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The objective is to determine the relation between severity of knee osteoarthritis (KOA) and levels of Collagen type II metabolite (C2C) and trace elements in the urine. The urine sample and knee joint films (anteroposterior and lateral) from the KOA patients and control subjects were collected. The KOA patients were divided into five groups (controls and grades I-IV) according to the Kellgren-Lawrence radiographic grading standards. Urine levels of C2C and trace elements were detected by enzyme-linked immunosorbent assay and inductively coupled plasma atomic emission spectrometry, respectively. Urine C2C levels in the KOA subjects (261.235 ± 39.944 pg/ml) were higher than those of the control group (218.341 ± 22.270 pg/ml). The Fe content in KOA groups was significantly lower than that of control group (group IV > group III > group II > group I or controls). The contents of Cu and Zn were also significantly higher in the KOA patients than in the control group (p < 0.05). However, Cr, Al, Cd, Ni, and Se levels of KOA patients were not significantly different from those of the controls (p > 0.05). Determination of the urine levels of C2C and trace elements may prove to be informative for an early diagnosis of KOA. It can also assist in the prognosis judgment of the disease and selecting an appropriate therapeutic regimen.