Cytotoxicity and Bioactivity of Mineral Trioxide Aggregate and Bioactive Endodontic Type Cements: A Systematic Review.

BACKGROUND: Knowledge of the cytotoxicity and bioactivity of endodontic materials may assist in understanding their ability to promote dental pulp stem cell activity and pulp healing in primary teeth. MATERIALS AND METHODS: This systematic review was carried out by searching the electronic databases such as PubMed, Google Scholar, and Cochrane reviews for the articles published between January 2000 and December 2018 using the appropriate MeSH keywords. An independent investigator evaluated the abstracts and titles for possible inclusion, as per the stipulated inclusion and exclusion criteria. The topics considered for extracting data from each study were: cell lineage, cytotoxicity assay used, and type of material tested. RESULTS: Seven eligible studies were selected for assessing the quality of evidence on the bioactivity of bioactive endodontic cements (BECs) (1 human cell line, 2 animal cell lines, and 4 in vitro, animal, and human studies) and 13 studies were selected for reviewing the quality of evidence on cytotoxicity (7 human cell lines, 4 animal cell lines, and 2 animal model studies). Very limited studies had been conducted on the bioactivity of materials other than mineral trioxide aggregate (MTA). With regards to cytotoxicity, the studies were diverse and most of the studies were based on MTT assay. Mineral trioxide aggregate is the most frequently used as well as studied root-end filling cement, and the literature evidence corroborated its reduced cytotoxicity and enhanced bioavailability. CONCLUSION: There was a lack of sufficient evidence to arrive at a consensus on the ideal material with minimal cytotoxicity and optimal bioactivity. More focused human/cell line-based studies are needed on the available root filling materials. CLINICAL SIGNIFICANCE: The present systematic review provides an update on the available literature evidence on the cytotoxicity and bioactivity of various BECs including MTAs and their influence on the different cells with respect to their composition and strength. HOW TO CITE THIS ARTICLE: Maru V, Dixit U, Patil RSB, et al. Cytotoxicity and Bioactivity of Mineral Trioxide Aggregate and Bioactive Endodontic Type Cements: A Systematic Review. Int J Clin Pediatr Dent 2021;14(1):30-39.

A Systematic Review Comparing Mineral Trioxide Aggregate to Other Commercially Available Direct Pulp Capping Agents in Dogs.

Vital pulp therapy (VPT) and direct pulp capping (DPC) are procedures regularly performed in dogs for the management of acute tooth fractures and as part of management for traumatic malocclusions. The purpose of this review is to apply an evidence-based medicine approach to systematically review and evaluate the scientific literature evaluating the efficacy of mineral trioxide aggregate (MTA) to other commercially available materials used for VPT in the permanent teeth of dogs. The 9 studies meeting inclusion criteria were reviewed and each studies evidence was classified using a grading system modified from the Oxford Centre for Evidence-Based Medicine. For the studies meeting inclusion criteria, MTA consistently performed as well or better than other commercially available products in terms of calcific barrier formation and biocompatibility. This review found a lack of consistency between the studies making a direct comparison of the results unreliable. Future studies would benefit from the implementation of a standard scoring system for histology, equivalent and longer study duration times and the correlation of histological and radiographic data.

Biological Effects of New Hydraulic Materials on Human Periodontal Ligament Stem Cells.

Background: The aim of this study was: to evaluate the biological properties of new hydraulic materials: Bio-C Repair and Bio-C Sealer. Methods: Periodontal ligament stem cells were exposed to several dilutions of Bio-C Repair and Bio-C Sealer. The ion release profile and pH were determined. Metabolic activity, cell migration and cell survival were assessed using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), wound-healing assays and Annexin assays, respectively. Cells were cultured in direct contact with the surface of each material. These were then analyzed via scanning electron microscopy (SEM) and energy dispersive X-ray (EDX). Statistical differences were assessed using a two-way ANOVA (α < 0.05). Results: Similar pH was observed in these cements. Bio-C Sealer released significantly more Ca and Si ions (p < 0.05) in comparison with Bio-C Repair. Undiluted Bio-C Sealer induced a significant reduction on cellular viability, cell survival and cell migration when compared to the control (p < 0.05). Moreover, SEM showed abundant cells adhered on Bio-C Repair and a moderate number of cells attached on Bio-C Sealer. Finally, EDX analysis identified higher percentages of Ca and O in the case of Bio-C repair than with Bio-C sealer, while other elements such as Zr and Si were more abundant in Bio-C sealer. Conclusions: Bio-C Repair displayed higher cell viability, cell adhesion and migration rates than Bio-C Sealer.

