IADR Abstract Archives
Synergistic Effect of HBSS and DMEM for Avulsed Teeth Preservation
Objectives: An avulsed tooth should be placed in storage media until replantation. The formulation of the storage medium must provide viability to the periodontal ligament cells. The objective of this study was to evaluate the synergistic effect of Hank’s Balanced Salt Solution (HBSS) and Dulbecco’s Modified Eagle’s Medium (DMEM) on maintaining viability of human periodontal ligament fibroblasts (HPDLFs).
Methods: The storage media assessed were HBSS, DMEM (containing 4mM L-Glutamine) and equivolume mixture of both (DMEM-HBSS). Deionized water was selected as negative control. HPDLFs underwent trypsinization and counting by the trypan blue exclusion method. The cells were seeded into 96-well cell culture plates and underwent 24-hour incubation at 37°. The cell culture medium in each well was replaced with 100 μl of one of the storage media. The cells were incubated at 25°C or 4°C while in contact with the media up to 48 hours. Cell viability was measured in increments using MTT assay.
Results: For 25°C and 4°C, viability decreased overtime for all samples and water was detrimental to the cells. Both temperatures also showed that DMEM and DMEM-HBSS maintained more than 60% of viable cells up to 10 hours. Only DMEM-HBSS kept more than 50% of viable cells after 24 hours, and more than 25% after 48 hours for 25°C. In 4°C, 25% were viable after 24 hours for DMEM-G-HBSS. Cell viability dropped to 50% for HBSS after 2 hours, but the remaining cells survived up to 10 hours in 25°C. DMEM maintained around 50% viable cells in 25°C and 40% viable cells in 4°C for 10 hours, but after that viability decreased substantially.
Conclusions: Overall, temperature differences did not have a significant effect on the preservation capability of the media tested. DMEM-HBSS showed the longest cell preservation time of 48 hours supporting the synergistic effect of two media.
Methods: The storage media assessed were HBSS, DMEM (containing 4mM L-Glutamine) and equivolume mixture of both (DMEM-HBSS). Deionized water was selected as negative control. HPDLFs underwent trypsinization and counting by the trypan blue exclusion method. The cells were seeded into 96-well cell culture plates and underwent 24-hour incubation at 37°. The cell culture medium in each well was replaced with 100 μl of one of the storage media. The cells were incubated at 25°C or 4°C while in contact with the media up to 48 hours. Cell viability was measured in increments using MTT assay.
Results: For 25°C and 4°C, viability decreased overtime for all samples and water was detrimental to the cells. Both temperatures also showed that DMEM and DMEM-HBSS maintained more than 60% of viable cells up to 10 hours. Only DMEM-HBSS kept more than 50% of viable cells after 24 hours, and more than 25% after 48 hours for 25°C. In 4°C, 25% were viable after 24 hours for DMEM-G-HBSS. Cell viability dropped to 50% for HBSS after 2 hours, but the remaining cells survived up to 10 hours in 25°C. DMEM maintained around 50% viable cells in 25°C and 40% viable cells in 4°C for 10 hours, but after that viability decreased substantially.
Conclusions: Overall, temperature differences did not have a significant effect on the preservation capability of the media tested. DMEM-HBSS showed the longest cell preservation time of 48 hours supporting the synergistic effect of two media.