Some biosynthetic activities of normal and otosclerotic temporal bone cultures have been studied. Bone cells were cultured for 24 hrs. in medium containing 3H-glucosamine, 35SO4 or 3H-proline. Labelled glycosaminoglycans (GAG) and proteins were precipitated from cells and media. In otosclerotic bone cells there was an evident reduction in the synthesis and secretion of radiolabelled macromolecules. The inhibitory effect was always greater in the extracellular than in the intracellular compartment. Some glycosidases were also studied. Otosclerosis decreased the activity of all enzymes examined, indicating that the lower GAG synthesis and secretion in otosclerotic bone cells were not due to an increased degradation.