Cytotoxicity of two concentrations of a dentine bonding agent on mouse 3T3 and human pulp fibroblast cell-lines.

The objective of this study was to investigate the cytotoxicity and effect of concentration of a recent dentine bonding agent on a mouse fibroblast cell-line (3T3) and four different human pulp fibroblast cell-lines. A mouse fibroblast 3T3 cell-line and 4 different human pulp fibroblast cell lines were used. The cells were grown and subcultured using standard conditions. For the testing of the cytotoxicity of the bonding agent (Prime & Bond NT) it was extracted with DMEM medium and the different cell-lines were exposed to the extraction. The MTT assay was used to establish the number of viable cells. For all 5 pairs (control vs test sample) of cell-lines for 4 microl bonding agent, the mouse and only two of the four human cell-lines showed a statistical significant difference (p < 0.05; Mann-Whitney) due to the cytotoxicity of the bonding agent. However, all 5 pairs showed significant differences (p < 0.05) at the 8 microl concentration level. To investigate the relative effect of concentration (4 microl compared to 8 microl) of a chemical agent (we used a bonding agent, Prime & Bond NT) on the cytotoxicity of these 5 cell-lines, the Kruskal-Wallis Multiple-Comparison test was used on the standardised medians. Statistically significant differences (p < 0.05) were found for various cell-lines between both concentration levels. In general, the tested bonding agent, at a higher concentration level, has an increased cytotoxic effect on all 5 cell-lines. About 69% (median) of 3T3 cells stayed viable with 4 microl (0.94cm2/ml) bonding agent and 61% when 8 microl was used. For the pulp cells the percentages were between 91% and 72% for 4 microl and between 77% and 24% for 8 microl. These findings indicated that the bonding agent Prime and Bond NT was cytotoxic and that different human pulp fibroblast cell-lines should be used in the cytotoxic testing of dental materials and secondly, if only one concentration is to be used it should be in the higher range of that suggested by the ANSI/ADA document (2000, Revision) for recommended standard practices for biological evaluation of dental materials.