The growth of engineered cartilage tissue in vitro is often impaired by the problem of insufficient oxygen and nutrient supply to cells seeded in 3D constructs. Despite its central role in controlling most cell functions, the scaffolding material has generally been thought of only as a transport barrier and its potential active role in controlling oxygen uptake has never been addressed. In this work the role of cell-material interaction on oxygen metabolism in 3D in vitro cultures was surveyed. To this aim bovine chondrocytes, at a cell density of 400,000 and 4,000,000 cells/mL, respectively, were seeded in collagen type I and in agarose, while keeping all other culture conditions constant. A unidirectional oxygen gradient was induced in the culture through the application of a "sandwich" model and the oxygen concentration at the pericellular level was measured by phosphorescence quenching microscopy. Results show that the oxygen consumption rate is two-fold higher in agarose than in collagen, which indicates that the nature of the material strongly influences cell metabolic behaviour. Moreover, since different oxygen consumption rates are linked to different cell biosynthetic activity, our findings will prove beyond any doubt the active role played by materials in tissue regeneration.