Journal: Annals of Translational Medicine

Article Title: Delay in articular cartilage degeneration of the knee joint by the conditional removal of discoidin domain receptor 2 in a spontaneous mouse model of osteoarthritis

doi: 10.21037/atm-20-5786

Figure Lengend Snippet: Immunohistostaining of Ddr2, MMP-13, and degraded collagen type II in the articular cartilage of mouse knee joints at 9 months of age. The protein expression of Ddr2, MMP-13, and degraded collagen type II were increased in oil-treated Acan+/CreERT2;Ddr2flox/flox;Col11a1+/− mice (as indicated by the brown-color–stained cells in the middle column of the figure). However, the protein expression of Ddr2, MMP-13, and degraded collagen type II were not detected in the articular cartilage of tamoxifen-treated Acan+/CreERT2;Ddr2flox/flox; Col11a1+/− mice (Acan+/CreERT2;Ddr2Δ/Δ;Col11a1+/−; right column) and wild-type mice (left column). The dashed line represents the boundary between articular cartilage (AC) and subchondral bone (SB). Bar =100 µm.

Article Snippet: The sections were incubated with the antibody against Ddr2 (1:300 dilution, Abcam, cat. no. ab203219), the antibody against MMP-13 (1:300 dilution, Abcam, cat. no. ab84594), or the antibody against degraded collagen type II (1:400 dilution, IBEX Pharmaceuticals Inc, cat. no. 50–1053) at 4 °C, overnight.

Techniques: Expressing, Staining

Journal: Arthritis Research & Therapy

Article Title: Ipriflavone attenuates the degeneration of cartilage by blocking the Indian hedgehog pathway

doi: 10.1186/s13075-019-1895-x

Figure Lengend Snippet: Ipriflavone reduced the degeneration of cartilage by inhibiting Ihh signaling in cultured human cartilage explants. a Safranin O staining results showed that cartilage samples were taken from “relatively normal” cartilage samples of the tibia obtained during total knee arthroplasty (Mankin score 0–2). Then, the cartilage samples were cut into 4-mm 3 pieces weighing 6–9 mg each and randomly subdivided into three groups: 0.1% DMSO group, 50 μM IP treatment group, and 100 μM IP treatment group. b Real-time PCR results showed that the mRNA expression of key genes in Ihh pathway, Smo, Gli-1,Gli-2,Gli-3, and Runx-2 was decreased in the IP treatment group, especially the reduction of Gli-2. Simultaneously, the mRNA levels of type II collagen was increased while MMP-13 and type X collagen were decreased in both IP treatment groups. c Western blotting results indicated that the expression of key proteins in Ihh signaling (Smo and Runx-2) were decreased in the IP treatment groups compared with the DMSO control group. MMP-13 and type X collagen protein were also reduced, while the expression of type II collagen protein was increased compared with the DMSO control group. Values are the mean ± SEM. n = 3, * P < 0.05, ** P < 0.01 versus the DMSO and IP treatment group

Article Snippet: The sections were incubated with specific antibodies against rat type II collagen, MMP-13, types X collagen, or type II collagen-C fragment (IBEX Technologies, Mont-Royal, QC, Canada) at 4 °C overnight.

Techniques: Cell Culture, Staining, Real-time Polymerase Chain Reaction, Expressing, Western Blot

Journal: Arthritis Research & Therapy

Article Title: Ipriflavone attenuates the degeneration of cartilage by blocking the Indian hedgehog pathway

doi: 10.1186/s13075-019-1895-x

Figure Lengend Snippet: Primers used in this paper with their species, name, orientation, and sequence used in the RT-PCR protocol

Article Snippet: The sections were incubated with specific antibodies against rat type II collagen, MMP-13, types X collagen, or type II collagen-C fragment (IBEX Technologies, Mont-Royal, QC, Canada) at 4 °C overnight.

Techniques: Sequencing

Journal: Arthritis Research & Therapy

Article Title: Ipriflavone attenuates the degeneration of cartilage by blocking the Indian hedgehog pathway

doi: 10.1186/s13075-019-1895-x

Figure Lengend Snippet: Chondroprotective effect of ipriflavone (IP) in human chondrocytes. a Real-time PCR results showed reduced mRNA expression of key genes in the Ihh pathway, Smo, Gli-1,Gli-2,Gli-3, and Runx-2, at 48 h after IP treatment, and among the three kinds of Glis, the reduction of Gli-2 was especially significant. The MMP-13 and type X collagen mRNA levels were decreased, and the type II collagen mRNA level was significantly increased in human chondrocytes. b Western blot results indicated that in chondrocytes, the expression of Smo and Runx-2 protein was decreased at 48 h after IP treatment, MMP-13 and type X collagen expression was decreased in the IP treatment group, and type II collagen expression was increased in the IP treatment group. The gray value of the Western blot bands was semiquantified using Image Analysis Software (Image Lab 3.0). Values are the mean ± SEM. n = 3, * P < 0.05, ** P < 0.01, *** P < 0.001 versus the DMSO group

Article Snippet: The sections were incubated with specific antibodies against rat type II collagen, MMP-13, types X collagen, or type II collagen-C fragment (IBEX Technologies, Mont-Royal, QC, Canada) at 4 °C overnight.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Software

Journal: Arthritis Research & Therapy

Article Title: Ipriflavone attenuates the degeneration of cartilage by blocking the Indian hedgehog pathway

doi: 10.1186/s13075-019-1895-x

Figure Lengend Snippet: Ipriflavone (IP) reduced the degeneration of cartilage by blocking the Indian hedgehog pathway in vivo. a The results of Safranin O staining indicated that there was less surface damage with stronger Safranin O staining in articular cartilage specimens from IP-treated animals compared with the ACLT group at 12 weeks after the operation. Cartilage destruction was quantified and compared with the ACLT group, and the summed OARSI scores were significantly decreased in the IP treatment group. Data are expressed as means ± SDs. ( n = 10), * P < 0.05 ** P < 0.01. b Type II collagen expression in articular cartilage was higher in IP-treated and sham-operated rats than in rats that underwent ACLT without treatment. In contrast, degraded collagen II (type II collagen-C fragment), type X collagen, and matrix metalloproteinase 13 (MMP-13) staining levels were elevated in rats that underwent ACLT without treatment with respect to the IP-treated and sham-operated rats, which is consistent with reduced OA damage. c Real-time PCR results indicated that the expression of key genes in the Ihh pathway (Smo, Gli-1, Gli-2, Gli-3) was increased in rats that underwent ACLT without IP treatment and reduced in IP-treated rats at 12 weeks after the ACLT operation, especially Smo and Gli-2. Concomitantly, type II collagen and Agg expression in articular cartilage was elevated in IP-treated and sham-operated rats compared with rats that underwent ACLT without treatment. In contrast, type X collagen and matrix metalloproteinase 13 (MMP-13) were elevated in rats that underwent ACLT without IP treatment, and their expressions were reduced in IP-treated rats at 12 weeks after the ACLT operation, consistent with the reduced OA damage. Values are the mean ± SEM. P < 0.05 versus the ACLT and IP treatment group

Article Snippet: The sections were incubated with specific antibodies against rat type II collagen, MMP-13, types X collagen, or type II collagen-C fragment (IBEX Technologies, Mont-Royal, QC, Canada) at 4 °C overnight.

Techniques: Blocking Assay, In Vivo, Staining, Expressing, Real-time Polymerase Chain Reaction