Comparative intra-articular gene transfer of seven adeno-associated virus serotypes reveals that AAV2 mediates the most efficient transduction to mouse arthritic chondrocytes

Figures

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  Fig 1. Animal treatment and study design of the project.

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  Fig 2. eGFP was expressed in mouse cartilage tissue 30 days after intra-articular injection of different AAV serotypes. (A) The expression of eGFP in mouse cartilage tissue was observed with an inverted fluorescence microscope. Yellow arrowhead, eGFP expression in articular cartilage; Yellow arrow, non-specific fluorescence in the subchondral bone. n = 5, Scale bar = 200 μm. (B) The density of eGFP in arthritic cartilage in A. Data are expressed as mean ± SD. Bars in B sharing a common letter are not significantly different (P ≥ 0.05) between groups.

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  Fig 3. Gross appearance and histological changes of knee joints. (A) After intra-articular injection, the knee joints were directly dissected and then tibial plateau (A1), meniscus (A2), femoral condyles (A3), femoral trochlea (A4) were observed under a stereomicroscope. n = 5, Scale bar = 0.8 mm. (B) The knee joints were dissected 30 days after AAV injection, and the cartilage area of the medial tibial plateaus was stained with toluidine blue. n = 5, Scale bar = 200 μm. (C) Degeneration of articular cartilage was evaluated using the classical Mankin scoring system. Black arrowhead, positive staining of MMPs in articular cartilage. Data are expressed as mean ± SD. *P < 0.05, ** P < 0.01.

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  Fig 4. Immunohistochemical analysis of Col2, MMP-3, MMP-13 expression in the cartilage of knee joints. (A-C) Immunohistochemical staining results of Col2 (A), MMP-3 (B) and MMP-13 (C) in articular cartilage. (D-F) The relative optical density of positive staining in A-C. Col2, Type II collagen; MMP-3, Matrix metalloproteinase-3; MMP-13, Matrix metalloproteinase-3. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01. n = 5. Scale bar = 50 μm.

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  Fig 5. Immunohistochemical analysis of Aggrecan, ADAMTS-4, ADAMTS-7 and MMP-19 expressions in the articular cartilage. (A-D) Immunohistochemical staining of Aggrecan (A), ADAMTS-4 (B), ADAMTS-7 (C), MMP-19 (D) expressions in articular cartilage. (E-H) The relative optical density of positive staining in A-D. ADAMTS, A disintegrin and metalloproteinase with thrombospondin motifs; MMP-19, Matrix metalloproteinase-19. Black arrowhead, positive staining of Aggrecanases in articular cartilage. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01. n = 5. Scale bar = 50 μm.

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  Fig 6. Immunohistochemical analysis of pyroptosis of the articular cartilage. (A-B) Immunohistochemical staining of Cryopyrin (A) and Caspase-1 (B). (C-D) The relative optical density of positive staining in A-B. Cryopyrin (NLRP3), Nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3; Caspase-1, Cysteinyl aspartate specific proteinase-1. Data are expressed as mean ± SD. *P < 0.05, **P < 0.01. n = 5. Scale bar = 50 μm.