A multiparametric assay to compare the cytotoxicity of different storage media for avulsed teeth

Aim: To evaluate the cytotoxicity of several storage media (coconut water, whole milk, isotonic solution, Hank ́s Balanced Salt Solution and tap water) using a multiparametric cytotoxicity analysis employing 3T3 cells. Methods: Plates containing confluent 3T3 fibroblasts were exposed to the various media for 24h, at 37°C with 5% CO2, and cell viability was evaluated by a multiparametric assay assessing sequentially, on the same cells, mitochondrial activity (XTT), membrane integrity (neutral red test), and total cell density (crystal violet dye exclusion test). Results from each test were compared by one-way analysis of variance (ANOVA). Results: Statistical analysis showed that whole milk and HBSS were the most effective media in maintaining cell viability at all tested times (p<0.05). Isotonic and tap water showed the highest cytotoxicity effects. Conclusions: This study shows that whole milk and HBSS are more efficient in maintaining the viability of 3T3 fibroblasts as demonstrated by three different cell viability tests.