human il1β elisa kit  (Elabscience Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: Intra-Articular Delivery of Nanoemulsified Curcumin Ameliorates Joint Degeneration in a Chemically Induced Model of Osteoarthritis

doi: 10.3390/ijms262211212

Figure Lengend Snippet: Effect of curcumin nanoemulsion on cell viability and chemotactic migration of macrophages towards IL1β-induced chondrocytes. ( A , B ) Cell viability by CHON-001 and RAW 264.7 cells, respectively, exposed to different concentrations of curcumin and nanoemulsion (0.1–100 μM) for 24 h. Cell viability was evaluated using the MTT assay and calculated by comparing optical density values between the control and treatment groups after normalizing them with blank values, and expressed as a percentage of the control. Each value is represented as mean ± SEM ( n = 8). ( C ) Transwell co-culture model. ( D ) Representative images of calcein AM (green)-stained macrophages on the underside of 8 μm insert after 24 h of co-culture. Chondrocytes were cultured in the lower chamber and exposed to stimulus, while macrophages were incubated in the insert with plain assay media. Images (magnification, 10×; scale bar, 100 μm) represent macrophage migration towards ( i ) unstimulated chondrocytes, ( ii ) IL1β-induced chondrocytes, ( iii ) IL1β-induced chondrocytes treated with curcumin (10 μM), and ( iv ) IL1β-induced chondrocytes treated with nanoemulsion (10 μM). ( E ) Quantification of migrated cells using mean fluorescence intensity per unit area. Data were analyzed using one-way ANOVA with Tukey’s multiple comparisons, and p < 0.05 was considered statistically significant compared to the control. Unlike letters denote significant differences.

Article Snippet: IL1β (#201-LB-005) was procured from R&D Systems (Minneapolis, MN, USA).

Techniques: Migration, MTT Assay, Control, Co-Culture Assay, Staining, Cell Culture, Incubation, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Intra-Articular Delivery of Nanoemulsified Curcumin Ameliorates Joint Degeneration in a Chemically Induced Model of Osteoarthritis

doi: 10.3390/ijms262211212

Figure Lengend Snippet: Expression of inflammatory mediator NF-κB in RAW 264.7 macrophages following IL1β stimulation in the presence of curcumin/nanocurcumin (5–10 μM). ( A ) Immunoblot of NF-κB and ( B ) its relative protein expression normalized with β-actin (mean ± SEM). ( C ) Nuclear and cytosolic expression of NF-κB in RAW 264.7 macrophages using immunocytochemistry after treating with IL1β and curcumin nanoemulsion (10 μM). Cells were stained with Alexa Fluor 594-tagged NF-κB (red) and DAPI (blue) to stain nuclei with original magnification of 40×; scale bar 100 μm. Images were captured using a confocal laser scanning microscope with an oil immersion objective (TCS SP5, Leica Microsystems, Germany). ( D ) Corrected total cell fluorescence (CTCF) was calculated, analyzed statistically and expressed in arbitrary units [CTCF = integrated density—(area of selected cell × mean fluorescence of background readings)]. Data were analyzed using one-way ANOVA with post hoc Tukey’s multiple comparison test, with p < 0.05 considered significant, and results depicted using different letters.

Article Snippet: IL1β (#201-LB-005) was procured from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Western Blot, Immunocytochemistry, Staining, Laser-Scanning Microscopy, Fluorescence, Comparison