Analysis of the total proteins secreted by cultured dental follicle cells revealed that transforming growth factor-beta 1 (TGF-beta 1) stimulated them to secrete more extracellular matrix proteins into a serum-free medium than did follicle cells not exposed to the growth factor. Electrophoresis and scanning densitometry showed that secretion of all the major proteins was increased by exposure to the growth factor but the amounts ranged from a 66% increase for one of the procollagen chains to a 7% increase for fibronectin. Immunofluorescence using anti-type I collagen and anti-fibronectin showed that the intracellular concentration and intracellular localization of the antibodies was not changed by incubating the cells with the growth factor. The growth factor did not cause an increase in cell number but did modify the association of the cells in the culture, causing them to aggregate into clusters whereas the control cells formed a confluent monolayer. These results suggest that TGF-beta 1 may signal the fibroblasts of the dental follicle to secrete the extracellular matrix needed for its development into a periodontal ligament.