The microdialysis technique was evaluated as a possible method to obtain local measurements of biochemical markers from knee joints with degenerative changes. Seven patients scheduled for arthroscopy of the knee due to minor to moderate degenerative changes had microdialysis catheters inserted under ultrasonographic guidance, intraarticularly and in the synovium-close tissue. Catheters were perfused at a rate of 2 µl/min for approximately 100 min with a Ringer solution containing radioactively labeled glucose, and the positions of the catheters were later visualized during arthroscopy. All intraarticular catheters and 6/7 subsynovial catheters were positioned correctly. Relative recovery (RR) was intraarticularly 0.64 ± 0.02 (mean ± SEM) and synovium-close 0.54 ± 0.06. Mean values of cartilage oligomeric matrix protein (COMP), aggrecan and glucosyl-galactosyl-pyridinoline in the intraarticular dialysates were 18.1 ± 7.0 U/l, 243.6 ± 108.6 ng/ml and 108.0 ± 29.0 pmol/ml, respectively. COMP and glucosyl-galactosyl-pyridinoline concentrations were significantly higher intraarticularly compared to perisynovial tissue (P <0.05), whereas for aggrecan, no significant difference was found (P = 0.06). The microdialysis method can be used for intraarticular and subsynovial determination of metabolites in human knees at these sites. The present methodology displays a potential for future studies of simultaneous biochemical changes within and around joints.