Human periodontal ligament (PDL) cells were derived from healthy premolars extracted for orthodontic treatment and were utilized for in vitro experiments in passages 4-6. Human PDL cells were seeded in tissue culture tubes and incubated with interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), indomethacin, parathyroid hormone (PTH), or their combinations, for 1 h. The medium was then replaced with serum-free BGJb medium and incubated for 24 h without further additions. Prostaglandin E (PGE) concentrations in the conditioned media (CM) were measured by radioimmunoassay, and bone-resorbing activity was measured using 45Ca-labeled neonatal mouse calvariae. The results of this study indicated that (1) unstimulated cultured PDL cells produced PGE, and PDL CM stimulated bone resorption; (2) cytokine-treated (IL-1 alpha, IL-1 beta, and TNF-alpha) PDL cells had increased production of PGE and bone-resorbing activity compared to unstimulated PDL cells; (3) indomethacin completely inhibited PGE production from unstimulated PDL cells but only partially inhibited bone-resorbing activity, indicating that PDL cells produced nonprostaglandin bone-resorbing factor(s); (4) IFN-gamma did not change PGE or bone-resorbing activity production by cytokine-stimulated PDL cells; and (5) PTH treatment of PDL cells in addition to cytokines (IL-1 alpha, IL-1 beta, and TNF-alpha) had additive effects on the production of bone-resorbing activity and synergistic effects on PGE production compared to cytokine treatment alone.