Histological and immunohistological analysis of degenerative changes in the cranial cruciate ligament in a canine model of excessive tibial plateau angle

T. Ichinohe; N. Kanno; Y. Harada; T. Yogo; M. Tagawa; Y. Hara

doi:10.3415/VCOT-14-08-0128

Veterinary and Comparative Orthopaedics and Traumatology, Table of Contents

Vet Comp Orthop Traumatol 2015; 28(04): 240-249
DOI: 10.3415/VCOT-14-08-0128

Original Research

Schattauer GmbH

Histological and immunohistological analysis of degenerative changes in the cranial cruciate ligament in a canine model of excessive tibial plateau angle

Authors

T. Ichinohe

¹Laboratory of Veterinary Surgery, Nippon Veterinary and Life Science University, Musashino, Tokyo, Japan
N. Kanno

¹Laboratory of Veterinary Surgery, Nippon Veterinary and Life Science University, Musashino, Tokyo, Japan
Y. Harada

¹Laboratory of Veterinary Surgery, Nippon Veterinary and Life Science University, Musashino, Tokyo, Japan
T. Yogo

¹Laboratory of Veterinary Surgery, Nippon Veterinary and Life Science University, Musashino, Tokyo, Japan
M. Tagawa

¹Laboratory of Veterinary Surgery, Nippon Veterinary and Life Science University, Musashino, Tokyo, Japan
Y. Hara

¹Laboratory of Veterinary Surgery, Nippon Veterinary and Life Science University, Musashino, Tokyo, Japan

Abstract

Summary

Objective: To create a canine model of excessive tibial plateau angle (eTPA) and assess the chondroid metaplasia and extracellular matrix alteration in the cranial cruciate ligament.

Methods: Seven mature female Beagles were included. Cylindrical osteotomy was performed bilaterally in the proximal tibia. The TPA was increased to approximately 40° in the left tibia (eTPA stifle) and left unchanged in the right tibia (control stifle). Exercise stress was started at three months postoperatively, and at 12 months postoperatively the dogs were euthanatized and the cranial cruciate ligaments were collected. The specimens were subjected to haematoxylin and eosin staining to assess the ligamentocyte morphology and immunostaining to assess the type I (COLI), type II (COLII), and type III (COLIII) collagen, and the sry-type HMG box 9 (SOX9) staining.

Results: Macroscopic cranial cruciate ligament injury was absent in six dogs but present in the eTPA stifle of one dog, which was excluded from the analysis. The ligamentocyte density decreased and the percentage of round ligamentocytes increased in the eTPA stifles. The COLII, COLIII, and SOX9 staining increased significantly and COLI deposition decreased in the eTPA stifles compared to the control stifle.

Clinical significance: The extracellular matrix changed, COLI deposition decreased, and COLIII and SOX9 staining increased in the cranial cruciate ligament of the eTPA stifles. SOX9 may contribute to COLII synthesis in the extracellular matrix of the cranial cruciate ligament in eTPA stifles, and eTPA may promote chondroid metaplasia and extra -cellular matrix alteration.

Keywords

Cranial cruciate ligament - dog - excessive tibial plateau angle - extracellular matrix - chondroid metaplasia

