Biocompatibility, bioactivity and immunomodulatory properties of three calcium silicate-based sealers: an in vitro study on hPDLSCs.

OBJECTIVES: To assess the biocompatibility, bioactivity, and immunomodulatory properties of three new calcium silicate cement-based sealers: Ceraseal (CS), Totalfill BC Sealer (TFbc) and WellRoot ST (WR-ST) on human periodontal ligament stem cells (hPDLSCs). MATERIALS AND METHODS: HPDLSCs were isolated from extracted third molars from healthy patients. Eluates (1:1, 1:2, and 1:4 ratio) and sample discs of CS, TFbc and WR-ST after setting were prepared. A series of assays were performed: cell characterization, cell metabolic activity (MTT assay) cell attachment and morphology (SEM assay), cell migration (wound-healing assay), cytoskeleton organization (phaloidin-based assay); IL-6 and IL-8 release (ELISA); differentiation marker expression (RT-qPCR assay), and cell mineralization (Alizarin Red S staining). HPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media were used as a comparison. Statistical significance was established at p < 0.05. RESULTS: All the tested sealers exhibited similar results in the cytocompatibility assays (cell metabolic activity, migration, attachment, morphology, and cytoskeleton organization) compared with a negative control group. CS and TFbc exhibited an upregulation of at least one osteo/cementogenic marker compared to the negative and positive control groups. CS and TFbc also showed a significantly higher calcified nodule formation than the negative and positive control groups. Both the marker expression and calcified nodule formation were significantly higher in CS-treated cells than TFbc treated cells. WR-ST exhibited similar results to the control group. CS and TFbc-treated cells exhibited a significant downregulation of IL-6 after 72 h of culture compared to the negative control group (p < 0.05). CONCLUSION: All the tested sealers exhibited an adequate cytocompatibility. CS significantly enhances cell differentiation by upregulating the expression of key genes associated with bone and cementum formation. Additionally, CS was observed to facilitate the mineralization of the extracellular matrix effectively. In contrast, the effects of TFbc and WR-ST on these processes were less pronounced compared to CS. Furthermore, both CS and TFbc exhibited an anti-inflammatory potential, contributing to their potential therapeutic benefits in regenerative endodontics. CLINICAL RELEVANCE: This is the first study to compare the biological properties and immunomodulatory potential of Ceraseal, Totalfill BC Sealer, and WellRoot ST. The results act as supporting evidence for their use in root canal treatment.

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.

Prognosis in home dental bleaching: a systematic review.

OBJECTIVES: The aim of this systematic review was to evaluate the prognosis of at-home dental bleaching using low concentration bleaching products. MATERIALS AND METHODS: This review was conducted was performed following the recommendations of the 2020 PRISMA statement and was registered in the International Prospective Register of Systematic Reviews (PROSPERO-CRD42022360530). The PICO question was "What is the prognosis of home teeth whitening treatment?". An advanced electronic search was made in three databases: PubMed, Web of Science, and Embase. RESULTS: The database search led to the retrieval of 225 articles. After elimination of duplicate references, the titles and abstracts of the articles were analyzed with respect to the eligibility criteria, and 24 studies were included for the development of the systematic review. CONCLUSIONS: Most authors state that the color remains stable between 1 and 2.5 years regardless of the type of bleaching agent or the forms of administration, and color stability in cases of severe discolorations presents a higher degree of recurrence. CLINICAL RELEVANCE: Given the growing demand for dental cosmetic treatments, the following systematic review may aid the clinician's continuing education and evidence-based practice by providing knowledge on the field of at-home dental bleaching agents and their long-term effects.

Biological properties of Ceraputty as a retrograde filling material: an in vitro study on hPDLSCs.

