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Chondrocyte flow cytometry

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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
#1
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
#2
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...
February 13, 2017: Experimental Cell Research
https://www.readbyqxmd.com/read/28177757/microrna-181-inhibits-proliferation-and-promotes-apoptosis-of-chondrocytes-in-osteoarthritis-by-targeting-pten
#3
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
#4
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
#5
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
#6
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
#7
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
#8
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
#9
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
#10
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
#11
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
#12
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
https://www.readbyqxmd.com/read/27864696/generation-and-differentiation-of-induced-pluripotent-stem-cells-reveal-ankylosing-spondylitis-risk-gene-expression-in-bone-progenitors
#13
Gerlinde Layh-Schmitt, Shajia Lu, Fatemeh Navid, Stephen R Brooks, Emily Lazowick, Kathryn M Davis, Cristina Montagna, Massimo Gadina, Robert A Colbert
Axial spondyloarthritis (axSpA), which encompasses ankylosing spondylitis, is a complex genetic disease. Aberrant bone formation is a key feature of pathogenesis that can lead to ankylosis of the spine. Our objective is to determine, whether genes whose variants confer susceptibility to AS are expressed in bone progenitors like mesenchymal stem cells (MSCs). Since MSCs from bone marrow is difficult to obtain, we first examined, whether MSCs can be derived from induced pluripotent stem cells (iPSCs). Dermal fibroblasts of two axSpA patients and one healthy control were reprogrammed into iPSCs using a Sendai virus vector encoding pluripotency genes...
January 2017: Clinical Rheumatology
https://www.readbyqxmd.com/read/27864279/nutrition-deficiency-promotes-apoptosis-of-cartilage-endplate-stem-cells-in-a-caspase-independent-manner-partially-through-upregulating-bnip3
#14
Zhiliang He, Luqiao Pu, Chao Yuan, Min Jia, Jian Wang
Nutrition deficiency is reported to induce apoptosis of chondrocytes and degeneration of cartilage endplate (CEP) in rabbit. Cartilage endplate stem cells (CESCs) are important for the integrity of structure and function of CEP. Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (BNIP3) has been reported to regulate apoptosis, autophagy, and cytoprotection. In this study, we aimed to determine whether nutrition deficiency induces apoptosis of CESCs, and whether or not the BNIP3-related pathway is activated in CESCs during nutrition deficiency...
January 2017: Acta Biochimica et Biophysica Sinica
https://www.readbyqxmd.com/read/27851782/alpha-b-crystallin-protects-rat-articular-chondrocytes-against-casein-kinase-ii-inhibition-induced-apoptosis
#15
Sung Won Lee, Jee Hyun Rho, Sang Yeob Lee, Seung Hee Yoo, Hye Young Kim, Won Tae Chung, Young Hyun Yoo
Although alpha (α)B-crystallin is expressed in articular chondrocytes, little is known about its role in these cells. Protein kinase casein kinase 2 (CK2) inhibition induces articular chondrocyte death. The present study examines whether αB-crystallin exerts anti-apoptotic activity in articular chondrocytes. Primary rat articular chondrocytes were isolated from knee joint slices. Cells were treated with CK2 inhibitors with or without αB-crystallin siRNA. To examine whether the silencing of αB-crystallin sensitizes rat articular chondrocytes to CK2 inhibition-induced apoptosis, we assessed apoptosis by performing viability assays, mitochondrial membrane potential measurements, flow cytometry, nuclear morphology observations, and western blot analysis...
2016: PloS One
https://www.readbyqxmd.com/read/27778036/-biological-characteristics-of-mesenchymal-stem-cell-and-hematopoietic-stem-cell-in-the-co-culture-system
#16
Wei Wei, Chao Xu, Zhi-Yong Ye, Xiao-Jun Huang, Jia-En Yuan, Tian-Bao Ma, Han-Biao Lin, Xiu-Qiong Chen
The aim of the present study was to obtain the qualified hematopoietic stem/progenitor cells (HSC/HPC) and human umbilical cord-mesenchymal stem cells (MSC) in vitro in the co-culture system. Cord blood mononuclear cells were separated from umbilical cord blood by Ficoll lymphocyte separation medium, and then CD34(+) HSC was collected by MACS immunomagnetic beads. The selected CD34(+) HSC/HPC and MSC were transferred into culture flask. IMDM culture medium with 15% AB-type cord plasma supplemented with interleukin-3 (IL-3), IL-6, thrombopoietin (TPO), stem cell factor (SCF) and FMS-like tyrosine kinase 3 ligand (Flt-3L) factors were used as the co-culture system for the amplification of HSC/HPC and MSC...
