Preparation and evaluation of gene-transfected cultured skin as a novel drug delivery system for severely burned skin.
OBJECTIVE
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.


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.


CONCLUSIONS
Gene transfection to cultured skin may produce several dermal patches that release potent endogenous bioactive peptides.
Hada
Nobuko
N
Faculty of Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado, Saitama 350-0295, Japan.
Todo
Hiroaki
H
Komada
Fusao
F
Sugibayashi
Kenji
K
D000818:Animals; D002056:Burns / Q000473:pathology / Q000628:therapy*; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D016503:Drug Delivery Systems / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051381:Rats; D015203:Reproducibility of Results; D012867:Skin / Q000293:injuries*; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection / Q000379:methods*
D049452:Green Fluorescent Proteins