fgf8b  (Miltenyi Biotec)

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b protects cartilage in DMM mouse model. A. Immunohistochemical analysis of FGF8b expression on the surface of tibial articular cartilage at different time points during osteoarthritis progression, n = 3. Scale bar: 50 μm. B. Endogenous FGF8b expression gradually decreased after DMM surgery. C. Schematic diagram of intra-articular injection of exogenous FGF8b recombinant protein for the treatment of traumatic OA. They received intra-articular injections of either the solvent PBS (10 μl) or exogenous FGF8b recombinant protein (2 μg/10 μl) once per week. D-E. Representative safranin O-fast green-stained sagittal sections of knee joints 8 and 12 weeks after Sham or DMM surgery. The black dashed boxes indicate the magnified regions. Scale bar: low magnification 200 μm, high magnification 50 μm. F-G. OARSI scores for femoral and tibial cartilage in mice treated with PBS or FGF8b 8 and 12 weeks after DMM or Sham surgery, n = 6. MFC: medial femoral condyle; MTP: medial tibial plateau. Data are presented as mean ± 95 % confidence interval. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Immunohistochemical staining, Expressing, Injection, Recombinant, Solvent, Staining

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: Continuous administrated FGF8b causes osteophyte formation. A. X-ray images showing osteophyte formation around the knee joint in Sham and DMM surgery groups treated with intra-articular injections of FGF8b. The white arrows indicate the location of osteophytes in the X-ray images. B. Three-dimensional CT reconstruction images displaying extensive osteophyte formation around the knee joint following FGF8b treatment. The red arrows indicate the location of FGF8b-induced osteophytes in the CT images. Scale bar: 1 mm. C. Safranin O-fast green stained sagittal sections of knee joints 8 weeks after DMM surgery. The black dashed lines indicate osteophyte regions near the femoral growth plate. Scale bar: 500 μm. n = 6. D-E. Statistical assessment of osteophyte size and maturity following FGF8b induction in femur. ns: not significant.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Staining

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b attenuates articular cartilage degeneration in a dose-dependent manner. A-B. Radiographic (X-ray and CT) evaluation of the overall condition of the knee joint in mice treated with 10 μl of different concentrations of FGF8b (once weekly) starting one week after DMM surgery. Scale bar: 1 mm. The white arrows indicate the location of osteophytes in the X-ray images. The red arrows indicate the location of FGF8b-induced osteophytes in the CT images. C. Representative safranin O-fast green stained images of knee joints treated with a gradient of FGF8b concentrations 12 weeks after DMM surgery. The black dashed box indicates the viewing area at high magnification. Scale bar: low magnification 200 μm, high magnification 50 μm. D. OARSI scoring system was used to evaluate the effects of different doses of FGF8b on knee joint cartilage 12 weeks after Sham or DMM surgery. MFC: medial femoral condyle; MTP: medial tibial plateau. n = 6. Data are presented as mean ± 95 % confidence interval. ns: not significant, ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Staining

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: Intermittent administration of FGF8b protects cartilage in DMM mouse model. A. Mice underwent sham or DMM surgery, and subsequently received intra-articular injections of PBS or FGF8b (2 μg in 10 μl) every four weeks until tissue harvest at 12 weeks for analysis. B-C. X-ray and CT imaging were used to observe the effects of intermittent FGF8b injections on the knee joint. Scale bar: 1 mm. D. Representative safranin O-fast green stained images of knee joints from the Sham or DMM groups treated with intermittent PBS or FGF8b injections. The black dashed boxes indicate the regions magnified. Scale bar: low magnification 200 μm, high magnification 50 μm. E. OARSI scoring system was used to statistically evaluate the degeneration of knee joint cartilage in traumatic OA treated with intermittent FGF8b. n = 6. Data are presented as mean ± 95 % confidence interval. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Imaging, Staining

