J.G Costouros, M.D., A.C Dang, H.T Kim, M.D., Ph.D.

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Inhibition of chondrocyte apoptosis in vivo following acute osteochondral injury J.G Costouros, M.D., A.C Dang, H.T Kim, M.D., Ph.D. Osteoarthritis and Cartilage Volume 11, Issue 10, Pages 756-759 (October 2003) DOI: 10.1016/S1063-4584(03)00157-2

Fig. 1 Representative cartilage sections after TUNEL analysis (A and B) and DAPI staining (C and D). Control specimens are shown on the left (A and C), and Z-VAD-fmk treated specimens are shown on the right (B and D). Arrows indicate the experimental injury sites. TUNEL analysis demonstrates decreased levels of PCD in the treated specimen compared to control. The inhibitory effect is more pronounced moving away from the site of injury. (E) Immunohistochemistry using anti-ssDNA staining of heat/formamide treated specimens to confirm PCD. Positive cells show dark brown nuclear staining; negative cells show only light blue-green nuclear staining from the counterstain. Osteoarthritis and Cartilage 2003 11, 756-759DOI: (10.1016/S1063-4584(03)00157-2)

Fig. 2 Inhibition of chondrocyte apoptosis in vivo with Z-VAD-fmk treatment four days following osteochondral injury. The percentage of chondrocytes undergoing PCD was assessed by TUNEL analysis and calculated as: (number TUNEL-positive cells/number DAPI positive cells×100). The area around the injury site was subdivided into three concentric ‘zones’ for independent analysis. Each zone encompasses the full thickness of articular cartilage for a defined linear distance from the edge of the injury site (Zone 1=0–0.5mm; Zone 2=0.5–1.0mm; Zone 3=1.0–1.5mm). Osteoarthritis and Cartilage 2003 11, 756-759DOI: (10.1016/S1063-4584(03)00157-2)