OBJECTIVES: To assess the cytocompatibility and bioactive potential of the new calcium silicate-based cement Ceraputty on human periodontal ligament stem cells (hPDLSCs) compared to Biodentine and Endosequence BC root repair material (ERRM). MATERIALS AND METHODS: hPDLSCs were isolated from extracted third molars from healthy donors. 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, osteo/cemento/odontogenic marker expression via RT-qPCR, and cell calcified nodule formation via Alizarin Red S staining. hPDLSCs cultured in unconditioned or osteogenic media were used as negative and positive control groups, respectively. Statistical analysis was performed using one-way ANOVA or two-way ANOVA and Tukey's post hoc test. Statistical significance was established at p < 0.05. RESULTS: The highest Ca2+ peak was detected from Biodentine samples, followed by ERRM and Ceraputty. hPDLSC viability was significantly reduced in Ceraputty samples (p < 0.001), while 1:2 and 1:4 Biodentine and ERRM samples similar results to that of the negative control (p > 0.05). Biodentine and ERRM exhibited an upregulation of at least one cemento/odonto/osteogenic marker compared to the negative and positive control groups. Cells cultured with Biodentine produced a significantly higher calcified nodule formation than ERRM and Ceraputty (p < 0.001), which were also higher than the control groups (p < 0.001). CONCLUSION: Ceraputty evidenced a reduced cytocompatibility towards hPDLSCs on its lowest dilutions compared to the other tested cements and the control group. Biodentine and ERRM promoted a significantly higher mineralization and osteo/cementogenic marker expression on hPDLSCs compared with Ceraputty. Further studies are necessary to verify the biological properties of this new material and its adequacy as a retrograde filling material. CLINICAL RELEVANCE: This is the first study to elucidate the adequate biological properties of Ceraputty for its use as a retrograde filling material.

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.

Scientific production on silicate-based endodontic materials: evolution and current state: a bibliometric analysis.

OBJECTIVES: To perform a bibliometric analysis on silicate-based biomaterials in endodontics; to elucidate the evolution and distribution of scientific production regarding research on these biomaterials, the authors and institutions involved, and the most used descriptors/keywords in this field. MATERIALS AND METHODS: A general advanced broad search was performed in Web of Science Core Collection, using the terms "Silicate" and "Endod*." By means of the "Analyze Results" and "Citation Analysis" tools from Web of Science, bibliometric data were extracted. Bibliometric networks on co-authorship and keyword co-occurrence were illustrated using VOSviewer software tool. RESULTS: Research in the field of silicate-based endodontic materials in endodontics has followed an exponential crescent tendency over the period between 1995 and 2020. Two major co-authorship networks lead the research production in the field, headed by Gandolfi MG and Prati C, and Rodríguez-Lozano FJ and Forner L; respectively. "Mineral trioxide aggregate," "MTA", "Biocompatibility," "Biodentine," and "In vitro" were the terms with the highest occurrence. The modal study type in this research area are laboratory studies, representing 77.7% of the total number of records. The relationship between keywords illustrates the recent tendency to assess biological properties of MTA and Biodentine, which could be extrapolated to other silicate-based materials. CONCLUSIONS: The results from the present study highlight the exponential increase and distribution of scientific production on silicate-based materials in endodontics, along with the modal keywords used as descriptors and the major authorship networks involved. The predominance of laboratory studies in this research field reinforces the need for clinical studies, to extrapolate the reported performance and characteristics of silicate-based materials into the clinical setting. CLINICAL RELEVANCE: The present study may aid the clinician's continuing education and evidence-based practice by providing knowledge and facilitating the literature search on the field of silicate-based biomaterials in endodontics.

Comparative cytocompatibility of the new calcium silicate-based cement NeoPutty versus NeoMTA Plus and MTA on human dental pulp cells: an in vitro study.

OBJECTIVES: The aim of the present in vitro study is to determine the cytocompatibility of the recently introduced NeoPutty in contact with human dental pulp cells compared with its precursor NeoMTA Plus and the classic gold standard MTA Angelus. MATERIALS AND METHODS: Sample disks were obtained for each of the tested materials (5 mm diameter; 2 mm thickness; n = 30), along with 1:1, 1:2, and 1:4 material eluents. HDPCs were extracted and cultured with the tested materials (test groups) or in unconditioned medium (control group), and the following biocompatibility assays were performed: MTT assay, scratch wound assay, cell cytoskeleton staining assays, and cell attachment assessment via SEM. Additionally, material ion release and surface element composition were evaluated via ICP-MS and SEM-EDX, respectively. Each experimental condition was carried out three times and assessed in three independent experiments. Statistical significance was established at p < 0.05. RESULTS: 1:2 dilutions of all the tested materials exhibited a comparable cell viability to that of the control group at 48 and 72 h of culture (p < 0.05). The same was observed for 1:4 dilutions of the tested materials at 24, 48, and 72 h of culture (p > 0.05). All the tested materials exhibited adequate cytocompatibility in the remaining biocompatibility assays. MTA exhibited a significantly higher calcium ion release compared to NeoPutty and NeoMTA Plus (p < 0.05). CONCLUSION: The results from the present work elucidate the adequate cytocompatibility of NeoPutty, NeoMTA Plus, and MTA Angelus towards human dental pulp cells. CLINICAL RELEVANCE: Within the limitations of the present in vitro study, our results may act as preliminary evidence for its use in vital pulp therapy as a pulp capper. However, results need to be interpreted with caution until further clinical supporting evidence is reported.

