Creation and Transplantation of an Adipose-derived Stem Cell (ASC) Sheet in a Diabetic Wound-healing Model.

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients with impeded blood flow or with large wounds might be prolonged. Cell-based therapies have appeared as a new technique for the treatment of diabetic ulcers, and cell-sheet engineering has improved the efficacy of cell transplantation. A number of reports have suggested that adipose-derived stem cells (ASCs), a type of mesenchymal stromal cell (MSC), exhibit therapeutic potential due to their relative abundance in adipose tissue and their accessibility for collection when compared to MSCs from other tissues. Therefore, ASCs appear to be a good source of stem cells for therapeutic use. In this study, ASC sheets from the epididymal adipose fat of normal Lewis rats were successfully created using temperature-responsive culture dishes and normal culture medium containing ascorbic acid. The ASC sheets were transplanted into Zucker diabetic fatty (ZDF) rats, a rat model of type 2 diabetes and obesity, that exhibit diminished wound healing. A wound was created on the posterior cranial surface, ASC sheets were transplanted into the wound, and a bilayer artificial skin was used to cover the sheets. ZDF rats that received ASC sheets had better wound healing than ZDF rats without the transplantation of ASC sheets. This approach was limited because ASC sheets are sensitive to dry conditions, requiring the maintenance of a moist wound environment. Therefore, artificial skin was used to cover the ASC sheet to prevent drying. The allogenic transplantation of ASC sheets in combination with artificial skin might also be applicable to other intractable ulcers or burns, such as those observed with peripheral arterial disease and collagen disease, and might be administered to patients who are undernourished or are using steroids. Thus, this treatment might be the first step towards improving the therapeutic options for diabetic wound healing.

[1]  Tomoyuki Nishikawa,et al.  Novel Autologous Cell Therapy in Ischemic Limb Disease Through Growth Factor Secretion by Cultured Adipose Tissue–Derived Stromal Cells , 2005, Arteriosclerosis, thrombosis, and vascular biology.

[2]  H. Lorenz,et al.  Multilineage cells from human adipose tissue: implications for cell-based therapies. , 2001, Tissue engineering.

[3]  R. Reis,et al.  Human adipose stem cells cell sheet constructs impact epidermal morphogenesis in full-thickness excisional wounds. , 2013, Biomacromolecules.

[4]  M. Řezáčová,et al.  Zucker diabetic fatty rat: A new model of impaired cutaneous wound repair with type II diabetes mellitus and obesity , 2011, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[5]  C. Toulouse.  Adipose-derived stromal cells: Their identity and uses in clinical trials, an update , 2011 .

[6]  T. Okano,et al.  Allogeneic transplantation of an adipose-derived stem cell ( ASC ) sheet combined with artificial skin accelerates wound healing in a rat wound model of type 2 diabetes and obesity Running title : Allogeneic ASC sheet therapy for diabetic ulcer , 2015 .

[7]  Hidezo Mori,et al.  Monolayered mesenchymal stem cells repair scarred myocardium after myocardial infarction , 2006, Nature Medicine.

[8]  Hermann Eichler,et al.  Comparative Analysis of Mesenchymal Stem Cells from Bone Marrow, Umbilical Cord Blood, or Adipose Tissue , 2006, Stem cells.

[9]  T. Okano,et al.  A novel recovery system for cultured cells using plasma-treated polystyrene dishes grafted with poly(N-isopropylacrylamide). , 1993, Journal of biomedical materials research.

[10]  D. Peterson,et al.  Human Mesenchymal Stem Cell Grafts Enhance Normal and Impaired Wound Healing by Recruiting Existing Endogenous Tissue Stem/Progenitor Cells , 2013, Stem cells translational medicine.

[11]  T. Okano,et al.  Cell sheet engineering: a unique nanotechnology for scaffold‐free tissue reconstruction with clinical applications in regenerative medicine , 2010, Journal of internal medicine.

[12]  T. Okano,et al.  Thermo-responsive culture dishes allow the intact harvest of multilayered keratinocyte sheets without dispase by reducing temperature. , 2001, Tissue engineering.

[13]  Tim Nolan,et al.  International Diabetes Federation. , 2013, Diabetes research and clinical practice.

[14]  P. Zuk.  The ASC: Critical Participants in Paracrine-Mediated Tissue Health and Function , 2013 .

[15]  A. Herling,et al.  Intramyocellular Lipid and Insulin Resistance a Longitudinal in Vivo H-spectroscopic Study in Zucker Diabetic Fatty Rats , 2022 .

[16]  A. Boulton,et al.  The global burden of diabetic foot disease , 2005, The Lancet.

[17]  Masayuki Yamato,et al.  Cell sheet engineering and its application for periodontal regeneration , 2015, Journal of tissue engineering and regenerative medicine.

[18]  B. Cousin,et al.  Adipose-derived stromal cells: Their identity and uses in clinical trials, an update. , 2011, World journal of stem cells.

[19]  G. Gurtner,et al.  Quantitative and reproducible murine model of excisional wound healing , 2004, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[20]  T. Okano,et al.  Genetically modified adipose tissue-derived stem/stromal cells, using simian immunodeficiency virus-based lentiviral vectors, in the treatment of hemophilia B. , 2013, Human gene therapy.

[21]  Masayuki Yamato,et al.  Cardiac cell sheet transplantation improves damaged heart function via superior cell survival in comparison with dissociated cell injection. , 2011, Tissue engineering. Part A.

[22]  J. Isner,et al.  VEGF contributes to postnatal neovascularization by mobilizing bone marrow‐derived endothelial progenitor cells , 1999, The EMBO journal.

[23]  J. Park,et al.  Wound healing effect of adipose-derived stem cells: a critical role of secretory factors on human dermal fibroblasts. , 2007, Journal of dermatological science.

[24]  T. Okano,et al.  Recovery course of full-thickness skin defects with exposed bone: an evaluation by a quantitative examination of new blood vessels. , 2007, The Journal of surgical research.

[25]  Y. Ikada,et al.  Re-freeze dried bilayer artificial skin. , 1993, Biomaterials.

[26]  J. Dunlap,et al.  Changes in endoneurial blood flow, motor nerve conduction velocity and vascular relaxation of epineurial arterioles of the sciatic nerve in ZDF‐obese diabetic rats , 2002, Diabetes/metabolism research and reviews.

[27]  E. Davidson,et al.  Progression of vascular and neural dysfunction in sciatic nerves of Zucker diabetic fatty and Zucker rats. , 2005, American journal of physiology. Endocrinology and metabolism.

[28]  T. Okano,et al.  Allogeneic Transplantation of an Adipose-Derived Stem Cell Sheet Combined With Artificial Skin Accelerates Wound Healing in a Rat Wound Model of Type 2 Diabetes and Obesity , 2015, Diabetes.

[29]  J. Gimble,et al.  Multipotential human adipose‐derived stromal stem cells exhibit a perivascular phenotype in vitro and in vivo , 2008, Journal of cellular physiology.

[30]  Kacey G. Marra,et al.  The use of adipose-derived stem cells as sheets for wound healing , 2013, Organogenesis.

[31]  Zhenxing Si,et al.  Locally Administered Adipose-Derived Stem Cells Accelerate Wound Healing through Differentiation and Vasculogenesis , 2011, Cell transplantation.

[32]  G. Zambruno,et al.  Diabetes impairs adipose tissue–derived stem cell function and efficiency in promoting wound healing , 2013, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[33]  J. Rubin,et al.  Evaluation of a multi-layer adipose-derived stem cell sheet in a full-thickness wound healing model. , 2013, Acta biomaterialia.