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 silicatebased 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-2thiazolyl)-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 2dimensional 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.