Bleaching efficacy of at home 16% carbamide peroxide. A long-term clinical follow-up study.

OBJECTIVE: To assess the whitening efficacy of a 16% carbamide peroxidase (CP) gel after 42 months of clinical follow-up. MATERIALS AND METHODS: A retrospective clinical study was carried out involving 95 participants. CP 16% was applied for 90 minutes a day for 4 weeks using individualized trays. Tooth color was assessed using spectrophotometry at baseline, 1 week after the end of treatment, and every 6 months until completing 42 months of follow-up. Color parameters were compared using analysis of variance for repeated measures. The analysis by teeth groups was performed using two-way ANOVA. RESULTS: The mean baseline L* was 81.7 ± 5.2 versus 83.6 ± 8.8 after 42 months (P<.01). L* showed a decrease of 2.3 after 30 months. The values corresponding to a* before treatment were 0.4 ± 1.1 versus -1.0 ± 1.0 after 42 months (P<.01). The initial values corresponding to b* were 23.4 ± 3.3 versus 15.8 ± 1.3 after 42 months (P<.01). Values of ΔE00 and ΔE, at the end of the treatment were 4.6 ± 1.2 and 7.8 ± 1.8, respectively. Teeth groups remained stable after 42 months of follow-up. CONCLUSIONS: The home application of 16% CP gel for 90 minutes a day for 4 weeks using individualized trays resulted in whitening that remained stable over the 42 months of follow-up. CLINICAL SIGNIFICANCE: Treatment with 16% CP is an effective and safe tooth whitening procedure, and the color obtained remains stable over the long term.

Biological effects of acid-eroded MTA Repair HP and ProRoot MTA on human periodontal ligament stem cells.

OBJECTIVE: The aim of this study was to analyze the biological effects of MTA Repair HP and ProRoot MTA on human periodontal ligament stem cells (hPDLSCs) after exposure to acidic and neutral environments. MATERIALS AND METHODS: Discs of each material (n = 30) were exposed to phosphate buffered saline (pH = 7.4) or butyric acid (pH = 5.2) for 7 days, and biological testing was carried out in vitro on hPDLSCs. Cell viability and apoptosis assays were performed using eluates of each root-end filling material. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. The chemical composition of the root-end filling materials was determined by energy-dispersive x-ray and eluates were analyzed by inductively coupled plasma-mass spectrometry. Statistical differences were assessed by ANOVA and Tukey test (p < 0.05). RESULTS: Under an acidic environment, both materials displayed similar ion release abilities, with the increased release of Si and Ca ions. Substantial changes in microstructure were observed for both materials after exposure to acidic pH. In addition, material exposure to an acidic environment showed a similar degree of cell adherence, and, surprisingly, MTA Repair HP exhibited higher cell viability rates at pH 5.2 than ProRoot MTA. CONCLUSIONS: Exposure to an acidic environment promoted Si and Ca ion release from ProRoot MTA and MTA Repair HP. Moreover, we observed optimal biological properties of ProRoot MTA and MTA Repair HP in terms of cell viability, cell death, and cell attachment in both environments. CLINICAL RELEVANCE: These results may suggest that MTA Repair HP and ProRoot exhibited optimal biological properties in terms of cell viability, cell death and cell attachment in acidic environment, being considered as materials for root-end filling and perforations.

Color stability of nanohybrid resin-based composites, ormocers and compomers.

