Evaluation of the genotoxicity, cytotoxicity, and bioactivity of calcium silicate-based cements.

BACKGROUND: As calcium silicate-based cements (CSCs) have found success in various vital pulp therapy applications, several new CSC products have emerged. This study aimed to assess the genotoxicity, cytotoxicity, and bioactivity of four CSCs by comparing the newly introduced materials Bio MTA+ and MTA Cem with previously studied materials, Biodentine and NeoMTA. METHODS: Genotoxicity was evaluated using the micronucleus (MN) assay in human peripheral blood lymphocyte cells, measuring MN frequency and nuclear division index (NDI). Cytotoxicity was assessed in human dental pulp stem cells through the Water-Soluble Tetrazolium Salt-1 (WST-1) colorimetric assay. Bioactivity was determined by ELISA, measuring the levels of angiogenic and odontogenic markers (BMP-2, FGF-2, VEGF, and ALP). Statistical analyses included ANOVA, Dunnet and Sidak tests, and Wald chi-square test. (p < .05). RESULTS: The MN frequency in the groups was significantly lower than that in the positive control group (tetraconazole) (p < .05). NDI values decreased with increasing concentration (p < .05). Bio MTA+ and NeoMTA showed decreased cell viability at all concentrations in 7-day cultures (p < .01). All materials increased BMP-2, FGF-2, and VEGF levels, with Biodentine and NeoMTA showing the highest levels of BMP-2 and FGF-2 on day 7. Biodentine displayed the highest VEGF levels on day 7. Biodentine and NeoMTA groups exhibited significantly higher ALP activity than the Bio MTA+ and MTA Cem groups by day 7. CONCLUSION: Bio MTA+ and MTA Cem demonstrated no genotoxic or cytotoxic effects. Moreover, this study revealed bioactive potentials of Bio MTA+ and MTA Cem by enhancing the expression of angiogenic and osteogenic growth factors.

The effect of three additives on properties of mineral trioxide aggregate cements: a systematic review and meta-analysis of in vitro studies.

BACKGROUND: Several efforts have been made to improve mechanical and biological properties of calcium silicate-based cements through changes in chemical composition of the materials. This study aimed to investigate the physical (including setting time and compressive strength) and chemical (including calcium ion release, pH level) properties as well as changes in cytotoxicity of mineral trioxide aggregate (MTA) after the addition of 3 substances including CaCl2, Na2HPO4, and propylene glycol (PG). METHODS: The systematic review was conducted in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Electronic searches were performed on PubMed, Embase, and Scopus databases, spanning from 1993 to October 2023 in addition to manual searches. Relevant laboratory studies were included. The quality of the included studies was assessed using modified ARRIVE criteria. Meta-analyses were performed by RevMan statistical software. RESULTS: From the total of 267 studies, 24 articles were included in this review. The results of the meta-analysis indicated that addition of PG increased final setting time and Ca2+ ion release. Addition of Na2HPO4 did not change pH and cytotoxicity but reduced the final setting time. Incorporation of 5% CaCl2 reduced the setting time but did not alter the cytotoxicity of the cement. However, addition of 10% CaCl2 reduced cell viability, setting time, and compressive strength. CONCLUSION: Inclusion of 2.5% wt. Na2HPO4 and 5% CaCl2 in MTA can be advisable for enhancing the physical, chemical, and cytotoxic characteristics of the admixture. Conversely, caution is advised against incorporating elevated concentrations of PG due to its retarding effect. TRIAL REGISTRATION: PROSPERO registration number: CRD42021253707.

The cytotoxic and oxidative effects of restorative materials in cultured human gingival fibroblasts.

