The use of photooxidized, mushroom-structured osteochondral grafts for cartilage resurfacing – a comparison to photooxidized cylindrical grafts in an.

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The use of photooxidized, mushroom-structured osteochondral grafts for cartilage resurfacing – a comparison to photooxidized cylindrical grafts in an experimental study in sheep1 1 The studies included in this publication were financed by a fellowship granted by Centerpulse Biologics, Winterthur, Switzerland B von Rechenberg, PD Dr.med.vet., Dipl. ECVS, M.K Akens, Dr.med.vet., PhD, D Nadler, P Bittmann, Dr.sc.techn., K Zlinszky, K Neges, med.vet., J.A Auer, Prof. Dr.med.vet., MS, Dipl. ACVS/ECVS Osteoarthritis and Cartilage Volume 12, Issue 3, Pages 201-216 (March 2004) DOI: 10.1016/j.joca.2003.11.001

Fig. 1 Structure of osteochondral grafts after photooxidation. The diameter of the mushroom-structured (a) and the cylindrical (b) grafts is 6mm and the depth is 7mm. Note bluish coloration and empty pores of the bone matrix of the grafts after photooxidation. (c=cartilage, tm=tide mark, sc=subchondral bone area). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 2 Surgical site of the stifle joints, where (a) depicts the preparation of the graft bed, and (b) shows the graft after insertion using the pressfit technique (lat=lateral, med=medial, p=patella, t=common tendon of m.ext. digit. pedis long. and m.fibularis tertius. Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 3 Macroscopic appearance of grafted cartilage site at the time of sacrifice. The interface between host and grafts was still visible in both, the mushroom structured grafts (a) and the cylindrical grafts (b) at 2 and 6 months after surgery. The original bluish color has faded, but still is easily detected in both types of grafts. Note the macroscopic appearance of the adjacent cartilage that is still glossy, whitish and does not reveal major signs of cartilage degeneration. Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 4 Cylindrical photooxidized graft at 6 months: cells from the subchondral area (red arrow) had penetrated the calcified cartilage zone (cc) including the tide mark (tm) and had already differentiated into chondroid type of cells. The old, photooxidized cartilage matrix (cm) is still easily distinguished from the newly synthesized cartilage due to its diminished metachromatic staining properties (ground section, 30–40μ, surface staining with toluidine blue). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 5 Cylindrical photooxidized graft at 2 months: Remodeling of the calcified cartilage zone (cc) is already visible. Columns of chondroid cells stemming from the subchondral bone area (sc) (white arrow head) penetrate the calcified cartilage zone and tide mark (tm). Repopulation of the old photooxidized cartilage matrix (cm) with new cells (red arrow) was noticed mainly in the deep zone of the cartilage (5μm section, staining with toluidine blue). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 6 Cylindrical photooxidized graft at 6 months: The interface (black arrows) between the host (h) and graft (g) cartilage is still not fused. Cluster formation is present mainly in the host cartilage, but no other signs of cartilage degeneration, such as fibrillation or loss of matrix staining are noticed (ground section, 30–40μm, surface staining with toluidine blue). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 7 Mushroom-shaped graft at 2 months: endochondral ossification (eo) already occurred at the base of the mushroom head, where the subchondral bone area (sc) and the stem (s) of the graft are already intensively remodeling. The interface (if) between the graft and host (h) bone is not united yet; however, synovial fluid penetration into the subchondral bone area is severely inhibited due to the mushroom structure of the graft (ground section, 30–40μm, surface staining with toluidine blue). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 8 Mushroom-shaped graft at 6 months: The bony part of the graft has almost completely remodeled. Although, at the left side (white arrow) a small cyst-like lesion has developed, the graft had not dislocated. At the base of the graft (red arrow), no cyst formation was noticed (ground section, 30–40μm, surface staining with toluidine blue). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 9 Mushroom-shaped graft at 6 months: Fusion between the host (h) and graft (g) cartilage is noticed. The interface (if) is united (red arrow) and chondroid cells are invading the old photooxidized cartilage matrix of the graft (black arrow) (5μm section, staining with toluidine blue). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 10 Graft-host cartilage interface of mushroom-shaped graft at 6 months: The host matrix (h) kept its viability and matrix staining properties at the edge of the cartilage (black arrow) throughout the study. The cells and chondrocyte clusters are visible in columns within the host matrix (g=graft, sc=subchondral bone area of the host) (5μm section, staining with toluidine blue). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 11 Mushroom-shaped graft at 2 months: An overview of the graft (a) shows that the deep and the superficial zone are already repopulated. Cells (b) migrate from the subchondral bone area (sc) through the calcified cartilage zone including the tide mark (red arrow) and repopulate the old photooxidized cartilage matrix of the graft (g). (b) shows a magnification of the area outlined in (a), where cells (red arrow) have already invaded some of the empty lacunae (black arrow) in the middle cartilage zone (5μm section, staining with von Kossa/McNeal). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 12 Mushroom-shaped graft at 2 months: The cartilage surface of the graft depicted was covered with a thin soft tissue layer (black arrow) and cells had invaded the superficial zone (sz) of the old photooxidized cartilage matrix (red arrow), while the middle zone (mz) was still mostly acellular (5μm section, staining with von Kossa/McNeal). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 13 Box plots representing the median percentage of fibrous and other non-bony tissue formation (a) in both types of grafts at 2 and 6 months. Note that the median percentage is higher in the cylindrical grafts and there, mostly in the 6 months specimens. The median percentage of bone matrix is recorded in both types of grafts at 2 and 6 months in (b). The median percentage is relatively stable in the mushroom structured grafts at 2 and 6 months, while in the cylindrical grafts, this value is decreased in the 6 months specimens. In (c&d) an overview of the cylindrical and mushroom-shaped grafts at 2 and 6 months are presented from where the histomophometrical data was obtained. The coloring of the cartilage (pink) and background (yellow) was made to avoid mistakes during detection of fibrous tissue and bone matrix. Note that cyst-like lesions are more pronounced in the cylindrical grafits (c) compared to the mushroom-shaped grafts, where only one graft had collapsed into the original defect at 6 months (c). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)

Fig. 13 Box plots representing the median percentage of fibrous and other non-bony tissue formation (a) in both types of grafts at 2 and 6 months. Note that the median percentage is higher in the cylindrical grafts and there, mostly in the 6 months specimens. The median percentage of bone matrix is recorded in both types of grafts at 2 and 6 months in (b). The median percentage is relatively stable in the mushroom structured grafts at 2 and 6 months, while in the cylindrical grafts, this value is decreased in the 6 months specimens. In (c&d) an overview of the cylindrical and mushroom-shaped grafts at 2 and 6 months are presented from where the histomophometrical data was obtained. The coloring of the cartilage (pink) and background (yellow) was made to avoid mistakes during detection of fibrous tissue and bone matrix. Note that cyst-like lesions are more pronounced in the cylindrical grafits (c) compared to the mushroom-shaped grafts, where only one graft had collapsed into the original defect at 6 months (c). Osteoarthritis and Cartilage 2004 12, 201-216DOI: (10.1016/j.joca.2003.11.001)