Incorporation of basic fibroblast growth factor into preconfluent cultured skin substitute to accelerate neovascularisation and skin reconstruction after transplantation.

Cultured skin substitutes (CSS) with both epidermal and dermal components seem to be ideal, but they have not been widely used clinically, partly because it takes several weeks to produce them. Decreasing the number of seeding cells may reduce the period required for production, but it still takes a long time before the cells become confluent and neovascularisation is completed in CSS after grafting. As we have already succeeded in reducing the number of seeded keratinocytes in this study, we first attempted to reduce the number of seeded fibroblasts. Consequently, preconfluent CSS with 10 x 10(3) cells/cm2 of fibroblasts combined with 100 x 10(3) cells/cm2 of keratinocytes could be successfully grafted on to full-thickness wounds. bFGF-impregnated gelatin microspheres were then added to the preconfluent CSS before grafting. Incorporation of bFGF significantly accelerated neovascularisation and increased epidermal thickness, cellular components, and thickness of the dermis. The incorporation of bFGF makes CSS a potential therapeutic approach for management of skin wounds.