Skip to main content

Possible role of Dickkopf-1 protein in the pathogenesis of tympanosclerosis in a rat model

  • Y Zhang (a1), S Wang (a1), Y Zheng (a2) and A Liu (a1)

This study aimed to investigate the expression of DKK1 protein in an experimental model of tympanosclerosis and its possible role in the pathogenesis of this disorder.


Forty Sprague Dawley rats were included in the study: 20 in the control group (which received no treatment) and 20 in the experimental group (which received an incision to induce tympanosclerosis). Otomicroscopy was performed to observe the development of myringosclerosis. Haematoxylin and eosin staining was performed to observe the morphological changes. Western blot analysis and immunohistochemistry were performed to assess the expression of DKK1 protein.


At day 15, sclerotic lesions were observed in 70 per cent of the tympanic membranes. Inflammatory infiltration and hyaline degeneration markedly appeared in the tympanic membranes and middle-ear mucosa. DKK1 protein was mainly distributed in the cytoplasm of epithelial cells, which were widely distributed in the tympanic membranes and middle-ear mucosa. The expression of DKK1 protein was significantly decreased in the calcified experimental ears.


DKK1 protein is involved in the pathogenesis of tympanosclerosis by regulating the Wnt/β-catenin signalling pathway.

Corresponding author
Address for correspondence: Dr Aiguo Liu, Department of Otolaryngology – Head and Neck Surgery, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, China E-mail:
Hide All
1 Wielinga, EW, Kerr, AG. Tympanosclerosis. Clin Otolaryngol Allied Sci 1993;18:341–9
2 Asiri, S, Hasham, A, al Anazy, F, Zakzouk, S, Banjar, A. Tympanosclerosis: review of literature and incidence among patients with middle-ear infection. J Laryngol Otol 1999;113:1076–80
3 Mattsson, C, Magnuson, K, Hellstrom, S. Myringosclerosis caused by increased oxygen concentration in traumatized tympanic membranes. Experimental study. Ann Otol Rhinol Laryngol 1995;104:625–32
4 Kinis, V, Ozbay, M, Alabalik, U, Gul, A, Yilmaz, B, Ozkurt, FE et al. Effect of caffeic acid phenethyl ester on myringosclerosis development in the tympanic membrane of rat. Eur Arch Otorhinolaryngol 2015;272:2934
5 Russell, JD, Giles, JJ. Tympanosclerosis in the rat tympanic membrane: an experimental study. Laryngoscope 2002;112:1663–6
6 Forseni, M, Eriksson, A, Bagger-Sjoback, D, Nilsson, J, Hultcrantz, M. Development of tympanosclerosis: can predicting factors be identified? Am J Otol 1997;18:298303
7 Koc, A, Uneri, C. Genetic predisposition for tympanosclerotic degeneration. Eur Arch Otorhinolaryngol 2002;259:180–3
8 Ferlito, A. Histopathogenesis of tympanosclerosis. J Laryngol Otol 1979;93:2537
9 Makishima, K, Toriya, Y, Inoue, S, Nakashima, T, Igarashi, Y. Clinicopathologic studies in tympanosclerosis. Am J Otol 1982;3:260–5
10 Tos, M, Stangerup, SE. Hearing loss in tympanosclerosis caused by grommets. Arch Otolaryngol Head Neck Surg 1989;115:931–5
11 Olsson, M, Dalsgaard, CJ, Haegerstrand, A, Rosenqvist, M, Ryden, L, Nilsson, J. Accumulation of T lymphocytes and expression of interleukin-2 receptors in nonrheumatic stenotic aortic valves. J Am Coll Cardiol 1994;23:1162–70
12 Mann, W, Beck, C, Schaefer, HE. The significance of calcium antagonists in rat experimental tympanosclerosis. Arch Otorhinolaryngol 1987;243:382–6
13 Haynes, KR, Pettit, AR, Duan, R, Tseng, HW, Glant, TT, Brown, MA et al. Excessive bone formation in a mouse model of ankylosing spondylitis is associated with decreases in Wnt pathway inhibitors. Arthritis Res Ther 2012;14:R253
14 Rossini, M, Gatti, D, Adami, S. Involvement of WNT/beta-catenin signaling in the treatment of osteoporosis. Calcif Tissue Int 2013;93:121–32
