Real-time atomic force microscopy of root dentine during demineralization when subjected to chelating agents

AIM: To explore the potential of atomic force microscopy (AFM) for the examination of changes to dentine surfaces during demineralization and evaluate qualitatively the effect of EDTA, EDTAC and citric acid. METHODOLOGY: Nine canine teeth were sectioned transversely at the cemento-enamel junction, and the crowns discarded. Subsequently, each root was embedded in an epoxy cylinder and discs approximately 5 mm thick were cut. A standard metallographic procedure was then used to prepare the surfaces for observation. From the central portion of these samples, two specimens were symmetrically prepared per tooth so that a total number of 18 samples was produced. To allow the use of a liquid cell during AFM, the samples were embedded in silicone rubber and were then randomly divided into three groups, as follows: group 1: 17% EDTA (pH 7.7), group 2: 17% EDTAC (pH 7.7) and group 3: 10% citric acid (pH 1.4). Topographical images were acquired during the demineralization process, allowing real-time observation of the dentine surface. Two operators assigned scores to the AFM images using a double-blind method. anova analysis with random effects (P < 0.05) was used to compare the results. RESULTS: The average scores were 6.13 +/- 0.35 for EDTAC, 7.36 +/- 0.23 for EDTA and 14.55 +/- 1.21 for citric acid. Citric acid was statistically different from EDTA and EDTAC while EDTA and EDTAC were not statistically different. CONCLUSIONS: The most effective demineralizing substance was citric acid. The methodology developed for real-time observation of dentine surfaces is a valuable method to evaluate demineralization.