Sports Medicine

, Volume 37, Issue 10, pp 837–856 | Cite as

Neutrophil Infiltration in Exercise-Injured Skeletal Muscle

How Do We Resolve the Controversy?
  • Barbara St. Pierre Schneider
  • Peter M. Tiidus
Leading Article


Neutrophils have not consistently been detected in exercise-injured skeletal muscle and, therefore, neutrophil infiltration in this muscle has become a controversial issue. Thirty-eight animal and human studies that assessed injured muscle for neutrophils and employed acute exercise (e.g. level, uphill or downhill running, eccentric contractions, or swimming) were analysed to help clarify the relationship between neutrophil infiltration and exercise-induced muscle injury. Findings from nearly three-quarters of the reviewed studies suggest that neutrophil accumulation follows exercise-induced muscle injury. Intramuscular neutrophil infiltration was present in 85% and 55% of the animal and human studies, respectively. However, no consistent relationship between the potential damaging effect of the exercise type and neutrophil infiltration can be conclusively established from these studies. Specific animal-related factors that could influence these results include age, animal strain, catecholamines, corticosterone, acute stressors and muscle type, whereas a specific human-related influencing factor is physical activity status. Factors affecting both animal and human studies could include sex hormones, muscle sampling techniques and neutrophil detection approaches. General categories of methods that have been used to detect neutrophil infiltration are microscopy, myeloperoxidase (MPO) biochemical assay, antibody staining and white blood cell radionuclide imaging. Only studies employing white blood cell radionuclide imaging have consistently detected neutrophil infiltration. However, antibody staining with a quantitative analysis is currently the most feasible, valid and sensitive method. Research recommendations, therefore, are warranted to resolve the neutrophil infiltration controversy. We propose two approaches for animal studies. The first approach encompasses (i) studying or measuring factors that could influence neutrophil infiltration; (ii) using quantitative antibody staining analysis (in all studies and employing a panel of anti-neutrophil antibodies); (iii) examining the relationship between fibre morphological changes and neutrophil antigen expression; and (iv) developing a neutrophil antibody-radionuclide imaging technique. The second approach will yield animal findings complementing or addressing the gaps from the human exercise studies. For human studies, we suggest that (i) physical activity status is investigated; (ii) quantitative antibody staining analysis is performed (including staining injured muscle with a panel of antibodies such as anti-elastase, anti-MPO, anti-CD11b and anti-CD15 or assessing injured muscle using both immunohistochemistry and the MPO biochemical assay); and (iii) the relationship between fibre morphological changes and neutrophil antigen expression is examined. Studies that incorporate these recommendations could lead to an increased understanding of whether neutrophils are essential for the recovery from an exerciseinduced muscle injury.


Extensor Digitorum Longus Neutrophil Infiltration Muscle Injury Eccentric Contraction Neutrophil Accumulation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The authors have received the following grants: NIH NINR NR05258 (BSS); Discovery grant from NSERC, Canada (PMT). The aforementioned funding agencies had no role in the preparation and decision to submit this article for publication. The authors have no conflicts of interest that are directly relevant to the content of this article. The authors wish to thank Kathleen Corbett Freimuth for her editing assistance.


  1. 1.
    Tidball JG. Inflammatory cell response to acute muscle injury. Med Sci Sports Exerc 1995; 27: 1022–32PubMedCrossRefGoogle Scholar
  2. 2.
    Tiidus PM. Radical species in inflammation and overtraining. Can J Physiol Pharmacol 1998; 76: 533–8PubMedCrossRefGoogle Scholar
  3. 3.
    Tidball JG. Inflammatory processes in muscle injury and repair. Am J Physiol Regul Integr Comp Physiol 2005; 288: R345–53PubMedCrossRefGoogle Scholar
  4. 4.
    Pizza FX, McLoughlin TJ, McGregor SJ, et al. Neutrophils injure cultured skeletal myotubes. Am J Physiol Cell Physiol 2001; 281: C335–41PubMedGoogle Scholar
  5. 5.
    Armstrong RB, Ogilvie RW, Schwane JA. Eccentric exerciseinduced injury to rat skeletal muscle. J Appl Physiol 1983; 54: 80–93PubMedGoogle Scholar
  6. 6.
    Lowe DA, Warren GL, Ingalls CP, et al. Muscle function and protein metabolism after initiation of eccentric contractioninduced injury. J Appl Physiol 1995; 79: 1260–70PubMedGoogle Scholar
  7. 7.
    Lapointe BM, Frenette J, Cote CH. Lengthening contraction induced inflammation is linked to secondary damage but devoid of neutrophil invasion. J Appl Physiol 2002; 92: 1995–2004PubMedCrossRefGoogle Scholar
  8. 8.
    Pizza FX, Koh TJ, McGregor SJ, et al. Muscle inflammatory cells after passive stretches, isometric contractions, and lengthening contractions. J Appl Physiol 2002; 92: 1873–8PubMedCrossRefGoogle Scholar
  9. 9.
    Brickson S, Ji LL, Schell K, et al. M1/70 attenuates blood-borne neutrophil oxidants, activation, and myofiber damage following stretch injury. J Appl Physiol 2003; 95: 969–76PubMedGoogle Scholar
  10. 10.
    Pizza FX, Peterson JM, Baas JH, et al. Neutrophils contribute to muscle injury and impair its resolution after lengthening contractions in mice. J Physiol 2005; 562: 899–913PubMedCrossRefGoogle Scholar
  11. 11.
    Kuipers H, Drukker J, Frederik PM, et al. Muscle degeneration after exercise in rats. Int J Sports Med 1983; 4: 45–51PubMedCrossRefGoogle Scholar
  12. 12.
    Duarte JA, Carvalho F, Bastos ML, et al. Do invading leucocytes contribute to the decrease in glutathione concentrations results. indicating oxidative stress in exercised muscle, or are they important for its recovery? Eur J Appl Physiol Occup Physiol 1994; 68: 48–53PubMedCrossRefGoogle Scholar
  13. 13.
    Belcastro AN, Arthur GD, Albisser TA, et al. Heart, liver, and skeletal muscle myeloperoxidase activity during exercise. J Appl Physiol 1996; 80: 1331–5PubMedGoogle Scholar
  14. 14.
    Ohishi S, Kizaki T, Ookawara T, et al. The effect of exhaustive exercise on the antioxidant enzyme system in skeletal muscle from calcium-deficient rats. Pflugers Arch 1998; 435: 767–74PubMedCrossRefGoogle Scholar
  15. 15.
    Raj DA, Booker TS, Belcastro AN. Striated muscle calciumstimulated cysteine protease (calpain-like) activity promotesmyeloperoxidase activity with exercise. Pflugers Arch 1998; 435: 804–9PubMedCrossRefGoogle Scholar
  16. 16.
    Best TM, Fiebig R, Corr DT, et al. Free radical activity, antioxi-dant enzyme, and glutathione changes with muscle stretch injury in rabbits. J Appl Physiol 1999; 87: 74–82PubMedGoogle Scholar
  17. 17.
    Tiidus PM, Bombardier E. Oestrogen attenuates post-exercise myeloperoxidase activity in skeletal muscle of male rats. Acta Physiol Scand 1999; 166: 85–90PubMedCrossRefGoogle Scholar
  18. 18.
    Blank SE, Tiidus PM, Hoffman-Goetz L. Neutrophil response to inflammaprolonged exercise in immune-competent and RAG2/gamma c null mice. Can J Physiol Pharmacol 2001; 79: 490–5PubMedCrossRefGoogle Scholar
  19. 19.
    Brickson S, Hollander J, Corr DT, et al. Oxidant production and adaptaimmune response after stretch injury in skeletal muscle. MedSci Sports Exerc 2001; 33: 2010–5PubMedCrossRefGoogle Scholar
  20. 20.
