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Chondrocyte flow cytometry

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https://www.readbyqxmd.com/read/28626114/thyroid-stimulating-hormone-stimulation-downregulates-autophagy-and-promotes-apoptosis-in-chondrocytes
#1
Wei Xin, Yue Yu, Yuan Ma, Yuan Gao, Ying Xu, Liyong Chen, Qiang Wan
Subclinical hypothyroidism (SCH) patients have normal thyroid hormone levels but increased thyroid stimulating hormone (TSH) level in serum. It has been reported that high TSH is related to abnormal skeletal development in mice with hypothyroidism. However, the cellular mechanism is not fully understood. In the present study, we aim to investigate the direct effects of TSH stimulation on chondrocytes, and the putative role of autophagy in this process. By using EdU incorporation assay and flow cytometry for mitochondrial membrane potential assay, we demonstrated deceased proliferation and promoted apoptosis in TSH stimulated primary mouse chondrocytes...
June 13, 2017: Endocrine Journal
https://www.readbyqxmd.com/read/28618867/human-articular-chondrocytes-induce-interleukin-2-nonresponsiveness-to-allogeneic-lymphocytes
#2
Satomi Abe, Hitoshi Nochi, Hiroshi Ito
Introduction We previously showed that articular chondrocytes (ACs) have immune privilege and immunomodulatory functions like those of mesenchymal stem cells. To elucidate these mechanisms, we focused on interleukin-2 (IL-2), which plays critical roles in lymphocyte mitogenic activity. The purpose of this study was to explore whether ACs affect the role of IL-2 underlying immunomodulatory functions. Material and Methods Irradiated human ACs from osteoarthritis donors were used. Third-party ACs were added to the mixed lymphocyte reaction (MLR) with or without recombinant human IL-2 (rhIL-2), and the levels of IL-2 and the soluble form of the IL-2 receptor α (sIL-2Rα) protein in supernatant were measured by enzyme-linked immunosorbent assay...
July 2017: Cartilage
https://www.readbyqxmd.com/read/28617559/blockade-of-il-17-alleviated-inflammation-in-rat-arthritis-and-mmp-13-expression
#3
X-L Shui, W Lin, C-W Mao, Y-Z Feng, J-Z Kong, S-M Chen
OBJECTIVE: Rheumatoid arthritis (RA) is one systemic auto-immune disorder featured as chronic synovitis and can destruct joint cartilage. Fibroblast-like synoviocyte (FLS) secretes various factors affecting chondrocyte matrix and degradation. This study thus investigated the effect of interleukin-17A (IL-17A) on FLS and osteoclast. MATERIALS AND METHODS: Type II collagen-induced arthritis (CIA) rats were assigned to CIA model, CIA + IgG1 isotype, and CIA + Anti-Rat IL-17A groups...
May 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/28616752/effects-of-c-myc-gene-silencing-on-interleukin-1%C3%AE-induced-rat-chondrocyte-cell-proliferation-apoptosis-and-cytokine-expression
#4
Jian Zou, Xiao-Lin Li, Zhong-Min Shi, Jian-Feng Xue
This study explores the effects of C-myc gene silencing on cell proliferation, apoptosis and cytokine expression in interleukin (IL)-1β-induced rat chondrocytes. Primary chondrocytes were obtained from 40 Sprague-Dawley rats. For in vitro C-myc3-shRNA transfection, chondrocytes were assigned to a blank 1, model 1, IL-1β + C-myc3-shRNA, C-myc3-shRNA, (IL-1β + C-myc3-shRNA) + C-myc overexpression, C-myc3-shRNA + C-myc overexpression or IL-1β + C-myc-Con group. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to detect C-myc, PCNA and cyclin D1 mRNA and protein expression...
June 14, 2017: Journal of Bone and Mineral Metabolism
https://www.readbyqxmd.com/read/28548679/cyclin-d1-gene-silencing-promotes-il-1%C3%AE-induced-apoptosis-in-rat-chondrocytes
#5
Peng-Fei Zan, Zhong Wu, Yong Yang, Zhi Zhao, Min Wu, Guo-Dong Li
This study investigated the effects of cyclin D1 gene silencing on cell proliferation and apoptosis of interleukin-1β (IL-1β)-induced osteoarthritis (OA) chondrocytes. Chondrocytes from healthy sprague-dawley rats were divided into blank, OA model (chondrocytes underwent IL-1β inducement), OA trial (chondrocytes underwent IL-1β inducement with cyclin D1-shRNA treatment), and negative control (NC; chondrocytes underwent IL-1β inducement and control-shRNA treatment) groups. Cell proliferation was assessed by CCK-8 assay, and cell cycle and apoptosis by flow cytometry...
