs. BrF also promoted the osteo/cementogenic differentiation of hPDLSCs. Both calcium silicate-based sealers favored the downregulation of the inflammatory cytokine IL-6 and the calcified nodule formation from hPDLSCs. BrF exerted a significantly higher influence on cell mineralization than AHPbcs. CLINICAL RELEVANCE: This is the first study to elucidate the biological properties and immunomodulatory potential of Bioroot Flow and AH Plus Bioceramic Sealer. The results act as supporting evidence for their use in root canal treatment.

Characterisation and evaluation of physical properties of AH-Plus sealer with and without the incorporation of petasin, pachymic acid, curcumin and shilajit-an invitro study.

BACKGROUND: AH Plus, an epoxy resin-based sealer, is widely used in endodontic practice, owing to its good physical properties that confers longstanding dimensional stability and good adhesion to dentin. Nevertheless, its propensity to trigger inflammation, especially in its freshly mixed state, has been extensively documented. Phytochemicals such as Petasin, Pachymic acid, Curcumin, and Shilajit are known for their anti-inflammatory and analgesic effects. This study aimed to analyze and determine the effect of these natural products on the physical properties of AH Plus sealer when incorporated with the sealer. METHODS: AH Plus (AHR) sealer was mixed with 10% petasin, 0.75% pachymic, 0.5% and 6%shilajit to obtain AHP, AHA, AHC and AHS in the ratio of 10:1 and 5:1 respectively. Five samples of each material were assessed for setting time, solubility, flow, and dimensional stability in accordance with the ISO 6876:2012 standardization. Sealers were characterized through scanning electron microscopy (SEM), X-ray energy dispersive spectroscopy, X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Statistical evaluation involved the Kolmogorov-Smirnov and Shapiro-Wilks tests for normality and the one-way ANOVA test for analysis. RESULTS: In this investigation, the characterisation analysis revealed a relatively similar microstructure in all the experimental root canal sealers. All experimental groups, excluding the control group, exhibited an increase in flow ranging from 11.9 to 31.4% at a 10:1 ratio. Similarly, for the 5:1 ratio, the increase ranged from 12.02 to 31.83%. In terms of dimensional stability, all groups at the 10:1 ratio showed a decrease compared to the control group. The addition of natural agents to AHR in 10:1 ratio led to a reduction in setting time by 8.9-31.6%, and at a 5:1 ratio, the reduction ranged from 8.1 to 31.5%. However, regarding solubility, the addition of natural agents did not induce any significant alterations. CONCLUSION: Based on the results of this study, it can be concluded that all tested root canal sealers exhibited properties that met the acceptable criteria outlined in the ISO 6876:2012 standardization.

The Effect of Adding Various Antibiotics on the Push-out Bond Strength of a Resin-based Sealer: An In Vitro Study.

AIM: This study aimed to compare the bond strength of AH Plus sealer to root canal dentin when used with or without various antibiotics including amoxicillin, clindamycin, and triple antibiotic mixture (TAM). MATERIALS AND METHODS: A total of 80 single-rooted extracted human teeth were instrumented and obturated with gutta-percha and four different sealer-antibiotic combinations (n = 20). Group I: AH Plus without any antibiotics, Group II: AH Plus with amoxicillin, Group III: AH Plus with clindamycin, and Group IV: AH Plus with TAM. After seven days, the roots were sectioned perpendicular to their long axis and 1 mm thick slices were obtained from the midroots. The specimens were subjected to a push-out bond strength test and failure modes were also evaluated. Data was analyzed using Kruskal-Wallis and Dunn's post hoc tests. RESULTS: Group IV had significantly higher bond strength compared to other groups (p ≤ 0.05). No significant differences were found between other groups. While the sealer-antibiotic groups predominantly showed cohesive failure modes, the control group displayed both cohesive and mixed failure modes. CONCLUSION: Within the limitations of this study, the addition of TAM increased the push-out bond strength of AH Plus. CLINICAL SIGNIFICANCE: Amoxicillin, clindamycin, or TAM can be added to AH Plus for increased antibacterial efficacy without concern about their effects on the bond strength of the sealer. How to cite this article: Adl A, Shojaei NS, Ranjbar N. The Effect of Adding Various Antibiotics on the Push-out Bond Strength of a Resin-based Sealer: An In Vitro Study. J Contemp Dent Pract 2024;25(3):231-235.

