Cellular origin of neocartilage formed at wound edges of articular cartilage in a tissue culture experiment P.K. Bos, M.D., Ph.D., N. Kops, B.Sc., J.A.N.

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Cellular origin of neocartilage formed at wound edges of articular cartilage in a tissue culture experiment P.K. Bos, M.D., Ph.D., N. Kops, B.Sc., J.A.N. Verhaar, M.D., Ph.D., G.J.V.M. van Osch, Ph.D. Osteoarthritis and Cartilage Volume 16, Issue 2, Pages 204-211 (February 2008) DOI: 10.1016/j.joca.2007.06.007 Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Graph representing outgrowth of tissue from immature and mature bovine full-thickness articular cartilage explants cultured for 4 weeks. Outgrowth at wound edges was semiquantified and expressed as area of outgrowth divided by height of explants at wound edges (μm2/μm). Osteoarthritis and Cartilage 2008 16, 204-211DOI: (10.1016/j.joca.2007.06.007) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Wound edge of full-thickness explant. Paraffin embedded tissue. Immature (a and b) and mature (c and d) bovine articular cartilage with newly formed cartilage covering the lesion edge after 4 weeks in culture. Thionin staining (a and c); immunohistochemical staining for collagen type II (b and d); bar=200μm. Osteoarthritis and Cartilage 2008 16, 204-211DOI: (10.1016/j.joca.2007.06.007) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Collagen immunostained cryosections of immature deep explants following 28 days of culture. (a) Collagen type I; (b) type II; (c) type III. Cartilage was immunopositive for collagen type II and very weak for collagen types I and III. Outgrowth was immunopositive for both collagen types II and I, very weak for type III. Osteoarthritis and Cartilage 2008 16, 204-211DOI: (10.1016/j.joca.2007.06.007) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 The location of cell proliferation by immunohistochemical staining for proliferation marker Ki67. In full-thickness immature explants (a) a positive immunoreaction was observed in superficial zone (b) and deep zone (d) chondrocytes, as well as in cells covering the lesion edge (e). No proliferation activity was observed in the middle zone (c) chondrocytes. Bar=200μm. Osteoarthritis and Cartilage 2008 16, 204-211DOI: (10.1016/j.joca.2007.06.007) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Graph representing tissue formation at wound edges and surrounding immature bovine articular cartilage explants cultured for 4 weeks. Superficial and deep cartilage was explanted separately before culture. After 14 days very little outgrowth was observed at lesion edges of superficial cartilage, whereas at lesion edges of deep cartilage a significant larger amount of neocartilage is formed. Significant more outgrowth was observed after 28 days. Outgrowth at wound edges was semiquantified and expressed as area of outgrowth divided by height of explants at wound edges (μm2/μm). Osteoarthritis and Cartilage 2008 16, 204-211DOI: (10.1016/j.joca.2007.06.007) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Wound edges of superficial and deep explants of immature bovine articular cartilage. (a) Superficial cartilage, 14 days of culture, H&E; (b) superficial cartilage, 14 days of culture, thionin; (c) superficial cartilage, 28 days of culture, H&E; (d) superficial cartilage, 28 days of culture, thionin; (e) superficial cartilage, 28 days of culture, collagen type II immunostaining; (f) deep cartilage, 14 days of culture, H&E; (g) deep cartilage, 14 days of culture, thionin; (h) deep cartilage, 28 days, H&E; (i) deep cartilage, 28 days, thionin; (j) deep cartilage, 28 days, collagen type II immunostaining. Bar=200μm. Osteoarthritis and Cartilage 2008 16, 204-211DOI: (10.1016/j.joca.2007.06.007) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions