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Chondrocyte flow cytometry

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https://www.readbyqxmd.com/read/28548679/cyclin-d1-gene-silencing-promotes-il-1%C3%AE-induced-apoptosis-in-rat-chondrocytes
#1
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
#2
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
#3
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
#4
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
#5
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
#6
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...
May 10, 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
#7
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
#8
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
#9
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...
March 15, 2017: Oncotarget
https://www.readbyqxmd.com/read/28391163/peroxiredoxin-4-inhibits-il-1%C3%AE-induced-chondrocyte-apoptosis-via-pi3k-akt-signaling
#10
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
#11
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
#12
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
#13
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
#14
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
#15
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
#16
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
https://www.readbyqxmd.com/read/28279188/comparison-of-exosomes-secreted-by-induced-pluripotent-stem-cell-derived-mesenchymal-stem-cells-and-synovial-membrane-derived-mesenchymal-stem-cells-for-the-treatment-of-osteoarthritis
#17
Yu Zhu, Yuchen Wang, Bizeng Zhao, Xin Niu, Bin Hu, Qing Li, Juntao Zhang, Jian Ding, Yunfeng Chen, Yang Wang
BACKGROUND: Osteoarthritis (OA) is the most common joint disease worldwide. In the past decade, mesenchymal stem cells (MSCs) have been used widely for the treatment of OA. A potential mechanism of MSC-based therapies has been attributed to the paracrine secretion of trophic factors, in which exosomes may play a major role. In this study, we aimed to compare the effectiveness of exosomes secreted by synovial membrane MSCs (SMMSC-Exos) and exosomes secreted by induced pluripotent stem cell-derived MSCs (iMSC-Exos) on the treatment of OA...
March 9, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28259429/effects-of-xylazine-and-dexmedetomidine-on-equine-articular-chondrocytes-in%C3%A2-vitro
#18
Francesco Mancini, Sara Nannarone, Sandra Buratta, Giuseppina Ferrara, Anna Maria Stabile, Matteo Vuerich, Isabella Santinelli, Alessandra Pistilli, Elisabetta Chiaradia
OBJECTIVE: To assess the effects of xylazine and dexmedetomidine on equine chondrocytes, in vitro. STUDY DESIGN: Prospective, experimental study. STUDY MATERIAL: Equine articular chondrocytes from five male horses. METHODS: Chondrocytes were isolated from healthy equine articular cartilage of the metacarpo/metatarsophalangeal joints. Cell viability was assessed using the WST-8 assay by exposing chondrocytes to xylazine (0...
January 17, 2017: Veterinary Anaesthesia and Analgesia
https://www.readbyqxmd.com/read/28229417/a-simplified-and-systematic-method-to-isolate-culture-and-characterize-multiple-types-of-human-dental-stem-cells-from-a-single-tooth
#19
Mohammed Bakkar, Younan Liu, Dongdong Fang, Camille Stegen, Xinyun Su, Murali Ramamoorthi, Li-Chieh Lin, Takako Kawasaki, Nicholas Makhoul, Huan Pham, Yoshinori Sumita, Simon D Tran
This chapter describes a simplified method that allows the systematic isolation of multiple types of dental stem cells such as dental pulp stem cells (DPSC), periodontal ligament stem cells (PDLSC), and stem cells of the apical papilla (SCAP) from a single tooth. Of specific interest is the modified laboratory approach to harvest/retrieve the dental pulp tissue by minimizing trauma to DPSC by continuous irrigation, reduction of frictional heat from the bur rotation, and reduction of the bur contact time with the dentin...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28214835/up-regulation-of-tgf-%C3%AE-promotes-tendon-to-bone-healing-after-anterior-cruciate-ligament-reconstruction-using-bone-marrow-derived-mesenchymal-stem-cells-through-the-tgf-%C3%AE-mapk-signaling-pathway-in-a-new-zealand-white-rabbit-model
#20
Rui Wang, Bin Xu, Hong-Gang Xu
BACKGROUND/AIMS: This study aimed to explore the role of TGF-β in tendon-to-bone healing after anterior cruciate ligament (ACL) reconstruction using bone marrow-derived mesenchymal stem cells (BMSCs) through the TGF-β/MAPK signaling pathway in a New Zealand white rabbit model. METHODS: A total of 72 healthy male New Zealand white rabbits were selected for these experiments. Flow cytometry and immunofluorescence were used to detect the expression of BMSC surface markers, and qRT-PCR was performed to detect TGF-β mRNA expression...
2017: Cellular Physiology and Biochemistry
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