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Chondrocyte flow cytometry

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https://www.readbyqxmd.com/read/28343378/adipose-derived-stem-cells-suppress-inflammation-induced-by-il-1beta-through-down-regulation-of-p2x7r-mediated-by-mir-373-in-chondrocytes-of-osteoarthritis
#1
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-1beta (IL-1beta) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and IL-1beta 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-1beta mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery...
March 27, 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
#2
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
#3
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)...
March 16, 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-signaling-pathway
#4
Jun-Hua Zhong, Jing Li, Cui-Fang Liu, Ning Liu, Rui-Xiang Bian, Shou-Mei Zhao, Shu-Yi Yan, Yong-Bing Zhang
This study aims to investigate the effects of microRNA-146a (miR-146a) on the proliferation and apoptosis of human osteoarthritis (OA) chondrocytes by targeting TRAF6 through NF-κB signaling 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 polymerase chain reaction (qRT-PCR) was applied to detect the expressions of miR-146a, TRAF6 mRNA and NF-κB mRNA...
March 17, 2017: Bioscience Reports
https://www.readbyqxmd.com/read/28291471/chondrotoxicity-of-liposomal-bupivacaine-in-articular-chondrocytes-preliminary-findings
#5
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
#6
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
#7
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
#8
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
#9
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...
January 20, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28202394/platelet-rich-plasma-prp-induces-chondroprotection-via-increasing-autophagy-anti-inflammatory-markers-and-decreasing-apoptosis-in-human-osteoarthritic-cartilage
#10
Mayssam Moussa, Daniel Lajeunesse, George Hilal, Oula El Atat, Gaby Haykal, Rim Serhal, Antonio Chalhoub, Charbel Khalil, Nada Alaaeddine
OBJECTIVES: Autophagy constitutes a defense mechanism to overcome aging and apoptosis in osteoarthritic cartilage. Several cytokines and transcription factors are linked to autophagy and play an important role in the degradative cascade in osteoarthritis (OA). Cell therapy such as platelet rich plasma (PRP) has recently emerged as a promising therapeutic tool for many diseases including OA. However, its mechanism of action on improving cartilage repair remains to be determined. The purpose of this study is to investigate the effect of PRP on osteoarthritic chondrocytes and to elucidate the mechanism by which PRP contributes to cartilage regeneration...
March 1, 2017: Experimental Cell Research
https://www.readbyqxmd.com/read/28177757/microrna-181-inhibits-proliferation-and-promotes-apoptosis-of-chondrocytes-in-osteoarthritis-by-targeting-pten
#11
Xiao-Feng Wu, Zi-Hui Zhou, Jian Zou
OBJECTIVE: To investigate the effects of microRNA-181 (miR-181) on proliferation and apoptosis of chondrocytes in osteoarthritis (OA) by targeting PTEN. METHODS: The chondrocytes in logarithmic growth phase were selected and divided into 6 groups: the normal, blank, negative control, miR-181 mimics, miR-181 inhibitors and miR-181 inhibitors + PTEN-siRNA groups. RT-qPCR was used to detect the expressions of miR-181 and PTEN mRNA. MTT assay and flow cytometry were performed to detect cell proliferation and apoptosis...
January 25, 2017: Biochemistry and Cell Biology, Biochimie et Biologie Cellulaire
https://www.readbyqxmd.com/read/28124102/in-vitro-chondrogenic-potential-of-synovial-stem-cells-and-chondrocytes-allocated-for-autologous-chondrocyte-implantation-a-comparison-synovial-stem-cells-as-an-alternative-cell-source-for-autologous-chondrocyte-implantation
#12
Eva Johanna Kubosch, Emanuel Heidt, Philipp Niemeyer, Anke Bernstein, Norbert P Südkamp, Hagen Schmal
PURPOSE: The use of passaged chondrocytes is the current standard for autologous chondrocyte implantation (ACI). De-differentiation due to amplification and donor site morbidity are known drawbacks highlighting the need for alternative cell sources. METHODS: Via clinically validated flow cytometry analysis, we compared the expression of human stem cell and cartilage markers (collagen type 2 (Col2), aggrecan (ACAN), CD44) of chondrocytes (CHDR), passaged chondrocytes for ACI (CellGenix™), bone marrow derived mesenchymal stem cells (BMSC), and synovial derived stem cells (SDSC)...