The aim of this study was to evaluate the cytotoxic and oxidative effects of the most commonly used dental restorative materials on human gingival fibroblast cells (HGFCs). HGFCs were obtained from healthy individuals. The tested restorative materials were a microhybrid resin based composite, a compomer resin, a glass ionomer cement, and an amalgam alloy. One hundred eight cylindirical samples, 10 mm in diameter and 2 mm in height, were prepared according to ISO 10993-12:2002 specifications (n = 9 in the tested subgroups). Freshly prepared and aged samples in artificial saliva at 37 °C (7 and 21 d) were placed into well plates and incubated. Wells without dental materials were constituted as the control group. After 72 h incubation period, cytotoxicity was determined using the neutral red (NR) assay. Oxidative alterations were assessed using total antioxidant capacity (TAC) and total oxidant status (TOS) assay kits. Data were analyzed using the ANOVA and LSD post hoc tests. All tested materials led to significant decreases in the cell viability rates (33-73%) compared to the control group. Glass ionomer and resin composite were found to be more cytotoxic than amalgam alloy and compomer. The highest TAC level was observed in glass ionomer after seven-day aging and these changes prevented an increase in TOS levels. Increases in TAC levels after seven-day aging in all groups exhibited significant differences with freshly prepared samples (p < 0.05). In all material groups, TOS levels of freshly prepared samples differed statistically and significantly from samples aged for 7 and 21 d (p < 0.05). The data obtained suggested that all the tested materials exhibited cytotoxic and pro-oxidant features. Freshly prepared samples caused higher TOS levels. However, oxidant status induced by materials decreased over time.

Genotoxic and cytotoxic potential of methacrylate-based orthodontic adhesives.

OBJECTIVES: The biocompatibility of methacrylate-based adhesives is a topic that is intensively discussed in dentistry. Since only limited evidence concerning the cyto- and genotoxicity of orthodontic adhesives is available, the aim of this study was to measure the genotoxic potential of seven orthodontic methacrylate-based adhesives. MATERIALS AND METHODS: The XTT assay was utilized to determine the cytotoxicity of Assure Plus, Assure Bonding Resin, ExciTE F, OptiBond Solo Plus, Scotchbond Universal Adhesive, Transbond MIP, and Transbond XT after an incubation period of 24 h on human gingival fibroblasts. We also performed the γH2AX assay to explore the genotoxic potential of the adhesives within cytotoxic dose ranges after an incubation period of 6 h. RESULTS: The XTT assay showed a concentration-dependent reduction in cell viability. The decrease in cellular viability was in the same dose range most significant for Assure Plus, rendering it the adhesive material with the highest cytotoxicity. Employing the γH2AX assay, a concentration-dependent increase in H2AX phosphorylation was detected, indicating induction of DNA damage. CONCLUSIONS: For most products, a linear correlation between the material concentration and γH2AX foci was observed. The most severe effect on γH2AX focus induction was found for Transbond MIP, which was the only adhesive in the test group containing the co-initiator diphenyliodonium hexafluorophosphate (DPIHP). CLINICAL RELEVANCE: The data indicate that orthodontic adhesives, notably Transbond MIP, bear a genotoxic potential. Since the study was performed with in vitro cultivated cells, a direct translation of the findings to in vivo exposure conditions should be considered with great diligence.

Biocompatibility of self-adhesive resin cement with fibroblast cells.

STATEMENT OF PROBLEM: Dental cements that release monomers that negatively impact adjacent oral soft tissues may adversely affect clinical outcomes. However, in vitro studies evaluating the cytotoxic and genotoxic potential of substances released from dental cements are lacking. PURPOSE: The purpose of this in vitro study was to define and compare the cytotoxicity and genotoxicity of the eluates of a self-adhesive resin cement (RelyX Unicem 2 Automix) autopolymerized and light polymerized with 2 other types of luting cements: a glass ionomer cement (Ketac Cem Easymix) and a resin-modified glass ionomer cement (Ketac Cem Plus). MATERIAL AND METHODS: The eluates were prepared, and 3T3 mouse fibroblast cells were exposed for 24 hours to serial eluate dilutions of the 3 types of cement. Cytotoxicity was determined by using a cell viability assessment through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet assays. Genotoxic effects were determined by using the cytokinesis-block micronucleus assay. RESULTS: Cell viability was higher in the presence of the glass ionomer cement eluate than of the resin-modified glass ionomer cement and resin cement eluates. A pronounced decrease in viability was found when the cells were exposed to undiluted samples of resin-modified glass ionomer cement (around 50%) or resin cement (around 80% to 90%). No significant difference in cell viability was found between autopolymerized and light-polymerized resin cements. All cements induced a dose-dependent response of mononucleated cell formation. However, only the resin cements showed double strand breaks significant differences in the deoxyribonucleic acid (DNA) molecules against the basal DNA lesions that occurred spontaneously. CONCLUSIONS: The glass ionomer cement was not found to be cytotoxic or genotoxic, whereas the eluates derived from the resin-modified glass ionomer cement and resin cement, independently of the polymerization method, were cytotoxic in fibroblast cells. Maximum cytotoxicity was observed in the presence of resin cement, which also showed genotoxicity, independently of being light polymerized.

