Effect of two different storage media of avulsed tooth on viability and apoptosis of human gingival fibroblast: An in vitro study.

Abstract

Objective:This study aimed to compare the effect of HBSS & almond on viability and apoptosis of human gingival fibroblast in vitro.Materials and Methods:The storage media were initially prepared and diluted in cell culture Dulbecco’s modified eagle medium (DMEM) and IC50 was calculated. After incubation and detaching of fibroblast with 0.25% trypsin without EDTA, cells were distributed on the used siliconized tubes and centrifuged at 335 x g for 10 minutes then suspended in 2 mL 1 x phosphate buffered saline. Apoptosis was determined using Annexin V-FITC apoptosis with 2 fluorescent channels flowcytometry. After staining, cells were injected via ACEA flowcytometer and analyzed for FITC and PI fluorescent signals using FL1 and FL2 signal detector, respectively for each sample, 12,000 events were acquired and positive FITC and/or PI cells was quantified using ACEA NovoExpress™ software.Results:According to Duncan multiple range tests, in necrotic phase, early apoptosis phase and late apoptosis phase, HBSS was highly significant to control and almond milk, while in normal intact cells phase almond milk showed the highest significant Q3 apoptotic cells against both control and HBSS. Moreover, HBSS also showed a significant to control, and almond milk as revealed by DMRTs.Conclusion:Within the limitations of this study, it was concluded that almond milk has the ability to maintain viability of PDL cells, so may be considered as a viable option when other storage mediums as HBSS are not available at the site of accident.