The aim of the present contribution was to develop a functional three-dimensional tissue construct to study ocular permeation of pilocarpine hydrochloride from different formulations. The in vitro model was compared to excised bovine cornea. Modified Franz cells were used to study the transcorneal permeability. Analysis was performed by reversed-phase high-performance liquid chromatography. Comparisons of the permeation rates through excised bovine cornea and the in vitro model show the same rank order for the different formulations. The permeation coefficient, K(P), obtained with the cornea construct, is about 2-4-fold higher than that from excised bovine cornea. It is possible to reconstruct bovine cornea as an organotypic culture and also to use this construct as a substitute for excised bovine cornea in drug permeation studies in vitro.