In vitro analysis of the cytotoxicity and the antimicrobial effect of four endodontic sealers

<p>Abstract</p> <p>Introduction</p> <p>The aim of this study was to investigate <it>in vitro </it>the cytotoxicity and antibacterial properties of four different endodontic sealers using human periodontal ligament fibroblast cell proliferation and visual analysis of growth inhibition.</p> <p>Methods</p> <p>A silicone (GuttaFlow), silicate (EndoSequence BC), zinc oxide eugenol (Pulp Canal Sealer EWT) and epoxy resin (AH Plus Jet) based sealer were incubated with PDL fibroblasts (10⁴cells/ml, n = 6) up to 96 h. Cell proliferation (RFU) was determined by means of the Alamar Blue assay. Cell growth and morphology was visualized by means of fluorescent dyes. Possible antibacterial properties of the different sealers were visualized by means of SEM (<it>Enterococcus faecalis; Parvimonas micra</it>).</p> <p>Results</p> <p>Fibroblast proliferation depended on sealer and cultivation time. After 72 and 96 h GuttaFlow and EndoSequence BC showed relatively non-cytotoxic reactions, while Pulp Canal Sealer EWT and AH Plus Jet caused a significant decrease of cell proliferation (p < 0.001). Visualization of cell growth and morphology with various fluorescent dyes supplemented the results. No antibacterial effect of EndoSequence BC to <it>P. micra </it>was found, whereas GuttaFlow showed a weak, Pulp Canal Sealer EWT and AH Plus Jet extensive growth inhibition. Also, no antibacterial effect of GuttaFlow, EndoSequence BC or AH Plus Jet to <it>E. faecalis </it>could be detected.</p> <p>Conclusions</p> <p>These <it>in vitro </it>findings reveal that GuttaFlow and EndoSequence BC can be considered as biocompatible sealing materials. However, prior to their clinical employment, studies regarding their sealing properties also need to be considered.</p

Influence of light-curing mode on the cytotoxicity of resin-based surface sealants

Background Surface sealants have been successfully used in the prevention of erosive tooth wear. However, when multiple tooth surfaces should be sealed, the light-curing procedure is very time-consuming. Therefore, the aim of this study was to investigate whether reduced light-curing time (while maintaining similar energy density) has an influence on resin-based surface sealant cytotoxicity. Methods Bovine dentine discs were treated as follows: group 1: untreated, groups 2–5: Seal&Protect and groups 6–9: experimental sealer. Groups 2 and 6 were light-cured (VALO LED light-curing device) for 40 s (1000 mW/cm2), groups 3 and 7 for 10 s (1000 mW/cm2), groups 4 and 8 for 7 s (1400 mW/cm2) and groups 5 and 9 for 3 s (3200 mW/cm2). Later, materials were extracted in culture medium for 24 h, and released lactate dehydrogenase (LDH) activity as a measure of cytotoxicity was determined photometrically after cells (dental pulp cells and gingival fibroblasts) were exposed to the extracts for 24 h. Three independent experiments, for both sample preparation and cytotoxicity testing, were performed. Results Overall, lowest cytotoxicity was observed for the unsealed control group. No significant influence of light-curing settings on the cytotoxicity was observed (p = 0.537 and 0.838 for pulp cells and gingival fibroblasts, respectively). No significant difference in the cytotoxicity of the two sealants was observed after light-curing with same light-curing settings (group 2 vs. 6, 3 vs. 7, 4 vs. 8 and 5 vs. 9: p > 0.05, respectively). Conclusions Shortening the light-curing time, while maintaining constant energy density, resulted in no higher cytotoxicity of the investigated sealants

In vitro effect of low intensity laser on the cytotoxicity produced by substances released by bleaching gel

This in vitro study aimed to analyze the effect of different parameters of phototherapy with low intensity laser on the viability of human dental pulp fibroblasts under the effect of substances released by bleaching gel. Cells were seeded into 96 wells plates (1 x 10³ cells/well) and placed in contact with culture medium conditioned by a 35 % hydrogen peroxide bleaching gel for 40 minutes, simulating the clinical condition of the in-office bleaching treatment. Cells cultured in ideal growth conditions served as positive control group (PC), and the cells grown in conditioned medium and non-irradiated served as negative control group (NC). Cells grown in conditioned medium were submitted to a single irradiation with a diode laser (40 mW, 0.04 cm²) emitting at visible red (660 nm; RL) or near infrared (780 nm; NIR) using punctual technique, in contact mode and energy densities of 4, 6 or 10 J/cm². The cell viability was analyzed through the MTT reduction assay immediately and 24 hours after the irradiation. The data was compared by ANOVA followed by the Tukey's test (p < 0.05). The cell viability increased significantly in 24 hours within each group. The PC presented cell viability significantly higher than NC in both experimental times. Only the NIR/10 J/cm² group presented cell viability similar to that of PC in 24 hours. The phototherapy with low intensity laser in defined parameters is able to compensate the cytotoxic effects of substances released by 35 % hydrogen peroxide bleaching gel