Evaluation of changes in ion release and biological properties of NeoMTA-Plus and Endocem-MTA exposed to an acidic environment.

AIM: To analyse in vitro changes in ion release and biological properties of Endocem-MTA (Maruchi, Wonju, Korea) and NeoMTA-Plus (Avalon Biomed Inc, Bradenton, FL, USA) exposed to acidic or neutral environment on human dental periodontal ligament stem cells (hPDLSCs). METHODOLOGY: Cell viability and wound healing assays were performed using eluates of each material. Cell death and changes in phenotype induced by the set endodontic sealer eluates were evaluated through flow cytometry. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy. The chemical composition of the materials was determined by energy-dispersive X-ray (EDX), and ion release was evaluated by inductively coupled plasma-mass spectrometry. Statistical analysis was performed with analysis of variance and a Bonferroni or Tukey post-test (α < 0.05). RESULTS: The MTT assay revealed non-cytotoxic effects of NeoMTA-Plus and Endocem-MTA at pH 5.2 and 7.4. However, there were minor differences compared with the control, especially at pH 5.2, where both materials were associated with significantly greater cell viability (P < 0.05). In both environments, the materials stimulated hPDLSCs to migrate. hPDLSCs were attached to the bioactive cements, with multiple prolongations proliferated on the surface of the samples. Moreover, there were no changes to cell phenotype or apoptosis/necrosis rates, indicating that the acidic environment did not induce cell death. Prismatic crystalline structures were seen on the surface of the cements exposed to butyric acid and EDX analysis identified a marked peak of Ca2+ from NeoMTA-Plus and Endocem-MTA in acidic and physiological environments. CONCLUSIONS: An acidic environment favoured the release of Ca2+ ions from both bioactive cements, and the cytotoxicity of these bioactive cements was low in both environments studied.

Higher hydration performance and bioactive response of the new endodontic bioactive cement MTA HP repair compared with ProRoot MTA white and NeoMTA plus.

The aim of this study was to characterize the hydration performance and the bioactive response of the new bioactive endodontic cement MTA HP repair (HP), comparing its physicochemical parameters with those of ProRoot MTA White (Pro) and NeoMTA Plus (Neo). Un-hydrated precursor materials were characterized by X-ray fluorescence, laser diffraction, N2 physisorption and field emission gun scanning electron microscopy (FEG-SEM). Setting time was assessed according to ASTM specification C 266. Hydrated materials were analyzed by X-ray diffraction, Fourier transform infrared spectroscopy (FT-IR) and (FEG-SEM). Bioactivity evaluation in vitro was carried out, by soaking processed cement disk in simulated body fluid (SBF) during 168 h. The cements surface was studied by FT-IR, FEG-SEM, and energy dispersive X-ray. Release to the SBF media of ionic degradation products was monitored using inductively coupled plasma atomic emission spectroscopy. HP showed shorter initial setting time compared to Pro and Neo and produce a quick and effective bioactive response in vitro in terms of phosphate phase surface coating formation. This higher bioactive response for HP is correlated with increasing calcium aluminate content, increasing surface area of un-hydrated powder precursor and the increasing release capacity of Si ionic products of the final hydrated product. The higher bioactive response of MTA HP repair highlights this material, as very interesting to further investigate its performance to improve the outcome of vital pulp therapy procedures. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2109-2120, 2019.

Porosity analysis of mineral trioxide aggregate Fillapex and BioRoot cements for use in endodontics using microcomputed tomography.

