Effects of mature adipocyte-derived dedifferentiated fat (DFAT) cells on generation and vascularisation of dermis-like tissue after artificial dermis grafting

Abstract Although artificial dermis (AD) is effective for skin reconstruction, it requires two separate procedures, because the AD must be vascularised before skin grafts. To shorten the period of the dermis-like tissue generation before the secondary skin grafting must be beneficial. Dedifferentiated fat (DFAT) cells are isolated from mature adipose cell suspensions and have potential to differentiate into multiple cell types including endothelial cells. This study aimed to investigate effects of DFAT cells on dermal regeneration after AD grafts in rats. The effects of combination use of DFAT cells and basic fibroblast growth factor (bFGF) were also tested to mimic clinical situations. DFAT cells were isolated from SD rats. Full-thickness wounds were created on the back of rats followed by AD grafting. Five groups were established; Group I: control, Group II: treated with DFAT cells, Group III: treated with bFGF, Group IV: treated with both of DFAT cells and bFGF, and Group V: treated with Green fluorescent protein (GFP)-labelled DFAT cells and bFGF. Histological evaluation was serially performed. Group IV showed markedly promoted vascularisation of dermis-like tissue. In particular, capillary infiltration into the dermis was obtained within 2 days. Immunohistochemical examination revealed that the transplanted DFAT cells had differentiated into endothelial cells and participated in angiogenesis. Group IV also showed a marked increase in the thickness of the dermis like tissue. The present results suggest that the use of DFAT cells under bFGF treatment could be beneficial to shorten the period required for dermal regeneration and vascularisation and contribute to use AD more effectively and safely.

[1]  E. Yeong,et al.  Is Artificial Dermis an Effective Tool in the Treatment of Tendon-Exposed Wounds? , 2013, Journal of burn care & research : official publication of the American Burn Association.

[2]  Sepideh Heydarkhan‐Hagvall,et al.  Endothelial differentiation in multipotent cells derived from mouse and human white mature adipocytes. , 2012, Journal of molecular and cellular cardiology.

[3]  I. Yannas,et al.  Template for Skin Regeneration , 2011, Plastic and reconstructive surgery.

[4]  M. Longaker,et al.  Stem Cells , 2010, Plastic and reconstructive surgery.

[5]  S. E. James,et al.  A review of tissue-engineered skin bioconstructs available for skin reconstruction , 2010, Journal of The Royal Society Interface.

[6]  M Przybylski,et al.  A review of the current research on the role of bFGF and VEGF in angiogenesis. , 2009, Journal of wound care.

[7]  A. Hirayama,et al.  Dedifferentiated fat cells convert to cardiomyocyte phenotype and repair infarcted cardiac tissue in rats. , 2009, Journal of molecular and cellular cardiology.

[8]  S. Canonico,et al.  The Use of a Dermal Substitute and Thin Skin Grafts in the Cure of “Complex” Leg Ulcers , 2009, Dermatologic surgery : official publication for American Society for Dermatologic Surgery [et al.].

[9]  Olivera Stojadinovic,et al.  PERSPECTIVE ARTICLE: Growth factors and cytokines in wound healing , 2008, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[10]  K. Kano,et al.  Establishment of a preadipocyte cell line derived from mature adipocytes of GFP transgenic mice and formation of adipose tissue , 2008, Cell and Tissue Research.

[11]  N. Fukuda,et al.  Mature adipocyte‐derived dedifferentiated fat cells exhibit multilineage potential , 2008, Journal of cellular physiology.

[12]  A. Hirano,et al.  A basic fibroblast growth factor improves lower extremity wound healing with a porcine-derived skin substitute. , 2008, The Journal of trauma.

[13]  I. Ono,et al.  Basic fibroblast growth factor in an artificial dermis promotes apoptosis and inhibits expression of α‐smooth muscle actin, leading to reduction of wound contraction , 2007, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[14]  A. Utani,et al.  Incorporation of basic fibroblast growth factor into preconfluent cultured skin substitute to accelerate neovascularisation and skin reconstruction after transplantation , 2007, Scandinavian journal of plastic and reconstructive surgery and hand surgery.

[15]  Kristina D. O'Shaughnessy,et al.  Chronic Wound Pathogenesis and Current Treatment Strategies: A Unifying Hypothesis , 2006, Plastic and reconstructive surgery.