January 25, 2017: International Orthopaedics
https://www.readbyqxmd.com/read/28109162/odontoblastic-differentiation-of-dental-pulp-stem-cells-from-healthy-and-carious-teeth-on-an-original-pcl-based-3d-scaffold
#13
A Louvrier, E Euvrard, L Nicod, G Rolin, F Gindraux, L Pazart, C Houdayer, P Y Risold, F Meyer, C Meyer
AIMS: To isolate and characterize Dental Pulp Stem Cells (DPSCs) obtained from carious and healthy mature teeth extracted when conservative treatment was not possible or for orthodontic reasons; to evaluate the ability of DPSCs to colonize, proliferate, and differentiate into functional odontoblast-like cells when cultured onto a polycaprolactone cone made by jet-spraying and prototyped into a design similar to a gutta-percha cone. METHODOLOGY: DPSCs were obtained from 9 carious and 12 healthy mature teeth...
January 21, 2017: International Endodontic Journal
https://www.readbyqxmd.com/read/28024508/-reconstruction-of-humanized-bone-marrow-niche-in-immunodeficiency-mouse
#14
Huan Chen, Zheng Tian, Bin Li, Hai-Yan Xing, Huan Li, Ke-Jing Tang, Min Wang, Qing Rao
OBJECTIVE: To reconstruct a human bone marrow niche in immunodeficiency mouse (NOD/SCID) so as to provide a model for observing the effect of abnormal BM niche on the occurence and development of leukemia. METHODS: Human platelet lysate(HPL) was obtained by repeated freezing and thawing of concentrated platelet. Bone marrow-derived mesenchymal stem cells were cultured in α-minimal essential medium (α-MEM) containing 10% HPL or 10% FBS. The morphology, cell phenotype, multilineage differentiation potential in vitro and proliferation capacity between the mesenchymal stem cells cultured with HPL or FBS were compared...
December 2016: Zhongguo Shi Yan Xue Ye Xue za Zhi
https://www.readbyqxmd.com/read/28002898/tranexamic-acid-effects-on-cartilage-and-synovial-tissue-an-in-vitro-study-for-a-possible-safe-intra-articular-use
#15
A Marmotti, S Mattia, L Mangiavini, D E Bonasia, M Bruzzone, F Dettoni, F Rosso, D Blonna, R Rossi, F Castoldi, G M Peretti
The possible toxic effects of intra-articular tranexamic acid (TA) are still debated. The aim of this study was to evaluate TA effects on human cartilage fragments and synovial biopsies. Explant culture of minced articular cartilage underwent prolonged TA exposure. Histological analysis, immunofluorescence and colorimetric assay for quantification of s-GAG and DNA were performed at the end term. Synoviocytes were cultured for 48h in presence of TA. Light microscopy and flow cytometry analysis were performed at the end of the exposure to TA and one week after the treatment...
October 2016: Journal of Biological Regulators and Homeostatic Agents
https://www.readbyqxmd.com/read/27925477/relevance-of-meniscal-cell-regional-phenotype-to-tissue-engineering
#16
Shawn P Grogan, Chantal Pauli, Martin K Lotz, Darryl D D'Lima
PURPOSE: Meniscus contains heterogeneous populations of cells that have not been fully characterized. Cell phenotype is often lost during culture; however, culture expansion is typically required for tissue engineering. We examined and compared cell-surface molecule expression levels on human meniscus cells from the vascular and avascular regions and articular chondrocytes while documenting changes during culture-induced dedifferentiation. MATERIALS AND METHODS: Expressions of 16 different surface molecules were examined by flow cytometry after monolayer culture for 24 h, 1 week, and 2 weeks...
