Expression of interleukin-1alpha, interleukin-6, and basic fibroblast growth factor by cultured skin substitutes before and after grafting to full-thickness wounds in athymic mice.

OBJECTIVES Cultured skin substitutes (CSSs), consisting of human keratinocytes and human fibroblasts attached to collagen-glycosaminoglycan substrates, have been demonstrated to cover wounds, and may release detectable quantities of growth factors that promote wound healing. MATERIALS AND METHODS Basic fibroblast growth factor (bFGF), interleukin-1alpha (IL-1alpha), and interleukin-6 (IL-6) were assayed by enzyme linked immunosorbent assay and immunohistochemistry in CSSs in vitro and at days 1, 3, 7, 14, and 21 after grafting to full-thickness wounds in athymic mice. MEASUREMENTS AND MAIN RESULTS When isolated cells were tested, IL-1alpha was found to come primarily from the keratinocytes, whereas bFGF was from the fibroblasts. Combinations of both cell types in the CSSs resulted in a synergistic enhancement of IL-6 expression. Quantities of all three cytokines from CSSs were greater in vitro compared with in vivo levels at all time points after grafting. bFGF increased from day 1 to day 7, and then remained relatively constant until day 21. At day 3 maximal levels of IL-1alpha were observed. By day 7, IL-1alpha decreased to approximately 40% of maximal levels, and subsequently increased until day 21. IL-6 levels were highest at day 7 after grafting. All cytokines had reached elevated levels during the time of wound revascularization (days 3-7). CONCLUSIONS The sequence of cytokine synthesis in the wounds (i.e., rapid IL-1alpha increase followed by IL-6 expression) parallels serum levels reported after a septic challenge. These findings support the hypothesis that the wound is a source of systemic cytokines.

[1]  D. Greenhalgh,et al.  Comparative Assessment of Cultured Skin Substitutes and Native Skin Autograft for Treatment of Full‐Thickness Burns , 1995, Annals of surgery.

[2]  D. Greenhalgh,et al.  Pigmentation and microanatomy of skin regenerated from composite grafts of cultured cells and biopolymers applied to full-thickness burn wounds. , 1995, Transplantation.

[3]  F. Watt,et al.  Integrin expression by human epidermal keratinocytes can be modulated by interferon-γ, transforming growth factor-β, tumor necrosis factor-α, and culture on a dermal equivalent , 1995 .

[4]  M. Fartasch,et al.  Improved barrier structure formation in air-exposed human keratinocyte culture systems. , 1994, The Journal of investigative dermatology.

[5]  D. Heimbach,et al.  Basic fibroblast growth factor in the early human burn wound. , 1994, The Journal of surgical research.

[6]  S. Boyce,et al.  Lipid supplemented medium induces lamellar bodies and precursors of barrier lipids in cultured analogues of human skin. , 1993, The Journal of investigative dermatology.

[7]  S. Boyce,et al.  Pigmentation and inhibition of wound contraction by cultured skin substitutes with adult melanocytes after transplantation to athymic mice. , 1993, The Journal of investigative dermatology.

[8]  D. Greenhalgh,et al.  Skin anatomy and antigen expression after burn wound closure with composite grafts of cultured skin cells and biopolymers. , 1993, Plastic and reconstructive surgery.

[9]  M. van Deuren,et al.  Cytokines and the response to infection , 1992, The Journal of pathology.

[10]  M. Nakamura,et al.  Interleukin 6 promotes epithelial migration by a fibronectin‐dependent mechanism , 1992, Journal of cellular physiology.

[11]  L. Broemeling,et al.  The Safety and Effect of Topically Applied Recombinant Basic Fibroblast Growth Factor on the Healing of Chronic Pressure Sores , 1992, Annals of surgery.

[12]  G. Kratz,et al.  Conditioned medium from cultured human keratinocytes has growth stimulatory properties on different human cell types. , 1991, The Journal of investigative dermatology.

