Objective To investigate the effects of different puncture levels on bone cement distribution and effectiveness in bilateral percutaneous vertebroplasty for osteoporotic thoracolumbar compression fractures. Methods A clinical data of 274 patients with osteoporotic thoracolumbar compression fractures who met the selection criteria between December 2017 and December 2020 was retrospectively analyzed. All patients underwent bilateral percutaneous vertebroplasty. During operation, the final position of the puncture needle tip reached was observed by C-arm X-ray machine. And 118 cases of bilateral puncture needle tips were at the same level (group A); 156 cases of bilateral puncture needle tips were at different levels (group B), of which 87 cases were at the upper 1/3 layer and the lower 1/3 layer respectively (group B1), and 69 cases were at the adjacent levels (group B2). There was no significant difference in gender, age, fracture segment, degree of osteoporosis, disease duration, and preoperative visual analogue scale (VAS) score, and Oswestry disability index (ODI) between groups A and B and among groups A, B1, and B2 (P>0.05). The operation time, bone cement injection volume, postoperative VAS score, ODI, and bone cement distribution were compared among the groups. Results All operations were successfully completed without pulmonary embolism, needle tract infection, or nerve compression caused by bone cement leakage. There was no significant difference in operation time and bone cement injection volume between groups A and B or among groups A, B1, and B2 (P>0.05). All patients were followed up 3-32 months, with an average of 7.8 months. There was no significant difference in follow-up time between groups A and B and among groups A, B1, and B2 (P>0.05). At 3 days after operation and last follow-up, VAS score and ODI were significantly lower in group B than in group A (P<0.05), in groups B1 and B2 than in group A (P<0.05), and in group B1 than in group B2 (P<0.05). Imaging review showed that the distribution of bone cement in the coronal midline of injured vertebrae was significantly better in group B than in group A (P<0.05), in groups B1 and B2 than in group A (P<0.05), and in group B1 than in group B2 (P<0.05). In group A, 7 cases had postoperative vertebral collapse and 8 cases had other vertebral fractures. In group B, only 1 case had postoperative vertebral collapse during follow-up. ConclusionBilateral percutaneous vertebroplasty in the treatment of osteoporotic thoracolumbar compression fractures can obtain good bone cement distribution and effectiveness when the puncture needle tips locate at different levels during operation. When the puncture needle tips locate at the upper 1/3 layer and the lower 1/3 layer of the vertebral body, respectively, the puncture sites are closer to the upper and lower endplates, and the injected bone cement is easier to connect with the upper and lower endplates.