    Tiidus PM, Holden D, Bombardier E, et al. Estrogen effect on post-exercise skeletal muscle neutrophil infiltration and supplemencalpain activity. Can J Physiol Pharmacol 2001; 79: 400–6PubMedCrossRefGoogle Scholar
  21. 21.
    Dawson Jr BM, Messina S, Dominy J. The cytoprotective role of taurine in exercise-induced muscle injury. Amino Acids 2002; 22: 309–24PubMedCrossRefGoogle Scholar
  22. 22.
    St. Pierre Schneider B, Brickson S, Corr DT, et al. CD11b+ neutrophils predominate over RAM11+ macrophages in stretch-injured muscle. Muscle Nerve 2002; 25: 837–44CrossRefGoogle Scholar
  23. 23.
    Stauber W, Willems M. Prevention of histopathological changes from 30 repeated stretches of active rat skeletal muscles by recovlong inter-stretch rest times. Eur J Appl Physiol 2002; 88: 94–9PubMedCrossRefGoogle Scholar
  24. 24.
    Aoi W, Naito Y, Sakuma K, et al. Astaxanthin limits exercise-induced skeletal and cardiac muscle damage in mice. Antioxid Redox Signal 2003; 5: 139–44PubMedCrossRefGoogle Scholar
  25. 25.
    Koh TJ, Peterson JM, Pizza FX, et al. Passive stretches protect skeletal muscle of adult and old mice from lengthening contraction-induced injury. J Gerontol A Biol Sci Med Sci 2003; 58: 592–7PubMedCrossRefGoogle Scholar
  26. 26.
    Tsivitse SK, McLoughlin TJ, Peterson JM, et al. Downhill running in rats: influence on neutrophils, macrophages, and MyoD+ cells in skeletal muscle. Eur J Appl Physiol 2003; 90: 633–8PubMedCrossRefGoogle Scholar
  27. 27.
    Aoi W, Naito Y, Takanami Y, et al. Oxidative stress and delayed-onset muscle damage after exercise. Free Radic Biol Med 2004; 37: 480–7PubMedCrossRefGoogle Scholar
  28. 28.
    Baker W, Schneider BA, Kulkarni A, et al. P-selectin inhibition suppresses muscle regeneration following injury. J Leukoc Biol 2004; 76: 352–8PubMedCrossRefGoogle Scholar
  29. 29.
    Schneider BS, Fine JP, Tiidus PM. Indices of leukocyte infiltration and muscle recovery after eccentric contraction-induced injury in young and adult male mice. Orthop Nurs 2005; 24: 399–405PubMedCrossRefGoogle Scholar
  30. 30.
    Tiidus PM, Deller M, Liu XL. Oestrogen influence on myogenic satellite cells following downhill running in male rats: a preliminary study. Acta Physiol Scand 2005; 184: 67–72PubMedCrossRefGoogle Scholar
  31. 31.
    Morozov VI, Tsyplenkov PV, Golberg ND, et al. The effects of leuko high-intensity exercise on skeletal muscle neutrophil myeloperoxidase in untrained and trained rats. Eur J Appl Physiol 2006; 97: 716–22PubMedCrossRefGoogle Scholar
  32. 32.
    Aoi W, Naito Y, Nakamura T, et al. Inhibitory effect of fermented milk on delayed-onset muscle damage after exercise. J Nutr Biochem 2007; 18: 140–5PubMedCrossRefGoogle Scholar
  33. 33.
    Fielding RA, Manfredi TJ, Ding W, et al. Acute phase response in exercise III: neutrophil and IL-1ß accumulation in skeletal muscle. Am J Physiol 1993; 265: R166–72PubMedGoogle Scholar
  34. 34.
    MacIntyre DL, Reid WD, Lyster DM, et al. Presence of WBC, activadecreased strength, and delayed soreness in muscle after eccentric exercise. J Appl Physiol 1996; 80: 1006–13PubMedCrossRefGoogle Scholar
  35. 35.
    MacIntyre DL, Reid WD, Lyster DM, et al. Different effects of instrenuous eccentric exercise on the accumulation of neutrophils in muscle in women and men. Eur J Appl Physiol 2000; 81: 47–53PubMedCrossRefGoogle Scholar
  36. 36.
    Malm C, Nyberg P, Engstrom M, et al. Immunological changes in human skeletal muscle and blood after eccentric exercise and multiple biopsies. J Physiol 2000; 529: 243–62PubMedCrossRefGoogle Scholar
  37. 37.
    MacIntyre DL, Sorichter S, Mair J, et al. Markers of inflammation and myofibrillar proteins following eccentric exercise in humans. Eur J Appl Physiol 2001; 84: 180–6PubMedCrossRefGoogle Scholar
