Platelet-rich plasma (PRP) products may be useful for treatment of joint disease in horses, but may contain undesirable pro-inflammatory cytokines in addition to growth factors. This study investigated whether autologous PRP increases synovial fluid growth factor and cytokine concentrations when injected into normal equine metacarpophalangeal and metatarsophalangeal (fetlock) joints. Fetlock joints of seven healthy horses received one of four treatments: saline, resting PRP, CaCl2-activated PRP or thrombin-activated PRP. Synovial fluid was sampled prior to injection and at 6, 24, 48 and 96 h post-injection. Platelet-derived growth factor (PDGF-BB), transforming growth factor β1 (TGFβ1), interleukin (IL)-6 and tumor necrosis factor α (TNFα) concentrations in synovial fluid and PRP were measured by ELISA. Synovial fluid PDGF-BB, TGFβ1, IL-6, TNFα and IL-1 concentrations were also measured in vitro after incubation for 6 h with resting PRP only. Growth factor concentrations, but not cytokine concentrations, were significantly higher in activated PRP than in resting PRP samples. After intra-articular injection with resting or thrombin-activated PRP, synovial TGFβ1 increased significantly compared to baseline levels. TNFα and IL-6 were significantly increased in synovial fluid after thrombin-activated PRP injection. In vitro, growth factor concentrations increased significantly in synovial fluid after mixing with PRP, indicating that exogenous activation of PRP for intra-articular injection may be unnecessary, whereas cytokine levels did not. In conclusion, thrombin-activated PRP induced an inflammatory cytokine response in joints, whereas resting or CaCl2-activated PRP did not. Synovial growth factor levels were low overall; the reported benefits of intra-articular PRP may not be attributable to changes in local PDGF or TGFβ1 concentrations.