To evaluate the effectiveness of orthopedic robot with modified tracer fixation (short for modified orthopedic robot) assisted percutaneous kyphoplasty (PKP) in treatment of single-segment osteoporotic vertebral compression fracture (OVCF). The clinical data of 155 patients with single-segment OVCF who were admitted between December 2017 and January 2021 and met the selection criteria was retrospectively analyzed. According to the operation methods, the patients were divided into robot group (87 cases, PKP assisted by modified orthopedic robot) and C-arm group (68 cases, PKP assisted by C-arm X-ray fluoroscopy). There was no significant difference in gender, age, body mass index, T value of bone mineral density, therapeutic segment, grade of vertebral compression fracture, and preoperative visual analogue scale (VAS) score, midline vertebral height, and Cobb angle between the two groups (P>0.05). The effectiveness evaluation indexes of the two groups were collected and compared. The clinical evaluation indexes included the establishment time of working channel, dose of intraoperative fluoroscopy, the amount of injected cement, VAS score before and after operation, and the occurrence of complications. The imaging evaluation indexes included the degree of puncture deviation, the degree of bone cement diffusion, the leakage of bone cement, the midline vertebral height and the Cobb angle before and after operation. The establishment time of working channel in robot group was significantly shorter than that in C-arm group, and the dose of intraoperative fluoroscopy was significantly larger than that in C-arm group (P<0.001). There was no significant difference in the amount of injected cement between the two groups (t=1.149, P=0.252). The patients in two groups were followed up 10-14 months (mean, 12 months). Except that the intraoperative VAS score of the robot group was significantly better than that of the C-arm group (P<0.05), there was no significant difference between the two groups at other time points (P>0.05). No severe complication such as infection, spinal cord or nerve injury, and pulmonary embolism occurred in the two groups. Five cases (5.7%) in robot group and 7 cases (10.2%) in C-arm group had adjacent segment fracture, and the difference in incidence of adjacent segment fracture between the two groups was not significant (χ2=1.105, P=0.293). Compared with C-arm group, the deviation of puncture and the diffusion of bone cement at 1 day after operation, the midline vertebral height and Cobb angle at 1 month after operation and last follow-up were significantly better in robot group (P<0.05). Eight cases (9.1%) in the robot group and 16 cases (23.5%) in the C-arm group had cement leakage, and the incidence of cement leakage in the robot group was significantly lower than that in the C-arm group (χ2=5.993, P=0.014). There was no intraspinal leakage in the two groups. Compared with traditional PKP assisted by C-arm X-ray fluoroscopy, modified orthopedic robot-assisted PKP in the treatment of single-segment OVCF can significantly reduce intraoperative pain, shorten the establishment time of working channel, and improve the satisfaction of patients with operation. It has great advantages in reducing the deviation of puncture and improving the diffusion of bone cement.