Bone replaces articular cartilage in the rat knee joint after prolonged immobilization

Background: Lost joint range of motion (ROM) is common in chronic osteoarthritis, alters regional weight bearing across the articular surfaces, and contributes to loss of cartilage and bone alterations. Limited data exist on the regional effects on joints subjected to chronic losses of ROM. Objective: To characterize the regional replacement by bone as part of articular cartilage degeneration after prolonged immobilization. Methods: Eleven rat knees were rigidly-immobilized in flexion for 32 weeks with contralateral and sham operated (n = 6) knees as controls. Sagittal medial tibial epiphysis histological sections assessed the anterior (non-weight=bearing), middle and posterior (both weight-bearing) regions. We quantified the distribution of collagen I, collagen II, cartilage thickness, glycosaminoglycan (GAG) staining, Mankin scoring, and subchondral bone plate cross-sectional area. Using immunohistochemistry (IHC), we visualized blood vessels, osteoblasts, and mesenchymal stem cells (MSCs). Results: Immobilized cartilage had increased collagen I content in the anterior tibial region with picrosirius red staining (immobilized = 61 +/- 20%; contralateral = 43 +/- 12%, p = 0.033; sham = 20 +/- 10%, p = 0.028) and collagen I IHC (immobilized = 40 +/- 10%; contralateral = 11 +/- 4%, p = 0.003; sham = 5 +/- 3%, p = 0.043). Articular cartilage was thinner anteriorly (18 +/- 30 mu m) in immobilized knees versus contralateral (124 +/- 40 mu m, p < 0.001) and sham (125 43 mu m, p = 0.043). GAG staining covered 2 4% of the anterior articular area in immobilized knees versus 28 +/- 12% contralaterally (p = 0.003) and 26 +/- 7% in sham (p = 0.043). Mankin scores in immobilized knees were 4.7 +/- 1.7 versus 0.2 +/- 0.4 and 0 +/- 0 for contralateral and sham (p = 0.003, p = 0.042), respectively. The trabecular bone plate area of anterior and posterior regions showed relative loss of cross-sectional area in immobilized knees compared to controls (immobilized/contralateral area ratios of 0.67 and 0.46 respectively, both p = 0.003), while the area in the middle region was preserved. Movat's pentachrome stain and CD31 staining showed chondral vascular ingrowth from subchondral bone. Osteocalcin and CD90 MSC staining were decreased in immobilized knees versus contralateral (p = 0.003, p = 0.036 respectively). Conclusions: Bony replacement characterizes articular cartilage degeneration of knees immobilized for 32 weeks in the anterior, non-weight bearing region of the tibia. Replacement of cartilage by bone may have been mediated by chondral vascularization, suggesting irreversible changes. These findings stress the importance of weight bearing and joint motion to maintain cartilage structure. (C) 2017 Elsevier Inc. All rights reserved.