Abstract
The dynamics of inflammatory cytokines such as interleukin-1α (IL-lα, interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNFα) during the healing process of mouse skin wounds were examined using the enzyme-linked immunosorbent assay and immunostaining. The applicability of this examination for wound age estimation is discussed from the perspective of forensic pathology. After wound induction, mice were sacrificed at intervals ranging from 0 to 240 h. The levels of TNFα and IL-1β began to elevate rapidly after wounding and reached a peak at 3 h. The IL-lα level reached a peak at 6 h, and IL-6 peaked at 12 h. An infiltration of numerous leukocytes, indicatingacute inflammation, was observed at 3 and 6 h, and the main source of the cytokines was immunohistochemically identified as neutrophils. These results indicate that TNFα and IL-1β play an important role in the commencement of inflammation. “Rebound” of cytokine levels, i.e. a re-increase, was observed at 72 h after wounding. Histological examination of the 72-h-old wound showed migration of fibroblasts and the formation of new granulation tissues, indicating the proliferative stage of the wound healing process. These experimental findings indicate that these cytokines have a close relationship to wound remodeling as well as to inflammation. From the viewpoint of forensic pathology, it is considered that inflammatory cytokines may become one of the markers for wound age estimation, but further studies are needed, especially those involving the investigation using human wound specimens with known time intervals after injury.
Similar content being viewed by others
References
Auron PE, Webb AC, Rosenwasser LJ, Mucci SF, Rich A, Wolff SM, Dinarello CA (1984) Nucleotide sequence of human monocyte interleukin 1 precursor cDNA. Proc Natl Acad Sci USA 81: 7907–7911
Baumann H, Gauldie J (1994) The acute phase response. Immunol Today 15: 74–80
Besedovsky H, Del Rey A, Sorkin E, Dinarello CA (1986) Immunoregulatory feedback between interleukin 1 and glucocorticoid hormones. Science 233: 652–654
Bettinger DA, Pellicane JV, Tarry WC, Yager DR, Diegelmann RF, Lee R, Cohen IK, DeMaria EJ (1994) The role of inflammatory cytokines in wound healing: accelerated healing in endotoxin-resistant mice. J Trauma 36: 810–814
Betz P (1994) Histological and enzyme histochemical parameters for the age estimation of human skin wounds. Int J Legal Med 107:60–68
Betz P, Nerlich A, Wilske J, Tübel J, Wiest I, Penning R, Eisenmenger W (1992a) Immunohistochemical localization of fibronectin as a tool for the age determination of human skin wounds. Int J Legal Med 105: 21–26
Betz P, Nerlich A, Wilske J, Tübel J, Wiest I, Penning R, Eisenmenger W (1992b) The time-dependent rearrangement of epithelial basement membrane in human skin wounds- immunohistochemical localization of collagen IV and VII. Int J Legal Med 105: 93–97
Betz P, Nerlich A, Wilske J, Tübel J, Penning R, Eisenmenger W (1993a) Analysis of the immunohistochemical localization of collagen type III and V for the time-estimation of human skin wounds. Int J Legal Med 105: 329–332
Betz P, Nerlich A, Wilske J, Tübel J, Penning R, Eisenmenger W (1993b) Immunohistochemical localization of collagen types I and VI in human skin wounds. Int J Legal Med 106: 31–34
Betz P, Nerlich A, Wilske J, Tübel J, Penning R, Eisenmenger W (1993c) The time-dependent localization of Ki-67 antigen-positive cells in human skin wounds. Int J Legal Med 106: 35–40
Del Rey A, Besedovsky H, Sorkin E, Dinarello CA (1987) Interleukin-1 and glucocorticoid hormones integrate an immunoregulatory feedback circuit. Ann NY Acad Sci 496: 85–90
