scholarly journals AB0036 APPA (APOCYNIN AND PAEONOL) REDUCES ROS PRODUCTION AND SENESCENCE IN HUMAN ARTICULAR CHONDROCYTES

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 1051.1-1051
Author(s):  
M. Fernandez-Moreno ◽  
N. Larkins ◽  
A. Reynolds ◽  
T. Hermida Gómez ◽  
F. J. Blanco
Keyword(s):  
Gene Expression ◽  
Research And Development ◽  
Articular Chondrocytes ◽  
Human Articular Chondrocytes ◽  
Ros Production ◽  
Tnf Α ◽  
Inflammatory Stimuli ◽  
And Cartilage ◽  
Pro Inflammatory Cytokines ◽  
Inflammatory Effect

Background:Disease modification is not yet possible for osteoarthritis (OA). Mitochondrial ROS and pro-inflammatory cytokines are involved in the pathogenesis of OA and are potential therapeutic targets. APPA, a combination of apocynin (AP) and paeonol (PA), has the potential capacity to modulate synthesis of pro-inflammatory stimuli.Objectives:To investigate the anti-inflammatory effect of APPA in human articular chondrocytes and cartilage.Methods:Tissue and chondrocytes from human OA cartilage were isolated. The effect of APPA on chondrocyte viability was analyzed using MTT. IL-1β 10 ng/mL and LPS 10 ng/mL were used as pro-inflammatory stimuli. ROS production was evaluated by flow cytometry using DCFH-DA and MitoSoxRed. The percentage of senescent cells was evaluated through the quantification of Fluorescein di-β-D-galactopyranoside (FDG) by flow cytometry. The effect of APPA on gene expression of pro-inflammatory cytokines (IL-8 and TNF-α) and enzymes degrading cartilage (MMP-13 and MMP-3) were analyzed in chondrocyte and cartilage by RT-PCR. Quantification of Toluidine Blue (TB) staining in cartilage was performed to evaluate proteoglycans content using software ImageJ/Fiji. Release of Glycosaminoglycan (GAGs) into the supernatant was quantified using BlyscanTM Glycosaminoglycan assay. Statistical analyses were performed with GraphPad Prism v6.Results:Chondrocytes, incubated in presence of APPA 10 µg/mL for 24 h had viability >85%, reduced cytoplasmic ROS (p=0.028) and mitochondrial anion superoxide production induced by LPS 10 ng/mL (p=0.057). Chondrocytes incubated in presence of APPA 10 µg/mL for 2 hours contained significantly fewer senescent cells (p=0.0079). APPA significantly reduced the gene expression induced by IL-1β 10 ng/mL in chondrocytes of IL-8, TNF-α, MMP-13 and MMP-3. Cartilage incubated with APPA 60 and 100 µg/mL for 48 h showed decreased the MMP-3 gene expression induced by IL-1β (p=0.021 and p<0.0001 respectively). Quantification of TB showed that APPA 60 and 100 µg/mL during 48h increased the proteoglycans in intermedial layer, which had been decreased through the incubation with IL-1β (p=0.0018 and p=0.018 respectively). Quantification of release GAGs into the supernatant decreased significantly when the cartilage explants were incubated for 48h in presence of APPA 100 µg/mL (p=0.028).Conclusion:APPA has a clear anti-inflammatory effect on human articular chondrocytes, and could reduce extracellular matrix degradation of cartilage. This could be mediated by the capacity to modulate ROS production and reduce senescence.Disclosure of Interests:Mercedes Fernandez-Moreno: None declared, Nicholas Larkins Shareholder of: I am a shareholder in AKL Research and Development Ltd, Alan Reynolds Shareholder of: I have share options in AKL Research and Development Ltd, Speakers bureau: I have not been a paid speaker for a pharma company - at least not since 2008 whichI think is outside the scope of this, Consultant of: The last time I was a paid consultant was in 2017 when I acted as a consultant for Avillion and Norgine, Employee of: I am also an employee of AKL Research and Development Ltd, Tamara Hermida Gómez: None declared, Francisco J. Blanco Speakers bureau: LillyPfizerSanofiGalapagos, Consultant of: LillyPfizerSanofiGalapagos, Grant/research support from: LillyMSDMerck SeronoPfizerPierre-FabraRocheSanofiServierUCBAbbvieAmgenBioibericaBristol MayerCelgeneCelltrionCellerixGrunenthalGebro PharmaAKL Research and Development Ltd

scholarly journals Effect of Inflammatory Signaling on Human Articular Chondrocyte Hypertrophy: Potential Involvement of Tissue Repair Macrophages