[16]  M. Nozaki,et al.  Novel application method of artificial dermis: one-step grafting procedure of artificial dermis and skin, rat experimental study. , 2006, Burns : journal of the International Society for Burn Injuries.

[17]  Y. Okazaki,et al.  A novel preadipocyte cell line established from mouse adult mature adipocytes. , 2004, Biochemical and biophysical research communications.

[18]  N. Moiemen,et al.  Reconstructive Surgery with a Dermal Regeneration Template: Clinical and Histologic Study , 2001, Plastic and reconstructive surgery.

[19]  J. Ascherman,et al.  Artificial Skin for Closure and Healing of Wounds Created by Skin Cancer Excisions , 2001, Dermatologic surgery : official publication for American Society for Dermatologic Surgery [et al.].

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

[21]  Y. Ikada,et al.  Accelerated tissue regeneration through incorporation of basic fibroblast growth factor-impregnated gelatin microspheres into artificial dermis. , 2000, Biomaterials.

[22]  E. Dabelsteen,et al.  Proliferation and mitogenic response to PDGF-BB of fibroblasts isolated from chronic venous leg ulcers is ulcer-age dependent. , 1999, The Journal of investigative dermatology.

[23]  J. Menzoian,et al.  Fibroblasts cultured from distal lower extremities in patients with venous reflux display cellular characteristics of senescence. , 1998, Journal of vascular surgery.

[24]  K. Sasaki,et al.  Reconstruction of burn deformity using artificial dermis combined with thin split-skin grafting. , 1997, Burns : journal of the International Society for Burn Injuries.

[25]  D. Prockop Marrow Stromal Cells as Stem Cells for Nonhematopoietic Tissues , 1997, Science.

[26]  Yoshimitsu Kuroyanagi,et al.  Design of artificial skin , 1996 .

[27]  Lothar Schweigerer Basic fibroblast growth factor as a wound healing hormone. , 1988, Trends in pharmacological sciences.

[28]  D. Heimbach,et al.  Artificial Dermis for Major Burns: A Multi‐Center Randomized Clinical Trial , 1988, Annals of surgery.

[29]  J. Aubin,et al.  Differentiation of muscle, fat, cartilage, and bone from progenitor cells present in a bone-derived clonal cell population: effect of dexamethasone , 1988, The Journal of cell biology.

[30]  D. Gospodarowicz,et al.  Fibroblast growth factor , 1986, Molecular and Cellular Endocrinology.

[31]  M. Klagsbrun,et al.  Accelerated wound repair, cell proliferation, and collagen accumulation are produced by a cartilage-derived growth factor , 1985, The Journal of cell biology.

[32]  J. Burke,et al.  Successful Use of a Physiologically Acceptable Artificial Skin in the Treatment of Extensive Burn Injury , 1981, Annals of surgery.

[33]  Xiaofeng Zhou,et al.  Accelerated healing of diabetic wound using artificial dermis constructed with adipose stem cells and poly (L-glutamic acid)/chitosan scaffold. , 2013, Chinese medical journal.

[34]  H. Mizuno,et al.  Adipose-derived stem cells for skin regeneration. , 2011, Methods in molecular biology.

[35]  O. Lee,et al.  blood Isolation of multipotent mesenchymal stem cells from umbilical cord , 2010 .

[36]  K. Kano,et al.  Mature adipocyte-derived dedifferentiated fat cells can trans-differentiate into osteoblasts in vitro and in vivo only by all-trans retinoic acid. , 2008, Cell structure and function.

[37]  S. Canonico,et al.  The use of artificial dermis in the treatment of chronic and acute wounds: regeneration of dermis and wound healing. , 2005, Acta bio-medica : Atenei Parmensis.

[38]  I. Ono,et al.  Effects of a collagen matrix containing basic fibroblast growth factor on wound contraction. , 1999, Journal of biomedical materials research.

[39]  N. Isshiki,et al.  Clinical evaluation of a new bilayer "artificial skin" composed of collagen sponge and silicone layer. , 1990, British journal of plastic surgery.

[40]  M. Longaker,et al.  Histologic study of artificial skin used in the treatment of full-thickness thermal injury. , 1990, The Journal of burn care & rehabilitation.

[41]  I. Yannas,et al.  Design of an artificial skin. I. Basic design principles. , 1980, Journal of biomedical materials research.