The aim of this study was to measure omentin-1 concentrations in serum and synovial fluid (SF) of knee osteoarthritis (OA) patients and to investigate their correlation with patient-reported symptomatic severity. We enrolled 263 knee OA patients and 62 healthy controls. We collected Western Ontario and McMaster Universities Arthritis Index (WOMAC) scores from OA patients and measured omentin-1 concentrations in serum and SF by enzyme-linked immunosorbent assay (ELISA). Our results demonstrated that omentin-1 concentrations in SF but not serum were independently and negatively correlated with self-reported greater pain and physical disability in OA patients. Omentin-1 in SF might serve as a potential biomarker for reflecting the symptomatic severity of OA.