NEL-like molecule-1-modified bone marrow mesenchymal stem cells/poly lactic-co- glycolic acid composite improves repair of large osteochondral defects.

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NEL-like molecule-1-modified bone marrow mesenchymal stem cells/poly lactic-co- glycolic acid composite improves repair of large osteochondral defects in mandibular condyle S. Zhu, B. Zhang, C. Man, Y. Ma, J. Hu Osteoarthritis and Cartilage Volume 19, Issue 6, Pages 743-750 (June 2011) DOI: 10.1016/j.joca.2011.02.015 Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 (A) s.e.m. photographs of produced PLGA scaffold without cells. (B) The seeded BMMSCs attached on the PLGA scaffold. (C) Critical-size osteochondral defect created in the mandibular condyle. (D) Implantation of BMMSCs/PLGA composite in the defect. Bars=1mm in A and 50μm in B. Osteoarthritis and Cartilage 2011 19, 743-750DOI: (10.1016/j.joca.2011.02.015) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Gross appearance of specimens. At 6 weeks, the defects in both NELL-1-modified BMMSCs/PLGA (A) and BMMSCs/PLGA (B) groups were filled with shiny, white, semitransparent tissue, but the surface of the defects in the BMMSCs/PLGA-treated groups remained concave. The PLGA group (C) was rough with depression. In the empty defect group (D), the defects were filled with opaque tissue with a deep concavity. At 24 weeks, the defects in the NELL-1-modified BMMSCs/PLGA (E) and BMMSCs/PLGA (F) groups were substituted with smooth and translucent tissue just like host cartilage. In the PLGA group, the defects showed an irregular surface with firm cartilage-like tissues of similar color to the surrounding cartilage (G). In the empty defect group, arthritic changes were evident and defects remained depressed (H). Arrows showed the defect created in the mandibular condyle. Osteoarthritis and Cartilage 2011 19, 743-750DOI: (10.1016/j.joca.2011.02.015) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Histological photomicrographs of specimens 6 (A–H) and 24 (I–P) weeks after operation (Safranin-O staining). At 6 weeks, fibrocartilage was regenerated completely in the NELL-1-modified BMMSCs/PLGA group, but was thicker and more cellular than normal (A and E). Partial cartilage regeneration was observed in the BMMSCs/PLGA group (B and F, arrows in F showed cartilage disruption). In the PLGA group, the defects were filled with concave fibrous tissues containing small bone islands (C and G, arrows in G showed fibrous tissues in the defect). In the empty defect group the surgical sites contained fibrous tissue (D and H). At 24 weeks, a layer of well-organized fibrocartilage covering the subchondral bone was observed in the NELL-1-modified BMMSCs/PLGA group (I and M); however, the regenerated cartilage in the BMMSCs/PLGA had incomplete integration (J and N, arrow in N showed incomplete integration of cartilage). The PLGA group showed small portions of cartilaginous tissue formed adjacent to host cartilage with an irregular surface (K and O, arrows in O showed cartilaginous tissue formed adjacent to host cartilage). The empty defect group showed non-cartilaginous structure and was almost filled with fibrous tissue and inflammatory changes (L and P). Bars=500μm in A–D and I–L, and 100μm in E–H and M–P. Osteoarthritis and Cartilage 2011 19, 743-750DOI: (10.1016/j.joca.2011.02.015) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Histological scores of three groups at 6 and 24 weeks after operation (n=6). The data shown are the means±95% CI. Osteoarthritis and Cartilage 2011 19, 743-750DOI: (10.1016/j.joca.2011.02.015) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Immunostaining for type II collagen in the NELL-1-modified BMMSCs/PLGA (A and C) and BMMSCs/PLGA (B and D) groups. The matrix in the NELL-1-modified BMMSCs/PLGA group had more intense type II collagen staining compared with the BMMSCs/PLGA group at 6 (A and B) and 24 (C and D) weeks after operation. Bar=100μm. Osteoarthritis and Cartilage 2011 19, 743-750DOI: (10.1016/j.joca.2011.02.015) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Representative μ-CT images of the defects at 6 (A–D) and 24 (E–H) weeks after operation (n=6). (A and E) NELL-1-modified BMMSCs/PLGA group. (B and F) BMMSCs/PLGA group. (C and G) PLGA group. (D and H) Empty defect group. (I) BV/TV of different groups at 6 and 24 weeks. The data shown are the means±95% CI. Osteoarthritis and Cartilage 2011 19, 743-750DOI: (10.1016/j.joca.2011.02.015) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions

Fig. 7 X-gal staining for implanted cells tracing at 6 weeks after operation. (A) X-gal staining in Ad-NELL-1 transduced BMMSCs. (B) Blue color tissues filled within the defects. (C and D) Histological section showed the blue color tissues in the defect. Safranin-O counterstaining; bars=100μm in A, and 50μm in C and D. Osteoarthritis and Cartilage 2011 19, 743-750DOI: (10.1016/j.joca.2011.02.015) Copyright © 2011 Osteoarthritis Research Society International Terms and Conditions