Objective: In Vitro cytotoxicity tests are widely used to evaluate a large range of dental materials. This study compared the cytotoxicity of the bonding agents ScotchBond 1, Prime & Bond NT and Xeno III on a Balb/c 3T3 mouse fibroblast cell-line. Methods: Dulbecco's Modified Eagle's Medium was exposed to a concentration of the various bonding agents for 24 hours, and then transferred into well plates containing near confluent 3T3 fibroblasts. Cell viability was compared to a similar number of controls without exposure to bonding agents using the MTT assay. Results: All three bonding agents were found to have a cytotoxic effect on 3T3 cells. Prime & Bond NT showed a survival rate of 62.4%, ScotchBond 58.8%, while for Xeno III only 25.3% of cells survived. No significant difference (Kruskal-Wallis Test, p>0.05) in the survival rate was found between Prime & Bond NT and ScotchBond 1. However, the survival rate of Xeno III was significantly less (p<0.01) than that of the other two. Xeno III reacts with dentine and results were true even when a barrier of thin etched human dentine discs (~120µm) was used instead of artificial membranes. An evaluation of the morphological appearance of the cells exposed to Xeno III also shows a more severe cytotoxic reaction. Conclusion: Although all 3 dentine bonding agents are cytotoxic, Xeno III is the most toxic even through a thin dentine barrier. The clinical signifance of this study is that the single step self-etching bonding agent was found to be more cytotoxic than ScotchBond 1 and Prime & Bond NT, but none of them should be used in deep cavities without a liner.

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