Purpose: The purpose of this study is to prepare and evaluate gene-transfected cultured skin to establish a dermal patch consisting of cultured skin as a new and novel delivery system for severely burned skin.
Materials and methods: Plasmid DNA encoding the green fluorescent protein (GFP) gene was used as a model gene and transfected to rat and human cultured dermis models (CDMs) using the hemagglutinating virus of Japan envelope vector (HVJ-E) to prepare gene-transfected CDM and evaluate GFP expression in the CDM. Two kinds of transfection methods were evaluated. In pre-transfection, the gene was first transfected into fibroblasts and then CDM was prepared using these gene-transfected cells. In post-transfection, the gene was transfected directly into CDM.
Results: GFP expression was observed in both the pre- and post-transfected CDMs. The post-transfection method showed higher GFP expression in the CDM than pre-transfection, although no statistically significant difference was observed. The cell viability of these transfected CDMs was also examined with MTT assay. Slight decrease in viability was observed in these transfected CDMs. These methods could be useful in preparing gene-transfected cultured skins with low cell damage.
Conclusion: Gene transfection to cultured skin may produce several dermal patches that release potent endogenous bioactive peptides.