A Comparative Study of Cryogenic Lesions in Organ-Cultured Human Skin and in Reconstituted Human Skin Equivalent

Abstract

Cryosurgery is a technique that is widely used in the treatment of cutaneous tumors. However, there are still features of healing in cryosurgery wounds that are incompletely understood and necessitate further study. In the present paper, we describe twoin vitromodels that were developed to study the initial stages of development of the cryolesion: reconstituted human skin and organ-cultured human skin. Cryolesions were generated in both models by applying a 2-mm-diameter cryoprobe at −196°C for 35 s. Histological features were analyzed at days 0, 3, 5, and 14 following cryotreatment and showed epidermal detachment and keratinocyte necrosis very close to the findings reportedin vivo.Results were similar in the two models. Gross alteration of the dermal architecture was noticed beneath the cryolesion, particularly in the reconstituted skin model. Cell proliferation was investigated at days 0, 3, and 5 by [3H]thymidine incorporation and Ki-67 antigen immunolabeling. In the case of organ-cultured skin, a significant increase in keratinocyte and fibroblast proliferation was observed at day 3, compared to the controls. At day 5, a return to the basic level was noticed. This was not observed in the reconstituted skin model at either day 3 or day 5. These data led us to propose that organ-cultured skin may be a useful model for evaluating the response of human skin to freezing; reconstituted skin was not adequate for this purpose.