Experimental Brain Research

, Volume 191, Issue 3, pp 371–382 | Cite as

Spatial and temporal aspects of muscle hyperalgesia induced by nerve growth factor in humans

  • Helle Andersen
  • Lars Arendt-Nielsen
  • Peter Svensson
  • Bente Danneskiold-Samsøe
  • Thomas Graven-Nielsen
Research Article


Intramuscular injection of nerve growth factor (NGF) has been shown to induce long-term sensitisation and time-dependent hyperalgesia indicating potential involvement of both central and peripheral pain mechanisms. This double-blind placebo-controlled study was designed to describe the spatial distribution of muscle hyperalgesia over time (immediately after, 3 h, 1, 4, 7 and 21 days) after injecting NGF (5 μg) into the tibialis anterior (TA) muscle, to explore possibly involved central pain mechanisms and to investigate the effect of gender on development of hyperalgesia. Totally 20 healthy volunteers (10 men and 10 women) participated in the study. An isotonic saline injection into the contralateral TA muscle served as a control condition for the NGF injection. Pressure pain thresholds (PPT) were used to test for muscle hyperalgesia along the TA (seven sites) muscle at the extensor digitorum longus and at the web between 1st and 2nd metatarsal (central involvement). One day after the NGF/control injections, hypertonic saline (0.5 ml, 5.8%) was injected into the left and right TA to study the pain response to chemical stimulation of the hyperalgesic muscle tissue. Scores on a modified Likert scale were used to assess soreness during muscle function. An area of hyperalgesia was observed locally at the injected site 3 h after injection of NGF, which expanded both proximally and distally on day 1; this effect subsided on day 4. Decreased PPT was also found between 1st and 2nd metatarsal on day 1. Hypertonic saline evoked more pain in men when injected in the NGF treated TA compared to the control leg. Injection of NGF increased muscle soreness during muscle activity for 7 days. In this material there was no gender effect of NGF-induced muscle hyperalgesia. The expansion of muscle hyperalgesia to distant areas indicates that central mechanisms are involved.


Nerve growth factor (NGF) Muscle hyperalgesia Pressure algometry Gender Hypertonic saline 



The authors are grateful to Dr. John Hansen (Center for Sensory-Motor Interaction (SMI), Aalborg University, Denmark) for invaluable help with software development and hardness analysis. This study was supported by the Oak Foundation, Sundheds CVU Nordjylland and the CW Obel’s Foundation.