Dinarello CA (1994) Interleukin-1. In: Thomson A (ed) The cytokine handbook, 2nd edn. Academic Press, London, pp 31–56
Eisenmenger W, Nerlich A, Glück G (1988) Die Bedeutung des Kollagens bei der Wundaltersbestimmung. Z Rechtsmed 100: 79–100
Elias JA, Gustilo K, Baeder W, Freundlich B (1987) Synergistic stimulation of fibroblast prostaglandin production by recombinant interleukin 1 and tumor necrosis factor. J Immunol 138: 3812–3816
Fahey TJ III, Sherry B, Tracey KJ, van Deventer S, Jones WG II, Minei JP, Morgello S, Shires GT, Cerami A (1990) Cytokine production in a model of wound healing: the appearance of MIP-1, MIP-2, cachectin/TNF and IL-1. Cytokine 2: 92–99
Fahey TJ III, Sadaty A, Jones WG II, Barber A, Smoller B, Shires GT (1991) Diabetes impairs the late inflammatory response to wound healing. J Surg Res 50: 308–313
Ford HR, Hoffman RA, Wing EJ, Magee DM, McIntyre L, Simmons RL (1989) Characterization of wound cytokines in the sponge matrix model. Arch Surg 124: 1422–1428
Hirano T (1992) The biology of interleukin 6. Chem Immunol 51: 153–180
Kondo T, Ohshima T (1995) Experimental study on the estimation of skin wound age after injury by immunostaining interleukin lα, collagen type I and fibronectin. In: Bonte W, Oehmichen M, Geserick G, Saternus KS (eds) Rechtsmedizinische Forschungsergebnisse, Band 13. The wound healing processforensic pathological aspects. Schmidt-Römhild, Lübeck (in press)
Kurita Y, Tsuboi R, Ueki R, Rifkin DB, Ogawa H (1992) Immunchistochemical localization of basic fibroblast growth factor in wound healing sites of mouse skin. Arch Dermatol Res 284:193–197
Le J, Vilček J (1987) Tumor necrosis factor and interleukinl: cytokines with multiple overlapping biological activities. Lab Invest 56: 234–248
Lomedico PT, Gubler U, Hellmann CP, Dukovich M, Giri JG, Pan YCE, Collier K, Semionow R, Chua AO, Mizel SB (1984) Cloning and expression of murine interleukin-1 cDNA in Escherichia coli. Nature 312: 458–462
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275
Mateo RB, Reichner JS, Albina JE (1994) Interleukin-6 activity in wounds. Am J Physiol 266: R1840-R1844
Nishihara T, Ishihara Y, Noguchi T, Koga T (1989) Membrane IL-1 induces bone resorption in organ culture. J Immunol 143: 1881–1886
Oehmichen M (1990) Die Wundheilung. Springer, Berlin Heidelberg New York, pp 5–67
Oehmichen M, Cröpelin A (1995) Temporal course of intravital and postmortem proliferation of epidermal cells after injury — an immunohistochemical study using bromodeoxyuridine in rats. Int J Legal Med 107: 257–262
Oppenheim JJ, Kovacs EJ, Matsushima K, Durum SK (1986) There is more than one interleukin 1. Immunol Today 7: 45–56
Raekallio J (1972) Determination of the age of wounds by histochemical and biochemical methods. Forensic Sci Int 1: 3–16
Raekallio J (1976) Timing of wounds in forensic medicine. Jpn J Legal Med 30: 125–136
Tracey KJ (1994) Tumor necrosis factor-alpha. In: Thomson A (ed) The cytokine handbook, 2nd edn. Academic Press, London, pp 289–304
Wahl LM, Wahl SM (1992) Inflammation. In: Cohen K, Diegelmann RF, Lindbald WJ (eds) Wound healing: biochemical & clinical aspects. Saunders, Philadelphia, pp 40–62
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kondo, T., Ohshima, T. The dynamics of inflammatory cytokines in the healing process of mouse skin wound: A preliminary study for possible wound age determination. Int J Leg Med 108, 231–236 (1996). https://doi.org/10.1007/BF01369816
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01369816