Cartilage ◽  
2021 ◽  
pp. 194760352110219
Author(s):  
Mauricio N. Ferrao Blanco ◽  
Yvonne M. Bastiaansen-Jenniskens ◽  
Mark G. Chambers ◽  
Andrew A. Pitsillides ◽  
Roberto Narcisi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Inflammatory Cytokines ◽  
Tissue Repair ◽  
Articular Chondrocytes ◽  
Human Articular Chondrocytes ◽  
Human Chondrocytes ◽  
Articular Chondrocyte ◽  
Inflammatory Signaling ◽  
Chondrocyte Hypertrophy ◽  
Pro Inflammatory Cytokines

Objective In osteoarthritis, chondrocytes tend to acquire a hypertrophic phenotype, which contributes to the modification of the extracellular matrix, resulting in permanent cartilage changes. In mouse chondrocytes, pro-inflammatory macrophages and pro-inflammatory cytokines have been shown to stimulate hypertrophy via the activation of the nuclear factor kappa B (NF-κB) pathway. Whether or not this also occurs in human chondrocytes remains unclear. We therefore aimed to investigate whether hypertrophy-like responses in human cartilage are driven mainly by intrinsic inflammatory signaling or shaped by specific macrophage populations. Design Human articular chondrocytes were cultured with pro-inflammatory cytokines or medium conditioned by defined macrophage subsets. Furthermore, the effect of inhibition of NF-κB-dependent gene expression was evaluated using the NF-κB inhibitor SC-514. Hypertrophy was assessed by measuring the transcription level of alkaline phosphatase ( ALPL), type X collagen ( COL10A1), Indian hedgehog ( IHH), and runt-related transcription factor 2 ( RUNX2). Results The expression of hypertrophic genes was not promoted in human chondrocytes by pro-inflammatory cytokines neither pro-inflammatory M(IFNγ + TNFα) macrophages. Inhibition of the NF-κB-dependent gene expression did not affect human articular chondrocyte hypertrophy. However, tissue repair M(IL4) macrophages induced hypertrophy by promoting the expression of COL10A1, RUNX2, and IHH. Conclusion Intrinsic inflammatory signaling activation is not involved in the hypertrophic shift observed in human articular chondrocytes cultured in vitro. However, tissue repair macrophages may contribute to the onset of this detrimental phenotype in human osteoarthritic cartilage, given the effect observed in our experimental models.

scholarly journals Concentration-Dependent Effects of Cobalt and Chromium Ions on Osteoarthritic Chondrocytes

Cartilage ◽  
2019 ◽  
pp. 194760351988938
Author(s):  
Christoph Bauer ◽  
Christoph Stotter ◽  
Vivek Jeyakumar ◽  
Eugenia Niculescu-Morzsa ◽  
Bojana Simlinger ◽  
...  
Keyword(s):  
Inflammatory Cytokines ◽  
Metabolic Activity ◽  
Transcriptional Activity ◽  
Articular Chondrocytes ◽  
Inflammatory Responses ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Articular Chondrocytes ◽  
Tnf Α ◽  
Late Apoptosis ◽  
Pro Inflammatory Cytokines

Objective Cobalt and chromium (CoCr) ions from metal implants are released into the joint due to biotribocorrosion, inducing apoptosis and altering gene expression in various cell types. Here, we asked whether CoCr ions concentration-dependently changed viability, transcriptional activity, and inflammatory response in human articular chondrocytes. Design Human articular chondrocytes were exposed to Co (1.02-16.33 ppm) and Cr (0.42-6.66 ppm) ions and cell viability and early/late apoptosis (annexin V and 7-AAD) were assessed in 2-dimensional cell cultures using the XTT assay and flow cytometry, respectively. Changes in chondrocyte morphology were assessed using transmitted light microscopy. The effects of CoCr ions on transcriptional activity of chondrocytes were evaluated by quantitative polymerase chain reaction (qPCR). The inflammatory responses were determined by measuring the levels of released pro-inflammatory cytokines (interleukin-1β [IL-1β], IL-6, IL-8, and tumor necrosis factor–α [TNF-α]). Results CoCr ions concentration-dependently reduced metabolic activity and induced early and late apoptosis after 24 hours in culture. After 72 hours, the majority of chondrocytes (>90%) were apoptotic at the highest concentrations of CoCr ions (16.33/6/66 ppm). SOX9 expression was concentration-dependently enhanced, whereas expression of COL2A1 linearly decreased after 24 hours. IL-8 release was enhanced proportionally to CoCr ions levels, whereas IL-1β, IL-6, and TNF-α levels were not affected by the treatments. Conclusions CoCr ions showed concentration- and time-dependent effects on articular chondrocytes. Fractions of apoptotic articular chondrocytes were proportional to CoCr ion concentrations. In addition, metabolic activity and expression of chondrocyte-specific genes were decreased by CoCr ions. Furthermore, exposure to CoCr ions caused a release of pro-inflammatory cytokines.