  1. Allen SJ, Dawbarn D (2006) Clinical relevance of the neurotrophins and their receptors. Clin Sci (Lond) 110:175–191CrossRefGoogle Scholar
  2. Amano T, Yamakuni T, Okabe N, Sakimura K, Takahashi Y (1991) Production of nerve growth factor in rat skeletal muscle. Neurosci Lett 132:5–7PubMedCrossRefGoogle Scholar
  3. Andersen H, Arendt-Nielsen L, Danneskiold-Samsøe B, Graven-Nielsen T (2006) Pressure pain sensitivity and hardness along human normal and sensitized muscle. Somatosens Mot Res 23:97–109PubMedCrossRefGoogle Scholar
  4. Apfel SC, Schwartz S, Adornato BT, Freeman R, Biton V, Rendell M, Vinik A, Giuliani M, Stevens JC, Barbano R, Dyck PJ (2000) Efficacy and safety of recombinant human nerve growth factor in patients with diabetic polyneuropathy: a randomized controlled trial. rhNGF Clinical Investigator Group. JAMA 284:2215–2221PubMedCrossRefGoogle Scholar
  5. Ashina M, Bendtsen L, Jensen R, Sakai F, Olesen J (1999) Muscle hardness in patients with chronic tension-type headache: relation to actual headache state. Pain 79:201–205PubMedCrossRefGoogle Scholar
  6. Bajaj P, Arendt-Nielsen L, Bajaj P, Madsen H (2001) Sensory changes during the ovulatory phase of the menstrual cycle in healthy women. Eur J Pain 5:135–144PubMedCrossRefGoogle Scholar
  7. Craft RM (2007) Modulation of pain by estrogens. Pain 132(Suppl 1):S3–S12PubMedCrossRefGoogle Scholar
  8. Drewes AM, Helweg-Larsen S, Petersen P, Brennum J, Andreasen A, Poulsen LH, Jensen TS (1993) McGill pain questionnaire translated into Danish: experimental and clinical findings. Clin J Pain 9:80–87PubMedCrossRefGoogle Scholar
  9. Dyck PJ, Peroutka S, Rask C, Burton E, Baker MK, Lehman KA, Gillen DA, Hokanson JL, O’Brien PC (1997) Intradermal recombinant human nerve growth factor induces pressure allodynia and lowered heat-pain threshold in humans. Neurology 48:501–505PubMedGoogle Scholar
  10. Friess H (1999) Nerve growth factor and its high-affinity receptor in chronic pancreatitis. Ann Surg 230:615–624PubMedCrossRefGoogle Scholar
  11. Ge HY, Madeleine P, Arendt-Nielsen L (2004) Sex differences in temporal characteristics of descending inhibitory control: an evaluation using repeated bilateral experimental induction of muscle pain. Pain 110:72–78PubMedCrossRefGoogle Scholar
  12. Ge HY, Madeleine P, Cairns BE, Arendt-Nielsen L (2006) Hypoalgesia in the referred pain areas after bilateral injections of hypertonic saline into the trapezius muscles of men and women: a potential experimental model of gender-specific differences. Clin J Pain 22:37–44PubMedCrossRefGoogle Scholar
  13. Gibson W, Arendt-Nielsen L, Graven-Nielsen T (2006) Delayed onset muscle soreness at tendon-bone junction and muscle tissue is associated with facilitated referred pain. Exp Brain Res 174:351–360PubMedCrossRefGoogle Scholar
  14. Giovengo SL, Russell IJ, Larson AA (1999) Increased concentrations of nerve growth factor in cerebrospinal fluid of patients with fibromyalgia. J Rheumatol 26:1564–1569PubMedGoogle Scholar
  15. Graven-Nielsen T (2006) Fundamentals of muscle pain, referred pain, and deep tissue hyperalgesia. Scand J Rheumatol 35:1–43CrossRefGoogle Scholar
  16. Graven-Nielsen T, Jansson Y, Segerdahl M, Kristensen JD, Mense S, Arendt-Nielsen L, Sollevi A (2003) Experimental pain by ischaemic contractions compared with pain by intramuscular infusions of adenosine and hypertonic saline. Eur J Pain 7:93–102PubMedCrossRefGoogle Scholar
  17. Graven-Nielsen T, Mense S, Arendt-Nielsen L (2004) Painful and non-painful pressure sensations from human skeletal muscle. Exp Brain Res 159:273–283PubMedCrossRefGoogle Scholar
  18. Greenspan JD, Craft RM, LeResche L, Rendt-Nielsen L, Berkley KJ, Fillingim RB, Gold MS, Holdcroft A, Lautenbacher S, Mayer EA, Mogil JS, Murphy AZ, Traub RJ (2007) Studying sex and gender differences in pain and analgesia: a consensus report. Pain 132(Suppl 1):S26–S45PubMedCrossRefGoogle Scholar
  19. Hathway GJ, Fitzgerald M (2006) Time course and dose-dependence of nerve growth factor-induced secondary hyperalgesia in the mouse. J Pain 7:57–61PubMedCrossRefGoogle Scholar
  20. Hoheisel U, Unger T, Mense S (2005) Excitatory and modulatory effects of inflammatory cytokines and neurotrophins on mechanosensitive group IV muscle afferents in the rat. Pain 114:168–176PubMedCrossRefGoogle Scholar
  21. Hoheisel U, Unger T, Mense S (2007) Sensitization of rat dorsal horn neurons by NGF-induced subthreshold potentials and low-frequency activation. A study employing intracellular recordings in vivo. Brain Res 1169:34–43PubMedCrossRefGoogle Scholar
  22. Horikawa M, Ebihara S, Sakai F, Akiyama M (1993a) Non-invasive measurement method for hardness in muscular tissues. Med Biol Eng Comput 31:623–627PubMedCrossRefGoogle Scholar
  23. Horikawa M, Ebihara S, Sakai F, Akiyama M (1993b) Non-invasive measurement method for hardness in muscular tissues. Med Biol Eng Comput 31:623–627PubMedCrossRefGoogle Scholar
  24. Horikawa M, Ebihara S, Sakai F, Akiyama M (1993c) Non-invasive measurement method for hardness in muscular tissues. Med Biol Eng Comput 31:623–627PubMedCrossRefGoogle Scholar
  25. Isselee H, De Laat A, Bogaerts K, Lysens R (2001) Long-term fluctuations of pressure pain thresholds in healthy men, normally menstruating women and oral contraceptive users. Eur J Pain 5:27–37PubMedCrossRefGoogle Scholar