OBJECTIVE: The aim of this study is to evaluate the color stability of two nanohybrid resin-based composites, two organic modified ceramic resin composites (ormocers) and a compomer, following their immersion during 4 weeks in four usual drinks. MATERIAL AND METHODS: Forty discs of each of the following materials were prepared: GrandioSO (GR), Esthet X (EX), Dyract EXTRA (DY), Ceram X duo (CX), and Admira Fusion (AD). The discs were polished and stored in distilled water during 24 h at 37 °C. Color according to the CIEL*a*b* scale was recoded. Ten randomly selected samples of each material were immersed in red wine (RW), coffee (CF), cola (CK), and distilled water (DW). The color was recorded after 1, 7, 21 and 28 days of immersion. The ΔE values between the baseline color and each of the study timepoints were calculated. The data were analyzed based on the Kruskal-Wallis test, Wilcoxon test, and Mann-Whitney U test. RESULTS: After 1 day of immersion, all the materials showed ΔE > 3.3, except EX, CF, and CX in CK. RD was the most staining beverage (ΔE 15.36 to 31.09). EX and CX were significantly less stained than the rest of the materials (p < 0.05). CONCLUSIONS: All the staining solutions produced darkening beyond clinically acceptable limits. EX and CX were the materials who experimented less staining, followed by GD. AD and DY were the materials more stained. RW was the drink that produced more staining, followed by CF and CK. CLINICAL RELEVANCE: After 24 h of immersion in all the solutions, equivalent to 1 month of exposure in the mouth, the evaluated materials showed clinically unacceptable ΔE values.

Effect of Hydrogen and Carbamide Peroxide in Bleaching, Enamel Morphology, and Mineral Composition: In vitro Study.

AIM: The aim of the study was to evaluate the bleaching effect, morphological changes, and variations in calcium (Ca) and phosphate (P) in the enamel with hydrogen peroxide (HP) and carbamide peroxide (CP) after the use of different application regimens. MATERIALS AND METHODS: Four groups of five teeth were randomly assigned, according to the treatment protocol: HP 37.5% applied for 30 or 60 minutes (HP30, HP60), CP 16% applied for 14 or 28 hours (CP14, CP28). Changes in dental color were evaluated, according to the following formula: ΔE = [(La-Lb)2+(aa-ab)2 + (ba-bb)2]1/2. Enamel morphology and Ca and P compositions were evaluated by confocal laser scanning microscope and environmental scanning electron microscopy. RESULTS: ΔE HP30 was significantly greater than CP14 (10.37 ± 2.65/8.56 ± 1.40), but not between HP60 and CP28. HP60 shows greater morphological changes than HP30. No morphological changes were observed in the groups treated with CP. The reduction in Ca and P was significantly greater in HP60 than in CP28 (p < 0.05). CONCLUSION: Both formulations improved tooth color; HP produced morphological changes and Ca and P a gradual decrease, while CP produced no morphological changes, and the decrease in mineral component was smaller. CLINICAL SIGNIFICANCE: CP 16% applied during 2 weeks could be equally effective and safer for tooth whitening than to administer two treatment sessions with HP 37.5%.

Intratubular penetration in post cementing: A comparative study between a total etching system and a self-etching cement.

PURPOSE: To analyze the penetration depth and percentage perimeter with penetration of two fiber post cementing systems using confocal laser scanning microscopy (CLSM). METHODS: 20 maxillary incisors were shaped with the Mtwo system and filled using lateral condensation and TopSeal mixed with fluorescein. Fiber posts were cemented. The samples were divided into two groups of 10 teeth each, according to the post cementing technique used: Prime&Bond NT combined with Rebilda DC using a total dentin etching technique (Group 1); or BisCem a self-adhesive cement (Group 2). Rhodamine B was incorporated in the adhesive systems. Cross-sections were prepared, with the selection of three sections (coronal, middle and apical thirds). CLSM was used to measure the percentage perimeter of the root canal showing penetration of the endodontic cement and of the adhesive system in the dentin tubules, together with the maximum penetration depth. The nonparametric Mann-Whitney U-test was used to compare the data referred to each of the three tooth sections between the two study groups. The Friedman test was used to compare the variables by coronal, middle and apical thirds within each group. RESULTS: Greater penetration was recorded with the BisCem system in all thirds, with statistically significant differences in the case of the middle and apical thirds (P = 0.001). The percentage perimeter with penetration was also greater in all thirds with the BisCem system, though without significant differences between the two groups. Penetration depth and percentage were found to decrease in the coronal to apical direction in both groups.

A Literature Review of the Effect of Heat on the Physical-Chemical Properties of Calcium Silicate-Based Sealers.

