Cytotoxicity and genotoxicity evaluation of AH Plus, Well-Root ST, and AH Plus Bioceramic Sealer.

backgroundThis study aimed to evaluate the cytotoxicity and genotoxicity of AH Plus, Well-Root ST, and AH Plus Bioceramic Sealer on osteoblast cells.

methodsTo prepare AH Plus, equal amounts of paste A and paste B were mixed following the manufacturer's instructions. The AH Plus BCS and Well Root ST syringes were fitted with appropriate cannulas. Root canal sealers (RCSs) prepared in a laminar flow cabinet under aseptic conditions were placed in a single layer in 24-well plates with a circular shape of 13 mm diameter. To simulate contact between cells and RCSs, materials were diluted in culture media and the resulting extract was then applied to the cells. The initial extracts (1:1) were then serially diluted with culture medium at ratios of 1:2, 1:4, 1:8, and 1:16. To determine the cytotoxic effects of RCSs on SAOS-2 cells, 20,000 cells were seeded in each well of 24-well culture plates. The cells were incubated for periods of 0, 8, 12, 24, 48 and 72 h with or without different concentrations of RCSs (1:1, 1:2, 1:4, 1:8, and 1:16 dilutions). Each experimental group was replicated three times to ensure reliable results and to calculate the standard deviation. Cell viability was assessed using the trypan blue exclusion method, which differentiates live (unstained) and dead (blue-stained) cells. TUNEL assay for apoptosis assessment was performed to evaluate genotoxicity. All statistical analyses were performed using the GraphPad software (Prism ver.10.0 San Diego, California, USA). Differences between RCS groups were assessed using One-way ANOVA analysis. Tukey's test was used to compare at a significance level of p < 0.05.

resultsCell viability was affected with 1:1 and 1:2 dilutions of any of the RCSs from 24 to 72 h. 1:4, 1:8 and 1:16 concentrations were not cytotoxic at any time points. Cytotoxic effects of AH Plus were higher at 24, 48, and 72 h compared to the Well-Root ST and AH Plus BCS (p < 0.0001). In terms of genotoxicity, both 1:1 and 1:2 dilutions of all RCSs showed significant differences at 48th hours compared to the controls. The toxic effect at the genetic level was significantly increased in the AH Plus group compared to the bioceramic sealers at 1:1 dilution (p = 0,0188 and p = 0,0331 for Well-Root ST and AH Plus BCS, respectively).

conclusionAlthough genotoxic and cytotoxic effects were observed in all RCSs, the toxicity of AH Plus was higher than Well-Root ST and AH Plus Bioceramic Sealer.