Purpose:Todetermine the suitability of selected blood biomarkers of articular cartilage as mechanosensitive markers and to investigate the dose-response relationship between ambulatory load magnitude andmarkerkinetics in response to load. Methods:Serum samples were collected from 24 healthy volunteers before and at three time points after a 30-minute walking stress test performed on three test days.In eachexperimentalsession, one of three ambulatory loads was applied: 100% body weight (BW); 80%BW; 120%BW. Serum concentrations of COMP, MMP-3, MMP-9, ADAMTS-4, PRG-4, CPII, C2C and IL-6 were assessed usingcommercialenzyme-linked immunosorbent assays. A two-stage analytical approach was used to determine the suitability of a biomarkerby testing the response to the stress test (criterion I) and thedose-response relationship between ambulatory load magnitude and biomarker kinetics(criterion II). Results.COMP, MMP-3 and IL-6 at all three time pointsafter, MMP-9 at 30 and 60 minutes after, and ADAMTS-4 and CPII atimmediately afterthe stress testshowedan average response to load or an inter-individual variation in response to loadof up to 25% of pre-test levels.Therelation to load magnitude on average or an inter-individual variation in this relationshipwasup to8%from load level to load level.There was a positive correlation for the slopes of the change-load relationship between COMP and MMP-3, and a negative correlation for the slopes between COMP, MMP-3 and IL-6 with MMP-9, and COMP with IL6. Conclusions:COMP, MMP-3, IL-6, MMP-9, and ADAMTS-4 warrant further investigation in the contextofarticular cartilagemechanosensitivityand its role in joint degeneration and OA.WhileCOMP seems to be able to reflect a rapid response, MMP-3 seems to reflect a slightly longer lasting, but probably also more distinct response. MMP-3 showed also the strongest association with the magnitude of load.