  38. 38.
    Stupka N, Tarnopolsky MA, Yardley NJ, et al. Cellular adaptation to repeated eccentric exercise-induced muscle damage. J Appl Physiol 2001; 91: 1669–78PubMedGoogle Scholar
  39. 39.
    Beaton LJ, Allan DA, Tarnopolsky MA, et al. Contraction induced muscle damage is unaffected by vitamin E supplementation. Med Sci Sports Exerc 2002; 34: 798–805PubMedCrossRefGoogle Scholar
  40. 40.
    Beaton LJ, Tarnopolsky MA, Phillips SM. Contraction-induced muscle damage in humans following calcium channel blocker administration. J Physiol 2002; 544: 849–59PubMedCrossRefGoogle Scholar
  41. 41.
    Peterson JM, Trappe TA, Mylona E, et al. Ibuprofen and acetaminophen: effect on muscle inflammation after eccentric exercise. Med Sci Sports Exerc 2003; 35: 892–6PubMedCrossRefGoogle Scholar
  42. 42.
    Raastad T, Risoy BA, Benestad HB, et al. Temporal relation between leukocyte accumulation in muscles and halted recovery 10–20h after strength exercise. J Appl Physiol 2003; 95: 2503–9PubMedGoogle Scholar
  43. 43.
    Malm C, Sjodin TL, Sjoberg B, et al. Leukocytes, cytokines, growth factors and hormones in human skeletal muscle and blood after uphill or downhill running. J Physiol 2004; 556: 983–1000PubMedCrossRefGoogle Scholar
  44. 44.
    Esposito AL, Poirier WJ, Clark CA. In vitro assessment of chemotaxis by peripheral blood neutrophils from adult and senescent C57BL/6 mice: correlation with in vivo responses to pulmonary infection with type 3 Streptococcus pneumoniae. Gerontology 1990; 36: 2–11PubMedCrossRefGoogle Scholar
  45. 45.
    Mello SB, Farsky SH, Sannomiya P, et al. Inhibition of neutrophil chemotaxis and chemokinesis associated with a plasma protein in aging rats: selective depression of cell responses mediated by complement-derived chemoattractants. J Leukoc Biol 1992; 51: 46–52PubMedGoogle Scholar
  46. 46.
    Marley SB, Hadley CL, Wakelin D. Effect of genetic variation on induced neutrophilia in mice. Infect Immun 1994; 62: 4304–9PubMedGoogle Scholar
  47. 47.
    Doddo JM, Hristopoulos ML, Welsh-Servinsky LE, et al. Strain-specific differences in sensitivity to ischemia-reperfusion lung injury in mice. J Appl Physiol 2006; 100: 1590–5CrossRefGoogle Scholar
  48. 48.
    Stolc V. Genetic control of blood neutrophil concentration in the rat. J Immunogenet 1988; 15: 345–51PubMedCrossRefGoogle Scholar
  49. 49.
    Dhabhar FS. Acute stress enhances while chronic stress suppresses skin immunity: the role of stress hormones and leukocyte trafficking. Ann N Y Acad Sci 2000; 917: 876–93PubMedCrossRefGoogle Scholar
  50. 50.
    Irwin J, Ahluwalia P, Zacharko RM, et al. Central norepinephrine and plasma corticosterone following acute and chronic stressors: influence of social isolation and handling. Pharmacol Biochem Behav 1986; 24: 1151–4PubMedCrossRefGoogle Scholar
  51. 51.
    Fitchett AE, Collins SA, Mason H, et al. Urinary corticosterone measures: effects of strain and social rank in BKW and CD-1 mice. Behav Processes 2005; 70: 168–76PubMedCrossRefGoogle Scholar
  52. 52.
    Viswanathan K, Dhabhar FS. Stress-induced enhancement of leukocyte trafficking into sites of surgery or immune activation. Proc Natl Acad Sci U S A 2005; 102: 5808–13PubMedCrossRefGoogle Scholar
  53. 53.
    Beaton LJ, Tarnopolsky MA, Phillips SM. Variability in estimating eccentric contraction-induced muscle damage and inflammation in humans. Can J Appl Physiol 2002; 27: 516–26PubMedCrossRefGoogle Scholar
  54. 54.
    Frenette J, Chbinou N, Godbout C, et al. Macrophages, not neutrophils, infiltrate skeletal muscle in mice deficient in P/E selectins after mechanical reloading. Am J Physiol Regul Integr Comp Physiol 2003; 285: R727–32PubMedGoogle Scholar
  55. 55.
