On The Effect Of Acellular Dermal Matrix Combined With Bone Marrow Mesenchymal Stem Cells For Repairing Full-thickness Skin Defect In Diabetic Rat

Object:1. Isolation, culture and amplification BMSCs in vitro;2. Construction of tissue-engineered skins contains bone marrow mesenchymal stem cells and acellular dermal matrix build;3. To investigate the effect of repairing full-thickness skin wounds in diabetic rat by acellular dermal matrix combined with bone marrow mesenchymal stem cells.Methods:1. The bone marrow mesenchymal stem cells were harvested from rat,and they were isolated、culture and amplificated in vitro by whole bone marrow culture method;2. Extract rat tail collagen were harvested by acid extraction;3. Preparation of tissue-engineered skin:acellular dermal matrix were prepared by trypsin-detergent method and embedded in rat tail collagen then co-cultured with bone marrow mesenchymal stem cells labeled with5-Brdu(5-bromodeoxy-uridine) about12days;4. Once celiac streptozocin injection was used to establish diabetes model rat, then tissue-engineered skin was transplant to rat diabetic wound;5. Transparent membrane method was used for calculation and analysis of the wound healing rate in different groups;6. Samples of newly-grown skin in different groups were taken to make moutine paraffin sections for microscopical examination and immunohistochemical staining for the origin of epidermic cells. Results:1. Bone marrow mesenchymal stem cells were isolated, cultured and amplification successfully;2. Rat tail collagen was extracted successfully;3. Tissue-engineered skin contain of bone marrow mesenchymal stem cells and acellular dermal matrix successfully4Full skin loss wound was created on back of diabetes rat successfully. Using method of tissue engineering, Tissue-engineering skin contains acellular dermal matrix as supporter, labeled5-BrdU BMSCs derivative from rat as original have positive effects on healing skin wound. The result from the hematoxylin and eosin staining showed that the epidermis is thicker in treat group than that in control group. A few Brdu positive cell appeared on the rengenerate skin and the stratum basale and the stratum spinosum of the rengenerate epidermis post operation21days.Conclusion:1. Acellular dermal matrix with bone marrow mesenchymal stem cells can good co-cultured and do not affect BMSCs proliferation and differentiation ability, is a good skin tissue engineering supporter.。2. The data suggest that tissue-engineered skin containing bone marrow mesenchymal stem cells as seed cells combined with collagen embedded acellular dermal matrix as supporter in a skin loss wound of rat diabetic can accelerate wound healing and receive satisfactory effects. It’s an effective method for treatment of diabetes and other causes of refractory wound.