October 25, 2016: Sheng Li Xue Bao: [Acta Physiologica Sinica]
https://www.readbyqxmd.com/read/27702388/epidermal-growth-factor-induces-proliferation-of-hair-follicle-derived-mesenchymal-stem-cells-through-epidermal-growth-factor-receptor-mediated-activation-of-erk-and-akt-signaling-pathways-associated-with-upregulation-of-cyclin-d1-and-downregulation-of-p16
#17
Tingting Bai, Feilin Liu, Fei Zou, Guifang Zhao, Yixu Jiang, Li Liu, Jiahong Shi, Deshun Hao, Qi Zhang, Tong Zheng, Yingyao Zhang, Mingsheng Liu, Shilun Li, Liangchen Qi, Jin Yu Liu
The maintenance of highly proliferative capacity and full differentiation potential is a necessary step in the initiation of stem cell-based regenerative medicine. Our recent study showed that epidermal growth factor (EGF) significantly enhanced hair follicle-derived mesenchymal stem cell (HF-MSC) proliferation while maintaining the multilineage differentiation potentials. However, the underlying mechanism remains unclear. Herein, we investigated the role of EGF in HF-MSC proliferation. HF-MSCs were isolated and cultured with or without EGF...
January 15, 2017: Stem Cells and Development
https://www.readbyqxmd.com/read/27665780/platelet-rich-plasma-protects-rat-chondrocytes-from-interleukin-1%C3%AE-induced-apoptosis
#18
Jinjiang Yang, Ying Lu, Ai Guo
Interleukin (IL)-1β-induced chondrocyte apoptosis is associated with the pathogenesis of arthritis. Platelet‑rich plasma (PRP), which is derived from the patient's own blood and contains numerous growth factors, has the potential for arthritis treatment. Therefore, the present study aimed to determine the effects of PRP on chondrocyte apoptosis, under IL‑1β‑induced pathological conditions. Chondrocytes isolated from the knee joint of Sprague Dawley rats were used in the present study. Cell viability was determined using the Cell Counting kit‑8 assay, cell apoptosis was evaluated by flow cytometry, and the expression of apoptosis‑, anabolism‑ and catabolism-associated genes were detected by quantitative polymerase chain reaction; protein expression was detected by western blot analysis...
November 2016: Molecular Medicine Reports
https://www.readbyqxmd.com/read/27609477/comparative-characteristics-of-mesenchymal-stem-cells-derived-from-reamer-irrigator-aspirator-iliac-crest-bone-marrow-and-adipose-tissue
#19
COMPARATIVE STUDY
S Toosi, H Naderi-Meshkin, F Kalalinia, M T Peivandi, H Hossein Khani, A R Bahrami, A Heirani-Tabasi, M Mirahmadi, J Behravan
Mesenchymal stem cells (MSCs) have been considered promising tools for new clinical concepts in supporting cellular therapy and regenerative medicine. More recently, Ream/Irrigator/Aspirator (RIA) was introduced as a source of MSCs. In this study we compared MSCs derived from three different sources (iliac crest bone marrow (ICBM), adipose tissue (AT), and (RIA)) regarding the morphology, the success rate of isolating MSCs, colony frequency, expansion potential, osteogenic and chondrogenic differentiation capacity...
August 31, 2016: Cellular and Molecular Biology
https://www.readbyqxmd.com/read/27574269/alloreactivity-and-immunosuppressive-properties-of-articular-chondrocytes-from-osteoarthritic-cartilage
#20
S Abe, H Nochi, H Ito
PURPOSE: To determine whether articular chondrocytes derived from osteoarthritic knee joints could evoke alloreactive proliferation of peripheral blood mononuclear cells (PBMC) and inhibit mitogenic activity of polyclonally activated CD4+ major histocompatibility complex (MHC) class II- restricted T cells in vitro. METHODS: Osteoarthritic cartilages of 17 patients aged 61 to 85 years were harvested during total knee arthroplasty. Chondrocytes were cultured for experiments...
August 2016: Journal of Orthopaedic Surgery
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