Enamel surface microhardness was measured before treatment (baseline) and after pH cycling using a Vickers microhardness tester (HM112, Mitutoyo Corporation, Tokyo, Japan) with a load of 100 g. Three indentations at distances of 100 μm from each other, were performed in the center of enamel specimens, the average was calculated and recorded as the Vickers microhardness number (VHN) for each specimen. Baseline (VHN1) and post-pH cycling (VHN2) Vickers microhardness numbers were used to calculate the percentage of loss surface microhardness (%VHN) following acid exposure using the formula: 100× (VHN2-VHN1) ⁄ VHN1 [20].
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