Influence of long -term thermal cycling and masticatory loading simulation on bond strength of roots filled with epoxy resin and calcium silicate based sealers.

BACKGROUND: The aim of this study was to evaluate the effect of thermal and mechanical cyclic aging using a mastication simulator on push-out bond strength of mandibular premolars obturated with AH Plus and BioRoot RCS root canal sealers. METHODS: With REVO-S files up to SU/0.06 taper, 48 single-rooted premolar teeth were instrumented. The teeth were randomly divided into two main groups (n = 24) based on the two root canal sealers used (AH Plus and BioRoot RCS). All teeth were obturated with h matched-taper single-cone. Each main group was then subdivided into three subgroups (A, B, and C) (n = 8). Group A served as the negative control group (no-thermocycling aging). While groups B and C were subjected to thermal changes in a thermocycler machine (15,000 and 30,000 thermal cycles, respectively), followed by two different dynamic loading periods, 3 × 105 and 6 × 105 in a masticatory simulator with a nominal load of 5 kg at 1.2 Hz which represent roughly 1½ and 3 years of clinical function respectively. 2 mm slice at 3 levels, apical, middle, and coronal, to obtain 3 sections were prepared and subjected to push-out test using a universal testing machine. Statistical analysis was performed using analysis of variance (ANOVA) followed by a Tukey post hoc comparisons test and an independent T-test. A significance level of 5% was used. RESULTS: After thermal-mechanical cyclic aging, the two root canal sealers showed a significantly decreased in push-out bond strength (p < 0.05), however, AH Plus had significantly higher bond strength values than BioRoot RCS after cycling aging. CONCLUSIONS: It could be concluded that thermal-mechanical cyclic aging had a significant impact on the outcome of the dislodgment resistance of AH Plus and BioRoot RCS.

Biocompatibility of three different root canal sealers, experimental study.

OBJECTIVES: This study was assessed the biocompatibility of three different root canal sealers (Well-Root St, GuttaFlow Bioseal, and AH-Plus) following implantation in rat subcutaneous tissues, using histopathological immunohistochemical analysis. METHODS: Four groups of eighty-four male rats each underwent subcutaneous dorsal implantation of a polyethylene tube, either empty or filled. Tissues were collected, fixed, and processed for histological analysis after 7, 15, and 30 d. Slides were photographed and digitally processed to identify lymphocytes and macrophages using Cluster of differentiation 3 (CD3) and cluster of differentiation 68 (CD68) markers, respectively. P was set at 0.05, when lymphocyte and macrophage infiltration was compared between groups and observation times using one-way analysis of variance (ANOVA). RESULTS: Histopathological analysis of all groups revealed an inflammatory reaction followed by the emergence of a fibrous capsule after 7 days. After 30 days, the thickness of the fibrous capsule and the inflammatory response subsided. CD3 staining for immunohistochemical analysis revealed that the AH-Plus group had the highest mean percentage of lymphocyte infiltration at 7 and 15 days, followed by the Well-Root St, GuttaFlow Bioseal, and Control groups. After 30 days, no discernible difference was observed between the groups in terms of the mean percentage of lymphocyte infiltration. After 7, 15, and 30 days, there was a significant difference in the mean percentage of macrophage infiltration across the groups, as demonstrated by CD68 staining. After 7, 15, and 30 days, the AH-Plus group had the highest mean percentage of macrophage infiltration, followed by the Well-Root St. and GuttaFlow Bioseal groups, while the control group had the lowest mean percentage. CONCLUSION: All observational periods showed minimal inflammatory reactions to GuttaFlow Bioseal. After subcutaneous tissue implantation in a rat model, the initial inflammatory reactions to Well-Root St and AH-Plus had abated by day 30, and all tested sealers had outstanding biocompatibility.

Cytocompatibility and bioactive potential of AH Plus Bioceramic Sealer: An in vitro study.

