The present study describes for the first time the changes of both AEFC thickness and the numerical density of collagen fibers inserting into AEFC at specified levels and sites of human premolars at different stages of development. The investigation was based on 45 premolars (25 maxillary, 20 mandibular; 25 first and 20 second), extracted from adolescents and young adults. All teeth were free of disease and presented with roots developed from 30-100% of their final length. They were prefixed in Karnovsky's fixative, decalcified in EDTA and subdivided into about 14 slices each, cut from mesial and distal root surfaces, vertical to and along the root axis. The slices were postfixed in OsO4, embedded in Epon and cut for light-microscopic study. AEFC thickness (4086 measurements) and the density of the collagenous fiber fringe (454 counts) inserting in AEFC were measured at 1, 3, 5 and 7 mm apical to the cementoenamel junction. The data obtained showed: AEFC thickness increased with age and varied between 0 and 57.5 microns. Between 9 and 17 years, cervical AEFC thickness increased in maxillary first premolars from an average of 5 to 30 microns, and in mandibular second premolars from 6 to 20 microns, i.e., AEFC grew at approximately the same rate as later in life. Depending on the differences in tooth development, AEFC on maxillary first premolars became thicker than that on mandibular second premolars. Due to the corono-apically decreasing gradient of AEFC development, its increase in mid-root regions lagged behind that in cervical regions of all teeth in people younger than about 14 yr.(ABSTRACT TRUNCATED AT 250 WORDS)