Using keratinocytes differentiated from rat adipose tissue stem cells and PRP in experimental burn wound model of diabetic rats

Diabetes is one of the most important health problems with a rising prevalance each year all around the world. The problems seen in diabetic wound healing process lead to complications which can even end up with amputations in severe cases. Adipose tissue is a rich source of autologous mesenchymal stem cells (MSCs). Use of these cells in therapeutic applications has a great potential for effective treatment of diabetic patients. In this study, we used adipose tissue as the source of MSCs where we excised rat adipose tissue, cultured MSCs isolated from this tissue and then differentiated them into keratinocytes. The differentiated cells were then transferred to the thermal burn areas of STZ-induced diabetic rats. At the same time, we evaluated the effect of platelet rich-plasma (PRP) alone and in combination with keratinocytes in our thermal burn animal model. The experimental approach in our study included histologic examination of the wound area by H&E staining, immunohistochemical and RT-PCR analysis of the various growth factors (EGF, PDGF, VEGF, TGF-β1, FGF-2, FGF-7, FGF-10), MCP-1 and collagen-I. In addition, cytokeratins were analyzed by immunohistochemical analysis. The effect of cell therapy and/or PRP on secondary burn wound healing process was assessed in biopsy samples excised from the wound areas on the dorsal side of the treated/untreated rats on 3rd , 7th , 10th and 14th days after treatment. According to our results, re-epithelization started sooner in the treated groups in comparison with the untreated control group. In all treatment groups where we applied either keratinocyte-like cells or PRP, the level of growth factors and collagen-1 were found to be increased while MCP-1 levels decreased compared to the control group. Our data also indicated that the combined application of PRP and keratinocytes derived from MSCs showed the most accelerated healing process and was the best treatment for thermal wound healing model of diabetic rats. Keywords: wound healing, burnwound, mesenchimal stem cell, PRP, keratinocyte, diabetes, Rat