INTRODUCTION: Recently, calcium silicate-based sealers (CSSs) have gained popularity in endodontic practice due to their biocompatibility and antimicrobial properties. They are considered viable alternatives to epoxy resin-based sealers. With the increased use of CSSs and warm vertical compaction techniques in root canal treatment, evaluating the impact of heat on CSSs properties is essential, therefore this review aimed to present a qualitative synthesis of available in vitro studies assessing the impact of heat on the physical-chemical properties of CSSs. METHODS: Following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis 2020 guidelines, a systematic advanced electronic search was performed in Scopus, Embase, Medline (via PubMed), Web of Science, and Cochrane databases in November 2023 and updated in April 2024. In vitro studies that evaluated the physical-chemical properties of CSSs were eligible. PRILE 2021 guidelines were used for the assessment of the risk of bias-included studies. RESULTS: The search identified a total of 6421 preliminary results and 10 studies were included for qualitative assessment. Eleven different physiochemical properties were assessed by the included studies. Setting time and flow were the most evaluated property among the studies. A qualitative synthesis of the evidence on each property is presented. CONCLUSIONS: Based on the in vitro studies assessed in the present systematic review, results reveal that exposing CSSs to heat can accelerate their setting time, reduce their flow, and increase their film thickness. Concerns persist regarding solubility, viscosity, radiopacity, dimensional change, microhardness, porosity, and compressive strength, requiring further research. Certain CSSs, such as MTA Fillapex and Endosequence BC sealer HiFlow, show minimal changes under heat, making them potential candidates for warm filling techniques.

The Characterization of Normal Male and Female Voice from Surface Electromyographic Parameters.

Currently, there is no consensus on the characterization of the human voice. The objective of the present study is to describe the myoelectric behavior of the extrinsic musculature of the larynx in 146 people with normal voice (Spanish speakers), aged between 20 and 50 years old. Different vocal tasks were recorded using a surface electromyograph (SEMG). In all vocal tasks, it was observed that women had higher activation (µV) in the suprahyoid and sternocleidomastoid muscles than men, while men had higher activation in the infrahyoid muscles. SEMG is a valid procedure to help define normal vocal characteristics in the studied population, providing reference values during clinical examination. However, it is necessary to adopt a universal system of assessment tasks and standardized measurement techniques to allow for comparisons with future studies.

In vitro biocompatibility of ammonia-free silver fluoride products on human dental pulp stem cells.

OBJECTIVES: Silver fluoride (SF) is a preventive and therapeutic option for dental pathological processes involving structural alterations of the hard tissues, either during their formation or those caused by caries or other pathological reasons. This study aimed to compare the biological properties of two commercial SF products, one of them with ammonium (Riva Star; SDF) and the other ammonium-free (Riva Star Aqua; AgF), both with or without potassium iodide (KI), by the assessment of the cytotoxicity of the materials' eluates. METHODS: Human dental pulp stem cells (hDPSCs) were obtained from healthy 18-23-year-old donors. Three dilutions were prepared for the tested materials (0.005%, 0.0005%, and 0.0001%). The following groups were assessed: (AgF, AgF+KI, SDF, SDF+KI, KI, negative control). A series of cytocompatibility assays were performed: MTT assay, IC50 assay, wound healing (migration) assay, cell cytoskeleton staining, analysis of cell apoptosis and necrosis, and reactive oxygen species production. The normality in the distribution of the data was previously confirmed via a Q-Q plot. Statistical significance was tested using one way ANOVA and Tukey's post hoc test. RESULTS: The incorporation of KI improved the cytocompatibility of both SF products in terms of viability, migration, morphology, apoptosis, and reactive oxygen species production. This difference was higher in the AgF group. The lowest dilutions of SF+KI and AgF+KI showed a similar cytocompatibility to that of the control group (MTT assay (p > 0.05 after 24, 48, and 72 h of culture); migration assay (p > 0.05 after 24, 48, and 72 h of culture); reactive oxygen species production (p > 0.05 after 72 h of culture). SIGNIFICANCE: Riva Star Aqua shows lower cytotoxicity than Riva Star on hDPSCs. It can be considered as a good alternative in the conservative treatment of dental caries and in the preservation and remineralisation of viable dentine tissue.

Assessment of the anti-inflammatory and biological properties of Bioroot Flow: A novel bioceramic sealer.

