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J Reconstr Microsurg 2004; 20(3): 253-259
DOI: 10.1055/s-2004-823113
Copyright © 2004 by Thieme Medical Publishers, Inc., 333 Seventh Avenue, New York, NY 10001, USA.

Comparison of Six Methods for the Assessment of Ischemia-Reperfusion Injury in Skeletal Muscle following Composite Tissue Allotransplantation

Steffen P. Baumeister1 , Nina Ofer1 , Christian Kleist2 , Martin Rebel3 , Bernd Dohler2 , Peter Terness2 , Martha Maria Gebhard3 , Guenter Germann1
  • 1Department of Hand, Plastic, and Reconstructive Surgery, Burn Center, BG Trauma Center, Ludwigshafen, University of Heidelberg, Germany
  • 2Institute of Immunology, Department of Transplantation Immunology, University of Heidelberg, Germany
  • 3Institute of Kapital Pathology, Clinic of Ludwigshafen, Germany
Further Information

Publication History

Accepted: 23 September 2003

Publication Date:
16 April 2004 (online)

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Ischemia/reperfusion (I/R) injury is one of the factors determining tissue survival in replantation and transplantation surgery. However, more than 20 methods have been used to evaluate I/R injury in muscle. The aim of this study was to examine I/R injury in muscle tissue in a model of composite tissue allotransplantation (rat hindlimb transplantation), based on the analyses of six parameters: nitroblue tetrazolium staining (NBT); histology of the anterior tibial and extensor digitorum muscle; wet-to-dry weight ratio; serum potassium; and serum creatine kinase (CK)), in order to identify the most practicable and reliable outcome parameter. Results demonstrated that NBT staining and the wet/dry weight ratio are reliable tools for outcome measurement. The wet/dry weight ratio is the easiest to perform and the authors consider it to be useful for screening purposes. Histologic assessment shows areas of necrosis, but is not a reliable method for semi-quantitative evaluation. Serum potassium and CK levels were higher following transplantation, but they cannot be recommended for assessment purposes, as no significant correlation with other parameters was seen. These findings help further researchers in their selection of reliable outcome parameters to measure I/R injury in skeletal muscle.

