Background: Spinal tumors can progressively develop and compress the spinal cord or cauda equina, leading to neurological dysfunction, and even secondary pathological fractures, resulting in spinal instability, leading to intractable pain, and seriously affecting the quality of life. Aims:To discuss the clinical application of three-dimensional printing multifunctional biological scaffolds in spinal tumor surgery. Materials and methods:A retrospective analysis of 40 patients with spinal tumors from January 2021 to July 2022 was conducted, and they were divided into observation group (Transfer the 3D reconstruction data to the 3D printing equipment according to the CTA and MRI examination results, and print the 3D model according to the ratio of 1:1 for pre-surgery evaluation and judgment)and control group (According to the MRI imaging examination to determine the condition and implement the surgery)according to whether 3D printing technology was used or not, with 20 cases in each group. The differences in surgical conditions, VAS score, KPS score, ODI score, JOA score and adverse reactions between the two groups were compared and analyzed. Results: After operation, the postoperative drainage volume and extubation time of the observation group were significantly lower than those of the control group, and the difference was statistically significant (P<0.05). The VAS score and ODI score of the observation group were significantly lower than those of the control group, and the KPS and JOA scores of the observation group were significantly higher than those of the control group, and the difference was statistically significant (P<0.05). The two groups of patients were followed up after 3 months of treatment, and the quality of life of the patients was evaluated. The nerve root palsy, lower limb weakness, hypoesthesia, cerebrospinal fluid leakage and infection in the observation group were lower than those in the control group, and the difference was statistically significant (P<0.05). Conclusion: With the support of 3D printing technology, complex surgery can be precisely operated, which can effectively reduce the risk of surgery, improve the fusion rate of bone graft, and provide better stability.