To investigate the effects of JEZTC and GA on IL-1β-induced chondrocytes, the cells were grouped as follow: (1) normal control group, chondrocytes treated with culture medium only; (2) IL-1β (OA model) group, chondrocytes treated with culture medium contained 10 ng/ml IL-1β (Gibco, USA); (3) JEZTC-1 group, chondrocytes pre-incubated with 2.344 µg/ml JEZTC for 1 h followed by stimulation with IL-1β for 24 h; (4) JEZTC-2 group, chondrocytes pre-incubated with 4.688 µg/ml JEZTC for 1 h followed by stimulation with IL-1β for 24 h; (5) JEZTC-3 group, chondrocytes pre-incubated with 9.376 µg/ml JEZTC for 1 h followed by stimulation with IL-1β for 24 h; (6) GA group, chondrocytes pre-incubated with 4.688 µg/ml GA for 1 h followed by stimulation with IL-1β for 24 h. The concentrations of JEZTC were determined according to previous study [26] . The dose of GA was derived from the preliminary cytotoxicity assay, which was optimal for cell growth.
Il 1β
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, China, Austria, Italy, France, Japan, Canada, Sweden, Switzerland, Sao Tome and Principe, Israel
IL-1β is a lab equipment product that measures the levels of interleukin-1 beta, a pro-inflammatory cytokine, in biological samples. It is used for research purposes to quantify IL-1β expression and activity.
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Lab products found in correlation
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Il 10, by Thermo Fisher Scientific (95 mentions)
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Il 23, by Thermo Fisher Scientific (65 mentions)
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Interleukin 2 (il 2), by Thermo Fisher Scientific (35 mentions)
Il 12, by Thermo Fisher Scientific (35 mentions)
Tumor necrosis factor (tnf), by Thermo Fisher Scientific (34 mentions)
Tgf β1, by Thermo Fisher Scientific (34 mentions)
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Tnf α, by R&D Systems (27 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (27 mentions)
Gm csf, by Thermo Fisher Scientific (26 mentions)
Il 17, by Thermo Fisher Scientific (25 mentions)
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Il 18, by Thermo Fisher Scientific (22 mentions)
1 245 protocols using il 1β
1
Effect of JEZTC and GA on Chondrocytes
2
Investigating IL-1 Modulation by JEZ-C, GA, and SMZ
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To investigate the effects of JEZ-C, GA and SMZ on IL-1 on the induction of chondrocytes, five groups were divided: (1) control group, chondrocytes without any treatment; (2) OA model group, chondrocytes treated with 10 ng/ml IL-1β (10 ng/ml, Gibco, USA); (3) JEZ-C treatment groups, chondrocytes pre-incubated with various concentrations of JEZ-C (6.25×10−7, 6.25×10−6, 6.25×10−5 μg/ml) for 1 h followed by stimulation with IL-1β for 24 h; (4) SMZ treatment groups, chondrocytes pre-incubated with various concentrations of SMZ (6.25×10−6, 6.25×10−5, 6.25×10−4 μg/ml) for 1 h followed by stimulation with IL-1β for 24 h; (5) GA treatment groups, chondrocytes pre-incubated with various concentrations of GA (0.078, 0.125, 0.156 μg/ml) for 1 h followed by stimulation with IL-1β for 24 h. The concentrations of JEZ-C, GA and SMZ were derived from the cytotoxicity assay.
3
Chondrocyte Response to IL-1β Modulation
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Chondrocytes were divided into four groups: (1) Control group: chondrocytes treated with culture medium only; (2) IL-1β group: chondrocytes stimulated with 10 ng/mL IL-1β (Gibco, USA) for 24 h; (3) PCA group: chondrocytes pre-incubated with 6 μg/mL PCA for 1 h followed by stimulating with 10 ng/mL IL-1β for 24 h; (4) MOF@HA@PCA group: chondrocytes pre-incubated with MOF@HA@PCA (containing 6 μg/mL of PCA) for 1 h followed by stimulating with 10 ng/mL IL-1β for 24 h.
4
Investigating HAMDC, SD-Na, and GA Effects on IL-1 Stimulated Chondrocytes
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To investigate the effect of HAMDC, SD-Na and GA on Interleukin-1 (IL-1) stimulated chondrocytes, five groups were divided as following: (1) control group, chondrocytes without any treatment; (2) OA model group, chondrocytes treated with IL-1β (10 ng/mL, Gibco, USA); (3) HAMDC treatment groups, chondrocytes pre-incubated with three concentrations of HAMDC (3.125, 6.250 and 12.500 μg/mL) followed by stimulation with IL-1β for 24 h; (4) SD-Na treatment groups, chondrocytes pre-incubated with three concentrations of SD-Na (2.344, 4.688 and 9.375 μg/mL) followed by stimulation with IL-1β for 24 h; (5) GA treatment groups, chondrocytes pre-incubated with three concentrations of GA (3.125, 6.250 and 12.500 μg/mL) followed by stimulation with IL-1β for 24 h. The concentrations of HAMDC, SD-Na and GA were determined by cytotoxicity assay results.
