Scale-up of a Composite Cultured Skin Using a Novel Bioreactor Device in a Porcine Wound Model

Extensive deep-burn management with a two-stage strategy can reduce reliance on skin autografts; a biodegradable polyurethane scaffold to actively temporize the wound and later an autologous composite cultured skin (CCS) for definitive closure. The materials fulfilling each stage have undergone in vitro and in vivo pretesting in "small" large animal wounds. For humans, producing multiple, large CCSs requires a specialized bioreactor. This article reports a system used to close large porcine wounds. Three Large White pigs were used, each with two wounds (24.5 cm × 12 cm) into which biodegradable dermal scaffolds were implanted. A sample from discarded tissue allowed isolation/culture of autologous fibroblasts and keratinocytes. CCS production began by presoaking a 1-mm-thick biodegradable polyurethane foam in autologous plasma. In the bioreactor cassette, fibroblasts were seeded into the matrix with thrombin until established, followed by keratinocytes. The CCSs were applied onto integrated dermal scaffolds on day 35, alongside a sheet skin graft (30% of one wound). Serial punch biopsies, trans-epidermal water loss readings (TEWL), and wound measurements indicated epithelialization. During dermal scaffold integration, negligible wound contraction was observed (average 4.5%). After CCS transplantation, the control skin grafts were "taken" by day 11 when visible islands of epithelium were clinically observed on 2/3 CCSs. Closure was confirmed histologically, with complete epithelialization by day 63 post-CCS transplantation (CCS TEWL ~ normal skin average 11.9 g/m2h). Four of six wounds demonstrated closure with robust, stratified epithelium. Generating large pieces of CCS capable of healing large wounds is thus possible using a specialized designed bioreactor.