    Biermann H, Pietz B, Dreier R, et al. Murine leukocytes with ring-shaped nuclei include granulocytes, monocytes, and their precursors. J Leukoc Biol 1999; 65: 217–31PubMedGoogle Scholar
  56. 56.
    Zweiman B. Neutrophils. Clin Allergy Immunol 2002; 16: 77–95PubMedGoogle Scholar
  57. 57.
    van Furth R, Hirsch JG, Fedorko ME. Morphology and peroxidase cytochemistry of mouse promonocytes, monocytes, and macrophages. J Exp Med 1970; 132: 794–812PubMedCrossRefGoogle Scholar
  58. 58.
    Sugiyama S, Okada Y, Sukhova GK, et al. Macrophage myexpreseloperoxidase regulation by granulocyte macrophage colony stimulating factor in human atherosclerosis and implications in acute coronary syndromes. Am J Pathol 2001; 158: 879–91PubMedCrossRefGoogle Scholar
  59. 59.
    Sirsjö A, Lewis DH, Nylander G. The accumulation of polymorphonuclear leukocytes in post-ischemic skeletal muscle in the rat, measured by quantitating tissue myeloperoxidase. Int J Microcirc Clin Exp 1990; 9: 163–73PubMedGoogle Scholar
  60. 60.
    Rubin B, Tittley J, Chang G, et al. A clinically applicable method for long-term salvage of postischemic skeletal muscle. J Vasc Surg 1991; 13: 58–67PubMedCrossRefGoogle Scholar
  61. 61.
    Schlag MG, Harris KA, Potter RF. Role of leukocyte accumuladiagtion and oxygen radicals in ischemia-reperfusion-induced injury in skeletal muscle. Am J Physiol Heart Circ Physiol 2001; 280: H1716–21PubMedGoogle Scholar
  62. 62.
    Hellsten Y, Frandsen U, Orthenblad N, et al. Xanthine oxidase in human skeletal muscle following eccentric exercise: a role in inflammation. J Physiol 1997; 498: 239–48PubMedGoogle Scholar
  63. 63.
    Stocks SC, Albrechtsen M, Kerr MA. Expression of the CD15 differentiation antigen (3-fucosyl-N-acetyl-lactosamine, LeX) on putative neutrophil adhesion molecules CR3 and NCA-160. Biochem J 1990; 268: 275–80PubMedGoogle Scholar
  64. 64.
    Oehler L, Majdic O, Pickl WF, et al. Neutrophil granulocyte committed cells can be driven to acquire dendritic cell characteristics. J Exp Med Sci 1998; 187: 1019–28CrossRefGoogle Scholar
  65. 65.
    Hart SP, Ross JA, Ross K, et al. Molecular characterization of the surface of apoptotic neutrophils: implications for functional downregulation and recognition by phagocytes. Cell Death Differ 2000; 7: 493–503PubMedCrossRefGoogle Scholar
  66. 66.
    Walrand S, Guillet C, Boirie Y, et al. In vivo evidences that insulin regulates human polymorphonuclear neutrophil functions. J Leukoc Biol 2004; 76: 1104–10PubMedCrossRefGoogle Scholar
  67. 67.
    Barrat F, Lesourd BM, Louise A, et al. Surface antigen expression in spleen cells of C57B1/6 mice during ageing: influence of sex and parity. Clin Exp Immunol 1997; 107: 593–600PubMedCrossRefGoogle Scholar
  68. 68.
    Barrat F, Lesourd B, Boulouis HJ, et al. Sex and parity modulate cytokine production during murine ageing. Clin Exp Immunol 1997; 109: 562–8PubMedCrossRefGoogle Scholar
  69. 69.
    Rypins EB, Kipper SL, Weiland F, et al. 99mTc anti-CD 15 monoclonal antibody (LeuTech) imaging improves diagnostic accuracy and clinical management in patients with equivocal presentation of appendicitis. Ann Surg 2002; 235: 232–9PubMedCrossRefGoogle Scholar
  70. 70.
    Palestro CJ, Kipper SL, Weiland FL, et al. Osteomyelitis: diagnosis with 99mTc-labeled antigranulocyte antibodies compared with diagnosis with (111) In-labeled leukocytes-initial experience. Radiology 2002; 223: 758–64PubMedCrossRefGoogle Scholar

Copyright information

© Adis Data Information BV 2007

Authors and Affiliations

  • Barbara St. Pierre Schneider
    • 1
  • Peter M. Tiidus
    • 2
  1. 1.School of NursingUniversity of Nevada Las VegasLas VegasUSA
  2. 2.Department of Kinesiology and Physical EducationWilfrid Laurier UniversityWaterlooCanada

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