To study how crystal growth in dental enamel is controlled by the components of the extracellular matrix, we investigated the functional roles of amelogenins and fluoride ions in apatite formation occurring through an octacalcium phosphate (OCP)-precursor pathway. Using a cation selective membrane system as a model of tooth enamel formation, we evaluated the resulting mineral habit grown in native porcine amelogenins and fluoride ions. In the absence of amelogenin and in the presence of 1 or 2 ppm F, we obtained OCP + apatite and apatite, respectively. Without amelogenins, the crystals were hexagonal prisms and cones with diameters of approximately 100-200 nm. In the presence of 10% amelogenins and in the absence of fluoride, rod-like OCP with a diameter of 35 nm were obtained. Remarkably, a combination of amelogenin and fluoride created the formation of rod-like apatite crystals with dimensions similar to the former crystals. These observations indicate a cooperative role of amelogenin and fluoride in the regulation of habit, size orientation and phase of the calcium-phosphate crystals, resulting in the formation of fine rod-like apatite whose habit and orientation were similar to that of authentic tooth enamel crystals. The significant modulating effect of the amelogenin matrix combined with fluoride ions suggests the potential for this artificial system to contribute to the engineering of novel enamel-like biomaterials in vitro.