IL-10 overexpression differentially affects cartilage matrix gene expression in response to TNF-α in human articular chondrocytes in vitro

Cytokine ◽  
2008 ◽  
Vol 44 (3) ◽  
pp. 377-385 ◽  
Author(s):  
R.D. Müller ◽  
T. John ◽  
B. Kohl ◽  
A. Oberholzer ◽  
T. Gust ◽  
...  
Keyword(s):  
Gene Expression ◽  
Articular Chondrocytes ◽  
Cartilage Matrix ◽  
Human Articular Chondrocytes ◽  
Matrix Gene ◽  
Tnf Α

scholarly journals Dipterocarpus tuberculatus Roxb. Ethanol Extract Has Anti-Inflammatory and Hepatoprotective Effects In Vitro and In Vivo by Targeting the IRAK1/AP-1 Pathway

Molecules ◽  
2021 ◽  
Vol 26 (9) ◽  
pp. 2529
Author(s):  
Haeyeop Kim ◽  
Woo Seok Yang ◽  
Khin Myo Htwe ◽  
Mi-Nam Lee ◽  
Young-Dong Kim ◽  
...  
Keyword(s):  
Ethanol Extract ◽  
Serum Levels ◽  
Hepatoprotective Activity ◽  
Tnf Α ◽  
Anti Inflammatory ◽  
Related Proteins ◽  
Pro Inflammatory Cytokines ◽  
Inflammatory Effect

Dipterocarpus tuberculatus Roxb. has been used traditionally as a remedy for many diseases, especially inflammation. Therefore, we analyzed and explored the mechanism of the anti-inflammatory effect of a Dipterocarpus tuberculatus Roxb. ethanol extract (Dt-EE). Dt-EE clearly and dose-dependently inhibited the expression of pro-inflammatory cytokines such as IL-6, TNF-α, and IL-1β in lipopolysaccharide (LPS)-treated RAW264.7 cells. Also, Dt-EE suppressed the activation of the MyD88/TRIF-mediated AP-1 pathway and the AP-1 pathway related proteins JNK2, MKK4/7, and TAK1, which occurred as a result of inhibiting the kinase activity of IRAK1 and IRAK4, the most upstream factors of the AP-1 pathway. Finally, Dt-EE displayed hepatoprotective activity in a mouse model of hepatitis induced with LPS/D-galactosamine (D-GalN) through decreasing the serum levels of alanine aminotransferase and suppressing the activation of JNK and IRAK1. Therefore, our results strongly suggest that Dt-EE could be a candidate anti-inflammatory herbal medicine with IRAK1/AP-1 inhibitory and hepatoprotective properties.

scholarly journals Metabolic Syndrome Predisposes to Osteoarthritis: Lessons from Model System

Cartilage ◽  
2020 ◽  
pp. 194760352098016
Author(s):  
Sampath Samuel Joshua Pragasam ◽  
Vijayalakshmi Venkatesan
Keyword(s):  
Subchondral Bone ◽  
Fat Mass ◽  
Articular Chondrocytes ◽  
Bone Cyst ◽  
Primary Cultures ◽  
Abdominal Circumference ◽  
Enzyme Linked Immunosorbent Assay ◽  
Micro Ct ◽  
Tnf Α ◽  
And Cartilage

Objective The present study aims to assess for temporal changes in tibial subchondral bone and cartilage in WNIN/Gr-Ob rats (portraying obesity, insulin resistance, dyslipidemia, impaired glucose tolerance, hypertension) in comparison with Wistar controls (WNIN) using anthropometry, micro-computed tomography (micro-CT), scanning electron microscopy (SEM), histopathology, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence. Design Body weight, abdominal circumference, body mass index (BMI), lean/fat mass, serum tumor necrosis factor (TNF)-α levels were measured (ELISA), followed by ultrastructural analysis of tibial subchondral bone (micro-CT) and cartilage architecture (histopathology and SEM) in WNIN/Gr-Ob and WNIN rats with age (3, 6 and 9 months). Additionally, primary cultures of articular chondrocytes isolated from 6-month-old WNIN/Gr-Ob and WNIN rats were assessed for matrix metalloproteinase (MMP)-13 and Collagen type II (COL2A1) by immunofluorescence. Results WNIN/Gr-Ob rats exhibited frank obesity with increased BMI, lean and fat mass vis-à-vis significantly higher levels of serum TNF-α (6>9>3 months) as compared with the controls. With an increase in BMI, WNIN/Gr-Ob rats presented with tibial cartilage fibrillation, erosion, osteophyte formation (6 months) and subchondral bone cyst (9 months) confirmed by histology and SEM. An increase in subchondral trabecular bone volume (sclerosis with decreased plate porosity) was observed in all ages in WNIN/Gr-Ob rats compared to their Control. Gaining insights, primary cultures of articular chondrocytes complemented with altered cellular expressions of COL2A1 and MMP-13 from WNIN/Gr-Ob rats, indicating osteoarthritis (OA) progression. Conclusion Multiple metabolic perturbations featured in WNIN/Gr-Ob rats were effective to induce spontaneous OA-like degenerative changes affecting knee joints akin to human OA.

