Research Article| Volume 60, ISSUE 8, P864-875, August 2007

Perivascular cells in a skin graft are rapidly repopulated by host cells

Published:March 12, 2007DOI:


      Survival of grafted tissues is dependent upon revascularisation. This study investigated revascularisation in a murine skin graft model, using two methods. The first involved 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine (DiI) labelling of the wound bed, prior to replacing the skin graft, to allow tracking of host cells into the grafts. At time points between day 3 and day 14 post-surgery, DiI-labelled cells which had tracked into the grafts, were found to co-localise with CD31 positive endothelial cells and patent perfused vessels (fluorescein isothiocyanate (FITC)–dextran perfusion), to show possible association with the vasculature. To further differentiate between graft and host-derived cells, C57BL/6 wild-type grafts were placed on enhanced-green fluorescent protein (e-GFP) transgenic mouse hosts, and at set times post-grafting examined using confocal microscopy.
      Patent vessels were found at all depths of the graft by day 3. Host (DiI- or GFP-positive) cells were predominantly co-localised with graft vessels in grafts from day 3 onwards, with a similar morphology to control skin. Significantly more GFP labelled host cells were visualised in the superficial dermis at day 5 compared to day 3.
      Initial restoration of circulation appears to be due to linkage between existing graft and bed vessels, followed by an influx of host cells with a definite perivascular distribution. These findings have implications for skin autografts and tissue engineered skin substitutes.


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        • O'Ceallaigh S.
        • Herrick S.E.
        • Bluff J.E.
        • et al.
        Quantification of total and perfused blood vessels in murine skin autografts using a fluorescent double-labeling technique.
        Plast Reconstr Surg. 2006; 117: 140-151
        • McCluskey J.
        • Martin P.
        Analysis of the tissue movements of embryonic wound healing – DiI studies in the limb bud stage mouse embryo.
        Dev Biol. 1995; 170: 102-114
        • Okabe M.
        • Ikawa M.
        • Kominami K.
        • et al.
        ‘Green mice’ as a source of ubiquitous green cells.
        FEBS Lett. 1997; 407: 313-319
        • Patterson G.H.
        • Knobel S.M.
        • Sharif W.D.
        • et al.
        Use of the green fluorescent protein and its mutants in quantitative fluorescence microscopy.
        Biophys J. 1997; 73: 2782-2790
        • Boyce S.T.
        • Goretsky M.J.
        • Greenhalgh D.G.
        • et al.
        Comparative assessment of cultured skin substitutes and native skin autograft for treatment of full-thickness burns.
        Ann Surg. 1995; 222: 743-752
        • Braverman I.M.
        • Yen A.
        Ultrastructure of the human dermal microcirculation. II. The capillary loops of the dermal papillae.
        J Invest Dermatol. 1977; 68: 44-52
        • Braverman I.M.
        Ultrastructure and organization of the cutaneous microvasculature in normal and pathologic states.
        J Invest Dermatol. 1989; 93: 2S-9S
        • Clemmesen T.
        The early circulation in split skin grafts: restoration of blood supply to split-skin grafts.
        Acta Chir Scand. 1964; 127: 1-8
        • Stevens S.M.
        The restoration of the vasculature of skin autografts in the rabbit.
        Pathology. 1975; 7: 79-90
        • Birch J.
        • Branemark P.I.
        • Lundskog J.
        The vascularization of a free full thickness skin graft. II. A microangiographic study.
        Scand J Plast Reconstr Surg. 1969; 3: 11-17
        • Haller Jr., J.A.
        • Billingham R.E.
        Studies of the origin of the vasculature in free skin grafts.
        Ann Surg. 1967; 166: 896-901
        • Marckmann A.
        Autologous skin grafts in the rat: vital microscopic studies of the microcirculation.
        Angiology. 1966; 17: 475-482
        • Goretsky M.J.
        • Breeden M.
        • Pisarski G.
        • et al.
        Capillary morphogenesis during healing of full-thickness skin grafts: an ultrastructural study.
        Wound Repair Regen. 1995; 3: 213-220
        • Zarem H.A.
        • Zweifach B.W.
        • McGehee J.M.
        Development of microcirculation in full thickness autogenous skin grafts in mice.
        Am J Physiol. 1967; 212: 1081-1085
        • Plenat F.
        • Vignaud J.M.
        • Guerret-Stocker S.
        • et al.
        Host–donor interactions in healing of human split-thickness skin grafts onto nude mice: in situ hybridization, immunohistochemical, and histochemical studies.
        Transplantation. 1992; 53: 1002-1010
        • Oswald J.
        • Boxberger S.
        • Jorgensen B.
        • et al.
        Mesenchymal stem cells can be differentiated into endothelial cells in vitro.
        Stem Cells. 2004; 22: 377-384
        • Lin Y.
        • Weisdorf D.J.
        • Solovey A.
        • et al.
        Origins of circulating endothelial cells and endothelial outgrowth from blood.
        J Clin Invest. 2000; 105: 71-77
        • Wang H.J.
        • Wan H.L.
        • Yang T.S.
        • et al.
        Acceleration of skin graft healing by growth factors.
        Burns. 1996; 22: 10-14
        • Rinsch C.
        • Quinodoz P.
        • Pittet B.
        • et al.
        Delivery of FGF-2 but not VEGF by encapsulated genetically engineered myoblasts improves survival and vascularization in a model of acute skin flap ischemia.
        Gene Ther. 2001; 8: 523-533
        • Zhang F.
        • Oswald T.
        • Lin S.
        • et al.
        Vascular endothelial growth factor (VEGF) expression and the effect of exogenous VEGF on survival of a random flap in the rat.
        Br J Plast Surg. 2003; 56: 653-659
        • Lees V.C.
        • Fan T.P.
        A freeze-injured skin graft model for the quantitative study of basic fibroblast growth factor and other promoters of angiogenesis in wound healing.
        Br J Plast Surg. 1994; 47: 349-359
        • Gruss C.J.
        • Satyamoorthy K.
        • Berking C.
        • et al.
        Stroma formation and angiogenesis by overexpression of growth factors, cytokines, and proteolytic enzymes in human skin grafted to SCID mice.
        J Invest Dermatol. 2003; 120: 683-692
        • Khozani T.T.
        • Noorafshan A.
        • Nikeghbalian S.
        • et al.
        Stereologic study of the effects of prostaglandin E2 on the induction of angiogenesis in full-thickness skin autografts.
        Adv Skin Wound Care. 2004; 17 ([206]): 202-204