AIM: The purpose of this study is to compare the porosity of two sealant cements, mineral trioxide aggregate (MTA) Fillapex® and BioRoot® root canal sealer (RCS). These samples were analyzed using microcomputed tomography. MATERIALS AND METHODS: Sixteen samples were used in the study that were divided according to the composition of the materials used. MTA Fillapex® (n = 8) and BioRoot® RCS (n = 8) were the samples prepared according to the manufacturer's instructions. They were placed in silicone molds of 5 ± 0.1 mm in height and an internal diameter of 5 ± 0.1 mm; 24 h after its preparation, the samples were scanned through a microcomputed tomography, and the porosity results were analyzed statistically by independent t-tests. RESULTS: It is evident that MTA Fillapex® has better porosity properties than BioRoot® RCS. The results of the study quantify a smaller number of pores per surface, a smaller volume in each pore per mm3, and a lower total porosity present in samples of MTA Fillapex® unlike BioRoot® RCS samples which is larger in both. CONCLUSIONS: The results obtained in computerized microtomography endodontic biomaterial samples concluded that MTA Fillapex® has a lower porosity than BioRoot® RCS.

Properties of NeoMTA Plus and MTA Plus cements for endodontics.

AIM: To test a novel calcium silicate cement mixed with a water-based gel (NeoMTA Plus) with regard to chemical-physical properties and apatite-forming ability. METHODOLOGY: NeoMTA Plus (Avalon Biomed Inc. Bradenton, FL, USA; lot. 2014090301) and a commercial MTA-based material with similar properties (MTA Plus, Prevest Denpro Limited, Jammu, India; lot. 41001) were tested for ion-releasing ability, initial and final setting times, radiopacity, open and impervious porosity and apparent porosity, water sorption, weight loss, solubility, ability to nucleate calcium phosphates (CaP) after immersion in HBSS (Hank's Balanced Salt Solution) by ESEM-EDX and micro-Raman spectroscopy. The results were analysed statistically with the anova test (P  <  0.05). RESULTS: NeoMTA Plus had a prolonged setting time (315 min) and a satisfactory radiopacity (3.76 mm Al). Calcium and hydroxyl ion release was significantly greater and more prolonged in comparison with MTA Plus (P < 0.05). Both NeoMTA Plus and MTA Plus had high values of open porosity and solubility. ESEM-EDX and micro-Raman confirmed the ability to nucleate calcium phosphates on their surface after immersion in HBSS. CONCLUSION: NeoMTA Plus is a new calcium silicate-based cement for root filling with an adequate radiopacity and prolonged setting time. The ion release and CaP-forming ability could increase stability of the root filling and promote endodontic and periodontal tissue regeneration, enhancing the bioactivity and biocompatibility of the material.

Development of novel tricalcium silicate-based endodontic cements with sintered radiopacifier phase.

OBJECTIVES: All implants, bone and endodontic cements need to be sufficiently radiopaque to be able to be distinguished from neighbouring anatomical structures post-operatively. For this purpose, radiopacifying materials are added to the cements to render them sufficiently radiopaque. Bismuth oxide has been quite a popular choice of radiopacifier in endodontic materials. It has been shown to cause dental discoloration. The aim of this study was to develop, characterize and assess the properties of tricalcium silicate cement with alternative radiopacifiers, which are either inter-ground or sintered to the tricalcium silicate cement. METHODS: Custom-made endodontic cements based on tricalcium silicate and 20 % barium, calcium or strontium zirconate, which were either inter-ground or sintered at high temperatures, were produced. The set materials stored for 28 days in Hank's balanced salt solution were characterized by scanning electron microscopy and X-ray diffraction analysis. Assessment of pH, leaching, interaction with physiological solution, radiopacity, setting time, compressive strength and material porosity were investigated. Mineral trioxide aggregate (MTA) Angelus was used as control. RESULTS: Addition of radiopacifying materials improved the radiopacity of the material. The sintered cements exhibited the formation of calcium zirconate together with the respective radiopacifier phase. All materials produced calcium hydroxide on hydration, which interacted with tissue fluids forming hydroxyapatite on the material surface. The physical properties of the tricalcium silicate-based cements were comparable to MTA Angelus. CONCLUSIONS: A novel method of producing radiopaque tricalcium silicate-based cements was demonstrated. The novel materials exhibited properties, which were either comparable or else improved over the control. CLINICAL RELEVANCE: The novel materials can be used to replace MTA for root-end filling, perforation repair and other clinical applications where MTA is indicated.