Are Denture Adhesives Safe for Oral Cells?

PURPOSE: To compare the cytotoxicity of six commercially available denture adhesives on human gingival cells: Poligrip Flavour Free Fixative Cream, Fixodent Pro Duo Protection, Novafix cream, FittyDent, Polident Total Action, and Fixodent Pro Plus Duo Protection. MATERIAL AND METHODS: Eluates of denture adhesives were brought into contact with human gingival cells and compared to untreated cells (w/o any dental adhesive elute). Cell toxicity was assessed by measuring cell viability (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assays), cell morphology (immunofluorescence assays), induction of apoptosis/necrosis and production of reactive oxygen species (ROS) (flow cytometry assays). In addition, the pH of each sample was determined. Data were analyzed using one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparisons test. RESULTS: All denture adhesives tested led to a reduction in pH, especially Fixodent Pro Duo Protection and Fixodent Pro Plus Duo Protection. The cell viability assays showed that Fixodent Pro Duo Protection (1:1 72 hours, p = 3.04 × 10-6 ; 1:2 72 hours, p = 2.07 × 10-6 ; 1:4 72 hours, p = 2.04 × 10-6 ) and Fixodent Pro Plus Duo Protection (1:1 72 hours, p = 2.01 × 10-6 ; 1:2 72 hours, p = 3.03 × 10-6 ; 1:4 72 hours, p = 2.02 × 10-6 ) significantly decreased cell viability at all dilutions. Compared to the control group and the rest of the adhesives, Poligrip Flavour Free Fixative Cream (PFF 1:1 72 hours, p = 2.24 × 10-6 ; 1:2 72 hours, p = 2.44 × 10-6 ; 1:4 72 hours, p = 2.04 × 10-6 ) showed a significantly higher cell viability score at all dilutions. Fixodent Pro Duo Protection and Fixodent Pro Plus Duo Protection, both adhesives containing zinc salts in their composition, were responsible for necrosis, and the number of cells was much reduced, with aberrant morphology and pyknotic nucleus. Finally, Fixodent (1:2, p = 2.04 × 10-6 , 1:4, p = 0.00036; 1:2, p = 8.82 × 10-6 , 1:4, p = 2.30 × 10-6 ) products significantly promoted ROS production in gingival cells. CONCLUSIONS: The results suggest that denture adhesives containing zinc in their composition could be responsible of the decrease of cell viability, ROS production, aberrant cell morphology, and induction of apoptosis and cell death. However, other possible additional cytotoxic factors must be considered. Thus, more studies are necessary to confirm this hypothesis.

In Vivo Molecular Toxicity Profile of Dental Bioceramics in Embryonic Zebrafish ( Danio rerio).

The investigation of the biocompatibility of potential and commercially available dental material is a major challenge in dental science. This study demonstrates that the zebrafish model is a novel in vivo model for investigating the biocompatibility of dental materials. Two commercially available dental materials, mineral trioxide aggregate (MTA) and Biodentine, were assessed for their biocompatibility. The biocompatibility analysis was performed in embryonic zebrafish with the help of standard toxicity assays measuring essential parameters such as survivability and hatching. The mechanistic and comparative analysis of toxicity was performed by oxidative stress analysis by measuring ROS induction and apoptosis in zebrafish exposed to dental materials at different concentrations. The molecular investigation at the protein level was done by a computational approach using in silico molecular docking and pathway analysis. The toxicity analysis showed a significant reduction in hatching and survivability rates along with morphological malformations with an increase in the concentration of exposed materials. ROS and apoptosis assay results revealed a greater biocompatibility of Biodentine as compared to that of MTA which was concentration-dependent. In silico analysis showed the significant role of the tricalcium silicate-protein ( Sod1, tp53, RUNX2B) interaction in an exhibition of toxicity. The study provides a new vision and standard in dental material sciences for assessing the biocompatibility of potential novel and commercially available dental materials.

