BackgroundAs the frequency of spine surgery increases in the veterinary field, many studies have been conducted on minimally invasive spine surgery (MISS). Although many studies have been conducted on the thoracolumbar spine about MISS in animals, several problems and limitations have emerged regarding this method. Therefore, we developed a three-dimensional (3D) printed patient-specific surgical guide (3DPSSG) using 3D printing technology to overcome these problems. We aimed to evaluate the accuracy and safety of the 3DPSSG in minimally invasive mini-hemilaminectomy-corpectomy (MI-MHC).MI-MHC using 3DPSSG and an endoscopic system was performed at L1–L2 in 15 cadaveric dogs. The procedure of fixing the surgical guide to the vertebral body through screws and the surgical procedure using the guide were performed by two surgeons with different experiences. Postoperative computed tomography was used to measure planned and postoperative screw trajectories (angle, protruding from the far cortex) and to create 3D rendering images of vertebrae to evaluate the direction of bone window formation, corpectomy slot length, depth, and height ratio.ResultsThe two groups which performed by two surgeons with different experiences did not differ in terms of screw angle deviation and length of the screw protruded from the far cortex. The corpectomy slot-length ratio was not different between the two groups; however, the slot-depth and height ratios were different.ConclusionsNo differences were detected in screw trajectory and corpectomy slot-length ratio between the two groups. The 3DPSSG for MI-MHC is classified as accurate and safe; therefore, it can be an alternative to the conventional technique in dogs.