AbstractNearly nothing is known about the consequences of ultra-long-distance running on knee cartilage. In this mobile MRI field study, we analysed the biochemical effects of a 4,486 km transcontinental multistage ultra-marathon on femorotibial joint (FTJ) cartilage. Serial MRI data were acquired from 22 subjects (20 male, 18 finisher) using a 1.5 T MR scanner mounted on a 38-ton trailer, travelling with the participants of the TransEurope FootRace (TEFR) day by day over 64 stages. The statistical analyses focused on intrachondral T2* behaviour during the course of the TEFR as the main outcome variable of interest. T2* mapping (sagittal FLASH T2* weighted gradient echo) is a validated and highly accurate method for quantitative compositional cartilage analysis of specific weightbearing areas of the FTJ. T2* mapping is sensitive to changes in the equilibrium of free intrachondral water, which depends on the content and orientation of collagen and the proteoglycan content in the extracellular cartilage matrix. Within the first 1,100 km, a significant running load-induced T2* increase occurred in all joint regions: 44.0% femoral-lateral, 42.9% tibial-lateral, 34.9% femoral-medial, and 25.1% tibial-medial. Osteochondral lesions showed no relevant changes or new occurrence during the TEFR. The reasons for stopping the race were not associated with knee problems. As no further T2* elevation was found in the second half of the TEFR but a decreasing T2* trend (recovery) was observed after the 3,500 km run, we assume that no further softening of the cartilage occurs with ongoing running burden over ultra-long distances extending 4,500 km. Instead, we assume the ability of the FTJ cartilage matrix to reorganize and adapt to the load.