Background: Increase in expression of substance P (SP) and calcitonin gene-related peptide (CGRP) has been reported in clinical samples of tendinopathy.
Purpose: To examine the spatial-temporal expression of these neuropeptides as well as their association with activity-related tendon pain, matrix degeneration, failed healing, and pathologic calcification in an established collagenase-induced tendon injury rat model.
Study Design: Controlled laboratory study.
Methods: Collagenase or saline was injected into the patellar tendon of rats. At weeks 2, 4, 8, 12, and 16, just before the rats were sacrificed, the double-stance duration of rats was examined by gait analysis method. After sacrifice, the patellar tendons were harvested for histologic analysis and immunohistochemical staining of SP and CGRP.
Results: There was an increase of SP and CGRP immunopositivity in tendon fibroblasts at week 2. The immunopositive signals decreased at weeks 4 and 8 and were observed in chondrocyte-like cells. At weeks 12 and 16, the immunopositive staining increased again and was observed in cells embedded in calcific deposits in addition to tendon fibroblasts and chondrocyte-like cells. The expression pattern was consistent with matrix degeneration, calcification, and failed healing in the animal model. There were significant positive correlations of immunopositivity of SP (rho = .502, P = .002) and CGRP (rho = .483, P = .003) with double-stance duration after collagenase injection.
Conclusion: There was increased expression of SP and CGRP after collagenase-induced tendon injury, and their expression was positively associated with double-stance duration.
Clinical Relevance: Substance P and CGRP might be involved in the pathogenesis and origin of pain of tendinopathy and could be the targets for future intervention.
Ackermann PW , Ahmed M., Kreicbergs A.Early nerve regeneration after Achilles tendon rupture: a prerequisite for healing? A study in the rat. J Orthop Res. 2002;20:849-856.
2.
Ackermann PW , Li J., Lundeberg T., Kreicbergs A.Neuronal plasticity in relation to nociception and healing of rat Achilles tendon. J Orthop Res. 2003;21:432-441.
3.
Alfredson H. , Ljung BO, Thorsen K., Lorentzon R. In vivo investigation of ECRB tendons with microdialysis technique: no signs of inflammation but high amounts of glutamate in tennis elbow. Acta Orthop Scand . 2000;71:475-479.
4.
Alfredson H. , Ohberg L.Neovascularisation in chronic painful patellar tendinosis: promising results after sclerosing neovessels outside the tendon challenge the need for surgery. Knee Surg Sports Traumatol Arthrosc. 2005;13(2):74-80.
5.
Alfredson H. , Ohberg L.Sclerosing injections to areas of neovascularization reduce pain in chronic Achilles tendinopathy: a double-blind randomized controlled trial. Knee Surg Sports Traumatol Arthrosc . 2005;13(4):338-344.
6.
Andersson G. , Danielson P., Alfredson H., Forsgren S.Nerve-related characteristics of ventral paratendinous tissue in chronic Achilles tendinosis . Knee Surg Sports Traumatol Arthrosc. 2007 ;15(10): 1272-1279.
7.
Andersson G. , Danielson P., Alfredson H., Forsgren S.Presence of substance P and the neurokinin-1 receptor in tenocytes of the human Achilles tendon. Regul Pept. 2008;150:81-87.
8.
Brain SD, Williams TJ, Tippins JR, Morris HR, MacIntyre I.Calcitonin gene-related peptide is a potent vasodilator. Nature. 1985;313:54-56.
9.
Burssens P, Steyaert A, Forsyth R, van Ovost EJ, De Paepe Y, Verdonk R. Exogenously administered substance P and neutral endopeptidase inhibitors stimulate fibroblast proliferation, angiogenesis and collagen organization during Achilles tendon healing. Foot Ankle Int.2005; 26(10):832-839.
10.
Cuesta MC, Quintero L., Pons H., Suarez-Roca H.Substance P.and calcitonin gene-related peptide increase IL-1 beta, IL-6 and TNF alpha secretion from human peripheral blood mononuclear cells. Neurochem Int. 2002;40(4):301-306.
11.
Cury PR, Canavez VC, de Araujo C., Furuse NS, de Araujo NSSubstance P regulates the expression of matrix metalloproteinases and tissue inhibitors of metalloproteinase in cultured human gingival fibroblasts . J Periodont Res. 2008;43:255-260.
12.
