Evaluation of cytocompatibility of calcium silicate-based endodontic sealers and their effects on the biological responses of mesenchymal dental stem cells.

AIM: To investigate in vitro the cytocompatibility of the calcium silicate-containing endodontic sealers MTA Fillapex and TotalFill BC Sealer on human periodontal ligament stem cells (hPDLSCs) by assaying their biological responses and compare them with that observed when using an epoxy resin-based sealer (AH Plus). METHODOLOGY: Specimens from the three different endodontic sealers were eluated with culture medium for 24 h. The cytotoxicity of these eluates was evaluated using the MTT assay. In addition, an in vitro scratch wound healing model was used to determine their effects on cell migration. Cell adhesion to collagen type I after treatment with the different sealer eluates was also measured, whereas cytotoxicity was determined using the DNA-specific fluorochrome Hoechst 33342. Finally, to assess cell morphology and attachment to the different sealers, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). One-way analysis of variance (anova) followed by a Bonferroni post-test were performed (P < 0.05). RESULTS: hPDLSCs exposed to different dilutions of TotalFill BC Sealer eluates had significantly higher cell proliferation compared with that observed when cells were treated with AH Plus and MTA Fillapex eluates (P < 0.001). In addition, TotalFill eluates were associated with significantly increased cell adhesion to collagen type I and migration of hPDLSCs in a concentration-dependent manner than displayed after treatment with MTA Fillapex or AH Plus eluates (P < 0.001). Moreover, TotalFill BC Sealer-induced cytotoxicity was significantly lower than observed using AH Plus and MTA Fillapex eluates (P < 0.001). Finally, SEM studies revealed suitable proliferation, cell spreading and attachment, especially when using TotalFill BC Sealer discs. CONCLUSION: TotalFill BC Sealer exhibited a higher cytocompatibility than AH Plus and MTA Fillapex. Further investigations using in vivo animal models are required to validate the potential biological responses of TotalFill BC Sealer on hPDLSCs.

Biocompatibility of three new calcium silicate-based endodontic sealers on human periodontal ligament stem cells.

AIM: To evaluate the biocompatibility of three calcium silicate-based endodontic sealers, Bioroot BC Sealer (Septodont, Saint-Maur-des-Fosses, France), Endoseal MTA (EndoSeal, Maruchi, Seoul, Korea) and Nano-ceramic Sealer (B&L Biotech, Fairfax, VA, USA) (NCS), on human periodontal ligament stem cells (hPDLSCs). METHODOLOGY: Human periodontal ligament stem cells were cultured in the presence of various endodontic sealer eluates for 24 h. Cell viability was determined using the MTT assay. Cell death and changes in phenotype induced by the set endodontic sealer eluates were evaluated through flow cytometry. Also, an in vitro scratch wound-healing model was used to determine their effects in cell migration. Finally, to assess cell morphology and attachment to the different sealers, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). One-way analysis of variance (anova) followed by a Bonferroni post-test was performed (P < 0.05). RESULTS: At 24 h, cell spreading was evident in the presence of Bioroot BC Sealer (BR) and Nano-ceramic Sealer (NCS), but not Endoseal MTA (ES). At 72 h, BR and NCS exhibited high and moderate cell proliferation, respectively, whereas ES revealed low rates of cell proliferation (P < 0.05). Similar results were obtained in a cell death assay. In addition, hPDLSCs maintained their mesenchymal phenotype in all conditions although their capacity to migrate was higher in the presence of BR. Finally, SEM studies revealed a good degree of proliferation, cell spreading and attachment, especially when using BR and NCS discs. CONCLUSIONS: BR and NCS were associated with better cytocompatibility than ES. Further in vitro and in vivo investigations are required to confirm the suitability of these calcium silicate-based endodontic sealers for clinical application.

Cytokine Production and Cytotoxicity of Calcium Silicate-based Sealers in 2- and 3-dimensional Cell Culture Models.

INTRODUCTION: The aim of the present study was to assess the effects of different silicate-based sealers (ie, BioRoot RCS [Septodont, Saint Maur des Fosses, France], ProRoot ES [Dentsply Sirona, York, PA], and MTA Fillapex [Angelus, Londrina, PR, Brazil]) on cytokine production and viability of human periodontal ligament stem cells (PDLSCs). AH Plus (Dentsply DeTrey GmbH, Konstanz, Germany) was used as a reference material. METHODS: PDLSCs were cultured either in 2-dimensional or 3-dimensional conditions (in 0.15%-0.5% PuraMatrix [BD Biosciences, Bedford, MA]) for 24 hours with eluates from set endodontic sealers. Additionally, the toxicity of eluates from endodontic sealers was evaluated using an in vitro root model experimental procedure. PDLSC viability was determined using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. PDLSC culture medium was used for cytokine quantification (interleukin [IL]-6, IL-8, growth-regulated oncogene, IL,-4 and IL-10) using the HCYTMAG-60K-PX41 Milliplex kit (EMD Millipore, Burlington, MA). RESULTS: In 2-dimensional culture conditions, BioRoot RCS revealed a good PDLSC viability rate. ProRoot ES had no effect on PDLSC viability regardless of the dilution. MTA Fillapex was strongly cytotoxic even at the lowest extract dilutions (1:1, 1:2, and 1:4). Encapsulation of PDLSCs in PuraMatrix tended to decrease the cytotoxic effect of the sealers. In the 3-dimensional in vitro root model experimental procedure, BioRoot RCS, ProRoot ES, and MTA Fillapex revealed a cytocompatibility pattern. Different calcium silicate-based sealers exhibited different proinflammatory cytokine production. BioRoot RCS greatly stimulated the release of IL-10 and, to a lesser degree, IL-4 by PDLSCs (P < .05). CONCLUSIONS: BioRoot RCS and ProRoot ES did not induce proinflammatory cytokines and promoted anti-inflammatory cytokine secretion by PDLSCs that may have a positive local impact by attenuating an initial inflammatory response.

Cytotoxicity of GuttaFlow Bioseal, GuttaFlow2, MTA Fillapex, and AH Plus on Human Periodontal Ligament Stem Cells.

INTRODUCTION: The aim of the present study was to evaluate the in vitro cytotoxicity of endodontic sealers (GuttaFlow Bioseal, GuttaFlow2, and MTA Fillapex) on human periodontal ligament stem cells (hPDLSCs). As a reference, AH Plus was compared with the more recent endodontic sealers regarding cell viability and cell attachment. METHODS: Biological testing was carried out in vitro on hPDLSCs. Cell viability assay was performed by using eluates from each endodontic sealer. To assess cell morphology and attachment to the different sealers, the hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. Chemical composition of the sealers was determined by energy-dispersive x-ray, and eluates were analyzed by inductively coupled plasma mass spectrometry. Statistical differences were assessed by analysis of variance and Tukey test (P < .05). RESULTS: Cell viability was evident after 24 hours in the presence of GuttaFlow Bioseal and GuttaFlow 2 but not in the case of AH Plus or MTA Fillapex. At 168 hours, GuttaFlow Bioseal and GuttaFlow 2 exhibited high and moderate cell viability, respectively, whereas AH Plus and MTA Fillapex revealed low rates of cell cell viability (P < .001). Finally, scanning electron microscopy studies revealed a high degree of proliferation, cell spreading, and attachment, especially when using GuttaFlow Bioseal disks. CONCLUSIONS: GuttaFlow Bioseal and GuttaFlow2 showed lower cytotoxicity than MTA Fillapex and AH plus. Further in vitro and in vivo investigations are required to confirm the suitability of GuttaFlow Bioseal for clinical application.