An In Vitro Evaluation of Ice Apple as a Novel Storage Medium to Preserve the Viability of Human Periodontal Ligament Fibroblasts

Abstract

Aim: For a better prognosis of an avulsed tooth, it is important to store it in an appropriate storage medium prior to replantation. The purpose of this study was to evaluate the ability of ice apples to preserve the viability of periodontal ligament (PDL) fibroblasts. Materials and methods: Periodontal ligament (PDL) fibroblasts were procured from the roots of healthy premolars and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM). They were preserved with—ice apple water (IAW), 30 and 10% ice apple fruit pulp extract (IAFPE), DMEM, negative control without any agent, and positive control DMEM supplemented with fetal bovine serum (FBS). Culture plates were incubated with investigational media at 37°C for 1, 3, and 24 hours. Each experiment was repeated thrice. Assessment of cell viability was done using 3-(4,5-dimethylthiazol-2-yl)—2,5-diphenyltetrazolium bromide tetrazolium reduction (MTT) assay. After every test period, storage media was removed; 60 µL of MTT solution was added to each well and incubated for 3 hours at 37°C. The supernatant was aspirated and the formazan blue crystals formed were solubilized using dimethyl sulphoxide (DMSO) (100 µL). Optical density was measured at a wavelength of 490 nm. The effects of the test storage media at each time period were evaluated by a one-way analysis of variance (ANOVA) test, and this was followed by post hoc Tukey’s multiple comparison tests (p < 0.05). Results: A total of 10% IAFPE demonstrated the maximum capacity to maintain PDL cell viability in all three test periods (p = 0.001). Among the different forms of ice apple used in this study, IAFPE showed better results as compared to IAW (p = 0.001). Conclusion: Ice apple fruit pulp extract (IAFPE) at 10% concentration showed the maximum capacity to maintain PDL cell viability across all three test periods. Therefore, it can be considered a suitable alternative natural storage medium for avulsed teeth. However, further, more detailed studies are required in this field.