Effects of ethylenediaminetetraacetic acid on stem cells from the apical papilla: In vitro study

BACKGROUND/PURPOSE: Ethylenediaminetetraacetic acid (EDTA) is used as an irrigant in regenerative endodontic treatment. The present study aimed to investigate the effects of EDTA on stem cells from apical papilla (SCAPs) in vitro. MATERIALS AND METHODS: Human SCAPs were isolated and characterised. The cells were treated with media supplemented with EDTA at concentrations ranging from 1.25% to 17%. Cell proliferation and apoptosis were examined using MTT assay and annexin V/propidium iodide staining. Cell migration was determined by a scratch assay. Gene expression was evaluated using a real-time polymerase chain reaction. Mineral deposition, a hallmark of osteogenesis in vitro, was determined using alizarin red s staining. RESULTS: Overall, SCAPs exhibited mesenchymal stem cell characteristics. EDTA treatment at 2.50% and 1.25% did not significantly exhibit cytotoxicity and alter cell morphology. However, EDTA attenuated cell proliferation and reduced MKI67 mRNA expression in SCAPs. Further, EDTA significantly induced early cell apoptosis at 48 h. Cell migration was delayed with EDTA treatment. After maintaining SCAPs in an osteogenic induction medium, EDTA diminished mineral deposition by SCAPs on day 14. CONCLUSION: EDTA treatment exhibits adverse effects on SCAPs in vitro. Hence, EDTA exposure to periapical tissues should be avoided to minimise the negative impacts on SCAPs cells in regenerative processes.