BackgroundOsteosarcoma (OSA) should be differentiated from other less frequent primary bone neoplasms, metastatic disease, and tumor‐like lesions, as treatment and prognosis can vary accordingly. Hence, a preoperative histologic diagnosis is generally preferred. This requires collection of multiple biopsies under general anesthesia, with possible complications, including pathological fractures. Fine‐needle aspiration cytology would allow an earlier diagnosis with a significant reduction of discomfort and morbidity.Hypothesis/ObjectivesThe aim of this study was to compare the accuracy of cytological and histologic biopsies in the diagnosis of canine osteodestructive lesions.AnimalsSixty‐eight dogs with bone lesions.MethodsRetrospective study. Accuracy was assessed by comparing the former diagnosis with the final histologic diagnosis on surgical or post‐mortem samples or, in the case of non‐neoplastic lesions, with follow‐up information.ResultsThe study included 50 primary malignant bone tumors (40 OSAs, 5 chondrosarcomas, 2 fibrosarcomas, and 3 poorly differentiated sarcomas), 6 carcinoma metastases, and 12 non‐neoplastic lesions. Accuracy was 83% for cytology (sensitivity, 83.3%; specificity, 80%) and 82.1% for histology (sensitivity, 72.2%; specificity, 100%). Tumor type was correctly identified cytologically and histologically in 50 and 55.5% of cases, respectively.Conclusions and Clinical ImportanceThe accuracy of cytology was similar to histology, even in the determination of tumor type. In no case was a benign lesion diagnosed as malignant on cytology. This is the most important error to prevent, as treatment for malignant bone tumors includes aggressive surgery. Being a reliable diagnostic method, cytology should be further considered to aid decisions in the preoperative setting of canine bone lesions.