An experimental study on costal osteochondral graft

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Presentation transcript:

An experimental study on costal osteochondral graft K. Sato, O.J. Moy, C.A. Peimer, T. Nakamura, C. Howard, S.H. Ko, T.C. Lee, Y. Nishiwaki Osteoarthritis and Cartilage Volume 20, Issue 2, Pages 172-183 (February 2012) DOI: 10.1016/j.joca.2011.12.001 Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 (A) Harvested costal osteochondral transit site. The arrow shows costal cartilage. (B) Costal cartilage (arrow) was shaped to fit the defect with a scalpel and osseous portion (rib) was trimmed with bone rongeur forceps. (C) The costal osteochondral plug press-fitted into the femoral defect. Osteoarthritis and Cartilage 2012 20, 172-183DOI: (10.1016/j.joca.2011.12.001) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Illustration of the experimental design. Osteoarthritis and Cartilage 2012 20, 172-183DOI: (10.1016/j.joca.2011.12.001) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Osteochondral defects after surgery. (A) Costal osteochondral graft at 6weeks after surgery. (B) Control at 6weeks after surgery. The defect was filled with semitransparent tissue but it was still in a concave shape. (C) Costal osteochondral graft at 12weeks after surgery. The graft had white and shiny surface but the margin between the graft and surrounding articular cartilage was still distinguishable. (D) Control at 12weeks after surgery. The surface of the defect was mostly flat and looked white and shiny. (E) Costal osteochondral graft at 24weeks after surgery. Although the margin was still distinguishable, the surface of the graft looked like normal articular cartilage. (F) Control at 24weeks after surgery. The surface of the defect was rough and irregular. (G) Costal osteochondral graft at 48weeks after surgery. The surface of the graft was still white and shiny. There was no sign of degenerative change. (H) Control at 48weeks after surgery. The defect had white but irregular surface. Osteoarthritis and Cartilage 2012 20, 172-183DOI: (10.1016/j.joca.2011.12.001) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Histological findings of osteochondral defect at 6 (A–D), 12 (E–H), 24 (I–L) and 48 (M–P) weeks after surgery. (A) Histology of grafted costal cartilage at 6weeks. Bone union was achieved at the subchondral bone level and chondrocytes in the graft looked viable in costal osteochondral graft (Hematoxylin-eosin; magnification ×40). (B) Histology of grafted costal cartilage at 6weeks. The matrix in the grafted costal cartilage was stained red which suggests the existence of glycosaminoglycans (Safranin-O; magnification ×40). (C) Histology of untreated control at 6weeks. The repair tissue was more cellular than adjacent articular cartilage and it possessed a population of smaller-sized cell (Hematoxylin-eosin; magnification ×40). (D) Histology of untreated control at 6weeks. The matrix of regenerated cartilage-like tissue was stained red (Safranin-O; magnification ×40). (E) Histology of grafted costal cartilage at 12weeks. Chondrocytes in the graft looked healthy but the gap between the cartilage layer of the graft and the surrounding articular cartilage had continuity to fibrocartilage-like tissue, but not hyaline cartilage (Hematoxylin-eosin; magnification ×40). (F) Histology of grafted costal cartilage at 12weeks. Although the matrix in the grafted cartilage was stained red at high levels with Safranin-O, the gap between the cartilage layer of the graft and the surrounding articular cartilage was not stained well (Safranin-O; magnification ×40). (G) Histology of untreated control at 12weeks. The surface of the regenerated cartilage was irregular (Hematoxylin-eosin; magnification ×40). (H) Histology of untreated control at 12weeks. Regenerated cartilage-like tissue was stained red but not so intense (Safranin-O; magnification ×40). (I) Histology of grafted costal cartilage at 24weeks. Chondrocytes in the grafted cartilage looked healthy (Hematoxylin-eosin; magnification ×40). (J) Histology of grafted costal cartilage at 24weeks. The gap between the cartilage layer of the graft and the surrounding articular cartilage was slightly stained red (Safranin-O; magnification ×40). (K) Histology of untreated control at 24weeks. Regenerated cartilage-like tissue showed fibrillation (Hematoxylin-eosin; magnification ×40). (L) Histology of untreated control at 24weeks. The intensity of the Safranin-O staining of the matrix was diminished compared with the adjacent cartilage (Safranin-O; magnification ×40). (M) Histology of grafted costal cartilage at 48weeks. There was no evidence of degenerative changes (Hematoxylin-eosin; magnification ×40). (N) Histology of grafted costal cartilage at 48weeks. There were no remarkable differences compared with 24weeks (Safranin-O; magnification ×40). (O) Histology of untreated control at 48weeks. Regenerated cartilage-like tissue showed remarkable fibrillation (Hematoxylin-eosin; magnification ×40). (P) Histology of untreated control at 48weeks. The intensity of the Safranin-O staining of the matrix was completely diminished (Safranin-O; magnification ×40). Bars=500μm. Osteoarthritis and Cartilage 2012 20, 172-183DOI: (10.1016/j.joca.2011.12.001) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Histological findings of osteochondral defect at 6 (A–D), 12 (E–H), 24 (I–L) and 48 (M–P) weeks after surgery. (A) Histology of grafted costal cartilage at 6weeks. Bone union was achieved at the subchondral bone level and chondrocytes in the graft looked viable in costal osteochondral graft (Hematoxylin-eosin; magnification ×40). (B) Histology of grafted costal cartilage at 6weeks. The matrix in the grafted costal cartilage was stained red which suggests the existence of glycosaminoglycans (Safranin-O; magnification ×40). (C) Histology of untreated control at 6weeks. The repair tissue was more cellular than adjacent articular cartilage and it possessed a population of smaller-sized cell (Hematoxylin-eosin; magnification ×40). (D) Histology of untreated control at 6weeks. The matrix of regenerated cartilage-like tissue was stained red (Safranin-O; magnification ×40). (E) Histology of grafted costal cartilage at 12weeks. Chondrocytes in the graft looked healthy but the gap between the cartilage layer of the graft and the surrounding articular cartilage had continuity to fibrocartilage-like tissue, but not hyaline cartilage (Hematoxylin-eosin; magnification ×40). (F) Histology of grafted costal cartilage at 12weeks. Although the matrix in the grafted cartilage was stained red at high levels with Safranin-O, the gap between the cartilage layer of the graft and the surrounding articular cartilage was not stained well (Safranin-O; magnification ×40). (G) Histology of untreated control at 12weeks. The surface of the regenerated cartilage was irregular (Hematoxylin-eosin; magnification ×40). (H) Histology of untreated control at 12weeks. Regenerated cartilage-like tissue was stained red but not so intense (Safranin-O; magnification ×40). (I) Histology of grafted costal cartilage at 24weeks. Chondrocytes in the grafted cartilage looked healthy (Hematoxylin-eosin; magnification ×40). (J) Histology of grafted costal cartilage at 24weeks. The gap between the cartilage layer of the graft and the surrounding articular cartilage was slightly stained red (Safranin-O; magnification ×40). (K) Histology of untreated control at 24weeks. Regenerated cartilage-like tissue showed fibrillation (Hematoxylin-eosin; magnification ×40). (L) Histology of untreated control at 24weeks. The intensity of the Safranin-O staining of the matrix was diminished compared with the adjacent cartilage (Safranin-O; magnification ×40). (M) Histology of grafted costal cartilage at 48weeks. There was no evidence of degenerative changes (Hematoxylin-eosin; magnification ×40). (N) Histology of grafted costal cartilage at 48weeks. There were no remarkable differences compared with 24weeks (Safranin-O; magnification ×40). (O) Histology of untreated control at 48weeks. Regenerated cartilage-like tissue showed remarkable fibrillation (Hematoxylin-eosin; magnification ×40). (P) Histology of untreated control at 48weeks. The intensity of the Safranin-O staining of the matrix was completely diminished (Safranin-O; magnification ×40). Bars=500μm. Osteoarthritis and Cartilage 2012 20, 172-183DOI: (10.1016/j.joca.2011.12.001) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Matrix staining score. The score of the grafted cartilage below the tidemark group at 48weeks was significantly lower than that at 6weeks (P =0.013). The scores of the control group sections 24 and 48weeks after the surgery were significantly lower than that of 6weeks (P =0.016, 0.001). Data expressed as mean with 95% CIs (n=5). *P <0.05 vs 6weeks in each group. Osteoarthritis and Cartilage 2012 20, 172-183DOI: (10.1016/j.joca.2011.12.001) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 (A) Confocal laser microscopic findings at 48weeks after surgery. Multiple green fluorescing living cells were seen in the graft as well as surrounding articular cartilage. (B) Relative cell density in the grafted costal cartilage and in the control. Data expressed as mean with 95% CIs (n=15: three coordinates from one animal, five animals in each group). Osteoarthritis and Cartilage 2012 20, 172-183DOI: (10.1016/j.joca.2011.12.001) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 7 Expression of type I collagen, type II collagen and aggrecan mRNA were assessed by RT-PCR. The band for type II collagen was more intense in costal osteochondral graft than the control. Type I collagen was not detected in costal osteochondral graft. The band for aggrecan was more intense than the control. (A) costal osteochondral graft, (B) control, (C) normal articular cartilage, (D) normal costal cartilage. Osteoarthritis and Cartilage 2012 20, 172-183DOI: (10.1016/j.joca.2011.12.001) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions