Objective To analyze the correlation between the morphology of tibial intercondylar eminence and non-contact anterior cruciate ligament (ACL) injury, and provide a theoretical basis for the prevention and risk identification of ACL injury. Methods A retrospective analysis was conducted on the knee radiographs of 401 patients admitted to the Chengdu Second People’s Hospital between January 2017 and October 2021, including 219 males and 182 females. Non-contact rupture of ACL was observed in 180 patients and confirmed by arthroscopy or surgery, while the remained 221 patients were confirmed to have normal ACL by physical examination and MRI. The heights of medial and lateral tibial intercondylar eminence and the width of tibial intercondylar eminence of the 401 patients were measured, and the risk factors of ACL injury were analyzed. Results The height of medial tibial intercondylar eminence was lower and the width of tibial intercondylar eminence was smaller in male patients with ACL fracture than those in the male control group with statistical significance (P<0.05). Logistic regression analysis showed that a narrow width of tibial intercondylar eminence was a risk factor of ACL injury in males (P<0.05). The receiver operating characteristic (ROC) curve showed that the diagnostic threshold was 11.40 mm, the area under the curve (AUC) was 0.851 [95% confidence interval (CI) (0.797, 0.896)], the sensitivity was 72.81%, and the specificity was 84.76%. The height of medial tibial intercondylar eminence was lower and the width of tibial intercondylar eminence was smaller in female patients than those in the female control group with statistical significance (P<0.05). Logistic regression analysis showed that both a low height of medial tibial intercondylar eminence and a narrow width of tibial intercondylar eminence were risk factors of ACL injury in females (P<0.05). For the width of medial tibial intercondylar eminence, the ROC curve showed that the diagnostic threshold was 8.30 mm, and the AUC was 0.684 [95%CI (0.611, 0.751)], the sensitivity and specificity were 63.64% and 72.41%, respectively; for the height of medial tibial intercondylar eminence, the diagnostic threshold was 11.30 mm, and the AUC was 0.699 [95%CI (0.627, 0.756)], the sensitivity was 89.39%, and the specificity was 47.41%. Conclusions The reduced width of tibial intercondylar eminence is a risk factor and effective predictor of non-contact ACL injury in males. Both the reduced height of the medial tibial intercondylar eminence and the reduced width of tibial intercondylar eminence are risk factors and may be predictors for non-contact ACL injury in females.