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Monolithic zirconia crowns fabricated using computer-aided design and computer-aided manufacturing (CAD-CAM) technology have recently become an alternative dental prosthetic treatment. The marginal fit of monolithic zirconia crown may be affected by different stages of the fabrication procedures in the laboratory and cementation. Information regarding the accuracy of fit of monolithic zirconia crowns at different stages of fabrication and cementation is limited.The purpose of this in vitro study was to evaluate the effect of different stages of fabrication and cementation on the vertical marginal discrepancy (VMD) of CAD-CAM fabricated monolithic zirconia crowns.Six ivorine right maxillary first molar typodont teeth with standardized anatomic preparations for complete coverage ceramic crowns were scanned with a 3-dimensional laboratory scanner. Crowns were designed using CAD software and milled from presintered monolithic zirconia blocks in a 5-axis dental milling machine. A cement space of 25 μm for the margins and a 50-μm space starting 1 mm above the finish lines of the teeth were virtually set in the CAD software. A total of 144 measurements were performed on 6 specimens with 8 measurement locations in 3 different stages using stereoscopic zoom microscopy; after initial production of the crowns (post-sintering group), after glazing (post-glazing group), and after cementation (post-cementation group). The VMD values were statistically analyzed with 1-way repeated measures ANOVA and the Holm-Sidak method (α=.05).Different stages of fabrication and cementation significantly affected the VMD of tested crowns (P=.003). The mean VMD was 38 μm for post-sintering group, 38 μm for post-glazing group, and 60 μm for post-cementation group, with statistical differences between the post-sintering group, the post-cementation group (P<.002), and the post-glazing group and post-cementation group (P<.003); there were no statistical differences between the post-sintering group and the post-glazing group (P=.966).Within the limitations of this in vitro study, glazing did not significantly change the VMD of CAD-CAM monolithic zirconia crowns. Cementation significantly increased the VMD values.