Experimental Study on Promotion of Wound Healing with Artificial Skin Constructed with Epidermal Stem Cells and Acellular Dermal Matrix

Objective To construct the artificial skin with epidermal stem cells and acellular dermal matrix and to repair the rabbit full thickness dermal wounds.Methods(1)Human fetal epidermis was isolated and then dispersed by Trypsin-EDTA,and the human epidermal stem cells were isolated and purified by type Ⅳ collagen adhering method.The cells were cultured in keratinocyte serum free medium supplemented with 10% fetal bovine serum and 10 μg/L epidermal growth factor.The non-attaching keratinocytes were cultured as the control.The colony forming efficiency was examined by Giemsa staining.(2)The human epidermal stem cells were inoculated into the acellular dermal matrix to construct tissue-engineered skin,which was used for transplantation in treatment of rabbit full thickness skin wounds.(3)The wounds were produced on the back of New Zealand white rabbits.The rabbits were randomly divided into four groups:group A(tissue-engineered skin graft with epidermal stem cells);group B(tissue-engineered skin graft with keratinocytes);group C(simple acellular dermal matrix graft) and group D(vacant wounds as the control).The repairing effect of wounds,the local inflammatory response and the wound healing time were determined.Results The human epidermal stem cells proliferated steadily in vitro and the colony forming efficiency was significantly higher than that in cultured keratinocyte group(P0.05).After the artificial skin transplantation,the woulds in group A healed well,accompanied by slight local inflammatory response without bleeding,empyema and necrosis.The wound healing time in group A significantly shortened compared with group B,C and D,respectively.(P0.05 or P0.01).Conclusion The construction of artificial skin with epidermal stem cells and acellular dermal matrix can be used to repair and treat the skin defect wounds.