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Chondrocyte flow cytometry

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https://www.readbyqxmd.com/read/28801211/parkin-clearance-of-dysfunctional-mitochondria-regulates-ros-levels-and-increases-survival-of-human-chondrocytes
#1
Mohammad Y Ansari, Nazir M Khan, Imran Ahmad, Tariq M Haqqi
OBJECTIVE: Mitochondrial dysfunction, oxidative stress and chondrocyte death are important contributors to the development and pathogenesis of osteoarthritis (OA). In this study, we determined the expression and role of Parkin in the clearance of damaged/dysfunctional mitochondria, regulation of ROS levels and chondrocyte survival under pathological conditions. METHODS: Human chondrocytes were from the unaffected area of knee OA cartilage (n=12) and were stimulated with IL-1β to mimic pathological conditions...
August 8, 2017: Osteoarthritis and Cartilage
https://www.readbyqxmd.com/read/28761446/characteristics-of-human-endometrium-derived-mesenchymal-stem-cells-and-their-tropism-to-endometriosis
#2
Yan Cheng, Liru Li, Dejun Wang, Qiuyan Guo, Yanan He, Tian Liang, Liyuan Sun, Xiaojun Wang, Yulei Cheng, Guangmei Zhang
Human endometrial tissue has become an attractive source of mesenchymal stem cells (MSCs) for cell-based therapies because these MSCs can be easily harvested and have tumour tropism as well as reduced immunogenic and inflammatory properties. Our study aimed to obtain and characterise human endometrial mesenchymal stem cells (EMSCs) and assess their endometriosis tropism. EMSCs were successfully isolated from the endometrium of women undergoing laparoscopy for idiopathic infertility. The EMSCs presented a fibroblast-like morphology during culture...
2017: Stem Cells International
https://www.readbyqxmd.com/read/28744712/vacuolar-protein-sorting-4b-vps4b-regulates-apoptosis-of-chondrocytes-via-p38-mitogen-activated-protein-kinases-mapk-in-osteoarthritis
#3
Libin Xu, Leilei Zhai, Qingtian Ge, Zhongbing Liu, Ran Tao
To aim of this study is to investigate the expression of VPS4B (vacuolar protein sorting 4B) in articular cartilage with osteoarthritis (OA) and to analyze the relationship between VPS4B and chondrocyte apoptosis. We established an OA rat model by the MLI (meniscal/ligamentous injury) modeling method, and we observed the expression of VPS4B in articular cartilage through immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). Human SW1353 chondrosarcoma cells were treated with IL-1β to mimic the OA-like chondrocyte injury in vitro, and Western blot was employed to examine the IL-1β-induced expression of VPS4B, phosphorylated p38, and apoptotic markers, namely active caspase-3 and cleaved poly (ADP-ribose) polymerase (PARP)...
July 25, 2017: Inflammation
https://www.readbyqxmd.com/read/28685122/nano-engineered-skin-mesenchymal-stem-cells-potential-vehicles-for-tumour-targeted-quantum-dot-delivery
#4
Liga Saulite, Dominyka Dapkute, Karlis Pleiko, Ineta Popena, Simona Steponkiene, Ricardas Rotomskis, Una Riekstina
Nanotechnology-based drug design offers new possibilities for the use of nanoparticles in imaging and targeted therapy of tumours. Due to their tumour-homing ability, nano-engineered mesenchymal stem cells (MSCs) could be utilized as vectors to deliver diagnostic and therapeutic nanoparticles into a tumour. In the present study, uptake and functional effects of carboxyl-coated quantum dots QD655 were studied in human skin MSCs. The effect of QD on MSCs was examined using a cell viability assay, Ki67 expression analysis, and tri-lineage differentiation assay...
2017: Beilstein Journal of Nanotechnology
https://www.readbyqxmd.com/read/28670366/inhibitory-effect-of-dihydroartemisinin-on-chondrogenic-and-hypertrophic-differentiation-of-mesenchymal-stem-cells
#5
Zhen Cao, Chuan Liu, Yun Bai, Ce Dou, Jian-Mei Li, Duo-Wei Shi, Shi-Wu Dong, Qiang Xiang
Chondrocytes located in hyaline cartilage may maintain phenotype while the chondrocytes situated in calcified cartilage differentiate into hypertrophy. Chondrogenic and hypertrophic differentiation of mesenchymal stem cells (MSCs) are two subsequent processes during endochondral ossification. However, it is necessary for chondrocytes to hold homeostasis and to inhibit hypertrophic differentiation in stem cell-based regenerated cartilage. Dihydroartemisinin (DHA) is derived from artemisia apiacea which has many biological functions such as anti-malarial and anti-tumor...
2017: American Journal of Translational Research
https://www.readbyqxmd.com/read/28664842/difference-between-biomarkers-of-tibial-bone-marrow-and-adipose-tissue
#6
Ersin Kuyucu, Mehmet Erdil, Adnan Kara, Murat Bülbül
BACKGROUND: Stem cells, with their regeneration capacity, long-term viability, and differentiation characteristics, have indispensable biological properties. As described by Hauner and Grigoradis et al., mesenchymal stem cell originating from adipose or bone marrow can be differentiated into many tissues such as adipocyte, chondrocyte, myeloblast, and osteoblast. The aim of our study is to compare the use of adipose and tibial bone marrow derived stem cells for therapeutic purposes in orthopedic surgery, which has not been clearly evaluated in the literature to our knowledge and to also evaluate their use...
2017: SICOT-J
https://www.readbyqxmd.com/read/28656222/activation-of-the-toll-like-receptor-8-pathway-increases-the-immunogenicity-of-mesenchymal-stem-cells-from-umbilical-cord
#7
Yu Yang, Yanwen Wang, Li Li, Fei Chen, Peng Zhang
Mesenchymal stem cells (MSCs) are now widely used in clinical cell‑based therapy due to their characteristics of low immunogenicity, multiple differentiation potency and the capability to modulate immune responses. However, accumulated research has indicated the absence of engrafted MSCs because of the increased immunogenicity of MSCs. Toll‑like receptors (TLRs) are essential for the innate immune response and regulating the biological function of MSCs. The present study used human umbilical cord‑derived MSCs (UCMSCs) and activated the TLR8 pathway of UCMSCs to study the role of TLR8 in mediating the immune status of UCMSCs...
August 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28626114/thyroid-stimulating-hormone-stimulation-downregulates-autophagy-and-promotes-apoptosis-in-chondrocytes
#8
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
#9
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
#10
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
#11
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
#12
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
#13
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
#14
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
#15
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
#16
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
#17
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
#18
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...
July 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
#19
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
#20
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
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