May 26, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28536528/rescued-chondrogenesis-of-mesenchymal-stem-cells-under-interleukin-1-challenge-by-foamyviral-interleukin-1-receptor-antagonist-gene-transfer
#6
Nicole Armbruster, Jennifer Krieg, Manuel Weißenberger, Carsten Scheller, Andre F Steinert
Background: Mesenchymal stem cells (MSCs) and their chondrogenic differentiation have been extensively investigated in vitro as MSCs provide an attractive source besides chondrocytes for cartilage repair therapies. Here we established prototype foamyviral vectors (FVV) that are derived from apathogenic parent viruses and are characterized by a broad host range and a favorable integration pattern into the cellular genome. As the inflammatory cytokine interleukin 1 beta (IL1β) is frequently present in diseased joints, the protective effects of FVV expressing the human interleukin 1 receptor antagonist protein (IL1RA) were studied in an established in vitro model (aggregate culture system) of chondrogenesis in the presence of IL1β...
2017: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/28529022/effect-of-microgravity-on-the-mesenchymal-stem-cell-characteristics-of-limbal-fibroblasts
#7
Shu-I Pao, Ke-Hung Chien, Hsin-Ting Lin, Ming-Cheng Tai, Jiann-Torng Chen, Chang-Min Liang
BACKGROUND: Mesenchymal stem cells (MSCs) are important for regenerative medicine. Limbal fibroblasts (LFs), present in the corneal limbus, have been shown to possess MSC characteristics, and can differentiate into other cell types. The current study sought to investigate the effect of microgravity on the proliferation and differentiation of LFs, and identify culture conditions to obtain a high proportion of LFs possessing MSC characteristics. METHODS: A rotary cell culture system was used to generate microgravity...
May 18, 2017: Journal of the Chinese Medical Association: JCMA
https://www.readbyqxmd.com/read/28521800/immunogenicity-and-immunomodulatory-effects-of-the-human-chondrocytes-hchonj
#8
Chae-Lyul Lim, Yeon-Ju Lee, Jong-Ho Cho, Heonsik Choi, Bumsup Lee, Myung Chul Lee, Sujeong Kim
BACKGROUND: Invossa™ (TissueGene-C) is a cell and gene therapy for osteoarthritis. It is composed of primary human chondrocytes (hChonJ cells) and irradiated human chondrocytes modified to express TGF-β1 (hChonJb#7 cells). The hChonJ cells were isolated from a polydactyly donor, and TGF-β1 cDNA was delivered to the cells, generating hChonJb#7 cells. Since the cells are allogeneic, the concern of immune response against cells has been raised. In this study, we investigated the immunogenicity of allogenic human chondrocyte, hChonJ cells...
May 18, 2017: BMC Musculoskeletal Disorders
https://www.readbyqxmd.com/read/28504144/low-molecular-weight-xanthan-gum-suppresses-oxidative-stress-induced-apoptosis-in-rabbit-chondrocytes
#9
Qixin Chen, Xintian Shao, Peixue Ling, Fei Liu, Huarong Shao, Aibin Ma, Jixu Wu, Wei Zhang, Fuyan Liu, Guanying Han, Fengshan Wang
We have previously reported the application of low molecular weight XG(LM-XG), with molecular weights ranging from 1×10(6)Da to 1.5×10(6)Da for treating osteoarthritis. In this study, we investigated the anti-apoptotic activity of LM-XG under oxidative stress conditions, activated by hydrogen peroxide (H2O2)-treated chondrocytes in vitro. Chondrocytes were pretreated with various doses of LM-XG (0, 10, 100, 500, or 1000μg/mL) or 1000μg/mL sodium hyaluronate for 12h, and then exposed to 0.5mmol/L H2O2 for another 12h...
August 1, 2017: Carbohydrate Polymers
https://www.readbyqxmd.com/read/28501006/liquid-chromatography-mass-spectrometry-based-quantitative-proteomics-analysis-reveals-chondroprotective-effects-of-astragaloside-iv-in-interleukin-1%C3%AE-induced-sw1353-chondrocyte-like-cells
#10
Huali Luo, Ling Yao, Yudi Zhang, Rongheng Li
OBJECTIVE: Chondrocyte apoptosis played a key role on the progression of Osteoarthritis (OA). Safe and effective drugs are urgently needed for the treatment of OA. Previous study reported that Astragaloside IV (ASG-IV) had exerted a protective effect against articular cartilage degeneration by promoting rapid proliferation of chondrocyte. Therefore, the aim of our study is to explore the effects and mechanisms of ASG-IV in chondrocyte apoptosis. METHODS: Isobaric Tags For Relative And Absolute Quantitation (iTRAQ)-based quantitative proteomics was used to quantitatively detect and map proteins in SW1353 chondrocyte-like cells pre-treated with ASG-IV or interleukin-1β (IL-1β) or ASG-IV+IL-1β...
July 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28498443/silencing-of-both-atf4-and-perk-inhibits-cell-cycle-progression-and-promotes-the-apoptosis-of-differentiating-chondrocytes
#11
Zhimeng Wu, Meiling Li, Wei Zheng, Qin Hu, Zhi Cheng, Fengjin Guo
In the current study, we demonstrate that the silencing of protein kinase R (PKR)-like endoplasmic reticulum (ER) kinase (PERK) and activating transcription factor 6 (ATF4) (using small interfering RNA expression constructs) inhibits the chondrocyte cell cycle and proliferation in vitro and ex vivo. The silencing of PERK alone using siRNA against PERK (siPERK) led to arrest in the G1 phase, it decreased the number of cells in the S phase, and delayed progressoin to the G2‑M phase. Co-transfection with siRNA against ATF (siATF4) led to a more profound inhibitory effect on cell cycle progression...
May 10, 2017: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/28436754/5-7-3-4-tetramethoxyflavone-protects-chondrocytes-from-er-stress-induced-apoptosis-through-regulation-of-the-ire1%C3%AE-pathway
#12
Longhuo Wu, Haiqing Liu, Linfu Li, Daohua Xu, Yun Gao, Yingjie Guan, Qian Chen
AIM OF THE STUDY: To investigate the roles of endoplasmic reticulum (ER) transmembrane sensor inositol-requiring enzyme-1 (IRE1)α signaling in ER stress-induced chondrocyte apoptosis, and to determine the molecular mechanisms underlying chondroprotective activity of 5,7,3',4'-tetramethoxyflavone (TMF) from Murraya exotica. MATERIALS AND METHODS: IRE1α was knocked down by siRNA transfection in chondrocytes, which were harvested from rats' knee cartilages. Chondrocytes with IRE1α deficiency were administrated with tunicamycin (TM) and TMF...
April 24, 2017: Connective Tissue Research
https://www.readbyqxmd.com/read/28418842/effects-of-tlr-2-nf-%C3%AE%C2%BAb-signaling-pathway-on-the-occurrence-of-degenerative-knee-osteoarthritis-an-in-vivo-and-in-vitro-study
#13
Yi-Xun Liu, Guo-Dong Wang, Xiao Wang, Yong-Le Zhang, Tian-Lun Zhang
The study aims to explore the effects of TLR-2/NF-κB signaling pathway on the occurrence of degenerative knee osteoarthritis (OA). Degenerative knee OA and normal cartilage samples were collected from patients with degenerative knee OA receiving total knee arthroplasty and amputation. Expressions of TLR-2, NF-κB and MMP-13 were determined by qRT-PCR and immunochemistry. The chondrocytes were divided into control, IL-1β, IL-1β + anti-TLR-2 and IL-1β + PDTC groups. MTT assay and flow cytometry were performed to determine proliferation and apoptosis of the chondrocytes...
June 13, 2017: Oncotarget
https://www.readbyqxmd.com/read/28391163/peroxiredoxin-4-inhibits-il-1%C3%AE-induced-chondrocyte-apoptosis-via-pi3k-akt-signaling
#14
Zhitao Rao, Shuqing Wang, Jiaqi Wang
BACKGROUND: Chondrocytes apoptosis induced by reactive oxygen species (ROS) plays a critical role in the pathogenesis of osteoarthritis (OA). Peroxiredoxin 4 (PRDX4), a member of the PRDX family, is essential for removing metabolic free radicals and reducing intracellular ROS. In this study, we sought to investigate the roles of PRDX4 on interleukin 1β (IL-1β)-induced chondrocyte apoptosis. METHODS: Primary chondrocytes were isolated from the articular cartilage of Sprague-Dawley rats, infected with PRDX4 overexpressing lentivirus and treated with IL-1β (10ng/mL)...
June 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28372298/effect-of-nicotine-on-the-proliferation-and-chondrogenic-differentiation-of-the-human-wharton-s-jelly-mesenchymal-stem-cells
#15
Xu Yang, Yongjian Qi, Léonore Avercenc-Leger, Jean-Baptiste Vincourt, Sébastien Hupont, Céline Huselstein, Hui Wang, Liaobin Chen, Jacques Magdalou
BACKGROUND: Osteoarthritis (OA) is a chronic joint disease characterized by a progressive and irreversible degeneration of articular cartilage. Among the environmental risk factors of OA, tobacco consumption features prominently, although, there is a great controversy regarding the role of tobacco smoking in OA development. Among the numerous chemicals present in cigarette smoke, nicotine is one of the most physiologically active molecules. OBJECTIVE: The aim of the study was (i) to measure the impact of nicotine on the proliferation and chondrogenic differentiation of mesenchymal stem cells from the human Wharton's jelly (hWJ-MSCs) into chondrocytes, (ii) to investigate whether the α7 nicotinic acetylcholine receptors (nAChRs) was expressed in hWJ-MSCs and could play a role in the process...