INTRODUCTION: BioRoot Flow (BRF) is a novel premixed bioceramic sealer indicated for endodontic treatments, but the biological and immunomodulatory effects of this endodontic sealer on human periodontal ligament stem cells (hPDLSCs) have not been elucidated. METHODS: To ascertain the biological impact of BRF, TotalFill BC Sealer (TFbc), and AH Plus (AHP) on human Periodontal Ligament Stem Cells (hPDLSCs), assessments were conducted to evaluate the cytocompatibility, cellular proliferation, migratory capacity, osteo/cementogenic differentiation potential, the ability to form mineralized nodules, and the immunomodulatory characteristics of hPDLSCs following treatment with these endodontic sealers. RESULTS: Biological assays showed adequate cell metabolic activity and cell migration in BRF, while SEM assay evidenced that TFbc and BRF groups demonstrated a superior cell adhesion process, including substrate adhesion, cytoskeleton development, and spreading on the niche-like structures of the cement as compared to the AHP group. TFbc and BRF-treated groups exhibited a significantly lower IL6 and IL8 production than AHP (* p <.05). The bioceramic sealers stimulated heightened expression of BSP, CEMP-1, and CAP genes within a 7-14 day period. Notably, BRF and TFbc demonstrated a significant enhancement in the mineralization of hPDLSCs when compared to the negative control. Among these, cells treated with BRF showed a more substantial accumulation of calcium (*** p < .001). CONCLUSIONS: Taken together, these findings indicate that BRF can potentially enhance cell differentiation by promoting the expression of essential genes related to bone and cement formation. In addition, BRF and TFbc displayed anti-inflammatory effects.

Comparative Cytotoxicity of Menthol and Eucalyptol: An In Vitro Study on Human Gingival Fibroblasts.

The aim of this study was to assess the influence of eucalyptol and menthol on the cell viability, migration, and reactive oxygen species production of human gingival fibroblasts (GFs) in vitro. Three different concentrations of eucalyptol and menthol were prepared following ISO 10993-5 guidelines (1, 5, and 10 mM). GFs were isolated from extracted teeth from healthy donors. The following parameters were assessed: cell viability via MTT, Annexin-V-FITC and 7-AAD staining, and IC50 assays; cell migration via horizontal scratch wound assay; and cell oxidative stress via reactive oxygen species assay. Data were analyzed using one-way ANOVA and Tukey's post hoc test. Statistical significance was established at p < 0.05. Eucalyptol and Menthol exhibited high cytotoxicity on gingival fibroblasts, as evidenced by cytotoxicity assays. Eucalyptol showed lower levels of cytotoxicity than menthol, compared to the control group. The cytotoxicity of the tested substances increased in a concentration-dependent manner. The same occurred in a time-dependent manner, although even 10 min of exposure to the tested substances showed a high cytotoxicity to the GFs. Commercially available products for oral application with these substances in their composition should be tested for cytotoxicity before their use.

Premixed calcium silicate-based ceramic sealers promote osteogenic/cementogenic differentiation of human periodontal ligament stem cells: A microscopy study.

To evaluate the effects of premixed calcium silicate based ceramic sealers on the viability and osteogenic/cementogenic differentiation of human periodontal ligament stem cells (hPDLSCs). The materials evaluated were TotalFill BC Sealer (TFbc), AH Plus Bioceramic Sealer (AHPbc), and Neosealer Flo (Neo). Standardized discs and 1:1, 1:2, and 1:4 eluates of the tested materials were prepared. The following in vitro experiments were carried out: ion release, cell metabolic activity 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell migration, immunofluorescence experiment, cell attachment, gene expression, and mineralization assay. Statistical analyses were performed using one-way ANOVA followed by Tukey's post hoc test (p < .05). Increased Ca2+ release was detected in TFbc compared to AHPbc and Neo (*p < .05). Biological assays showed a discrete cell metabolic activity and cell migration in Neo-treated cell, whereas scanning electronic microscopy assay exhibited that TFbc group had a better cell adhesion process of substrate attachment, spreading, and cytoskeleton development on the niche-like structures of the cement than AHPbc and Neo. The sealers tested were able to induce overexpression of the CEMP-1, ALP, and COL1A1 genes in the first days of exposure, particularly in the case of TFbc (***p < .001). All materials tested significantly increased the mineralization of hPDLSCs when compared to the negative control, although more pronounced calcium deposition was observed in the TFbc-treated cells (***p < .001). Our results suggested that TFbc promotes cell differentiation, both by increasing the expression of key osteo/odontogenic genes and by promoting mineralization of the extracellular matrix, whereas this phenomenon was less evident in Neo and AHPbc. RESEARCH HIGHLIGHTS: TFbc group had a better cell adhesion process of substrate attachment, spreading, and cytoskeleton development on the niche-like structures of the cement than AHPbc and Neo. The sealers tested were able to induce overexpression of the CEMP-1, ALP, and COL1A1 genes in the first days of exposure, particularly in the case of TFbc. All materials tested significantly increased the mineralization of hPDLSCs when compared to the negative control, although more pronounced calcium deposition was observed in the TFbc-treated cells.