Qualitative and Quantitative Evaluation of Cytotoxicity of Five Different One-Step Self-Etching Adhesives.

PURPOSE: To qualitatively and quantitatively compare the cytotoxic potentials of five different one-step self-etching adhesives: Prime&Bond One-Select (PB-OS), Optibond All-in-One (OB-AIO), G-Bond (GB), Clearfil Universal Bond (CUB), Single Bond Universal (SBU). MATERIALS AND METHODS: During the first stage of the study, the cytotoxic activities of the test materials were evaluated qualitatively using the direct contact method. In this method, the test materials were placed directly into a monkey kidney epithelial cell culture medium. Reaction zones which occurred in the culture medium were evaluated, in addition to the density and changes in the morphology of the cells. During the second stage, the cytotoxic potential of four different dilutions (1%, 0.1%, 0.01%, 0.001%) of the test materials on L929 rat fibroblast cells was quantitatively evaluated at three different time periods (24 h, 48 h, 72 h) with the MTT tetrazolium-based assay. RESULTS: In the first stage, a zone exceeding 1 cm was observed around or below SBU, CUB, GB and OB-AIO. In PB-OS, the zone borders were approximately 1 cm. In the second stage after the MTT assay, CUB was the most cytotoxic after 24 h, GB and SBU after 48 h, and OB-AIO after 72 h. CONCLUSION: All adhesives tested showed different degrees of cytotoxicity, which statistically significantly increased with dose. Changes were seen related to time.

Cytotoxicity of modified glass ionomer cement on odontoblast cells.

Recently a modified glass ionomer cement (GIC) with enhanced bioactivity due to the incorporation of wollastonite or mineral trioxide aggregate (MTA) has been reported. The aim of this study was to evaluate the cytotoxic effect of the modified GIC on odontoblast-like cells. The cytotoxicity of a conventional GIC, wollastonite modified GIC (W-mGIC), MTA modified GIC (M-mGIC) and MTA cement has been evaluated using cement extracts, a culture media modified by the cement. Ion concentration and pH of each material in the culture media were measured and correlated to the results of the cytotoxicity study. Among the four groups, conventional GIC showed the most cytotoxicity effect, followed by W-mGIC and M-mGIC. MTA showed the least toxic effect. GIC showed the lowest pH (6.36) while MTA showed the highest (8.62). In terms of ion concentration, MTA showed the largest Ca(2+) concentration (467.3 mg/L) while GIC showed the highest concentration of Si(4+) (19.9 mg/L), Al(3+) (7.2 mg/L) and Sr(2+) (100.3 mg/L). Concentration of F(-) was under the detection limit (0.02 mg/L) for all samples. However the concentrations of these ions are considered too low to be toxic. Our study showed that the cytotoxicity of conventional GIC can be moderated by incorporating calcium silicate based ceramics. The modified GIC might be promising as novel dental restorative cements.

Transdentinal cytotoxicity of resin-based luting cements to pulp cells.

OBJECTIVES: The aim of this study was to evaluate the transdentinal cytotoxicity of components released from different resin-based luting cements to cultured MDPC-23 odontoblast-like cells and human dental pulp cells (HDPCs). MATERIALS AND METHODS: Artificial pulp chamber (APC)/dentin disc sets were distributed into four groups according to the materials tested (n = 10), as follows: G1, control (no treatment); G2, resin-modified glass-ionomer cement (RelyX Luting 2); G3, self-adhesive resin cement (RelyX U200); and G4, conventional resin cement (RelyX ARC). The materials were applied to the occlusal surfaces (facing up) of the dentin discs adapted to the APCs. The pulpal surfaces of the discs were maintained in contact with culture medium. Then, an aliquot of 400 µL from the extract (culture medium + resin-based components that diffused through dentin) of each luting cement was applied for 24 h to HDPCs or MDPC-23 cells previously seeded in wells of 24-well plates. Cell viability analysis was performed by the MTT assay (1-way ANOVA/Tukey test; α = 5 %). RESULTS: For MDPC-23 cells, RelyX ARC (G4) and RelyX Luting 2 (G2) caused greater reduction in cell viability compared with the negative control group (P < 0.05). Only the HDPCs exposed to RelyX ARC (G4) extract showed a tendency toward viability decrease (9.3 %); however, the values were statistically similar to those of the control group (G1) (P > 0.05). CONCLUSIONS: In accordance with the safe limits of ISO 10993-5:1999 (E) recommendations, all resin-based luting cements evaluated in this study can be considered as non-toxic to pulp cells. CLINICAL RELEVANCE: Cytotoxicity of resin-based luting cements is material-dependent, and the different protocols for the application of these dental materials to dentin may interfere with their cytotoxicity.

