Porcine buccal mucosa is frequently used for in vitro drug absorption studies as its structure and permeability characteristics are close to those of human tissue. However, this tissue model suffers from practical disadvantages, including a limited surface area, damage caused by mastication, and a fastidious and time-consuming excision procedure. It has been hypothesized that such limitations may be overcome by replacing the buccal tissue with the pig esophageal mucosa. The latter has a very similar structure and is easier to separate from the underlying tissue; furthermore, its surface area is greater and is generally intact. The aims of this work, therefore, were (i) to perform histological studies on the two membranes; (ii) to compare the transport of fentanyl citrate across buccal and esophageal mucosae; and (iii) to evaluate the effects of freezing on the tissue permeability. The results show that their histology is comparable, that the permeability of fentanyl citrate across the two epithelial barriers is similar, and that freezing the tissues did not alter their permeability.