1. The effects of zinc on bone metabolism and tooth formation was examined in organ cultures of calvaria and tooth germ, and in cell cultures of osteoblast-like cells, MC3T3-E1. 2. Treatment of calvaria with zinc (10, 100 microM) for 4 days both increased alkaline phosphatase (ALP) activity in bone and reduced the secretion of N-acetyl beta-glucosaminidase from bone, without affecting bone mineral or collagen content. The increase in ALP activity produced by zinc (10 microM) was inhibited neither by actinomycin D (5 micrograms/ml) nor by cycloheximide (0.5 micrograms/ml). 3. Treatment of MC3T3-E1 cells with zinc (50, 100 microM) for 25 days also increased ALP activity, but reduced calcium content in cells and in the matrix layer. 4. These results indicate that zinc increases ALP activity in osteoblasts without affecting de novo enzyme synthesis, and that it inhibits bone mineralization, in accordance with the inhibition of osteoclastic activity. 5. Treatment of tooth germ with zinc (100 microM) for 7 days also produced an increase in ALP activity and inhibition of mineralization. These results indicate that the increased ALP activity produced by zinc is a common phenomenon in hard tissues, and, further, that zinc inhibits mineralization during tissue formation.