Real-time Analysis of Cytotoxicity of Current Root-Canal Sealers on Cementoblasts

Objective: The aim of this study was to investigate the effects of a newly developed sealer based on calcium-fluoro-silicate cement TECHBiosealer Endo;TBE (Isasan, Italy) and compare with Hybrid Root Seal;HRS (MetaSeal; Sun Medical Co. Japan), SimpliSeal;SS (DiscusDental, LLC), RealSeal;RS (SybronEndo, Orange, CA, USA) and AH plus;AH (Dentsply, Germany) on cell viability and morphology of cementoblasts. Methods: 2x10 mm cylinders of each material were fabricated, set for 2 days at 37ºC and sterilized with ethylene oxide. Materials were incubated in DMEM culture medium for 72 hrs according to ISO 10993-5 (surface area-to volume ratio of the specimen to cell-culture medium; 3cm2/ml). Immortalized cementoblasts (OCCM) were maintained with Dulbecco modified Eagle medium containing 10% fetal bovine serum. Real-time cell analyzer (RT-CES) was used to evaluate cell survival. After seeding 200µL of the cell suspensions into the wells (5000 cells/well) of the E-plate 96, OCCM.30 cells were treated with bioactive components released by the test sealers (1/2 and 1/4 dilutions) and were monitored every 15 min for a period for 96 hrs. Results: Statistical analysis (ANOVA-Tukey-Kramer/Multiple-Comparison tests) demonstrated that apparent increase in cell viability was observed in TBE(1/4) and AH(1/2,1/4) when compared to the control. TBE(1/2), RS(1/2,1/4) and HRS(1/2,1/4) groups showed a decrease in cell viability. SS(1/2,1/4) showed similar cell viability with control. Inverted microscopy examination revealed changed cell morphologies in the same groups TBE(1/2), RS(1/2 and 1/4), HRS(1/2 and 1/4) confirming real-time cell analysis results. Viability degrees for the materials were as follows: TBE(1/4)>AH(1/4)=AH(1/2)>control=SS(1/2)=SS(1/4)>HRS(1/4)>HRS(1/2)=TBE(1/2)=RS(1/4)=RS(1/2). Conclusions: Considering the close contact of root-canal sealers with cementoblasts at apical region, AH and SS can be used safely in the practice due to their good biocompatibility.