Produção de IL-1beta, IL-6 e IL-8 por fibroblastos estimulados por cimentos endodônticos e submetidos à terapia laser de baixa potência / IL-1beta, IL-6 and IL-8 fibroblasts production stimulated by endodontic sealers and submitted to Low-Level Laser Therapy

O sucesso do tratamento endodôntico depende da cuidadosa realização de todas as suas fases, terminando com uma obturação tridimensional que alcance todo o sistema de canais radiculares. Desta forma, os materiais obturadores, ou as substâncias liberadas, entrarão em contato com os tecidos perirradiculares, o que poderá influenciar a resposta inflamatória e o processo de reparo. A terapia laser de baixa potência (TLBP) tem sido estudada quanto à sua ação anti-inflamatória, favorecendo o reparo. O objetivo deste estudo foi investigar a produção das citocinas IL-1β, IL-6 e IL-8 por fibroblastos de gengiva humana (linhagem FMM1) como resposta à presença dos extratos dos cimentos endodônticos AH Plus, MTA Fillapex e EndoSequence BC Sealer, bem como a eficácia da TLBP, neste modelo. Para isto, extratos destes cimentos, recém-manipulados e após 24 h do endurecimento, foram preparados em meio de cultura DMEM fresco, conforme as normas ISO 10993-12. Inicialmente, a citotoxicidade dos cimentos foi avaliada, após a interação das células com a diluição seriada destes extratos (1:1 a 1:16), por meio do ensaio MTT. Para a análise da produção de citocinas, 106 células por poço foram cultivadas em placas de cultura de 24 poços, para a interação com os extratos dos cimentos, na diluição 1:4. Estabeleceram-se os grupos não irradiado e irradiado. No grupo irradiado, as culturas celulares receberam duas irradiações do laser InGaAlP (660 nm, 30 mW, 5 J/cm2 e área do feixe de 0,028 cm2), com intervalo de 12 h. O grupo não irradiado foi submetido às mesmas condições ambientais que o irradiado. Os sobrenadantes das culturas foram coletados, centrifugados, aliquotados e armazenados congelados, para a posterior análise pelo ensaio de ELISA. Todos os dados obtidos (médias ± erro padrão) foram tratados estatisticamente por ANOVA one-way, complementado pelo teste de Tuckey e ANOVA...

Endodontic treatment success depends on careful accomplishment of all steps, finishing with a three-dimensional obturation that reachs the entire root canal system. Thus, the obturation materials or the substances that are released will be in contact with perirradicular tissues, which may influence the inflammatory response and the repair process. Low-level laser therapy (LLLT) has been studied about its anti-inflammatory action, improving the repair. The aim of this study was to investigate the production of IL-1β, IL-6 and IL-8 cytokines by human gingival fibroblasts (FMM1 lineage) as a response to the extracts of the endodontic cements AH Plus, MTA Fillapex and EndoSequence BC Sealer, besides the effectiveness of LLLT in this model. For that, freshly mixed and 24 h-set elutes of these sealers were prepared, in fresh DMEM, in accordance with ISO 10993-12. Firstly, cytotoxicity was assessed after cells were treated with cements extracts serial dilution (1:1 to 1:16), by MTT assay. To evaluate the cytokines production, 106 cells per well were cultured at 24-well-plates to interact with the 1:4 dilution of sealers extracts. Non-irradiated and irradiated groups were established. On the irradiated group, the cell culture received two InGaAlP laser irradiation (660 nm, 30 mW, 5 J/cm2, 0.028 cm2 spot size) at 12h-interval. The non-irradiated group was submitted to the same environment conditions of the irradiated group. The culture supernatants were collected, centrifuged, aliquoted and stored frozen to further analyse by ELISA assay. Data obtained (media ± standard error) were treated by ANOVA one-way, complemented by Tuckey’s test and ANOVA two-way, with Bonferroni correction (p< 0.05). The cytotoxicity of AH Plus and EndoSequence BC Sealer was related to time/dose-dependent manner, whilst MTA Fillapex cytotoxicity was showed...