Faure G., Daculsi G. Calcifiedtendinitis: a review. Ann Rheum Dis. 1983;42(suppl 1):49-53.
13.
Fenwick S., Harrall R., Hackney R., et al. Endochondral ossification in Achilles and patella tendinopathy . Rheumatology (Oxford). 2002;41: 474-476.
14.
Fredberg U. , Stengaard-Pedersen K. Chronictendinopathy tissue pathology, pain mechanisms, and etiology with a special focus on inflammation . Scand J Med Sci Sports. 2008;18(1):3-15.
15.
Fu SC, Chan BP, Wang W., Pau HM, Chan KM, Rolf CGIncrease expression of matrix metalloproteinase 1 (MMP1) in 11 patients with patellar tendinosis. Acta Orthop Scand. 2002 ;73(6):658-662.
16.
Fu SC, Chan KM, Chan LS, Fong DT, Lui PYThe use of motion analysis to measure pain-related behaviour in a rat model of degenerative tendon injuries. J Neurosci Methods . 2009;179:309-318.
17.
Fu SC, Chan KM, Rolf CGIncreased deposition of sulfated glycosaminoglycans in human patellar tendinopathy . Clin J Sport Med. 2007; 17(2):129-131.
18.
Fu SC, Wang W., Pau HM, Wong YP, Chan KM, Rolf CGIncreased expression of transforming growth factor-beta1 in patellar tendinosis . Clin Orthop Relat Res. 2002;400:174-183.
19.
Gotoh M., Hamada K., Yamakawa H., Inoue A., Fukuda H.Increased substance P in subacromial bursa and shoulder pain in rotator cuff diseases. J Orthop Res. 1998;16(8):618-621.
20.
Hart D., Archambault J., Kydd A., Reno C., Frank CB, Herzog W.Gender and neurogenic variables in tendon biology and repetitive motion disorders . Clin Orthop Relat Res. 1998;351:44-56.
21.
Hart DA, Kydd A., Reno C.Gender and pregnancy affect neuropeptide responses of the rabbit Achilles tendon. Clin Orthop. 1999;365: 237-246.
22.
Hart DA, Reno C.Pregnancy alters the in vitro responsiveness of the rabbit medial collateral ligament to neuropeptides: effect on mRNA levels for growth factors, cytokines, iNOS, COX-2, metalloproteinases and TIMPs. Biochim Biophys Acta.1998;1408:35-43.
23.
Hecker-Kia A., Kolkenbrock H., Orgel D., Zimmermann B., Sparmann M., Uibrich N.Substance P induces the secretion of gelatinase A from synovial fibroblast. Eur J Clin Chem Clin Biochem. 1997;35:655-660.
24.
Ihara H., Nakanishi S.Selective inhibition of expression of the substance P receptor mRNA in pancreatic acinar AR42J cells by glucocorticoids. J Biol Chem. 1990;265:22441-22445.
Karahan S., Kincaid SA, Baird AN, Kammermann JRDistribution of β-endorphin and substance P in the shoulder joint of the dog before and after a low impact exercise programme. Anat Histol Embryol. 2002;31(2):72-77.
27.
Khan KM, Cook JL, Maffulli N., Kannus P.Where is the pain coming from in tendinopathy? It may be biochemical, not only structural, in origin. Br J Sports Med . 2000;34:81-83.
28.
Lai XN, Wang ZG, Zhu JM, Wang LLEffect of substance P on gene expression of transforming growth factor beta-1 and its receptors in rat’s fibroblasts. Chin J Traumatol . 2003;6:350-354.
29.
Lembeck F., Donnerer J., Colpaert FCIncrease in substance P in primary afferent nerves during chronic pain. Neuropeptides. 1981;1: 175-180.
30.
Lerner UHNeuropeptidergic regulation of bone resorption and bone formation . J Musculoskelet Neuronal Interact. 2002 ;2:440-447.
31.
Lian O., Dahl J., Ackermann PW, Frihagen F., Engebretsen I., Bahr R.Pronociceptive and antinociceptive neuromediators in patellar tendinopathy . Am J Sports Med. 2006;34:1801-1808.
32.
Ljung BO, Alfredson H., Forsgren S.Neurokinin 1-receptors and sensory neuropeptides in tendon insertions at the medial and lateral epicondyles of the humerus: studies on tennis elbow and medial epicondylalgia. J Orthop Res. 2004 ;22:321-327.