5
Evaluating pH-Sensitive Anti-Inflammatory Nanocarriers
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To better understand the pH-sensitive potential of nanocarriers, a partial acidic culture condition (pH = 6.8) was utilized to simulate the OA environment to study the anti-inflammatory effect of AG@MSNs-PAA on IL-1β-induced chondrocytes in vitro. Chondrocytes were divided into four groups: (1) Control group: with culture medium only; (2) IL-1β group: with IL-1β (PeproTech, USA, 10 ng/mL) for 24 h; (3) AG group: with 10 ng/mL IL-1β for 24 h after pre-incubating with 8 µM AG for 1 h; (4) AG@MSNs-PAA group: with 10 ng/mL IL-1β for 24 h after pre-incubating with AG@MSNs-PAA (containing 8 µM of AG) for 1 h.
6
Cytokine Modulation of ARPE-19 Cells
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Cytokines were detected in the supernatants of ARPE-19 cells, seeded at 50,000 cells/well in 24-well plates. ARPE-19 cells were pretreated with 5 ng/ml of different kinds of cytokines (TNF-α (Cat# 300-01A, PeproTech, NJ, USA), IL-6 (Cat# 200-06, PeproTech, NJ, USA), IL-1β (Cat# 200-01B, PeproTech, NJ, USA), TNF-α+IL-6, TNF-α+IL-1β, IL-6+IL-1β, and TNF-α+IL-6+IL-1β) for 10 mins, then stimulated with diacerein (10 and 100 μM/ml), and incubated for 2 hours. Cell-free supernatants were collected at 2 hours after culture and stored at -80°C until further use. Levels of IL-6, IL-8, and MCP-1 were determined using a human IL-6 (Cat# 88-7066-22, Thermo Fisher Scientific, MA, USA), IL-8 (Cat# 88-8086-22, Thermo Fisher Scientific, MA, USA), and MCP-1 (Cat# 88-7399-22, Thermo Fisher Scientific, MA, USA) ELISA Ready-Set-Go kit following the manufacturer's instructions.
7
ART Modulates IL-1β-Induced Chondrocytes
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To investigate the effects of ART on IL-1β-induced chondrocytes of rat, three groups were formed: (1) normal group, chondrocytes without any treatment; (2) IL-1β group, chondrocytes treated with IL-1β (10 ng/ml, Gibco, USA); and (3) IL-1β+ART group, chondrocytes pre-incubated with ART at the optimal dose for 1 h, followed by stimulation with IL-1β for 48 h. The OA patient-derived chondrocytes were treated with ART at the optimal dose for 2, 4, and 6 d, respectively. To study the association of ART with the Wnt/βcatenin signaling pathway, 10 mM lithium chloride (LiCl) (Shanghai Macklin Biochemical Co. Ltd, China), an activator of the Wnt signaling pathway, was introduced.
8
Chondrocyte Viability and Inflammation Assay
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To assess cell viability, chondrocytes were seeded in 96-well plates (5x104/ml, 100 µl/well), treated with 0, 0.3, 1, or 3 mM AG (Sigma-Aldrich; Merck KGaA) for 24, 48 and 72 h at 37˚C and evaluated using a Cell Counting Kit-8 (CCK-8) assay (Dojindo Molecular Technologies, Inc.), as described below. For the evaluation of iNOS and COX-2 expression, chondrocyte cells were exposed to the following conditions for 24 h at 37˚C: i) 10 ng/ml IL-1β (PeproTech, Inc.) alone or ii) co-treatment with different concentrations of AG (0.3, 1, 3 mM) and 10 ng/ml IL-1β for 24 h, after treatment with 10 ng/ml IL-1β for 24 h alone. The activity of the NF-κB signaling pathway was assessed in chondrocytes treated with either 10 ng/ml IL-1β alone for 0.5 h or co-treated with AG (0.3, 1 or 3 mM) and 10 ng/ml IL-1β at 37˚C for 2 h. Rat chondrocytes cultured without AG and IL-1β were used as controls.
9
Cytokine Stimulation Titration Analysis
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Titration analyses of cytokine stimulation were performed as described in the accompanying study (26 (link)). To follow up on those studies, recombinant human (rh) TNFα (#300-01A, PeproTech, Rocky Hill, NJ) and rhIL-1β (#200-01B, PeproTech) were used in the following concentrations: MDA-MB-231 cells, MCF-7 cells, MSCs and CAFs: TNFα 10 ng/ml, IL-1β 350 pg/ml; MDA-MB-468 cells: TNFα 50 ng/ml, IL-1β 500 pg/ml; BT-549 cells: TNFα 25 ng/ml, IL-1β 350 pg/ml.
10
Endplate Chondrocyte Viability Assay
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A Cell Counting Kit-8 (CCK8) assay (Dojindo, Kumamoto, Japan) was used to detect the viability of endplate chondrocytes. Endplate chondrocytes (5 × 103 cells per well) were seeded into 96-well plates. Subsequently, cells were treated with IL-1β (PeproTech, Rocky Hill, NJ, USA) IL-1β + HSYA (10 μM) or IL-1β + HSYA (10 μM) for 48 h. After that, cells were incubated with10 μL CCK-8 reagents at a 37°C incubator for another 2 h. Then, the optical density at 450 nm was measured with a microplate reader.
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