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.

The aim of this study was to assess the biological properties and mineralization potential of the new Theracal PT (Bisco Inc, Schaumburg, IL) compared with its predecessor Theracal LC (Bisco Inc) and the hydraulic silicate-based cement Biodentine (Septodont, Saint-Maur-des-Fossés, France) on human dental pulp stem cells (hDPSCs) in vitro. Standardized sample discs were obtained for each material (n = 30) together with 1:1, 1:2, and 1:4 material eluates. Previously characterized hDPSCs were cultured with the different materials in standardized conditions, and the following assays were performed: a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, a wound healing assay, Annexin-V-FITC and 7-AAD staining (BD Biosciences, San Jose, CA), reactive oxygen species production analysis, cell adhesion and morphology evaluation via scanning electron microscopy and immunofluorescence, quantification of the expression of osteo/odontogenic markers via real-time quantitative reverse-transcriptase polymerase chain reaction, and alizarin red S staining. Statistical significance was established at P < .05. All of the tested dilutions of Theracal LC exhibited a significantly higher cytotoxicity and reactive oxygen species production (P < .001) and a lower cell migration rate than the control group (hDPSCs cultured in growth medium without material extracts) at all of the measured time points (P < .001). Both 1:4 Theracal PT and Biodentine-treated hDPSCs exhibited similar levels of cytocompatibility to that of the control group, a significant up-regulation of at least 1 odontogenic marker (Biodentine: dentin sialophosphoprotein (P < .05); Theracal PT: osteonectin and runt-related transcription factor 2 [P < .001]), and a significantly higher mineralized nodule formation (P < .001). The newly introduced TheraCal PT offers an improved in vitro cytocompatibility and mineralization potential on hDPSCs compared with its predecessor, TheraCal LC, and comparable biological properties to Biodentine.

Background: Most recently, the biological interactions, that is cytocompatibility, cell differentiation and mineralization potential, between calcium silicate‐based biomaterials and periodontal ligament stem cells (PDLSCs) have been studied at an in vitro level, in order to predict their clinical behaviour during endodontic procedures involving direct contact with periodontal tissues, namely root canal treatment, endodontic surgery and regenerative endodontic treatment.Objective: The aim of the present systematic review was to present a qualitative synthesis of available in vitro studies assessing the biological interaction of PDLSCs and calcium silicate‐based biomaterials.Methodology: The present review followed PRISMA 2020 guidelines. An advanced database search was performed in Medline, Scopus, Embase, Web of Science and SciELO on 1 July 2020 and last updated on 22 April 2021. Studies assessing the biological interactions of PDLSCs with calcium silicate‐based sealers (CSSs) and/or cements (CSCs) at an in vitro level were considered for inclusion. The evaluation of the ‘biological interaction’ was defined as any assay or test on the cytotoxicity, cytocompatibility, cell plasticity or differentiation potential, and bioactive properties of PDLSCs cultured in CSC or CSS‐conditioned media. Quality (risk of bias) was assessed using a modified CONSORT checklist for in vitro studies of dental materials.Results: A total of 20 studies were included for the qualitative synthesis. CSCs and CSSs, as a group of endodontic materials, exhibit adequate cytocompatibility and favour the osteo/cementogenic differentiation and mineralization potential of PDLSCs, as evidenced from the in vitro studies included in the present systematic review.Discussion: The influence of the compositional differences, inclusion of additives, sample preparation, and varying conditions and manipulations on the biological properties of calcium silicate‐based materials remain a subject for future research.Conclusions: Within the limitations of the in vitro nature of the included studies, this work supports the potential use of calcium silicate‐based endodontic materials in stem cell therapy and biologically based regenerative endodontic procedures.Registration: OSF Registries; https://doi.org/10.17605/OSF.IO/SQ9UY.

Dentistry-applied bioceramic materials are ceramic materials that are categorized as bioinert, bioactive and biodegradable. They share a common characteristic of being specifically designed to fulfil their function; they are able to act as root canal sealers, cements, root repair or filling materials. Bioactivity is only attributed to those materials which are capable of inducing a desired tissue response from the host. The aim of this study is to present a systematic review of available literature investigating bioactivity of dentistry-applied bioceramic materials towards dental pulp stem cells, including a bibliometric analysis of such a group of studies and a presentation of the parameters used to assess bioactivity, materials studied and a summary of results. The research question, based on the PICO model, aimed to assess the current knowledge on dentistry-based bioceramic materials by exploring to what extent they express bioactive properties in in vitro assays and animal studies when exposed to dental pulp stem cells, as opposed to a control or compared to different bioceramic material compositions, for their use in the dentin-pulp complex therapy. A systematic search of the literature was performed in six databases, followed by article selection, data extraction, and quality assessment. Studies assessing bioactivity of one or more bioceramic materials (both commercially available or novel/experimental) towards dental pulp stem cells (DPSCs) were included in our review. A total of 37 articles were included in our qualitative review. Quantification of osteogenic, odontogenic and angiogenic markers using reverse transcriptase polymerase chain reaction (RT-PCR) is the prevailing method used to evaluate bioceramic material bioactivity towards DPSCs in the current investigative state, followed by alkaline phosphatase (ALP) enzyme activity assays and Alizarin Red Staining (ARS) to assess mineralization potential. Mineral trioxide aggregate and Biodentine are the prevalent reference materials used to compare with newly introduced bioceramic materials. Available literature compares a wide range of bioceramic materials for bioactivity, consisting mostly of in vitro assays. The desirability of this property added to the rapid introduction of new material compositions makes this subject a clear candidate for future research.

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.

Biocompatibility is an essential property for any vital pulp material that may interact with the dental pulp tissues. Accordingly, this study aimed to compare the chemical composition and ultrastructural morphology of Biodentine (Septodont, Saint Maur-des-Fosses, France), ProRoot MTA (Dentsply Tulsa Dental Specialties, Johnson City, TN, USA), and Bio-C Repair (Angelus, Londrina, PR, Brazil), as well as their biological effects on human dental pulp cells. Chemical element characterization of the materials was undertaken using scanning electron microscopy and energy dispersive X-ray analysis (SEM-EDX). The cytotoxicity was assessed by analyzing the cell viability (MTT assay), cell morphology (immunofluorescence assay), and cell attachment (flow cytometry assay). The results were statistically analyzed using ANOVA and Tukey’s test (p < 0.05). EDX revealed that ProRoot MTA and Biodentine were mostly composed of calcium, carbon, and oxygen (among others), whereas Bio-C Repair evidenced a low concentration of calcium and the highest concentration of zirconium. SEM showed adequate attachment of human dental pulp cells (hDPCS) to vital pulp materials and cytoskeletal alterations were not observed in the presence of material eluates. Remarkably, the undiluted Biodentine group showed higher viability than the control group cells (without eluates) at 24 h, 48 h, and 72 h (p < 0.001). Based on the evidence derived from an in vitro cellular study, it was concluded that Bio-C Repair showed excellent cytocompatibility that was similar to Biodentine and ProRoot MTA.