Opiate regulation of IL-1β and TNF-α in cultured human articular chondrocytes

2005 ◽  
Vol 333 (4) ◽  
pp. 1295-1299 ◽  
Author(s):  
Nenad Andjelkov ◽  
Jan Elvenes ◽  
James Martin ◽  
Oddmund Johansen
Keyword(s):  
Articular Chondrocytes ◽  
Human Articular Chondrocytes ◽  
Tnf Α

scholarly journals The effect of interleukin-1 on cytokine gene expression in cultured human articular chondrocytes analyzed by messenger rna phenotyping

1993 ◽  
Vol 36 (1) ◽  
pp. 35-43 ◽  
Author(s):  
Janet M. Seid ◽  
Shamim Rahman ◽  
Robert Graveley ◽  
Rowena A. D. Bunning ◽  
Rene Nordmann ◽  
...  
Keyword(s):  
Gene Expression ◽  
Messenger Rna ◽  
Articular Chondrocytes ◽  
Interleukin 1 ◽  
Human Articular Chondrocytes ◽  
Cytokine Gene Expression ◽  
Cytokine Gene

scholarly journals 94 Essential oils improve barrier function and attenuate inflammatory responses in porcine intestinal epithelial cells

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 78-79
Author(s):  
Qianru Hui ◽  
Faith Omonijo ◽  
Shangxi Liu ◽  
Hua Zhang ◽  
Ludovic Lahaye ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Essential Oils ◽  
Barrier Function ◽  
Inflammatory Responses ◽  
Mrna Abundance ◽  
Cell Permeability ◽  
Ros Production ◽  
Tnf Α ◽  
Pre Treatment

Abstract Thymol has been known as a functional phytochemical isolated from thyme essential oils and possesses antioxidant, antimicrobial, and anti-inflammatory properties. In this study, an in vitro lipopolysaccharide (LPS)-induced inflammation model using IPEC-J2 cell line was established to evaluate the inflammatory responses after thymol treatment. Cells were pre-treated with thymol for 1 h followed by LPS stimulation. Interleukin 8 (IL-8) secretion, reactive oxygen species (ROS) production, mRNA abundance of two pro-inflammatory cytokines, nutrient transporters, and tight junction proteins, transepithelial electrical resistance (TEER) and cell permeability were measured. The localization of zonula occludens-1 (ZO-1) and β-actin were also detected by immunofluorescent staining. The results showed that LPS stimulation increased IL-8 secretion, ROS production, and tumor necrosis factor alpha (TNF-α) mRNA abundance (P &lt; 0.05), but the mRNA abundance of sodium-dependent glucose transporter 1 (SGLT1), excitatory amino acid transporter 1 (EAAC1) and H+/peptide cotransporter 1 (PepT1) were decreased (P &lt; 0.05). However, thymol blocked ROS production (P &lt; 0.05) and tended to decrease the production of LPS-induced IL-8 secretion (P = 0.0766). The mRNA abundance of IL-8 and TNF-α was reduced by thymol pre-treatment (P &lt; 0.05), but thymol was unable to improve the gene expression of nutrient transporters (P &gt; 0.05). TEER was reduced and cell permeability was increased after LPS stimulation (P &lt; 0.05), but these effects were attenuated by thymol pre-treatment (P &lt; 0.05). Moreover, thymol boosted ZO-1 and β-actin staining in the cells, but the mRNA abundance of ZO-1 and occludin-3 was not affected by either LPS or thymol treatments. These results indicated that thymol can enhance gut barrier structure and functions by reducing ROS production and pro-inflammatory cytokine gene expression in porcine epithelial cells during inflammation. The regulation of barrier function by thymol may be at post-transcriptional or post-translational levels.

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