Produção de IL-1beta, IL-6 e IL-8 por fibroblastos estimulados por cimentos endodônticos e submetidos à terapia laser de baixa potência / IL-1beta, IL-6 and IL-8 fibroblasts production stimulated by endodontic sealers and submitted to Low-Level Laser Therapy

O sucesso do tratamento endodôntico depende da cuidadosa realização de todas as suas fases, terminando com uma obturação tridimensional que alcance todo o sistema de canais radiculares. Desta forma, os materiais obturadores, ou as substâncias liberadas, entrarão em contato com os tecidos perirradiculares, o que poderá influenciar a resposta inflamatória e o processo de reparo. A terapia laser de baixa potência (TLBP) tem sido estudada quanto à sua ação anti-inflamatória, favorecendo o reparo. O objetivo deste estudo foi investigar a produção das citocinas IL-1β, IL-6 e IL-8 por fibroblastos de gengiva humana (linhagem FMM1) como resposta à presença dos extratos dos cimentos endodônticos AH Plus, MTA Fillapex e EndoSequence BC Sealer, bem como a eficácia da TLBP, neste modelo. Para isto, extratos destes cimentos, recém-manipulados e após 24 h do endurecimento, foram preparados em meio de cultura DMEM fresco, conforme as normas ISO 10993-12. Inicialmente, a citotoxicidade dos cimentos foi avaliada, após a interação das células com a diluição seriada destes extratos (1:1 a 1:16), por meio do ensaio MTT. Para a análise da produção de citocinas, 106 células por poço foram cultivadas em placas de cultura de 24 poços, para a interação com os extratos dos cimentos, na diluição 1:4. Estabeleceram-se os grupos não irradiado e irradiado. No grupo irradiado, as culturas celulares receberam duas irradiações do laser InGaAlP (660 nm, 30 mW, 5 J/cm2 e área do feixe de 0,028 cm2), com intervalo de 12 h. O grupo não irradiado foi submetido às mesmas condições ambientais que o irradiado. Os sobrenadantes das culturas foram coletados, centrifugados, aliquotados e armazenados congelados, para a posterior análise pelo ensaio de ELISA. Todos os dados obtidos (médias ± erro padrão) foram tratados estatisticamente por ANOVA one-way, complementado pelo teste de Tuckey e ANOVA ...

Endodontic treatment success depends on careful accomplishment of all steps, finishing with a three-dimensional obturation that reachs the entire root canal system. Thus, the obturation materials or the substances that are released will be in contact with perirradicular tissues, which may influence the inflammatory response and the repair process. Low-level laser therapy (LLLT) has been studied about its anti-inflammatory action, improving the repair. The aim of this study was to investigate the production of IL-1β, IL-6 and IL-8 cytokines by human gingival fibroblasts (FMM1 lineage) as a response to the extracts of the endodontic cements AH Plus, MTA Fillapex and EndoSequence BC Sealer, besides the effectiveness of LLLT in this model. For that, freshly mixed and 24 h-set elutes of these sealers were prepared, in fresh DMEM, in accordance with ISO 10993-12. Firstly, cytotoxicity was assessed after cells were treated with cements extracts serial dilution (1:1 to 1:16), by MTT assay. To evaluate the cytokines production, 106 cells per well were cultured at 24-well-plates to interact with the 1:4 dilution of sealers extracts. Non-irradiated and irradiated groups were established. On the irradiated group, the cell culture received two InGaAlP laser irradiation (660 nm, 30 mW, 5 J/cm2, 0.028 cm2 spot size) at 12h-interval. The non-irradiated group was submitted to the same environment conditions of the irradiated group. The culture supernatants were collected, centrifuged, aliquoted and stored frozen to further analyse by ELISA assay. Data obtained (media ± standard error) were treated by ANOVA one-way, complemented by Tuckey’s test and ANOVA two-way, with Bonferroni correction (p< 0.05). The cytotoxicity of AH Plus and EndoSequence BC Sealer was related to time/dose-dependent manner, whilst MTA Fillapex cytotoxicity was showed ...