Abstract
INTRODUCTION: Collagen-glycosaminoglycan (CG) scaffolds, also known as dermal regeneration templates, are used for the reconstruction of full-thickness skin defect in patients with extensive resections due to burns, traumatic or inflammatory conditions.1 The natural process of revascularization typically lasts two to three weeks precluding immediate skin grafting. The usage of stem cells has demonstrated to accelerate wound healing. However, stem cell usage requires several steps of cell culturing that preclude using this method in the acute setting. Aldehyde dehydrogenase (ALDH) is an enzyme that plays an important role in retinoid metabolism and is highly expressed in stem cells.2 In this study, we isolated ALDH-expressing cells from subcutaneous adipose tissue and tested them for their potential to enhance healing in a full-thickness skin wound in rats by co-implanting them with CG scaffolds. MATERIALS AND METHODS: Stromal-Vascular-Fraction (SVF) was obtained from subcutaneous adipose tissue of syngeneic rats. ALDHhi cells were isolated using a fluorescence-activated cell sorting technique with ALDEFLUOR assay kitTM. Each recipient rat underwent four full-thickness wounds creation on the recipient rat’s back, each wound was treated differently. A total of four treatment groups were formed (n=11). Group 1 (control group) consisted of wounds treated with CG and 100 μL normal saline. Group 2 (SVF group) consisted of CG and 1 × 105 cells/cm2 SVF cells. Group 3 (ALDH group) consisted of CG and 1 × 105 cells/cm2 ALDHhi cells. Group 4 (ASCs group) consisted of CG and 1 × 105 cells/cm2 ASCs. Animals were evaluated by histology on day 7 after surgeries. RESULTS: Scaffolds seeded with ALDHhi cells histologically demonstrated remarkable enhancements in dermal regeneration, vascularization, and collagen growth, if compared to the wound treated with CG alone, CG with SVF, and CG with ASCs groups. Immunofluorescent staining with CD31 emphasized that transplanted ALDHhi cells differentiated into vascular endothelial cells. CONCLUSIONS: Composite transplantation of CG scaffolds and adipose-derived ALDHhi cells promoted dermal regeneration, not worse than cultured ASC, suggesting that ALDHhi cells could be used in an acute setting as a reliable alternative for cultured ASCs. DISCLOSURE/FINANCIAL SUPPORT:Supported by Toho Women’s Clinic Research Foundation, the Gillian Reny Stepping Strong Fund, and the Brigham Research Institute and the Center for Faculty Development and Diversity’s Office for Research Careers Microgrant Program. None of the authors has a financial interest in any of the products, devices, or drugs mentioned in this manuscript. REFERENCES: 1. Yannas IV, Orgill DP, Burke JF. Template for Skin Regeneration. Plastic and Reconstructive Surgery 2011;127(1):60S-70S 2. Cai JL, Weiss ML, Rao MS. In search of “stemness”. Experimental Hematology 2004;32(7):585–98
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