Background: This present study is aimed to retrospectively assess the efficacy of three-dimensional (3D) printing assisted osteotomy guide plate in accurate osteotomy of adolescent cubitus varus deformity. Material and methods: Twenty-five patients (15 males and 10 females) with the cubitus varus deformity from June 2014 to December 2017 were included in this study and were enrolled into the conventional group (n = 11) and 3D printing group (n = 14) according to the different surgical approaches. The operation time, intraoperative blood loss, osteotomy degrees, osteotomy end union time, and postoperative complications between the two groups were observed and recorded. Results: Compared with the conventional group, the 3D printing group has the advantages of shorter operation time, less intraoperative blood loss, higher rate of excellent correction, and higher rate of the parents' excellent satisfaction with appearance after deformity correction (P < 0.001, P < 0.001, P = 0.019, P = 0.023). Nevertheless, no significant difference was presented in postoperative carrying angle of the deformed side and total complication rate between the two groups (P = 0.626, P = 0.371). Conclusions: The operation assisted by 3D printing osteotomy guide plate to correct the adolescent cubitus varus deformity is feasible and effective, which might be an optional approach to promote the accurate osteotomy and optimize the efficacy.