Journal: Antimicrobial Agents and Chemotherapy

Article Title: Prophylactic Antiheparanase Activity by PG545 Is Antiviral In Vitro and Protects against Ross River Virus Disease in Mice

doi: 10.1128/AAC.01959-17

Figure Lengend Snippet: Prophylactic PG545 treatment protects chondrocytes from RRV-induced cartilage damage and breakdown. (A) Knee joints were fixed in 4% PFA and then decalcified in 14% EDTA and stained with Safranin O/fast green. (Ai to iii) The width of the articular cartilage was enumerated from 6 distinct regions of the tibiofemoral cartilage interphase. (Bi and ii) Serum was used to quantify cartilage breakdown proteins. COMP and CTX-II were used in commercial sandwich ELISA kits. PBS, mock-infected PBS control; PG545, mock infected and PG545 treated; RRV, RRV infected and PBS treated; RRV + PG545, RRV infected and PG545 treated. Values are expressed as mean width (μm) or mean protein concentration (ng or pg/ml) ± SEM for 4 to 16 mice per group. SB, subchondral bone; eGP, epiphyseal growth plate; ALM, anterior lateral meniscus; PLM, posterior lateral meniscus.

Article Snippet: The serum COMP (MBS451829; 1:5 dilution; MyBioSource) and CTX-II (MBS760453; 1:10 dilution; MyBioSource) concentrations and the muscle HPSE (MBS453434; 1:100 dilution; MyBioSource) level of each group were measured using commercial sandwich ELISA.

Techniques: Staining, Sandwich ELISA, Infection, Protein Concentration

Journal: Bioengineered

Article Title: Silence of TANK-binding kinase 1 (TBK1) regulates extracellular matrix degradation of chondrocyte in osteoarthritis by janus kinase (JAK)-signal transducer of activators of transcription (STAT) signaling

doi: 10.1080/21655979.2021.2018976

Figure Lengend Snippet: Silencing of TBK1 inhibited ECM degradation in OA mice. ACLT mice were infected with lentivirus shTBK1 or shNC. (a) TBK1 mRNA level was detected by qRT-PCR. (b) Safranin O-fast green staining and (c) OARSI score were used for elevating cartilage destruction. (d) Serum CTX-II and (e) COMP were analyzed by the corresponding ELISA kits. * p < 0.05.

Article Snippet: The serum levels of cartilage metabolic markers, including C-terminal crosslinking telopeptide of type II collagen (CTX-II) and cartilage oligomeric matrix protein (COMP), were measured using the corresponding enzyme-linked immuno sorbent assay (ELISA) kits (Wuhan Huamei Biotech Co. Ltd., Wuhan, China).

Techniques: Infection, Quantitative RT-PCR, Staining, Enzyme-linked Immunosorbent Assay

Journal: Aging Cell

Article Title: Transthyretin deposition promotes progression of osteoarthritis

doi: 10.1111/acel.12665

Figure Lengend Snippet: Mediators and markers of osteoarthritis ( OA ) in wild‐type ( WT ), mTTR ‐ KO , and hTTR ‐ TG mice. (A) RNA was isolated from knee articular cartilage from 6‐month‐old mice for RT ‐ qPCR . OA ‐related genes were significantly elevated in cartilage from TTR ‐ TG mice ( WT n = 6, mTTR ‐ KO n = 6, hTTR ‐ TG n = 5). The expression values were relative to Gapdh . ** P < 0.01 and * P < 0.05. (B) Immunohistochemistry for CTX ‐ II , IL ‐6, ADAMTS ‐4, and MMP 13 was performed on joint sections from 6‐month‐old mice ( WT n = 4; mTTR ‐ KO n = 4; hTTR ‐ TG n = 4). Cells positive for IL ‐6, ADAMTS ‐4, and MMP 13 were significantly elevated in 6‐month‐old hTTR ‐ TG mice (scale bars = 100 μm). The ratio of positive cells was counted using three sections from four mice for each genotype. * P < 0.05.

Article Snippet: Anti‐human TTR (Dako A0002; 1:400), anti‐mouse and human TTR (Abcam ab9015; 1:100), anti‐CTX‐II (Cloud‐Clone Corp. PAA686Hu01; 1:2000 dilution), anti‐ADAMTS‐4 (Abcam ab28285; 1:100 dilution), anti‐MMP‐13 (Acam ab39012; 1:100 dilution), anti‐IL‐6 (Abcam ab6672; 1:600 dilution), and negative control rabbit IgG (Vector I‐1000; 1 μg/mL) were applied with 0.1% Tween 20 and incubated overnight at 4°C, and followed by secondary antibody using Vectastain ABC‐AP alkaline phosphatase (Vector Laboratories, Burlingame, CA, USA).

Techniques: Isolation, Quantitative RT-PCR, Expressing, Immunohistochemistry

Journal: Bone Reports

Article Title: Concerted actions by MMPs, ADAMTS and serine proteases during remodeling of the cartilage callus into bone during osseointegration of hip implants

doi: 10.1016/j.bonr.2020.100715

Figure Lengend Snippet: MMPs, ADAMTS, serine proteases and endogenous inhibitors in the three groups of healthy. Mean ± SEM.

Article Snippet: PIICP (Nordic Biosite cat.no.EKH4286) and CTX-II (Nordic Biosite cat.no.EKH2012) were supplied as kits containing precoated plates, antibodies and hRP and were analyzed by use of colorimetric ELISA (Epoch®, Biotek, Vermont, USA).

Techniques:

Journal: Molecular Medicine Reports

Article Title: Mori Folium water extract alleviates articular cartilage damages and inflammatory responses in monosodium iodoacetate-induced osteoarthritis rats

doi: 10.3892/mmr.2017.7075

Figure Lengend Snippet: Effects of MF on the production and expression of COMP and CTX-II in MIA-induced OA rats. (A) COMP and (B) CTX-II production was measured in the serum of MIA-induced OA rats using commercial ELISA kits. (C) Immunohistochemical staining was used to identify the expression of COMP and CTX-II in the articular cartilage. Data are expressed as the mean ± standard deviation (n=8). *P<0.05, **P<0.01. MIA, monosodium iodoacetate; MF, Mori folium; CLX, celecoxib; COMP, cartilage oligomeric matrix protein; CTX-II, C-telopeptide of type II collagen.

Article Snippet: The primary antibodies against the following proteins were used: MMP-13 (cat no. ab39012; dilution, 1:50; Abcam, Cambridge, UK), cartilage oligomeric matrix protein (COMP; cat no. ab74524; dilution, 1:50; Abcam), C-telopeptide of type II collagen (CTX-II; cat no. PAA686Hu01; dilution, 1:50; Cloud-Clone Corp., Houston, TX, USA), inducible nitric oxide (NO) synthase (iNOS; cat no. sc-651; dilution 1:100; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), and cyclooxygenase-2 (COX-2; cat no. sc-514489; dilution 1:100; Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Standard Deviation