3D Graphene/silk fibroin scaffolds enhance dental pulp stem cell osteo/odontogenic differentiation.

OBJECTIVES: The current in vitro study aims to evaluate silk fibroin with and without the addition of graphene as a potential scaffold material for regenerative endodontics. MATERIAL AND METHODS: Silk fibroin (SF), Silk fibroin/graphene oxide (SF/GO) and silk fibroin coated with reduced graphene oxide (SF/rGO) scaffolds were prepared (n = 30). The microarchitectures and mechanical properties of scaffolds were evaluated using field emission scanning electron microscopy (FESEM), pore size and water uptake, attenuated total reflectance fourier transformed infrared spectroscopy (ATR-FTIR), Raman spectroscopy and mechanical compression tests. Next, the study analyzed the influence of these scaffolds on human dental pulp stem cell (hDPSC) viability, apoptosis or necrosis, cell adhesion, odontogenic differentiation marker expression and mineralized matrix deposition. The data were analyzed with ANOVA complemented with the Tukey post-hoc test (p < 0.005). RESULTS: SEM analysis revealed abundant pores with a size greater than 50 nm on the surface of tested scaffolds, primarily between 50 nm and 600 µm. The average value of water uptake obtained in pure fibroin scaffolds was statistically higher than that of those containing GO or rGO (p < 0.05). ATR-FTIR evidenced that the secondary structures did not present differences between pure fibroin and fibroin coated with graphene oxide, with a similar infrared spectrum in all tested scaffolds. Raman spectroscopy showed a greater number of defects in the links in SF/rGO scaffolds due to the reduction of graphene. In addition, adequate mechanical properties were exhibited by the tested scaffolds. Regarding biological properties, hDPSCs attached to scaffolds were capable of proliferating at a rate similar to the control, without affecting their viability over time. A significant upregulation of ALP, ON and DSPP markers was observed with SF/rGO and SF/GO groups. Finally, SF/GO and SF/rGO promoted a significantly higher mineralization than the control at 21 days. SIGNIFICANCE: Data obtained suggested that SF/GO and SF/rGO scaffolds promote hDPSC differentiation at a genetic level, increasing the expression of key osteo/odontogenic markers, and supports the mineralization of the extracellular matrix. However, results from this study are to be interpreted with caution, requiring further in vivo studies to confirm the potential of these scaffolds.

Microleakage of Class II restorations and microtensile bond strength to dentin of low-shrinkage composites.

PURPOSE: To evaluate the microleakage of Class II cavities restored with experimental low-shrinking resin composites proposed for bulk filling and to measure their microtensile bond strength (microTBS) to dentin and compare to those of previously marketed low-shrinkage composites. METHODS: Class II cavities (7 mm occluso-gingival height, 2 mm mesio-distal depth of the box, 4 mm occlusal depth, 4 mm bucco-lingual width) were prepared in 50 molars and randomly divided into five groups (n = 10), according to the material: SureFil SDR flow (SDR), Filtek Silorane (FS) and experimental materials (G-aenial Flo bulk fill, GF; G-aenial Universal Flo bulk fill, GUF; GC Kalore bulk fill, GK). Microleakage was separately assessed at enamel and dentin margins by scoring the depth of silver-nitrate penetration. Twenty teeth divided into five groups (n = 4) were selected for microTBS testing. The same materials as for microleakage assessment were placed in bulk to mid-coronal dentin. Microtensile beams were loaded in tension, and microTBS at failure was calculated in MPa. Microleakage and microTBS data were analyzed by ANOVA on ranks, followed Dunn's post hoc test (P< 0.05). RESULTS: Microleakage was not observed at the enamel interface in any of the groups. At the dentin interface, SDR recorded significantly higher microleakage than the other materials. microTBS of GF, GUF and GK did not differ among each other (33 +/- 12 MPa, 31 +/- 11 MPa, 30 +/- 9 MPa, respectively), while SDR (63 +/- 17 MPa) and FS (55 +/- 17MPa) achieved significantly higher microTBS values. No direct association between the sealing properties and the bond strength values was observed.