Porcine skin has frequently been used as a model in in vitro permeation studies being considered as an excellent alternative for human skin due to biochemical similarities.
The use of frozen skin after thawing is particularly more convenient for these studies since it is available whenever needed. Even though several researchers have noticed that freezing and then thawing the tissue result in enhancement of drug permeation, many published articles have still described studies using frozen pig skin for bioequivalence and transdermal evaluation of drug products. The aim of this commentary article is therefore to explicitly demonstrate, according to our experience, the low-reliability of the convenient protocol of freezing pig skin for in vitro percutaneous studies. It has been shown in three separate studies using three different drugs that frozen/thawed pig skin had a higher permeability to drugs compared to that obtained by using fresh skin. The results have further demonstrated that the abnormal permeability through frozen pig skin is independent of the effect of different formulations, placing a question mark on the validity of the not freshly-used porcine skin for in vitro penetration studies.