KEYWORDS

Ischemia-reperfusion injury - nitroblue tetrazolium - hindlimb transplantation

REFERENCES

  • 1 Wagh M, Pantazi G, Romeo R et al.. Cold storage of rat skeletal muscle free flaps and pre-ischemic perfusion with modified UW solution.  Microsurgery. 2000;  20 343-349
    CrossrefPubMedGoogle Scholar
  • 2 Shaw W W, Ko C Y, Ahn C Y et al.. Safe ischemia time in free-flap surgery: a clinical study of contact surface cooling.  J Reconstr Microsurg. 1996;  12 421-424
    PubMedGoogle Scholar
  • 3 Babajanian M, Zhang W X, Aviv J E et al.. Prolongation of secondary critical ischemia time of experimental skin flaps using UW solution as a normothermic perfusate.  Otolaryngol Head Neck Surg. 1993;  108 149-155
    PubMedGoogle Scholar
  • 4 van Alphen W A, Smith A R, ten Kate F J. Maximum hypothermic ischemia in replants containing muscular tissue.  J Hand Surg. 1988;  13A 415-422
    PubMedGoogle Scholar
  • 5 Wang B H, Ye C, Stagg C A et al.. Improved free musculocutaneous flap survival with induction of heat shock protein.  Plast Reconstr Surg. 1998;  101 776-784
    CrossrefPubMedGoogle Scholar
  • 6 Piza-Katzer H, Hussl H, Ninkovic M et al.. Beidseitige Handtransplantation.  Handchir Mikrochir Plast Chir. 2002;  34 75-83
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Beyersdorf F, Unger A, Wildhirt A et al.. Studies of reperfusion injury in skeletal muscle: preserved cellular viability after extended periods of warm ischemia.  J Cardiovasc Surg (Torino). 1991;  32 664-676
    PubMedGoogle Scholar
  • 8 Blaisdell F W. The pathophysiology of skeletal muscle ischemia and the reperfusion syndrome: a review.  Cardiovasc Surg. 2002;  10 620-630
    CrossrefPubMedGoogle Scholar
  • 9 Bushell A J, Klenerman L, Davies H M et al.. Damage to skeletal muscle induced by prolonged ischemia and reperfusion.  Transplant Proc. 1995;  27 2834-2835
    PubMedGoogle Scholar
  • 10 Carroll C M, Carroll S M, Overgoor M L et al.. Acute ischemic preconditioning of skeletal muscle prior to flap elevation augments muscle-flap survival.  Plast Reconstr Surg. 1997;  100 58-65
    CrossrefPubMedGoogle Scholar
  • 11 Cordeiro P G, Santamaria E, Hu Q Y. Use of a nitric oxide precursor to protect pig myocutaneous flaps from ischemia-reperfusion injury.  Plast Reconstr Surg. 1998;  102 2040-2048
    PubMedGoogle Scholar
  • 12 Gaines G C, Welborn III M B, Moldawer L L et al.. Attenuation of skeletal muscle ischemia/reperfusion injury by inhibition of tumor necrosis factor.  J Vasc Surg. 1999;  29 370-376
    PubMedGoogle Scholar
  • 13 Garramone Jr R R, Winters R M, Das D K et al.. Reduction of skeletal muscle injury through stress conditioning using the heat-shock response.  Plast Reconstr Surg. 1994;  93 1242-1247
    PubMedGoogle Scholar
  • 14 Hamada Y, Ishikawa S, Kanda T et al.. Sodium/hydrogen ion exchange inhibitor ameliorates ischaemia-reperfusion injuries in the rat hind limb.  Int Angiol. 1999;  18 320-326
    PubMedGoogle Scholar
  • 15 Kishi M, Tanaka H, Seiyama A et al.. Pentoxifylline attenuates reperfusion injury in skeletal muscle after partial ischemia.  Am J Physiol. 1998;  274(5 Pt 2) H1435-H1442
    PubMedGoogle Scholar
  • 16 Lazarus B, Messina A, Barker J E et al.. The role of mast cells in ischaemia-reperfusion injury in murine skeletal muscle.  J Pathol. 2000;  191 443-448
    CrossrefPubMedGoogle Scholar
  • 17 Lepore D A, Morrison W A. Ischemic preconditioning: lack of delayed protection against skeletal muscle ischemia-reperfusion.  Microsurgery. 2000;  20 350-355
    CrossrefPubMedGoogle Scholar
  • 18 Lille S, Su C Y, Schoeller T et al.. Induction of heat-shock protein 72 in rat skeletal muscle does not increase tolerance to ischemia-reperfusion injury.  Muscle Nerve. 1999;  22 390-393
    PubMedGoogle Scholar
  • 19 Mitrev Z, Beyersdorf F, Hallmann R et al.. Reperfusion injury in skeletal muscle: controlled limb reperfusion reduces local and systemic complications after prolonged ischaemia.  Cardiovasc Surg. 1994;  2 737-748
