X-rays were employed to observe the intervertebral fusion 1 week, 4 weeks, 8 weeks, and 12 weeks after surgery. During the detection, isoflurane-anesthetized rats were placed in the right lateral position and the X-ray settings were as follows: voltage 42 kV, electric current 320 mA, and exposure time of 8 ms. The relative new bone mass was analyzed by Mimics software. Then, the rats were euthanized at 12 weeks postoperation and the caudal vertebrae of the operated segment were soaked in 4% paraformaldehyde. The internal-fixation plates and the surrounding soft tissues were carefully removed prior to the computed tomography (CT) scanning. Micro-CT (vivaCT 40, Scanco 274 Medical, Switzerland) scanning was performed using the following conditions: 70 mA, 120 kV, and 15 μm. All 3D images were reconstructed by Mimics (a highly integrated and easy-to-use 3D image generation and editing software). The bone volume relative to total volume (BV/TV) and the bone mineral density (BMD) within the defect area were calculated by Mimics software (n = 6).

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