Dexamethasone transport across ocular epithelium was evaluated by means of permeability studies on a series of ester prodrugs with the aim of identifying the most promising candidates for the treatment of the ocular surface. Organotypic conjunctival bovine epithelial cell cultures were assumed representative of an average ocular epithelium and used to describe the mechanism of permeation. Permeability coefficients were also determined in excised rabbit corneas set up and in vivo pharmacokinetic experiments. All dexamethasone esters permeated through the transcellular route and their permeability coefficient rose with the increase of the molecules lipophilicity until a maximum was reached in correspondence of dexamethasone butyrate (Log P = 3.95). It was found that esters hydrolysis occurring in various extent along the transport process, affected the overall permeability rate. There was evidence that the permeation process can be confined at the ocular epithelium layer if the ester is highly hydrophobic and not susceptible of fast hydrolysis.