1Diabetes Center, Tokyo Women's Medical University, Tokyo, Japan; 2Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan
Impaired skin wound healing caused by reduced blood supply is a major complication especially in diabetic patients. Although artificial dermis is currently utilized for the treatment of ulcers with full-thickness skin defects in diabetic patients, it is difficult to cure because of blood flow obstruction.
Previous studies have shown that adipose derived stromal cells (ADSCs) possess angiogenic property, and ADSCs have been transplanted to wounded skin. However, it is hard to retain the transplanted ADSCs on the site and the outcome is limited. To resolve this problem, we take advantage of cell sheet technology using temperature responsive culture dishes and the efficacy of ADSC sheet was evaluated using rat full-thickness skin defect in head.
First, inguinal fat tissue from a normal rat was taken and extracted ADSCs (rADSCs) . Colony forming, osteogenic and adipogenic differentiation potential of the rADSCs were investigated in vitro. One thousand of rADSCs were seeded onto 10 cm dishes and cultured in growth, osteogenic, and adipogenic differentiation medium, respectively. rADSCs exhibited colony forming, osteogenic, and adipogenic capacities, suggesting that rADSCs used in this study possessed multipotent mesenchymal stromal cells-like properties. rADSCs from normal rats were seeded onto temperature responsive culture dishes to fabricate cell sheets.
Full-thickness skin defect was made on a normal rat calvariae, and periosteum was removed. The defect in the calvariae of rat was covered with artificial dermises (Pernac(R)) with or without transplantation of rADSC sheets. Two weeks after the transplantation, rats were sacrificed and the wound regions including the calvariae were excised and histologically investigated. Newly formed blood vessels were observed in the middle of wounds both in rats with or without the transplantation of the ADSC cell sheet. However, the density of blood vessels was 1.9-fold higher in the transplanted rat compared that of control rat.
These results suggest that rADSC sheet transplantation combined with artificial dermis accelerates the vascularization in full-thickness skin defect model, and is useful for the skin wound treatment of diabetic patients.
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