Comparative cell attachment, cytotoxicity and antibacterial activity of radiopaque dicalcium silicate cement and white-coloured mineral trioxide aggregate.

AIM: To comparatively examine the cell attachment, cytotoxicity, and antibacterial activity of radiopaque dicalcium silicate cement (RDSC) and ProRoot white-coloured mineral trioxide aggregate (WMTA). METHODOLOGY: AlamarBlue was used for real-time and repeated monitoring of MG63 cell attachment on freshly mixed and set cements. The pH changes in the growth medium at different time-points were also measured. Cytotoxicity evaluation was performed according to ISO 10993-5 specifications. The antibacterial activity of the cement specimens was evaluated using Enterococcus faecalis. RESULTS: There were no significant differences (P > 0.05) between the two cements for cell attachment either in the fresh groups or in the set groups at all culture times. Neither freshly mixed group nor set groups had significant pH differences. In the case of cytotoxicity, RDSC was significantly (P < 0.05) superior to WMTA at 12 and 24 h of incubation. RDSC and WMTA possessed similar antimicrobial activity, substantiated by the formation of growth inhibition zones and bacteriostasis ratio in E. faecalis strains. CONCLUSIONS: The cell attachment, cytotoxicity and antibacterial efficacy of RDSC were comparable to those reported for ProRoot WMTA. The results of the current study suggest that this RDSC could be used as a root-end filling material and root sealer.

Toxicity of a dental adhesive compared with ionizing radiation and zoledronic acid.

BACKGROUND: To determine the toxicity of aqueous dilutions of a universal self-priming dental adhesive (DA) and comparing these with those elicited by exposure to ionizing radiation (IR), Zoledronic acid (Z) treatment and the synergic effects of the combined treatment with IR+Z. MATERIAL AND METHODS: The genotoxic effect of DA was determined by the increase in the frequency of micronuclei in cytokinesis-blocked in cultured human lymphocytes before and after exposure to 2Gy of X-rays. The cytotoxic effect was studied by using the MTT cell viability test in normal prostate cell lines (PNT2) after exposure to different X-ray doses (0Gy-20Gy). The cell lines divided into different groups and treated with different test substances: DA in presence of O2, DA in absence of O2, Z-treated and control. RESULTS: An in vitro dose-dependent and time-dependent cytotoxic effect of DA, Z and IR on PNT2 cells (p>0.001) was demonstrated. DA without-O2, following the recommendations of manufacturers, had a more pronounced effect of increasing cell death than DA with-O2 (p<0.001). In the genotoxicity assay, DA at 25% of its original concentration significantly increased chromosome damage (p<0.001). The samples studied were found to be toxic, and the samples photo-polymerized in absence of O2 showed a bigger cytotoxic effect comparable to the additive toxic effect showed by the combined treatment of IR+Z. CONCLUSIONS: Additional effort should be carried out to develop adhesives, which would reduce the release of hazardous substances; since toxic effects are similar to that reported by other agents whose clinical use is controlled by the health authorities.

In vitro comparison of cytotoxicity of four root canal sealers on human gingival fibroblasts.