2017: Bio-medical Materials and Engineering
https://www.readbyqxmd.com/read/28343378/adipose-derived-stem-cells-suppress-inflammation-induced-by-il-1%C3%AE-through-down-regulation-of-p2x7r-mediated-by-mir-373-in-chondrocytes-of-osteoarthritis
#16
Rilong Jin, Miaoda Shen, Liedao Yu, Xuanwei Wang, Xiangjin Lin
Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-1β (IL-1β) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and IL-1β is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and IL-1β mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery...
March 2017: Molecules and Cells
https://www.readbyqxmd.com/read/28321270/autophagy-is-a-protective-response-to-the-oxidative-damage-to-endplate-chondrocytes-in-intervertebral-disc-implications-for-the-treatment-of-degenerative-lumbar-disc
#17
Ke Chen, Xiaohua Lv, Wei Li, Fei Yu, Jianjing Lin, Junxuan Ma, Deming Xiao
Low back pain (LBP) is the leading cause of disability in the elderly. Intervertebral disc degeneration (IDD) was considered as the main cause for LBP. Degeneration of cartilaginous endplate was a crucial harmful factor during the initiation and development of IDD. Oxidative stress was implicated in IDD. However, the underlying molecular mechanism for the degeneration of cartilaginous endplate remains elusive. Herein, we found that oxidative stress could induce apoptosis and autophagy in endplate chondrocytes evidenced by western blot analysis, flow cytometry, immunofluorescence staining, GFP-LC3B transfection, and MDC staining...
2017: Oxidative Medicine and Cellular Longevity
https://www.readbyqxmd.com/read/28320092/inhibition-of-pi3k-akt-mtor-signaling-pathway-promotes-autophagy-of-articular-chondrocytes-and-attenuates-inflammatory-response-in-rats-with-osteoarthritis
#18
Jian-Feng Xue, Zhong-Min Shi, Jian Zou, Xiao-Lin Li
OBJECTIVE: This study aims to explore the relationship between PI3K/AKT/mTOR signaling pathway and autophagy of articular chondrocytes in rats with osteoarthritis (OA). METHODS: Rat articular chondrocytes were isolated and cultured, and then induced by protein inhibitors of PI3K/AKT/mTOR signaling pathway. Chondrocytes were assigned into blank group, IL-1β induction group (IL-1β group), PI3K inhibitor+IL-1β induction group (PI3Ki+IL-1β group), AKT inhibitor+IL-1β induction group (AKTi+IL-1β group) and mTOR inhibitor+IL-1β induction group (mTORi+IL-1β group)...
May 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28314786/effects-of-microrna-146a-on-the-proliferation-and-apoptosis-of-human-osteoarthritis-chondrocytes-by-targeting-traf6-through-the-nf-%C3%AE%C2%BAb-signalling-pathway
#19
Jun-Hua Zhong, Jing Li, Cui-Fang Liu, Ning Liu, Rui-Xiang Bian, Shou-Mei Zhao, Shu-Yi Yan, Yong-Bing Zhang
The present study aims to investigate the effects of miR-146a on the proliferation and apoptosis of human osteoarthritis (OA) chondrocytes by targeting tumour necrosis factor receptor-associated factor 6 (TRAF6) through nuclear factor-κB (NF-κB) signalling pathway. Human normal and OA chondrocytes were selected and divided into the normal group, blank group, negative control (NC) group, miR-146a mimics group, miR-146a inhibitors, miR-146a inhibitor + si-TRAF6 group and si-TRAF6 group. Quantitative real-time PCR (qRT-PCR) was applied to detect the expressions of miR-146a, TRAF6 mRNA and NF-κB mRNA...
April 30, 2017: Bioscience Reports
https://www.readbyqxmd.com/read/28291471/chondrotoxicity-of-liposomal-bupivacaine-in-articular-chondrocytes-preliminary-findings
#20
K Aaron Shaw, Peter C Johnson, Steve Zumbrun, Augustine H Chuang, Craig D Cameron
OBJECTIVE: The chondrotoxicity of local anesthetics has been previously recognized. Recent introduction of a liposomal formulation of bupivacaine has been found to significantly improve postoperative pain control but its effect on chondrocyte viability has yet to be investigated with this new formulation. We sought to assess the in vitro chondrotoxicity of liposomal bupivacaine. METHODS: Chondrocytes were isolated from articular cartilage from fresh stifle joints and grown in culture medium...
March 2017: Military Medicine
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