The aim was to evaluate the chemical, morphological aspects and microhardness of root dentin after treatment with Triton™ solution. Twenty blocks of root dentin were distributed in two groups (n = 10): Control (C) (2.5% NaOCl+17% EDTA+2.5% NaOCl) and Triton™ solution (T). Morphological analysis was performed before and after treatments with confocal laser microscopy. Chemical composition and microhardness were analysed after the treatments using Raman spectroscopy and Knoop microhardness. Results were submitted to Student's t-test and Mann-Whitney test (p < 0.05). The C group had greater tubule number, area and perimeter (p < 0.001), besides a regular surface, while T showed an irregular surface with cracks and erosions, lower organic content intensity (p < 0.001) and higher inorganic/organic ratio (p = 0.003) than C, which had higher microhardness than T (p = 0.003). Triton™ exposed a lower number, area and perimeter of dentinal tubules, with cracks and erosions in root dentin. It also showed significant chemical alterations in the organic content, reducing it, resulting in lower microhardness.
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