    PubMedGoogle Scholar
  • 20 Mitrev Z, Ihnken K, Poloczek Y et al.. Controlled reperfusion of the extremities for preventing local and systemic damage after prolonged ischemia: an experimental study with the swine model.  Zentralbl Chir. 1996;  121 774-787
    PubMedGoogle Scholar
  • 21 Pang C Y, Yang R Z, Zhong A et al.. Acute ischaemic preconditioning protects against skeletal muscle infarction in the pig.  Cardiovasc Res. 1995;  29 782-788
    CrossrefPubMedGoogle Scholar
  • 22 Pudupakkam S, Harris K A, Jamieson W G et al.. Ischemic tolerance in skeletal muscle: role of nitric oxide.  Am J Physiol. 1998;  275(1 Pt 2) H94-H99
    PubMedGoogle Scholar
  • 23 Schroeder Jr C A, Lee H T, Shah P M et al.. Preconditioning with ischemia or adenosine protects skeletal muscle from ischemic tissue reperfusion injury.  J Surg Res. 1996;  63 29-34
    CrossrefPubMedGoogle Scholar
  • 24 Skjeldal S, Grogaard B, Reikeras O et al.. Model for skeletal muscle ischemia in rat hindlimb: evaluation of reperfusion and necrosis.  Eur Surg Res. 1991;  23 355-365
    PubMedGoogle Scholar
  • 25 Whetzel T P, Stevenson T R, Sharman R B et al.. The effect of ischemic preconditioning on the recovery of skeletal muscle following tourniquet ischemia.  Plast Reconstr Surg. 1997;  100 1767-1775
    CrossrefPubMedGoogle Scholar
  • 26 Hickey M J, Knight K R, Hurley J V et al.. Phosphoenolpyruvate/adenosine triphosphate enhances post-ischemic survival of skeletal muscle.  J Reconstr Microsurg. 1995;  11 415-422
    PubMedGoogle Scholar
  • 27 Iwahori Y, Ishiguro N, Shimizu T et al.. Selective neutrophil depletion with monoclonal antibodies attenuates ischemia/reperfusion injury in skeletal muscle.  J Reconstr Microsurg. 1998;  14 109-116
    PubMedGoogle Scholar
  • 28 Chachques J C, Fabiani J N, Perier P et al.. Reversibility of muscular ischemia: a histochemical quantification by the nitroblue tetrazolium (NBT) test.  Angiology. 1985;  36 493-499
    PubMedGoogle Scholar
  • 29 Labbe R, Lindsay T, Gatley R et al.. Quantitation of postischemic skeletal muscle necrosis: histochemical and radioisotope techniques.  J Surg Res. 1988;  44 45-53
    CrossrefPubMedGoogle Scholar
  • 30 Barker J E, Knight K R, Romeo R et al.. Targeted disruption of the nitric oxide synthase 2 gene protects against ischaemia/reperfusion injury to skeletal muscle.  J Pathol. 2001;  194 109-115
    PubMedGoogle Scholar
  • 31 Mowlavi A, Ghavami A, Song Y H et al.. Limited use of cyclosporin A in skeletal muscle ischemia-reperfusion injury.  Ann Plast Surg. 2001;  46 426-430
    CrossrefPubMedGoogle Scholar
  • 32 Hirose J, Yamaga M, Ide J et al.. Reduced ischemia-reperfusion injury in muscle. Experiments in rats with EPC-K1, a new radical scavenger.  Acta Orthop Scand. 1997;  68 369-373
    PubMedGoogle Scholar
  • 33 Sirsjo A, Soderkvist P, Gustafsson U et al.. The relationship between blood flow, development of edema and leukocyte accumulation in post-ischemic rat skeletal muscle.  Microcirc Endothelium Lymphatics. 1990;  6 21-34
    PubMedGoogle Scholar
  • 34 Crinnion J N, Homer-Vanniasinkam S, Hatton R et al.. Role of neutrophil depletion and elastase inhibition in modifying skeletal muscle reperfusion injury.  Cardiovasc Surg. 1994;  2 749-753
    PubMedGoogle Scholar
  • 35 Dahlback L O. Effects of temporary tourniquet ischemia on striated muscle fibers and motor end-plates. Morphological and histochemical studies in the rabbit and electromyographical studies in man.  Scand J Plast Reconstr Surg. 1970;  Suppl 7 1-91
    PubMedGoogle Scholar
  • 36 Harman J W. A histological study of skeletal muscle in acute ischemia.  Am J Path. 1947;  23 551-561
    PubMedGoogle Scholar
  • 37 Sanderson R A, Foley R K, McIvor G W et al.. Histological response on skeletal muscle to ischemia.  Clin Orth Rel Res. 1975;  113 27-35
    PubMedGoogle Scholar

Steffen P BaumeisterM.D. 

BG-Unfalklinik Ludwigshafen, Ludwig-Guttmanstr. 13

67071 Ludwigshafen, Germany

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