Knee Surgery, Sports Traumatology, Arthroscopy

, Volume 12, Issue 5, pp 497–502 | Cite as

Distribution of substance-P nerve fibers in intact and ruptured human anterior cruciate ligament: a semi-quantitative immunohistochemical assessment

  • Dariusz Witoński
  • Małgorzata Wągrowska-Danilewicz
Experimental Study


Numerous studies have reported qualitative and quantitative analysis of nerve supply in the anterior cruciate ligament; however, as yet relatively little is known about the distribution of substance-P nerve endings in the human anterior cruciate ligament. The objective of this work was to evaluate the distribution of substance-P nerve fibers in intact human anterior cruciate ligament, and determine if rupture of the ligament has any influence on occurrence of these receptors. The intact anterior cruciate ligament group (group 1) of osteoarthritis knee, undergoing total knee arthroplasty, consisted of nine patients (eight females) with a mean age of 65.3 years at surgery. The anterior cruciate ligament rupture group (group 2) consisted of 20 patients (18 males and 2 females) with a mean age of 27.8 years at reconstruction. Healing time of the torn ligament in vivo, determined by the time period between the rupture and reconstruction, lasted from 1 to 40 months and the patients were divided into 3 groups (I, II and III) embracing diverse time periods. All harvested anterior cruciate ligaments were sectioned in thirds so that there was a proximal, middle and distal third for each ligament. The distribution of nociceptive receptors was studied by immunohistochemistry with monoclonal antibody to substance-P, including the semi-quantitative assessment. No significant difference was found between the number of substance-P nerve fibers in the proximal, middle and distal third of the intact anterior cruciate ligament ( p >0.05). During the first 4 months after injury (group I) the mean number of neuropeptide-containing fibers was greater in the proximal than in the distal third ( p =0.048996). The number of SP-positive nerve fibers in the proximal third decreased between 5 and 12 months after rupture, in a statistically significant manner ( p =0.045864). This study showed that distribution of the nociceptive nerve supply, positively stained for substance-P, is equal among the intact anterior cruciate ligament. The substance-P nerve ending density was significantly affected by the injury as well as by the time since rupture. The results of this study provide immunohistochemical evidence suggesting that between 1 to 4 months after rupture the site of the injury undergoes neurogenic inflammation, which could have an influence on the healing course of the torn ligament.


Knee joint Human Anterior cruciate ligament Innervation Substance-P fibers 



This study was supported by University of Łódź School of Medicine, grant # 502–11–713(64). The experiments comply with the current laws of Poland.


