Effect of ethylenediaminetetraacetic acid and hyaluronic acid on the viability and cytokine expression of periodontal ligament fibroblasts.

Abstract

Dental root cell proliferation following tooth avulsion has not been well researched. Understanding the effects of dry time and dentin treatment influences on cell proliferation is essential to provide evidence-based guidelines for tooth replantation. The study evaluated the viability of periodontal ligament fibroblasts (PLF) in contact with roots, submitted to surface treatments with ethylenediaminetetraacetic acid (EDTA) and hyaluronic acid (HA) at different times, including to quantify inflammatory cytokines interleukin (IL)-6, IL-8, IL-1β and TNF-α expressed by PLF. The adhesion of fibroblasts to treated root surfaces was also evaluated. One hundred and eight cementum discs from bovine teeth were randomly divided into groups according to time periods of being dry (n=12) as follows: (i) fresh discs that were not kept dry, (WD); (ii) dry for 1hour (1hd); and (iii) dry for 24hours (24hd). The discs were subdivided into 3 subgroups (n=12) according to surface treatments: (iv) no treatment, (v) treatment with EDTA, (vi) treatment with HA. The discs were placed in 96-well plates, and PLF were seeded and kept in contact with the discs for 48hours. Cell viability on the surface of the discs was assessed by XTT, and PLF adhesion was evaluated using scanning electron microscopy (SEM). Quantification of cytokines was performed using enzyme-linked immunosorbent assay (ELISA). Data were submitted to ANOVA and Tukey's test (α=.05). Surface treatment had a statistically significant effect on the cell viability in groups WD (P=.03), 1hd (P=.01) and 24hd (P=.048). PLF in contact with dried root surfaces expressed more cytokines without treatment with IL-6 decreasing the expression when treated with HA for 24hours. SEM also showed adhesion of PLF to the surface of all discs at all time periods. EDTA+HA is an alternative treatment for cases of avulsed teeth as it promoted adhesion and increased viability of PLF.