The efficacy of commercial tooth storage media for maintaining the viability of human periodontal ligament fibroblasts

Abstract

Aim

To evaluate Save-A-Tooth (SAT), EMT Toothsaver (EMT), and Hank's Balanced Salt Solution (HBSS) for their influence on the viability and proliferative capacity of human periodontal ligament fibroblast (HPDLF).

Methodology

Primary HPDLFs were seeded on 96-well cell culture plates and exposed to SAT, EMT, HBSS and water (negative control) for 0.5, 1, 3, 6, 12 and 24 hours at room temperature (22°C). After each exposure time, cell viability was measured by quantifying adenosine triphosphate (ATP) using a luminescent dye. The proliferative capacity was also quantified using the prestoblue assay after 12 or 24 hours storage in each medium. The data was analyzed statistically by two-way ANOVA and post hoc Least Significant Difference (LSD) test (P < 0.05). The morphology of the cells after 12 hours storage was also investigated through live/dead viability/cytotoxicity kit together with fluorescence microscopy.

Results

There was no significant difference in cell viability among HBSS, SAT and EMT groups up to 6 hours. SAT was effective in maintaining cell viability only up to 12 hours and then became detrimental to HPDLF; after 24 hours, the effectiveness of SAT in maintaining cell viability was similar to that of water (p > 0.05). Among all the media, only EMT could maintain the proliferative capacity of HPDLFs significantly higher than the negative control, i.e. water (p <0.05) after 24 hours storage.

Conclusions

EMT maintained the proliferative capacity of HPDLFs after 24 hours storage.

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