Protein adsorption is known to direct biological response to biomaterials and is important in determining cellular response in tissue scaffolds. In this study we investigated the effect of the duration of protein adsorption to 3D polymer scaffolds on cell attachment and proliferation. 3D macro-porous polymer scaffolds were pre-aged in serum-containing culture medium for 5 min, 1 d or 7 d prior to seeding osteoblasts. The total amount of protein adsorbed was found to increase with pre-ageing time. Cell attachment and proliferation were measured 1 d and 14 d, respectively, after cell seeding. Osteoblast proliferation, but not attachment, increased with scaffold pre-ageing time and amount of adsorbed serum protein. These results demonstrate that the amount of time that scaffolds are exposed to serum-containing medium can affect cell proliferation and suggest that these effects are mediated by differences in the amount of protein adsorption.