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The differences in the optical properties of multi-layered zirconia with and without yttria-gradient are not fully understood. This study aimed to evaluate and compare the optical properties, related microstructures, and phase fractions of multi-layered zirconia with and without yttria-gradient. For this, multi-layered zirconia of 5 mol% yttria (5Y) stabilized (Katana STML) and 4Y/5Y stabilized (e.max MT Multi) were cut layerwise, sintered, and analyzed using the opalescence parameter (OP), average transmittance (AT%), translucency parameter (TP), and contrast ratio (CR). The average grain size and phase fractions were obtained from field-emission scanning electron micrographs and X-ray diffraction patterns, respectively. Although the TP values of Katana STML and e.max MT Multi did not show a significant difference (except for transition layer 1), the results of AT and CR showed that the translucency of e.max MT Multi was slightly higher than that of Katana STML (p < 0.05). The opalescence gradient was higher in Katana STML than in the e.max MT Multi. In both zirconia types, translucency increased from the dentin to enamel layer based on the AT, TP, and CR results, while OP decreased (p < 0.05). The higher translucency from the dentin to enamel layer in Katana STML was caused by the pigmentation gradient, while in e.max MT Multi, it was caused by the difference in phase fraction and the pigmentation gradient. [ABSTRACT FROM AUTHOR]