  26. Jensen K, Andersen HØ, Olesen J, Lindblom U (1986) Pressure-pain threshold in human temporal region. Evaluation of a new pressure algometer. Pain 25:313–323PubMedCrossRefGoogle Scholar
  27. LeResche L, Mancl L, Sherman JJ, Gandara B, Dworkin SF (2003) Changes in temporomandibular pain and other symptoms across the menstrual cycle. Pain 106:253–261PubMedCrossRefGoogle Scholar
  28. Lewin GR, Mendell LM (1993) Nerve growth factor and nociception. Trends Neurosci 16:353–359PubMedCrossRefGoogle Scholar
  29. Lewin GR, Ritter AM, Mendell LM (1992) On the role of nerve growth factor in the development of myelinated nociceptors. J Neurosci 12:1896–1905PubMedGoogle Scholar
  30. Lewin GR, Ritter AM, Mendell LM (1993) Nerve growth factor-induced hyperalgesia in the neonatal and adult rat. J Neurosci 13:2136–2148PubMedGoogle Scholar
  31. Liuzzi FJ, Scoville SA, Bufton SM (1999) Effects of short-term estrogen replacement on trkA mRNA levels in axotomized dorsal root ganglion neurons. Exp Neurol 159:433–440PubMedCrossRefGoogle Scholar
  32. Melzack R (1975) Mcgill pain questionnaire—major properties and scoring methods. Pain 1:277–299PubMedCrossRefGoogle Scholar
  33. Miller LJ (2002) Nerve growth factor and chronic prostatitis/chronic pelvic pain syndrome. Urology 59:603–608PubMedCrossRefGoogle Scholar
  34. Murayama M, Nosaka K, Yoneda T, Minamitani K (2000) Changes in hardness of the human elbow flexor muscles after eccentric exercise. Eur J Appl Physiol 82:361–367PubMedCrossRefGoogle Scholar
  35. Petruska JC, Mendell LM (2004) The many functions of nerve growth factor: multiple actions on nociceptors. Neurosci Lett 361:168–171PubMedCrossRefGoogle Scholar
  36. Petty BG, Cornblath DR, Adornato BT, Chaudhry V, Flexner C, Wachsman M, Sinicropi D, Burton LE, Peroutka SJ (1994) The effect of systemically administered recombinant human nerve growth factor in healthy human subjects. Ann Neurol 36:244–246PubMedCrossRefGoogle Scholar
  37. Pezet S, McMahon SB (2006) Neurotrophins: mediators and modulators of pain. Annu Rev Neurosci 29:507–38PubMedCrossRefGoogle Scholar
  38. Reinert A, Kaske A, Mense S (1998) Inflammation-induced increase in the density of neuropeptide-immunoreactive nerve endings in rat skeletal muscle. Exp Brain Res 121:174–180PubMedCrossRefGoogle Scholar
  39. Riley JLIII, Robinson ME, Wise EA, Price DD (1999) A meta-analytic review of pain perception across the menstrual cycle. Pain 81:225–235PubMedCrossRefGoogle Scholar
  40. Sarchielli P, Alberti A, Floridi A, Gallai V (2001) Levels of nerve growth factor in cerebrospinal fluid of chronic daily headache patients. Neurology 57:132–134PubMedGoogle Scholar
  41. Schumacher MA, Jong BE, Frey SL, Sudanagunta SP, Capra NF, Levine JD (2000) The stretch-inactivated channel, a vanilloid receptor variant, is expressed in small-diameter sensory neurons in the rat. Neurosci Lett 287:215–218PubMedCrossRefGoogle Scholar
  42. Scoville SA, Bufton SM, Liuzzi FJ (1997) Estrogen regulates neurofilament gene expression in adult female rat dorsal root ganglion neurons. Exp Neurol 146:596–599PubMedCrossRefGoogle Scholar
  43. Slater H, Arendt-Nielsen L, Wright A, Graven-Nielsen T (2003) Experimental deep tissue pain in wrist extensors—a model of lateral epicondylalgia. Eur J Pain 7:277–288PubMedCrossRefGoogle Scholar
  44. Stohler CS, Kowalski CJ, Lund JP (2001) Muscle pain inhibits cutaneous touch perception. Pain 92:327–333PubMedCrossRefGoogle Scholar
  45. Svensson P, Wang K, Arendt-Nielsen L, Cairns BE (2008) Effects of NGF-induced muscle sensitization on proprioception and nociception. Exp Brain Res (in press)Google Scholar
  46. Svensson P, Cairns BE, Wang K, Arendt-Nielsen L (2003) Injection of nerve growth factor into human masseter muscle evokes long-lasting mechanical allodynia and hyperalgesia. Pain 104:241–247PubMedCrossRefGoogle Scholar
  47. Unruh AM (1996) Gender variations in clinical pain experience. Pain 65:123–167PubMedCrossRefGoogle Scholar
  48. Ure DR, Campenot RB (1997) Retrograde transport and steady-state distribution of 125I-nerve growth factor in rat sympathetic neurons in compartmented cultures. J Neurosci 17:1282–1290PubMedGoogle Scholar
  49. Woolf CJ (1996) Phenotypic modification of primary sensory neurons: the role of nerve growth factor in the production of persistent pain. Philos Trans R Soc Lond B Biol Sci 351:441–448PubMedCrossRefGoogle Scholar
  50. Woolf CJ, Ma QP, Allchorne A, Poole S (1996) Peripheral cell types contributing to the hyperalgesic action of nerve growth factor in inflammation. J Neurosci 16:2716–2723PubMedGoogle Scholar
  51. Yamamoto M (2003) Pain-related differential expression of NGF, GDNF, IL-6, and their receptors in human vasculitic neuropathies. Intern Med 42:1100–1103PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Helle Andersen
    • 1
  • Lars Arendt-Nielsen
    • 1
  • Peter Svensson
    • 2
  • Bente Danneskiold-Samsøe
    • 3
  • Thomas Graven-Nielsen
    • 1
  1. 1.Laboratory for Musculoskeletal Pain and Motor Control, Center for Sensory–Motor Interaction (SMI)Aalborg UniversityAalborg EDenmark
  2. 2.Department of Clinical Oral Physiology, Dental SchoolUniversity of AarhusAarhusDenmark
  3. 3.The Parker InstituteFrederiksberg HospitalFrederiksbergDenmark

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