UNLABELLED: The goal of this in vitro study was to evaluate the relative biocompatibility of four endodontic sealers on the cell culture of the human fibroblast through cytotoxicity. MATERIALS AND METHODS: In this study four endodontics sealers was used GuttaFlow (Roeko)silicone based sealer, AH plus (De Tray-DENTSPLY) epoxy resin based, Apexit (Vivadent) calcium hydroxide based and Endorez (Ultradent) methacrylate based sealer. Sealers were tested on primary cell lines of human gingival fibroblasts. Experiments were preformed in laboratories of Hacettepe University of Ankara, Turkey and Faculty of Dentistry, University of Sarajevo, Bosnia and Herzegovina Cytotoxicity was determinate using WST-1 assay. RESULTS: Results were analyzed by SPSS 19 program. Kolgomorov-Smirnov test, Shapiro-Wilk and descriptive statistics also were used, as well as Kriskall-Wallis, ANOVA test and T- test. According to our results all four sealers showed different cytotoxicity effects on human gingival fibroblast cell culture, but all of them are slightly cytotoxic. CONCLUSIONS: According to results of this study it can be concluded: all four sealers showed different cytotoxicity effects on primary cell lines of human gingival fibroblasts, but all of them are slightly cytotoxicity.

Residual HEMA and TEGDMA release and cytotoxicity evaluation of resin-modified glass ionomer cement and compomers cured with different light sources.

The purpose of this study was first to evaluate the elution of 2-hydroxyethyl methacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) monomers from resin-modified glass ionomer cement (RMGIC) and compomers cured with halogen and light-emitting diode (LED) light-curing units (LCUs). The effect of cured materials on the viability of L929 fibroblast cells was also evaluated. One RMGIC (Ketac N100) and two compomers (Dyract Extra and Twinkystar) were tested. Materials were prepared in teflon disks and light-cured with LED or halogen LCUs. The residual monomers of resin materials in solution were identified using high-performance liquid chromatography. The fibroblast cells' viability was analyzed using MTT assay. The type of LCU did not have a significant effect on the elution of HEMA and TEGDMA. A greater amount of HEMA than TEGMDA was eluted. The amount of TEGDMA eluted from Twinkystar was greater than Dyract Extra (P < 0.05) when cured with a halogen LCU. All material-LCU combinations decreased the fibroblast cells' viability more than the control group (P < 0.01), except for Dyract Extra cured with a halogen LCU (P > 0.05). Curing with the LED LCU decreased the cells' viability more than curing with the halogen LCU for compomers. For Ketac N100, the halogen LCU decreased the cells' viability more than the LED LCU.

Cytotoxicity of resin-based luting cements to pulp cells.

PURPOSE: To evaluate the cytotoxicity of components released from different types of luting cements to two cell lines obtained from pulp tissue. METHODS: Three types of luting cements were evaluated, distributed into the following groups: G1--negative control (no treatment); G2--resin-modified glass-ionomer cement (Rely X Luting 2); G3--self-adhesive resin cement (Rely X U200); and G4--conventional resin cement (Rely X ARC). Standardized cylindrical specimens (14 mm diameter and 1 mm thick) prepared with the dental materials were immersed in culture medium (DMEM) for 24 hours to obtain the extracts (DMEM + components released from the cements). Then, the extracts were applied to cultured odontoblast-like MDPC-23 cells or human dental pulp cells (HDPCs). Finally, cell viability (MTT assay), cell death (Annexin/PI) (Kruskal-Wallis/Mann-Whitney; α = 5%) and cell morphology (SEM) were assessed. Cements' components in contact with cells (SEM/EDS) and pH of the extracts were also evaluated. RESULTS: The resin-modified glass-ionomer cement (G2) caused the most intense toxic effect to the two cell lines; the cell viability reduction was around 95.8% and 89.4% for MDPC-23 cells and HDPCs, respectively, which was statistically significantly different compared with that of the negative control group (G1). Also, a high quantity of particles leached from this ionomeric cement was found on the cells, which showed intense morphological alterations. In the G2 group, 100% necrosis was observed for both cell lines, and an acidic pH was detected on the extract. Conversely, Rely X U200 (G3) and Rely X ARC (G4), which presented low solubility and no alteration in pH, caused only slight cytotoxicity to the cultured cells.

In-vitro-cytotoxicity of self-adhesive dental restorative materials.

