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Objective: Preoperative imaging assessment influences the decision to perform mastoidectomy for the mastoid extension of middle ear cholesteatoma. This study compared the performance of temporal subtraction CT (TSCT) and non-echoplanar diffusion-weighted imaging (non-EP DWI) in evaluating such mastoid extensions.Methods: We retrospectively evaluated 239 consecutive patients with surgically proven middle ear cholesteatoma between April 2016 and April 2021. The diagnostic performance of TSCT, wherein the presence of black color indicated progressive bone erosion, and non-EP DWI, wherein high signal intensity in the mastoid region suggested mastoid extension, was compared using Fisher's exact test.Results: In 34 patients with evaluable TSCT images, black color was significantly more common in patients with mastoid extension than in those without; the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of TSCT were 1.00, 0.95, 0.94, 1.00, and 0.97, respectively. In 90 patients with evaluable non-EP DWI, high signal intensity was significantly more common in patients with mastoid extension than in those without; the sensitivity, specificity, PPV, NPV, and accuracy of non-EP DWI were 0.88, 0.85, 0.91, 0.81, and 0.87, respectively. In 16 patients with both evaluable TSCT and non-EP DWI, the diagnostic performance of the TSCT was slightly superior to that of the non-EP DWI for predicting mastoid extension, although the difference was not significant.Conclusions: TSCT images generated using consecutively acquired preoperative high-resolution CT images are useful for predicting mastoid extension of middle ear cholesteatoma, and the diagnostic performance of TSCT is non-inferior to that of non-EP DWI. [ABSTRACT FROM AUTHOR]