This study aims to evaluate the application value of the tibial tubercle-trochlear groove distance (TT-TG distance) and tibial tubercle-midepicondyle distance (TT-ME distance) on CT images in patellofemoral instability, and further investigate the association between knee joint rotation angles and patellofemoral instability. We retrospectively analyzed CT image data of 59 patients with patellar dislocation (case group) and 39 normal knee joints (control group). We measured the TT-TG distance, TT-ME distance, and knee joint rotation angle (KJRA) of both groups, and the related indicators were analyzed using single-factor/multi-factor binary logistic stepwise regression analysis. Two senior radiologists were assigned to assess the inter-rater reliability. Interclass correlation coefficients (ICC) were calculated. Finally, we used receiver operating characteristic (ROC) curves to compare the diagnostic efficiency of these indicators in patellofemoral instability. The results found significant differences between both groups in terms of TT-TG distance, TT-ME distance, KJRA angle, age, location, and gender (P < 0.05). In terms of inter-rater reliability, TT-TG distance and TT-ME distance ratios showed an excellent correlation between observers (TT-TG inter-rater ICC 0.969, TT-ME inter-rater ICC 0.955). Univariate logistic regression analysis indicated that except for location and gender, all other factors significantly affected patellofemoral instability (P < 0.05). The multivariate logistic regression analysis revealed that the TT-ME distance, age, and KJRA angle were statistically significant factors related to patellofemoral instability, with TT-ME distance being a risk factor for patellofemoral instability (OR value 1.572, P value 0.000). Moreover, the ROC curve analysis demonstrated that the diagnostic capability of the TT-ME distance for detecting patellofemoral instability was higher than that of the TT-TG distance and KJRA (AUC were 0.912, 0.851, and 0.735, respectively). The TT-ME distance, age, and knee joint rotation angle are factors that affect patellofemoral instability. The TT-ME distance has better diagnostic efficiency for patellofemoral instability compared to the TT-TG distance and knee joint rotation angle.