OBJECTIVES: Although the introduction of self-adhesive composites in restorative dentistry is very promising, the innovation of new materials also presents challenges and unknowns. Therefore, the aim of this study was to investigate the cytotoxicity of four different self-adhesive composites (SAC) in vitro and to compare them with resin-modified glass ionomer cements (RM-GIC), a more established group of materials. METHODS: Samples of the following materials were prepared according to ISO 7405/10993-12 and eluted in cell culture medium for 24 h at 37 °C: Vertise Flow, Fusio Liquid Dentin, Constic, Surefil One, Photac Fil and Fuji II LC. Primary human pulp cells were obtained from extracted wisdom teeth and cultured for 24 h with the extracts in serial dilutions. Cell viability was evaluated by MTT assay, membrane disruption was quantified by LDH assay and apoptosis was assessed by flow cytometry after annexin/PI staining. RESULTS: Two SAC (Constic and Vertise Flow) and one RM-GIC (Photac Fil) significantly reduced cell viability by more than 30% compared to the untreated control (p < 0.001). Disruptive cell morphological changes were observed and the cells showed signs of late apoptosis and necrosis in flow cytometry. Membrane disruption was not observed with any of the investigated materials. CONCLUSION: Toxic effects occurred independently of the substance group and need to be considered in the development of materials with regard to clinical implications. CLINICAL SIGNIFICANCE: SAC have many beneficial qualities, however, the cytotoxic effects of certain products should be considered when applied in close proximity to the dental pulp, as is often required.

Cytotoxic effect of dental luting cement on human gingival mesenchymal stem cell and evaluation of cytokines and growth factor release - An in vitro study.

AIM: In routine dental care, various dental luting cements are utilized to cement the dental prosthesis. Thus, the aim of the current study was to assess the Cytotoxic effect of three different dental luting cements on human gingival mesenchymal stem cell and evaluation of cytokines and growth factors release. SETTINGS AND DESIGN: Cytotoxicity of glass ionomer cement (GIC), resin modified glass ionomer cement (RMGIC) and resin cement (RC) on the human gingival mesenchymal stem cells (HGMSCs) was evaluated. Amongst the cements tested, least cytotoxic cement was further tested for the release of cytokines and growth factors. MATERIALS AND METHODS: MTT test was used to evaluate the cytotoxicity of the dental luting cements at 1 h, 24 h, and 48 h on HGMSCs. Cytokines such as interleukin (IL) 1α & IL 8 and growth factors such as platelet derived growth factor & transforming growth factor beta release from the least cytotoxic RC was evaluated using flow cytometry analysis. STATISTICAL ANALYSIS USED: The mean absorbance values by MTT assay and cell viability at various time intervals between four groups were compared using a one way analysis of variance test and Tukey's post hoc test. The least cytotoxic RC group and the control group's mean levels of cytokines and growth factors were compared using the Mann-Whitney test. RESULT: As exposure time increased, the dental luting cement examined in this study were cytotoxic. RC was the least cytotoxic, RMGIC was moderate and glass ionomer cement showed the highest cytotoxic effect. Concomitantly, a significant positive biological response of gingival mesenchymal stem cells with the release of ILs when exposed to the RC was observed. CONCLUSION: For a fixed dental prosthesis to be clinically successful over the long term, it is imperative that the biocompatibility of the luting cement be taken into account in order to maintain a healthy periodontium surrounding the restoration.

Apoptotic effects of biodentine, calcium-enriched mixture (CEM) cement, ferric sulfate, and mineral trioxide aggregate (MTA) on human mesenchymal stem cells isolated from the human pulp of exfoliated deciduous teeth.

