The effect of the ethylenediaminetetraacetic acid (EDTA) treatment on the hybrid layer durability of total-etch dentin adhesives was investigated. A flat dentin surface was exposed and treated with 37% phosphoric acid or 0.1M EDTA. Dentin adhesive was applied and a 4 mm thick composite resin was built-up. Twenty 0.9×0.9 mm² resin-dentin beams were fabricated in each experimental group. Artificial aging with 10% NaClO was performed in half of the experimental groups for 1 hour. A micro-tensile bond strength (μTBS) test was performed and the bonded interface was analyzed with transmission electron microscopy (TEM). The 37% phosphoric acid-treated group showed a lower μTBS after artificial aging, whereas the 0.1M EDTA-treated group did not. TEM analysis revealed collagen fibrils dissolved in the 37% phosphoric acid-treated group but not in the 0.1M EDTA-treated group. Undissolved minerals were observed in the 0.1M EDTA-treated group. In conclusion, a dentin treatment with 0.1M EDTA is effective in improving the hybrid layer durability.