The in vitro growth of pre-osteoblasts in multi-layer, three-dimensional scaffolds was determined from experimental measurements and was compared to a mathematical model. Immediately following cell seeding, the initial cell density was uniform throughout the scaffold. After 10 days, the cell density increased from 2.1 x 10(5) cells/cm(3) to 1.3 x 10(7) cells/cm(3) at the fluid-scaffold interface. The increase in cell density was largely confined to the outermost 200 microm from the fluid-scaffold interface. The cell density profile was in good agreement with a mathematical model that simulated the cell growth based on the local oxygen tension. The improved understanding derived from this mathematical model may be useful in the design of three-dimensional scaffolds that can support more uniform growth of cells.