1. Volume 12, Issue 2, Pages 229-238 (August 2005)
Improved Tissue Repair in Articular Cartilage Defects in Vivo by rAAV-Mediated Overexpression of Human Fibroblast Growth Factor 2 
Magali Cucchiarini, Henning Madry, Chunyan Ma, Tanja Thurn, David Zurakowski, Michael D. Menger, Dieter Kohn, Stephen B. Trippel, Ernest F. Terwilliger 
Molecular Therapy 
Volume 12, Issue 2, Pages (August 2005)
DOI: /j.ymthe
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

2. Fig. 1
Analysis of FGF-2 expression in vitro. (A) Western blotting of lysates from rabbit articular chondrocytes and rabbit bone marrow clots transduced by rAAV. Lane 1, extracts from rAAV-lacZ-transduced chondrocytes (10 μg); lane 2, extracts from rAAV-hFGF-2-transduced chondrocytes (10 μg); lane 3, extracts from rAAV-lacZ-transduced bone marrow clots (60 μg); lane 4, extracts from rAAV-hFGF-2-transduced bone marrow clots (60 μg). (B) Time course analysis of FGF-2 production in transduced alginate–chondrocyte constructs. Cells were transduced by rAAV-lacZ or rAAV-hFGF-2 and encapsulated in alginate 2 days after vector application. rAAV-lacZ- and rAAV-hFGF-2-transduced constructs were prepared and maintained in culture for 26 days. Conditioned medium was collected at the denoted time points after encapsulation (n = 9 per time point and condition) and FGF-2 production was measured by ELISA (±SD) with a detection limit of 3 pg/ml.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

3. Fig. 2
Histological sections of transduced alginate–chondrocyte constructs. rAAV-lacZ- (A and B) and rAAV-hFGF-2-transduced constructs (C and D) were histologically processed at day 26 after encapsulation (n = 6 per condition) and analyzed for HE staining (A and C) and for immunohistological detection of type-II collagen with a mouse anti-type-II antibody (1:100), using a biotinylated goat anti-mouse antibody (1:200). Revelation was performed by the ABC method using DAB as the chromogen. Samples were examined under light microscopy. Original magnification, ×20.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

4. Fig. 3
Representative analysis of lacZ expression in osteochondral cartilage defects in vivo. Transgene expression was detected by X-Gal staining in knees retrieved 10 days (A–D; C and D, original magnification, ×100) or 20 days (E and F, original magnification, ×100) after vector application and by immunohistochemistry using a mouse anti-β-gal antibody (1:50) in knees retrieved 4 months postadministration (G and H, original magnification, ×20), as described in the legend to Fig. 2 and under Materials and Methods. (A, C, E, and G) Application of rAAV-lacZ (10 μl); (B, D, F, and H) application of rAAV-hFGF-2 (10 μl).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

5. Fig. 4
Representative analysis of FGF-2 expression in osteochondral cartilage defects in vivo. Transgene expression was detected by immunohistochemistry in sections from knees retrieved 10 days (A and B), 20 days (C and D), and 4 months (E and F) after vector application using a mouse anti-FGF-2 antibody (1:100), as described in the legend to Fig. 2 and under Materials and Methods. (A, C, and E) Application of rAAV-lacZ (10 μl); (B, D, and F) application of rAAV-hFGF-2 (10 μl). Original magnification, ×20.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

6. Fig. 5
Analysis of type-II and type-I collagen expression in osteochondral cartilage defects in vivo. Immunostaining was performed in sections from knees retrieved 20 days (A, C, E, and G) and 4 months (B, D, F, and H) after vector application using a mouse anti-type-II collagen antibody (1:50) (A–D) and a mouse anti-type-I collagen antibody (1:100) (E–H), as described in the legend to Fig. 2 and under Materials and Methods. (A, B, E, and F) Application of rAAV-lacZ (10 μl); (C, D, G, and H) application of rAAV-hFGF-2 (10 μl). Original magnification, ×4.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

7. Fig. 6
Histological sections of osteochondral cartilage defects. Safranin O staining was performed on sections from knees retrieved 20 days (A and D, original magnification, ×2) and 4 months after vector application (B and E, original magnification, ×2; C and F, original magnification ×4). (A–C) Application of rAAV-lacZ (10 μl); (D–F) application of rAAV-hFGF-2 (10 μl).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2005 The American Society of Gene Therapy Terms and Conditions