Full Text

References

References
1 Wilke VL, Robinson DA, Evans RB. et al. Estimate of the annual economic impact of treatment of cranial cruciate ligament injury in dogs in the United States. J Am Vet Med Assoc 2005; 227: 1604-1607.
2 Arnoczky SP, Marshall JL. The cruciate ligaments of the canine stifle: an anatomical and functional analysis. Am J Vet Res 1977; 38: 1807-1814.
3 Slocum B, Devine T. Cranial tibial thrust: a primary force in the canine stifle. J Am Vet Med Assoc 1983; 183: 456-459.
4 Marshall JL, Olsson SE. Instability of the knee: a long term experimental study in dogs. J Bone Joint Surg 1971; 53: 1561-1570.
5 Duerr FM, Duncan CG, Savicky RS. et al. Risk factors for excessive tibial plateau angle in large-breed dogs with cranial cruciate ligament disease. J Am Vet Med Assoc 2007; 231: 1688-1691.
6 Bennett D, Tennant B, Lewis DG. et al. A reappraisal of anterior cruciate ligament disease in the dog. J Small Anim Pract 1988; 29: 275-297.
7 Hayashi K, Frank JD, Dubinsky C. et al. Histologic changes in ruptured canine cranial cruciate ligament. Vet Surg 2003; 32: 269-277.
8 Muir P, Schamberger GM, Manley PA. et al. Localization of cathepsin K and tartrate-resistant acid phosphatase in synovium and cranial cruciate ligament in dogs with cruciate disease. Vet Surg 2005; 34: 239-246.
9 Comerford EJ, Tarlton JF, Wales A. et al. Ultrastructural differences in cranial cruciate ligaments from dogs of two breeds with a differing predisposition to ligament degeneration and rupture. J Comp Pathol 2006; 134: 8-16.
10 Wang JH. Mechanobiology of tendon. J Biomech 2006; 39: 1563-1582.
11 Tsukamoto N, Maeda T, Miura H. et al. Repetitive tensile stress to rat caudal vertebrae inducing cartilage formation in the spinal ligaments: a possible role of mechanical stress in the development of ossification of the spinal ligaments. J Neurosurg Spine 2006; 5: 234-242.
12 Reif U, Probst CW. Comparison of tibial plateau angles in normal and cranial cruciate deficient stifles of Labrador Retrievers. Vet Surg 2003; 32: 385-389.
13 Wilke VL, Conzemius MG, Besancon MF. Comparison of tibial plateau angle between clinically normal Greyhounds and Labrador Retrievers with and without rupture of the cranial cruciate ligament. J Am Vet Med Assoc 2002; 221: 1426-1429.
14 Slocum B, Devine T. Tibial Plateau Leveling Osteotomy Seminar. 2002. June 7-8 Eugene, OR, USA: Slocum Enterprises Inc;
15 Dismukes DI, Tomlinson JL, Fox DB. et al. Radiographic measurement of the proximal and distal mechanical joint angles in the canine tibia. Vet Surg 2007; 36: 699-704.
16 Slocum B, Slocum TD. Tibial plateau leveling osteotomy for repair of cranial cruciate ligament rupture in the canine. Vet Clin North Am Small Anim Pract 1993; 23: 777-795.
17 Tallat MB, Kowaleski MP, Boudrieau RJ. Combination tibial plateau leveling osteotomy and cranial closing wedge osteotomy of the tibia for the treatment of cranial cruciate ligament-deficient stifles with excessive tibial plateau angle. Vet Surg 2006; 35: 729-739.
18 Moeller EM, Cross AR, Rapoff AJ. Change in tibial plateau angle after leveling osteotomy in dogs. Vet Surg 2006; 35: 460-464.
19 Busby ER, Soeta S, Sherwood NM. et al. Molecular analysis of the koala reproductive hormones and their receptors: gonadotrophin-releasing hormone (GnRH), follicle-stimulating hormone β and luteinising hormone β with localisation of GnRH. J Neuroendocrinol 2014; 26: 870-887.
20 Kumagai K, Sakai K, Kusayama Y. et al. The extent of degeneration of cruciate ligament is associated with chondrogenic differentiation in patients with osteoarthritis of the knee. Osteoarthritis Cartilage 2012; 20: 1258-1267.
21 Vasseur PB, Pool RR, Arnoczky SP. et al. Correlative biomechanical and histologic study of the cranial cruciate ligament in dogs. Am J Vet Res 1985; 46: 1842-1854.
22 Chen J, Wang A, Xu J. et al. In chronic lateral epicondylitis, apoptosis and autophagic cell death occur in the extensor carpi radialis brevis tendon. J Shoulder Elbow Surg 2010; 19: 355-362.
23 Bi W, Deng JM, Zhang Z. et al. Sox9 is required for cartilage formation. Nat Genet 1999; 22: 85-89.
24 Furumatsu T, Hachioji M, Saiga K. et al. Anterior cruciate ligament-derived cells have high chondrogenic potential. Biochem Biophys Res Commun 2010; 391: 1142-1147.
25 Takimoto A, Oro M, Hiraki Y. et al. Direct conversion of tenocytes into chondrocytes by Sox9. Exp Cell Res 2012; 318: 1492-1507.
26 Narama I, Masuoka M, Matsuura T. et al. Morphogenesis of degenerative changes predisposing dogs to rupture of the cranial cruciate ligament. J Vet Med Sci 1996; 58: 1091-1097.
27 Waggett AD, Kwan APL, Woodnutt DJ. et al. Collagens in fibrocartilages at the Achilles tendon insertion-A biochemical, molecular biological and immunohistochemical study. Trans Orthop Res Soc 1996; 25.
28 Kumagai J, Sarkar K, Uhthoff HK. et al. Immunohistochmical distribution of type I, II, and III collagens in the rabbit supraspinatus tendon insertion. J Anat 1994; 185: 279-284.
29 Riley GP, Harrall RL, Constant CR. et al. Tendon degeneration and chronic shoulder pain: changes in the collagen composition of the human rotator cuff tendons in rotator cuff tendinitis. Ann Rheum Dis 1994; 53: 359-366.
30 Frank C, Amiel D, Woo SL. et al. Normal ligament properties and ligament healing. Clin Orthop Relat Res 1985; 196: 15-25.
31 Comerford EJ, Innes JF, Tarlton JF. et al. Investigation of the composition, turnover, and thermal properties of ruptured cranial cruciate ligament rupture. Am J Vet Res 2004; 65: 1136-1141.
32 Breshears LA, Cook JL, Stoker AM. et al. The effect of uniaxial cyclic tensile load on gene expression in canine cranial cruciate ligamentocytes. Vet Surg 2010; 39: 433-443.
33 Furumatsu T, Matsumoto E, Kanazawa T. et al. Tensile strain increases expression of CCN2 and COL2A1 by activating TGF-β-Smad2/3 pathway in chondrocytic cells. J Biomech 2013; 46: 1508-1515.
34 Read RA, Robins GM. Deformity of the proximal tibia in dogs. Vet Rec 1982; 111: 295-298.
35 Hulse D, Beale B, Kerwin S. Second look arthroscopic findings after tibial plateau levelling osteotomy. Vet Surg 2010; 39: 350-354.
36 Zachos TA, Arnoczky SP, Lavagnino M. et al. The effect of cranial cruciate ligament insufficiency on caudal cruciate ligament morphology: An experimental study in dogs. Vet Surg 2002; 31: 596-603.