[13]  N C Krejci,et al.  In vitro reconstitution of skin: fibroblasts facilitate keratinocyte growth and differentiation on acellular reticular dermis. , 1991, The Journal of investigative dermatology.

[14]  B. Berman,et al.  Stimulation of collagen and glycosaminoglycan production in cultured human adult dermal fibroblasts by recombinant human interleukin 6. , 1991, The Journal of investigative dermatology.

[15]  I. Leigh,et al.  Epidermal cytokines and their roles in cutaneous wound healing , 1991, The British journal of dermatology.

[16]  Thomas A. Mustoe, MD, FACS,et al.  Growth factor-induced acceleration of tissue repair through direct and inductive activities in a rabbit dermal ulcer model. , 1991, The Journal of clinical investigation.

[17]  V. Dixit,et al.  Keratinocytes as initiators of inflammation , 1991, The Lancet.

[18]  T. Luger,et al.  Evidence for an epidermal cytokine network. , 1990, The Journal of investigative dermatology.

[19]  T. Kupper,et al.  The activated keratinocyte: a model for inducible cytokine production by non-bone marrow-derived cells in cutaneous inflammatory and immune responses. , 1990, The Journal of investigative dermatology.

[20]  D. Greenhalgh,et al.  PDGF and FGF stimulate wound healing in the genetically diabetic mouse. , 1990, The American journal of pathology.

[21]  H. Ford,et al.  Characterization of wound cytokines in the sponge matrix model. , 1989, Archives of surgery.

[22]  K. Tracey,et al.  Antibodies to cachectin/tumor necrosis factor reduce interleukin 1 beta and interleukin 6 appearance during lethal bacteremia , 1989, The Journal of experimental medicine.

[23]  H Green,et al.  Treatment of skin ulcers with cultured epidermal allografts. , 1989, Journal of the American Academy of Dermatology.

[24]  S. Boyce,et al.  Structure of a collagen-GAG dermal skin substitute optimized for cultured human epidermal keratinocytes. , 1988, Journal of biomedical materials research.

[25]  A. Sommer,et al.  Recombinant basic fibroblast growth factor accelerates wound healing. , 1988, The Journal of surgical research.

[26]  S. Boyce,et al.  Biologic attachment, growth, and differentiation of cultured human epidermal keratinocytes on a graftable collagen and chondroitin-6-sulfate substrate. , 1988, Surgery.

[27]  I. Leigh,et al.  Treatment of chronic venous ulcers with sheets of cultured allogenic keratinocytes , 1987, The British journal of dermatology.

[28]  J. Vilček,et al.  A cytokine network in human diploid fibroblasts: interactions of beta-interferons, tumor necrosis factor, platelet-derived growth factor, and interleukin-1 , 1987, Molecular and cellular biology.

[29]  T. Kupper,et al.  The human burn wound as a primary source of interleukin-1 activity. , 1986, Surgery.

[30]  D. Gospodarowicz,et al.  Molecular and biological characterization of fibroblast growth factor, an angiogenic factor which also controls the proliferation and differentiation of mesoderm and neuroectoderm derived cells. , 1986, Cell differentiation.

[31]  A. Balin,et al.  Grafting of skin ulcers with cultured autologous epidermal cells. , 1986, Journal of the American Academy of Dermatology.

[32]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[33]  F. Watt,et al.  Integrin expression by human epidermal keratinocytes can be modulated by interferon-gamma, transforming growth factor-beta, tumor necrosis factor-alpha, and culture on a dermal equivalent. , 1995, The Journal of investigative dermatology.

[34]  S. Boyce.  Epidermis as a secretory tissue. , 1994, The Journal of investigative dermatology.

[35]  O. Røkke,et al.  The synergistic effect of trauma and infection on interleukin-1 but not tumor necrosis factor liberation during posttraumatic gram-negative septicemia. , 1993, European surgical research. Europaische chirurgische Forschung. Recherches chirurgicales europeennes.

[36]  W. Eaglstein,et al.  Interleukin-1 enhances epidermal wound healing. , 1990, Lymphokine research.