We aim to determine serum dynamic thiol/disulphide homeostasis with novel methods in early-stage osteoarthritis and late-stage osteoarthritis patients and investigated whether it was associated with the progression of osteoarthritis risk or not. One hundred eighteen patients were included in this prospective study. Osteoarthritis patients were divided into five stages, according to the Kellgren-Lawrence scale. Dynamic thiol/disulphide homeostasis was determined with a novel spectrophotometric method. Late-stage osteoarthritis patients had significantly lower levels of native and total thiol than the patients of early-stage osteoarthritis. Disulphide, index-1, index-2 levels, and WOMAC score of late-stage osteoarthritis patients were significantly higher than the ones belonging to patients of early-stage osteoarthritis. Decreased native thiol and total thiol levels and increased WOMAC score and disulphide levels were independently associated with increased risk of late-stage osteoarthritis. We suggest that both WOMAC score and dynamic thiol/disulphide homeostasis may be implicated in the pathogenesis and progression of osteoarthritis. We also recommend that dynamic thiol/disulphide homeostasis may have clinical utility as possible markers of differential diagnosis of early-stage and late-stage osteoarthritis.