• chitosan skin graft;
  • keratinocyte;
  • fibroblast;
  • co-culture;
  • wound healing


This study evaluated the modulatory effect of chitosan-sponge co-cultured with keratinocyte and fibroblast on wound healing. Dermal fibroblasts and keratinocyte isolated from rabbit skin were co-cultured on chitosan sponge, to fabricate cell loaded chitosan tissue engineered construct. Full thickness excision wounds created on the rabbit dorsum were treated with three types of graft materials – a non-cellular chitosan graft, homologous keratinocyte fibroblast loaded chitosan and a commercial product. Post-graft skin-wound samples were examined histomorphologically at 7th, 14th and 28th day after staining with H and E, Picrosirius red and/ or immuno-histochemistry. Wound healing parameters considered were the extent of re-epithelialization, collagen deposition and neo-angiogenesis. The number of proliferating cells, vimentin positive cells and alpha smooth muscle actin cells were also quantified. The histology results suggested that the grafts aided wound healing but, the cell loaded graft induced a differential pattern of healing and had lower scarring tendency. The cell loaded tissue construct may be useful as a therapeutic graft for treating wounds where there is a total loss of tissue and cells as in burn injury.