The aim was to elucidate mechanisms for phenytoin-induced gingival overgrowth, using effects of type 1 human mast cell (HMC-1) supernatant and histamine on the expression of 5alpha-reductase in human gingival fibroblasts (HGF), and the effects of phenytoin on this activity. Duplicate incubations of HGF in Eagle's minimum essential medium (MEM) were performed with [14C] testosterone and serial concentrations of mast-cell histamine (1-100 microgram/ml)/HMC-1 culture supernatant at serial dilutions of 1-100 microl/ml for 24 h. Other experiments employed optimal concentrations of HMC-1 supernatant (10 microl/ml) or histamine (8 microgram/ml) and phenytoin (1 and 5 microgram/ml), alone and in combination, using two androgen substrates, [14C] testosterone and [14C] 4-androstenedione. At the end of a 24-h incubation the medium was solvent-extracted for steroid metabolites, analysed and quantified in a radioisotope scanner. HMC-1, histamine (n=3) and phenytoin (n=6) significantly increased the synthesis of dihydrotestosterone and 4-androstenedione by up to 80% (P<0.01); the combination of HMC-1 and phenytoin caused two-fold increases (n=6; P<0.01). The incubations with histamine alone and in combination with phenytoin showed significant stimulation of dihydroxytestosterone and the diols alone and in combination, which was less pronounced in combination. This investigation demonstrates significant stimulation of 5alpha-reductase activity in human gingival fibroblasts by mast-cell supernatant and a specific product histamine, alone and in combination with phenytoin. Androgen 5alpha-reductase-mediated anabolic actions in connective tissue are well documented. The findings suggest a novel hypothesis that mast-cell mediated androgen action in the gingiva in response to phenytoin could contribute to gingival overgrowth.