AIM: To assess the cytocompatibility and bioactive potential of the new calcium silicate cement-based sealer AH Plus Bioceramic Sealer (AHPbcs) on human periodontal ligament stem cells (hPDLSCs) compared with the epoxy resin-based sealer AH Plus (AHP) and the calcium silicate cement-based sealer Endosequence BC Sealer (ESbcs). METHODOLOGY: Standardized sample discs and 1:1, 1:2 and 1:4 eluates of the tested materials were prepared. The following assays were performed: surface element distribution via SEM-EDX, cell attachment and morphology via SEM, cell viability via a MTT assay, cell migration/proliferation via a wound-healing assay, osteo/cemento/odontogenic marker expression via RT-qPCR and cell mineralized nodule formation via Alizarin Red S staining. HPDLSCs were isolated from extracted third molars. Comparisons were made with hPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media. Statistical significance was established at p < .05. RESULTS: A higher peak of Ca2 + was detected from ESbcs compared with AHPbcs and AHP in SEM-EDX. Both AHPbcs and ESbcs showed significantly positive results in the cytocompatibility assays (cell viability, migration/proliferation, attachment and morphology) compared with a negative control group, whilst AHP showed significant negative results. Both AHPbcs and ESbcs exhibited an upregulation of at least one osteo/odonto/cementogenic marker compared with the negative and positive control groups. Both ESbcs and AHPbcs showed a significantly higher calcified nodule formation than the negative and positive control groups, indicative of their biomineralization potential and were also significantly higher than AHP group. CONCLUSION: AH Plus Bioceramic Sealer exhibited a significantly higher cytocompatibility and bioactive potential than AH Plus and a similar cytocompatibility to that of Endosequence BC Sealer. Endosequence BC Sealer exhibited a significantly higher mineralization potential than the other tested sealers. The results from this in vitro study act as supporting evidence for the use of AH Plus Bioceramic Sealer in root canal treatment.

Comparison of the effect of post space preparation time on the apical seal of two different sealers.

BACKGROUND: The present study compared the effect of post space preparation time on the apical seal of two different sealers. METHODS: In the in vitro study, 94 central incisors were used. After the samples' root canal preparation, they were randomly assigned to four experimental groups (n = 21). The samples in groups 1 and 2 were obturated with AH Plus sealer, gutta-percha, and in groups 3 and 4 with Endoseal MTA bioceramic sealer and single cone technique. The post spaces in groups 1 and 3 were prepared immediately and in groups 2 and 4 with a delay. The samples were evaluated at 7-, 30-, and 90-day intervals for apical microleakage using the fluid filtration technique. The data were analyzed with SPSS 25, using three-way ANOVA and independent t-test. RESULTS: The apical microleakage in groups 3 and 4, obturated with Endoseal MTA bioceramic sealer and prepared immediately and after a delay, respectively, was not significantly different between the interval times. In group 2, obturated with AH Plus sealer and prepared for post space with a delay, the apical microleakage was significantly less than all the other groups. Group 1, obturated with AH Plus sealer and prepared for post space immediately, exhibited the least microleakage after seven days, but its microleakage increased over time to reach the level of groups 3 and 4. CONCLUSION: According to the results, the apical microleakage in the AH + sealer group and the delayed post-space preparation method, was significantly less than all the other groups over time.

Evaluation of the Bond Strength of Epoxy Resin-based Sealer Following Different Calcium Hydroxide Paste Removal Methods in Oval-shaped Root Canal.

AIM: The study aimed to investigate the bond strength of epoxy resin-based sealer following five different calcium hydroxide paste removal methods in oval-shaped root canals. MATERIALS AND METHODS: Sixty single-rooted human mandibular premolars having an oval-shaped root canal as evidenced by radiographs were decoronated and instrumented to size 40/0.04, medicated with calcium hydroxide paste for 7 days, before being randomly assigned to the conventional needle irrigation (CNI), manual dynamic agitation (MDA), sonic irrigation (SI), passive ultrasonic irrigation (PUI), and XP-endo Finisher (XP) groups to be irrigated using different irrigation systems. All specimens were then obturated using an epoxy resin-based root canal sealer and warm vertical compaction obturation technique. After 7 days, each specimen was sectioned into 1 mm root slices at the coronal, middle, and apical third of the root canal and tested for the push-out bond strength using a universal testing machine. RESULTS: The XP group was the only group that had comparable bond strength to the control group at every level of the root canal following removal of calcium hydroxide medicament (p >0.05). The control group had higher bond strength than the CNI group at every level of the root canal (p < 0.05). CONCLUSION: Calcium hydroxide interfered with the bonding of epoxy resin-based sealer to root canal walls. Irrigation with the XP increased the bonding sealer at every level of the root canal. CLINICAL SIGNIFICANCE: The XP was efficacious as a final rinse agitation technique to promote the bonding of the epoxy resin-based sealer at every level of the root canal following calcium hydroxide medication.