December 7, 2016: Connective Tissue Research
https://www.readbyqxmd.com/read/27916780/mir-15a-5p-regulates-viability-and-matrix-degradation-of-human-osteoarthritis-chondrocytes-via-targeting-vegfa
#17
Hongwei Chen, Yun Tian
Previous studies demonstrated that miR-15a-5p was probably associated with human hepatocellular carcinoma, while the function of miR-15a-5p in OA (Osteoarthritis) still remains unknown. Here, we uncovered the potential role of miR-15a-5p on OA pathogenesis and confirmed its predicted target VEGFA (Vascular Endothelial Growth Factor A). Measured by RT-PCR, miR-15a-5p expression increased remarkably while VEGFA expression was significantly decreased in OA chondrocytes compared with normal conditions. According to Luciferase activity assay, miR-15a-5p directly targeted the 3'-UTR of VEGFA to inhibit its expression...
January 16, 2017: Bioscience Trends
https://www.readbyqxmd.com/read/27910811/ll-37-hnp-1-and-hbd2-3-modulate-the-secretion-of-cytokines-tnf-%C3%AE-il-6-ifn-%C3%AE-il-10-and-mmp1-in-human-primary-cell-cultures
#18
Carlos Erik Medina Santos, Carmen Nathaly López Hurtado, Bruno Rivas Santiago, Roberto Gonzalez-Amaro, Yolanda Guadalupe Cataño Cañizales, Margarita de la Luz Martínez Fierro, José Antonio Enciso-Moreno, Mariana Haydee García Hernández
The aim of this study was to evaluate the effects of the LL-37, HNP-1 and HBD2/3 peptides on cytokine and MMP production in human polymorphonuclear cells, mononuclear cells and chondrocytes. The levels of cytokines in supernatants from mononuclear and polymorphonuclear cell cultures were measured with a cytometric bead array by flow cytometry. Likewise, the levels of metalloproteinase/MMP-1, 3, and 13 were measured in supernatants from chondrocyte cultures using an ELISA. The expression of RANKL on lymphocytes was analyzed by flow cytometry...
September 1, 2016: European Cytokine Network
https://www.readbyqxmd.com/read/27907835/inhibition-of-the-p38-mapk-signaling-pathway-suppresses-the-apoptosis-and-expression-of-proinflammatory-cytokines-in-human-osteoarthritis-chondrocytes
#19
He-Yan Sun, Kong-Zu Hu, Zong-Sheng Yin
This study aims to investigate the effects of p38-MAPK signaling pathway on the apoptosis and expression of proinflammatory cytokines in human osteoarthritis (OA) chondrocytes. Human articular cartilage specimens were obtained from 57 OA patients and 31 patients with lower extremity traumatic amputations. The expressions of p38-MAPK pathway-related proteins in cartilage tissue were detected by immunochemistry. Cultured chondrocytes isolated from human OA cartilage were assigned into the blank group, the IL-1β group, the PD (PD980959, ERK pathway inhibitors)+IL-1β group, the SB (SB203580, p38 pathway inhibitors)+IL-1β group, and the SP (SP600125, JNK signaling pathway inhibitors)+IL-1β group...
November 28, 2016: Cytokine
https://www.readbyqxmd.com/read/27882184/phosphorylation-of-osteopontin-has-proapoptotic-and-proinflammatory-effects-on-human-knee-osteoarthritis-chondrocytes
#20
Shu-Guang Gao, Yang Yu, Chao Zeng, Shi-Tao Lu, Jian Tian, Chao Cheng, Liang-Jun Li, Guang-Hua Lei
The aim of the present study was to investigate the effects of phosphorylated osteopontin (p-OPN) on apoptosis and pro-inflammatory cytokine expression in human knee osteoarthritis (OA) chondrocytes. Human knee OA chondrocytes obtained from patients who underwent total knee arthroplasty were treated with p-OPN, OPN or buffer. Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) and western blot analysis were used to assess the expression levels of proinflammatory factors, including interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6 and nuclear factor (NF)-κB...
November 2016: Experimental and Therapeutic Medicine
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