  1. 1.
    Ackermann PW, Ahmed M, Kreicbergs A (2002) Early nerve regeneration after Achilles tendon rupture—a prerequisite for healing? A study in the rat. J Orthop Res 20:849–856CrossRefPubMedGoogle Scholar
  2. 2.
    Ackermann PW, Finn A, Ahmed M (1999) Sensory neuropeptidergic pattern in tendon, ligament and joint capsule. A study in the rat. Neuroreport 10:2055–2060PubMedGoogle Scholar
  3. 3.
    Arcand MA, Rhalmi S, Rivard CH (2000) Quantification of mechanoreceptors in the canine anterior cruciate ligament. Int Orthop 24:272–275CrossRefPubMedGoogle Scholar
  4. 4.
    Biedert RM, Stauffer E, Friederich NF (1992) Occurrence of free nerve endings in the soft tissue of the knee joint. A histologic investigation. Am J Sports Med 20:430–433PubMedGoogle Scholar
  5. 5.
    Denti M, Monteleone M, Berardi A, Panni AS (1994) Anterior cruciate ligament mechanoreceptors. Histologic studies on lesions and reconstruction. Clin Orthop 308:29–32PubMedGoogle Scholar
  6. 6.
    Gronblad M, Korkala O, Konttinen YT, Kuokkanen H, Liesi P (1991) Immunoreactive neuropeptides in nerves in ligamentous tissue. An experimental neuroimmunohistochemical study. Clin Orthop 265:291–296PubMedGoogle Scholar
  7. 7.
    Halata Z, Haus J (1989) The ultrastructure of sensory nerve endings in human anterior cruciate ligament. Anat Embryol (Berl) 179:415–421Google Scholar
  8. 8.
    Hanesch U, Heppelmann B, Schmidt RF (1997) Quantification of cat’s articular afferents containing calcitonin gene-related peptide or substance-P innervating normal and acutely inflamed knee joints. Neurosci Lett 233:105–108CrossRefPubMedGoogle Scholar
  9. 9.
    Hannafin JA, Attia ET, Warren RF, Bhargava MM (1999) Characterization of chemotactic migration and growth kinetics of canine knee ligament fibroblasts. J Orthop Res 17:398–404PubMedGoogle Scholar
  10. 10.
    Heppelmann B, Messlinger K, Neiss WF, Schmidt RF (1990) Ultrastructural three-dimensional reconstruction of group III and group IV sensory nerve endings (“free nerve endings”) in the knee joint capsule of the cat: evidence for multiple receptive sites. J Comp Neurol 292:103–116PubMedGoogle Scholar
  11. 11.
    Hirasawa Y, Okajima S, Ohta M (2000) Nerve distribution to the human knee joint: anatomical and immunohistochemical study. Int Orthop 24:1–4CrossRefPubMedGoogle Scholar
  12. 12.
    Kennedy JC, Alexander IJ, Hayes KC (1982) Nerve supply of the human knee and its functional importance. Am J Sports Med 10:329–335PubMedGoogle Scholar
  13. 13.
    Kolasinski SL, Haines KA, Siegel EL, Cronstein BN, Abramson SB (1992) Neuropeptides and inflammation. A somatostatin analog as a selective antagonists of neutrophil activation by substance-P. Arthritis Rheum 35:369–375PubMedGoogle Scholar
  14. 14.
    Krauspe R, Schmidt M, Schaible H (1992) Sensory innervation of the anterior cruciate ligament. An electrophysiological study of the response properties of single identified mechanoreceptors in the cat. J Bone Joint Surg Am 74:390–397PubMedGoogle Scholar
  15. 15.
    Marshall KW, Theriault E, Homonko DA (1994) Distribution of substance-P and calcitonin-related peptide immunoreactivity in the normal feline knee. J Rheumatol 21:883–889PubMedGoogle Scholar
  16. 16.
    Marui T, Niyibizi C, Georgescu HJ, Cao M, Kavalkovich KW, Levine RE, Woo SL-Y (1997) Effect of growth factors on matrix synthesis by ligament fibroblasts. J Orthop Res 15:18–23PubMedGoogle Scholar
  17. 17.
    Murray MM, Martin SD, Martin TL, Spector M (2000) Histological changes in the human anterior cruciate ligament after rupture. J Bone Joint Surg Am 82:1387–1397Google Scholar
  18. 18.
    Pernow B (1983 ) Substance-P. Pharmacol Rev 35:85–141PubMedGoogle Scholar
  19. 19.
    Sanchis-Alfonso V, Rosello-Sastre E (2000) Immunochistochemical analysis for neural markers of the lateral retinaculum in patients with isolated symptomatic patellofemoral malalignment. A neuroanatomic basis for anterior knee pain in the active young patients. Am J Sports Med 28:725–731PubMedGoogle Scholar
  20. 20.
    Schutte MJ, Dabezies EJ, Zimny ML, Happel L (1987) Neural anatomy of the human anterior cruciate ligament. J Bone Joint Surg Am 69:243–247PubMedGoogle Scholar
  21. 21.
    Scott DT, Lam FY, Ferrell WR (1994) Acute joint inflammation – mechanisms and mediators. Gen Pharmacol 25:1285–1296CrossRefPubMedGoogle Scholar
  22. 22.
    Sjolander P, Johansson H, Sojka P, Rehnholm A (1989) Sensory nerve endings in the cat cruciate ligaments: a morphological investigation. Neurosci Lett 102:33–38CrossRefPubMedGoogle Scholar
  23. 23.
    Tamura R, Hanesch U, Schmidt RF, Kumazawa T, Mizumura K (1998) Examination of co-localization gene-related peptide- and substance-P-like immunoreactivity in the knee joint of the dog. Neurosci Lett 18:53–56CrossRefGoogle Scholar
  24. 24.
    Wallengren J, Chen D, Sundler F (1999) Neuropeptide-containing C-fibers and wound healing in rat skin. Neither capsaicin nor peripheral neurotomy affect the rate of healing. Br J Dermatol 140:400–408CrossRefPubMedGoogle Scholar
  25. 25.
    Walsh DA, Salmon M, Mapp I, Garrett N, Blake D, Polak J (1994) Microvascular substance-P binding to normal and inflamed rat and human synovium. J Pharmacol Exp Ther 267:951–960Google Scholar
  26. 26.
    Witkowski J, Yang L, Wood DJ, Sung KL (1997) Migration and healing of ligament cells under inflammatory conditions. J Orthop Res 15:269–277PubMedGoogle Scholar
  27. 27.
    Witoński D, Wągrowska-Danilewicz M (1999) Distribution of substance-P nerve fibers in the knee joint in patients with anterior knee pain syndrome. A preliminary report. Knee Surg Sport Traumatol Arthrosc 7:177–183CrossRefGoogle Scholar
  28. 28.
    Wojtys EM, Beaman DN, Glover RA, Janda D (1990) Innervation of the human knee joint by substance-P fibers. Arthroscopy 6:254–263PubMedGoogle Scholar
  29. 29.
    Zimny ML, Schutte M, Dabezies E (1986) Mechanoreceptors in the human anterior cruciate ligament. Anat Rec 214:204–209PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Dariusz Witoński
    • 1
  • Małgorzata Wągrowska-Danilewicz
    • 2
  1. 1.Department of OrthopedicsUniversity of Łódź School of MedicineŁódźPoland
  2. 2.Department of PathomorphologyUniversity of Łódź School of MedicineŁódźPoland

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