The presence of smear layer affects the antimicrobial action of root canal sealers.

AIM: To assess the chemical and microstructural characteristics of dentine after the use of two irrigation protocols and correlate this with the antimicrobial properties of hydraulic calcium silicate cement (HCSC) sealers and changes to the dentine structure/chemistry after sealer placement. METHODOLOGY: Two irrigation protocols - Protocol A using 2% NaOCl used 5 mL/5 min and Protocol B with 2% NaOCl (5 mL/5 min) followed by 17% EDTA (5 mL/3 min) - were used to prepare dentine. The chemical and microstructural changes following irrigation were assessed by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and Fourier transform infrared (FT-IR) spectroscopy (n = 5) on dentine obtained from the mid-root and coronal parts of extracted human teeth. Four sealers (AH Plus, BioRoot, MTA Fillapex, TotalFill) were characterized by SEM/EDS (n = 3). The ability of the sealers to eradicate intratubular Enterococcus faecalis biofilms was assessed by live/dead dye and confocal laser scanning microscopy to measure the percentage of living cells. The effect of combined irrigation and root filling on dentine was assessed by SEM and EDS analysis (n = 5). Statistical analysis was undertaken using one-way anova and a number of post hoc tests to detect intergroup differences. The F-test was used for comparison of variances in the microbiology testing. RESULTS: The use of NaOCl alone left the smear layer intact, with traces of chlorine remaining on dentine. The use of BioRoot sealer restored the calcium levels of dentine which had been depleted by the irrigation with EDTA. BioRoot exhibited antimicrobial properties against intratubular bacteria even in the presence of smear layer (Protocol A). Smear layer removal improved the bactericidal effect of all sealers and Ca2+ leaching. The use of a chelating agent was important for the intratubular sealer penetration for AH Plus but not the other sealers. CONCLUSION: The removal of smear layer was necessary for penetration of AH Plus into the dentinal tubules. BioRoot was a more effective sealer in reducing the bacterial load in the dentinal tubules than the other materials tested and the presence of smear layer did not affect its activity.

Sealer penetration: effect of separated file's cross-section, taper and motion characteristics.

OBJECTIVES: The separated root canal instruments may affect the quality of root canal filling, hence the success of endodontic treatment. The aim of this study was to evaluate the effects of separated file fragments of nickel-titanium rotary systems with different cross-section, taper and motion characteristics on the apical sealer penetration in oval-shaped root canals via confocal laser scanning microscope (CLSM). MATERIALS AND METHODS: Distal roots of 60 mandibular molars with oval-shaped root canals were randomly divided into 4 groups as follows: group 1, FlexMaster Nickel Titanium Rotary File System (NTRFS) (separated instrument: 30/.06); group 2, ProTaper Next NTRFS (X3); group 3, ProTaper Universal NTRFS (F3); group 4, Revo-S NTRFS (AS30/.06). Root canals were filled with gutta-percha and AH plus labelled with 0.1% rhodamine B using a warm vertical compaction technique. Each specimen was horizontally sectioned at 1st, 3rd and 5th mm from apical foramen. Amount of maximum and average penetration depths, penetration percentage and sealer penetrated area were measured and analysed with one-way repeated measures of ANOVA and the Bonferroni post hoc tests. p < 0.05 was considered significant. RESULTS: The penetration depth, percentage and penetrated area of the sealer increased from apical to coronal in all systems. The maximum and average penetration depths and penetration areas were higher in FlexMaster and Revo-S groups at the 3rd mm (p < 0.05). At the 5th mm, the Revo-S group had a higher penetration percentage, when compared with ProTaper Next and ProTaper Universal groups (p < 0.05). CONCLUSIONS: In the 1st mm, separated fragments of any system did not allow the penetration of the sealer, while it was observed that the files with constant taper showed more positive results in terms of sealer penetration at apical 3rd and 5th mm. CLINICAL RELEVANCE: In the presence of a separated file, the taper of the file might significantly affect the amount of penetrated sealer into the dentinal tubules as compared with the cross-section and motion characteristics of the file.