Abstract Objective—To examine mRNA expression of cytokines in synovial fluid (SF) cells from dogs with cranial cruciate ligament (CrCL) rupture and medial patellar luxation (MPL) and determine mRNA expression for 3 joints (affected stifle, unaffected contralateral stifle, and left shoulder joints) in dogs with unilateral CrCL rupture. Sample Population—29 stifle joints with CrCL rupture (29 dogs), 8 stifle joints with MPL (7 dogs), and 24 normal stifle joints (16 clinically normal dogs). Procedures—Immediately before reconstructive surgery, SF was aspirated from the cruciate-deficient stifle joint or stifle joint with MPL. Fourteen of 29 dogs had unilateral CrCL rupture; SF was also aspirated from the unaffected contralateral stifle joint and left shoulder joint. Those 14 dogs were examined 6 and 12 months after reconstructive surgery. Total RNA was extracted from SF cells and reverse transcription–PCR assay was performed to obtain cDNA. Canine-specific cytokine mRNA expression was determined by use of a real-time PCR assay. Results—Interleukin (IL)-8 and -10 and interferon-G expression differed significantly between dogs with arthropathies and dogs with normal stifle joints. For the 14 dogs with unilateral CrCL rupture, a significant difference was found for IL-8 expression. Before reconstructive surgery, IL-8 expression differed significantly between the affected stifle joint and left shoulder joint or contralateral stifle joint. Six months after surgery, IL-8 expression was significantly increased in the unaffected contralateral stifle joint, compared with the shoulder joint. Conclusions and Clinical Relevance—No conclusions can be made regarding the role of the examined cytokines in initiation of CrCL disease.