BACKGROUND: Preservation of primary teeth in children is highly important. Pulpotomy is a commonly performed treatment procedure for primary teeth with extensive caries. Thus, biocompatibility of pulpotomy agents is highly important. Biodentine, calcium enriched mixture (CEM) cement, ferric sulfate, and mineral trioxide aggregate (MTA) Angelus are commonly used for this purpose. Thus, this study aimed to assess the apoptotic effects of Biodentine, CEM cement, ferric sulfate, and MTA on stem cells isolated from the human pulp of exfoliated deciduous teeth. METHODS: In this in-vitro, experimental study, stem cells isolated from the human pulp of exfoliated deciduous teeth were exposed to three different concentrations of Biodentine, CEM cement, ferric sulfate, and MTA for different time periods. The cytotoxicity of the materials was evaluated by flow cytometry using the annexin propidium iodide (PI) kit. Data were analyzed by ANOVA and Tukey's test at P<0.05 level of significance. RESULTS: All four tested materials induced significantly greater apoptosis compared with the control group. The difference in cell apoptosis caused by the first concentration of ferric sulfate and MTA was not significant at 24 hours. In other comparisons, the cytotoxicity of ferric sulfate was significantly lower than that of other materials. Biodentine showed higher cytotoxicity than MTA at first; but this difference faded over time. The cytotoxicity of CEM cement was comparable to that of MTA. The highest cell viability was noted at 24 hours in presence of the minimum concentration of ferric sulfate. The lowest cell viability was noted at 72 hours in presence of the maximum concentration of CEM cement. CONCLUSIONS: In comparison with other materials, ferric sulfate showed minimum cytotoxicity; the cytotoxicity of the three cements was comparable. It appears that the concentration of ferric sulfate and the composition of cements are responsible for different levels of cytotoxicity.

A novel root-end filling material based on hydroxyapatite, tetracalcium phosphate and polyacrylic acid.

AIM: To develop a novel root-end filling material (NRFM) based on hydroxyapatite, tetracalcium phosphate and polyacrylic acid, to determine its chemical composition and to compare its physical properties and cytotoxicity with those of glass-ionomer cement (GIC) and grey Portland cement (GPC). METHODOLOGY: The NRFM was prepared by blending distilled water with powders of hydroxyapatite, tetracalcium phosphate, solid polyacrylic acid, solid citric acid and sodium citrate. Chemical analysis was then performed by X-ray diffraction and Fourier transform infrared spectrophotometry. Physical properties were compared with GIC and GPC regarding setting time, compressive strength, pH value and washout resistance; cytotoxicity was assessed using a transwell cell culture system. RESULTS: (i) The NRFM was primarily composed of HA, calcium polyacrylate and calcium citrate. (ii) The mean and standard deviation setting time of NRFM was 11.0 (0.8) min; its compressive strength was 25.6 (2.7) MPa and 38.2 (5.7) MPa at 1 and 7 days respectively. Its pH value ranged from 6.14 to 8.28 and it remained intact after the washout test, whereas GIC and GPC disintegrated. (iii) Dimethyl-thiazol-diphenyl tetrazolium bromide (MTT) and crystal violet (CV) assay revealed that cell-viability on the NRFM was not significantly different form that of the negative control group after treatment for 24, 48 and 72 h. CONCLUSION: The novel root-end filling material (NRFM) is a promising root-end filling material with good physicochemical properties and low cytotoxicity.

Biocompatibility of a conventional glass ionomer, ceramic reinforced glass ionomer, giomer and resin composite to fibroblasts: in vitro study.

OBJECTIVE: This aim of this study was at compare the fibroblast cytotoxicicty of four restorative materials--a conventional glass ionomer cement (GC Fuji Type II GIC), a ceramic reinforced glass ionomer cement (Amalgomer), a giomer (Beautifl II) and a resin composite (Filtek Z350) at three different time periods (24, 48 and 72 hours). METHOD: The succinyl dehydrogenase (MTT) assay was employed Cylindrical specimens of each material (n = 15) were prepared and stored in Dulbecco's modified Eagle medium, following which L929 fibroblasts were cultured in 96 well plates. After 24 hours of incubation, the MTT assay was performed to detect the cell viability. The method was repeated after 48 and 72 hours. The impact of materials and exposure times on cytotoxicity of fibroblasts was statistically analyzed using two way ANOVA (P = 0.05). RESULTS: Both time and material had an impact on cell viability, with giomer demonstrating the maximum cell viability at all time periods. The cell viability in the giomer group was significantly different from all other materials at 24 and 72 hours (P < 0.05), while at 48 hours giomer was significantly different only with resin composite (P < 0.05). CONCLUSIONS: Giomers showed better biocompatibility than conventional and ceramic reinforced glass ionomer cements and, resin composite. Ceramic reinforced glass ionomer demonstrated superior biocompatibility compared to conventional glass ionomer.