33.
Lotz M., Carson DA, Vaughan JHSubstance P activation of rheumatoid synoviocytes: neural pathway in pathogenesis of arthritis. Science. 1987 ;235:893-895.
34.
Lotz M., Vaughan JH, Carson DAEffect of neuropeptides on production of inflammatory cytokine by human monocytes . Science. 1988;241:1218-1221.
35.
Lui PPY , Fu SC, Chan LS, Hung LK, Chan KMChondrocyte phenotype and ectopic ossification in collagenase-induced tendon degeneration. J Histochem Cytochem.2009;57(2):91-100.
36.
Maffulli N. , Testa V., Capasso G., et al. Similar histopathological picture in males with Achilles and patellar tendinopathy. Med Sci Sports Exerc. 2004;36:1470-1475.
37.
Maffulli N. , Wong J., Almekinders LCTypes and epidemiology of tendinopathy. Clin Sports Med. 2003;22(4):675-692.
38.
Matsumoto I. , Ito Y., Tomo H., Nakao Y., Takaoka K.Case reports: ossified mass of the rotator cuff tendon in the subacromial bursa. Clin Orthop Relat Res . 2005;437:247-250.
39.
Messner K., Wei Y., Andersson B., Gillquist J., Rasanen T.Rat model of Achilles tendon disorder: a pilot study. Cells Tissues Organs. 1999; 165:30-39.
40.
Millward-Sadler SJ, Mackenzie A., Wright MO, et al. Tachykinin expression in cartilage and function in human articular chondrocyte mechanotransduction . Arthritis Rheum. 2003;48(1):146-156.
41.
Nilsson J., von Euler AM, Dalsgaard C-J.Stimulation of connective tissue cell growth by substance P and substance K. Nature. 1985; 315:61-63.
42.
Ohberg L., Alfredson H.Sclerosing therapy in chronic Achilles tendon insertional pain-results of a pilot study. Knee Surg Sports Traumatol Arthrosc. 2003;11:339-343.
43.
Ohberg L., Lorentzon R., Alfredson H.Neovascularization in Achilles tendon with painful tendinosis but not in normal tendons: an ultrasonographic investigation. Knee Surg Sports Traumatol Arthrosc. 2001; 9(4):233-238.
44.
Pernow B.Role of tachykinins in neurogenic inflammation. J Immunol. 1985;135(suppl 2):812s-815s.
45.
Saria A., Lundberg JM, Skofitsch G., Lembeck F.Vascular protein linkage in various tissue induced by substance P, capsaicin, bradykinin, serotonin, histamine and by antigen challenge. Naunyn Schmiedebergs Arch Pharmacol. 1983;324:212-218.
46.
Schubert TE , Weidler C., Lerch K., Hofstadter F., Straub RHAchilles tendinosis is associated with sprouting of substance P positive nerve fibers. Ann Rheum Dis. 2005;64:1083-1086.
47.
Shih C., Bernard GWNeurogenic substance P stimulates osteogenesis in vitro. Peptides. 1997;18:323-326.
48.
Steyart AE, Burssens PJ, Vercruysse CW, Vanderstraeten GG, Verbeeck RMThe effects of substance P on the biomechanic properties of ruptured rat Achilles’s tendon. Arch Phys Med Rehabil. 2006;87(2): 254-258.
49.
Sullo A., Maffulli N., Capasso G., Testa V.The effects of prolonged peritendinous administration of PGE1 to the rat Achilles tendon: a possible animal model of chronic Achilles tendinopathy . J Orthop Sci. 2001;6:349-357.
50.
Tanabe T., Otani H., Mishima K., Ogawa R., Inagaki C.Mechanisms of oxyradical production in substance P stimulated rheumatoid synovial cells. Rheumatol Int. 1996;16:159-167.
51.
Woolf C., Wiesenfeld-Hallin Z.Substance P and calcitonin gene-related peptide synergistically modulate the gain of the nociceptive flexor withdrawal reflex in the rat. Neurosci Lett. 1986;66: 226-230.
52.
Ziche M., Morbidelli L., Pacini M., Geppetti P., Alessandri G., Maggi CASubstance P stimulates neovascularisation in vivo and proliferation of cultured endothelial cells. Microvasc Res. 1990;40: 264-278.