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b promotes anabolism and inhibits catabolism of articular cartilage in mice. A. Adult wild-type mice (12 weeks old) underwent DMM surgery. Intra-articular injection of PBS or FGF8b (2 μg/10 μl) was intermittent administrated. At 4 weeks post-DMM surgery, articular cartilage was extracted, subjected to RNA transcriptome sequencing. n = 4. B. Volcano plot of differentially expressed genes between the PBS or FGF8b-injected groups post-DMM surgery. C. GO enrichment analysis of upregulated genes in the FGF8b-injected group compared to the PBS-injected group post-DMM surgery. D. GO terms of downregulated genes in the FGF8b-injected group compared to the PBS-injected group. E-F. IHC staining of Sox9 and Acan on the surface of articular cartilage in mice treated with intermittent FGF8b. G-H. IHC images of ColⅩ and Mmp13 after FGF8b treatment. I-L. Relative quantitative statistics of gene expression related to cartilage homeostasis. Scale bar: 50 μm. n = 3. ∗∗P < 0.01.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Injection, Sequencing, Immunohistochemistry, Gene Expression

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b prevents human cartilage from IL-1β induced degeneration. A. Safranin O staining revealed increased proteoglycan content in the extracellular matrix of human cartilage sections after FGF8b treatment for Ex vivo OA model. B-D. IHC staining for Acan, Col X, and Mmp13 was performed on human articular cartilage explants treated with IL-1β and FGF8b. Scale bar: 100 μm, n = 3. E-G. Statistical analysis of relative expression of chondro-homeostasis-associated genes in cartilage explants. H. Human primary chondrocytes were treated with varying concentrations of FGF8b for 24 h, followed by western blotting analysis. I-J. The signal intensities of Sox9 and Runx2 were quantitatively analyzed. K. Human primary chondrocytes were treated with IL-1β (10 ng/ml) to induce an OA-like chondrocyte model, along with FGF8b (25 ng/ml) for 24 h. Western blotting was performed to assess the effects of FGF8b on chondrocyte homeostasis-related markers. L-M. Quantitative analysis of Mmp13 and Adamts5 gray value. n = 3. ns: not significant, ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: Staining, Ex Vivo, Immunohistochemistry, Expressing, Western Blot

Journal: Journal of Orthopaedic Translation

Article Title: Intermittent intra-articular delivery of FGF8b enhances cartilage homeostasis and attenuates osteoarthritis progression

doi: 10.1016/j.jot.2025.101037

Figure Lengend Snippet: FGF8b exerts a chondroprotective effect via the FGFR3-PI3K-AKT signaling pathway. A. Surface plasmon resonance analysis shows the high-affinity binding of FGF8b to FGFR3. B-C. FGF8b treatment of SW1353 chondrocytes for 15 min increased the expression of phosphorylated FGFR3. D-E. The knockdown efficiency of FGFR3 following transfection with FGFR3 siRNA in SW1353 cells was assessed by Western blotting. F-H. The chondroprotective effect of FGF8b for 24 h was partially attenuated after FGFR3 knockdown. I. KEGG enrichment analysis of transcriptome sequencing data suggested an upregulation of the PI3K-AKT signaling pathway following FGF8b treatment for osteoarthritis. J-L. The FGFR3-PI3K-AKT signaling pathway was downregulated in chondrocytes treated with IL-1β for 15 min, whereas FGF8b treatment activated the FGFR3-PI3K-AKT signaling pathway. M. The activation of the PI3K-AKT signaling pathway by FGF8b is dependent on FGFR3. N-P. The chondroprotective effect of FGF8b was partially abolished upon blockade of the PI3K-AKT signaling pathway using the LY294002 inhibitor under inflammatory conditions. n = 3, ns: not significant, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 and ∗∗∗∗P < 0.0001.

Article Snippet: The primary antibodies used were as follows: FGF8b (1:100, AF-423-NA, R&D Systems), Sox9 (1:200, Ab185230 , Abcam), Acan (1:200, AB1031, Invitrogen), Col X (1:200, Ab58632, Abcam) and Mmp13 (1:200, 18165-1-AP, Proteintech).

Techniques: SPR Assay, Binding Assay, Expressing, Knockdown, Transfection, Western Blot, Sequencing, Activation Assay