15 Walsh, NC, Gravallese, EM. Bone remodeling in rheumatic disease: a question of balance. Immunol Rev 2010;233:301–12
16 Huang, H, Wu, P, Cui, Y, Ge, R, Zhang, L. Expressions of OPG and RANKL in a rat model of tympanosclerosis [in Chinese]. Chinese Journal of Otology 2013;2:302–5
17 Boyce, BF, Xing, L. Functions of RANKL/RANK/OPG in bone modeling and remodeling. Arch Biochem Biophys 2008;473:139–46
18 Belibasakis, GN, Bostanci, N. The RANKL-OPG system in clinical periodontology. J Clin Periodontol 2012;39:239–48
19 Glass, DA 2nd, Karsenty, G. Canonical Wnt signaling in osteoblasts is required for osteoclast differentiation. Ann N Y Acad Sci 2006;1068:117–30
20 Glass, DA 2nd, Bialek, P, Ahn, JD, Starbuck, M, Patel, MS, Clevers, H et al. Canonical Wnt signaling in differentiated osteoblasts controls osteoclast differentiation. Dev Cell 2005;8:751–64
21 Baron, R, Rawadi, G. Targeting the Wnt/beta-catenin pathway to regulate bone formation in the adult skeleton. Endocrinology 2007;148:2635–43
22 Baron, R, Kneissel, M. WNT signaling in bone homeostasis and disease: from human mutations to treatments. Nat Med 2013;19:179–92
23 Fedi, P, Bafico, A, Nieto Soria, A, Burgess, WH, Miki, T, Bottaro, DP et al. Isolation and biochemical characterization of the human Dkk-1 homologue, a novel inhibitor of mammalian Wnt signaling. J Biol Chem 1999;274:19465–72
24 Kawano, Y, Kypta, R. Secreted antagonists of the Wnt signalling pathway. J Cell Sci 2003;116:2627–34
25 Sakalli, E, Baylancicek, S, Yuksel, M, Erdurak, SC, Dadas, B. Levels of reactive oxygen species in rat tympanic membranes after incisional versus radiofrequency myringotomy. Int J Pediatr Otorhinolaryngol 2013;77:792–5
26 Erdurak, SC, Coskun, BU, Sakalli, E, Tansuker, HD, Turan, F, Kaya, D. Does the use of radiofrequency myringotomy for insertion of a ventilation tube reduce the incidence of myringosclerosis? Eur Arch Otorhinolaryngol 2014;271:459–62
27 Mattsson, C, Magnuson, K, Hellstrom, S. Myringotomy: a prerequisite for the development of myringosclerosis? Laryngoscope 1998;108:102–6
28 Song, JJ, Kwon, SK, Cho, CG, Park, SW. The effect of caffeic acid phenethyl ester on the prevention of experimentally induced myringosclerosis. Int J Pediatr Otorhinolaryngol 2007;71:1287–91
29 NaderPour, M, Moghaddam, YJ, Peirovifar, A, Mollajavadi, R, Abbasi, MM, Mohajeri, D. Microscopic comparison of topical use of Mitomycin C and Fluorouracil on cold knife myringotomy. Int J Pediatr Otorhinolaryngol 2012;76:913
30 Dogan, E, Erdag, TK, Sarioglu, S, Ecevit, MC, Ikiz, AO, Guneri, EA. The preventive effect of N-nitro L-arginine methyl ester in experimentally induced myringosclerosis. Int J Pediatr Otorhinolaryngol 2011;75:1035–9
31 Mattsson, C, Hellstrom, S. Inhibition of the development of myringosclerosis by local administration of fenspiride, an anti-inflammatory drug. Eur Arch Otorhinolaryngol 1997;254:425–9
32 Li, J, Sarosi, I, Cattley, RC, Pretorius, J, Asuncion, F, Grisanti, M et al. Dkk1-mediated inhibition of Wnt signaling in bone results in osteopenia. Bone 2006;39:754–66
33 Ueland, T, Otterdal, K, Lekva, T, Halvorsen, B, Gabrielsen, A, Sandberg, WJ et al. Dickkopf-1 enhances inflammatory interaction between platelets and endothelial cells and shows increased expression in atherosclerosis. Arterioscler Thromb Vasc Biol 2009;29:1228–34
34 Yamaguchi, Y, Passeron, T, Hoashi, T, Watabe, H, Rouzaud, F, Yasumoto, K et al. Dickkopf 1 (DKK1) regulates skin pigmentation and thickness by affecting Wnt/beta-catenin signaling in keratinocytes. FASEB J 2008;22:1009–20
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

The Journal of Laryngology & Otology
  • ISSN: 0022-2151
  • EISSN: 1748-5460
  • URL: /core/journals/journal-of-laryngology-and-otology
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed