Effects of EDTA on Proliferation, Migration, and Differentiation of SCAP

Objectives: Ethylenediaminetetraacetic acid (EDTA), a chelating agent, is used as an irrigant in regenerative endodontic treatment. However, it is not known if EDTA has deleterious effects on stem cells from the apical papilla (SCAPs). The aim of the present study was to investigate the effects of EDTA on SCAPs.
Methods: SCAPs were isolated and characterized. The cells were treated at concentration of EDTA range from 1.25 to 17% for 30 and 60 seconds. Cell proliferation and apoptosis was examined using an MTT assay and annexin V/propidium iodide staining, respectively. Cell migration was determined using a scratch assay. Gene expression was evaluated using real-time polymerase chain reaction. Osteogenic differentiation was examined using alizarin red s staining.
Results: The isolated cells expressed CD44, CD90, CD105 and lacked CD45 expression. The cells differentiated into osteogenic and adipogenic lineages, confirming the stem cell characteristics of SCAPs. EDTA at 2.50% and 1.25% attenuated cell proliferation. Correspondingly, reduced MKI67 mRNA expression was observed in the EDTA-treated condition at 24 h. However, there was no significant change in cell apoptosis. EDTA-treated SCAPs demonstrated reduced cell migration in vitro at 24 and 48 hours. In addition, EDTA treatment resulted in decreased mineral deposition by SCAPs after osteogenic induction for 14 days.
Conclusions: Despite the beneficial use of EDTA in clinical regenerative endodontic treatment, EDTA exhibited a negative effect on cell proliferation, migration and osteogenic differentiation in SCAPs in vitro. Hence, further studies required to determine the mechanism underlying the effect of EDTA on SCAPs.