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Search Results: 1 - 10 of 103 matches for " chondrocytes "
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Proteomic Analysis of Celecoxib on Chondrocytes from Patients with Osteoarthritis  [PDF]
Kenji Takenouchi, Mitusmi Arito, Toshiyuki Sato, Kenji Takahashi, Manae S. Kurokawa, Kazuo Yudoh, Shinro Takai, Tomohiro Kato, Hiroshi Nakamura
Modern Research in Inflammation (MRI) , 2014, DOI: 10.4236/mri.2014.33011
Abstract:

Objective: To study a comprehensive proteomic analysis of celecoxib in oseteoarthritis (OA) chondrocytes. Methods: OA chondrocytes were stimulated with celecoxib, IL-1β and IL-1β together with celecoxib. Proteins were extracted from the cells and subjected to 2-dimensional differential image gel electrophoresis (2D-DIGE). Proteins of interest were identified by mass spectrometry. Results: Eighty-six protein spots showed significantly different intensities with each reagent or reagent combination. AAA+ protein, HSP47/Serpin, cAMP-dependent protein kinase type II-beta regulatory subunit, alpha-actin-4 and tubulin decreased with the addition of celecoxib, while apolipoprotein A-V, glutamate carboxipeptide 2, mitochondrial stress-70 protein, sorting nexin-9 and GRP78 increased with the addition of celecoxib. GRP78 is a stress protein and may be chondroprotective. Celecoxib modulated IL-1β stimulated chondrocytes, and CD200R and moesin were identified as such resulting proteins. Conclusion: Protein profiles of OA chondrocytes changed after administration of celecoxib. Further investigation is needed to elucidate the function of each protein in OA chondrocytes.

Calcium Homeostasis in Articular Chondrocytes of Two Different Animal Species  [PDF]
Rachel White, John Stanley Gibson
Open Journal of Veterinary Medicine (OJVM) , 2018, DOI: 10.4236/ojvm.2018.88012
Abstract: Introduction: Intracellular calcium concentration ([Ca2+]i) is a critical parameter in cellular homeostasis, including articular chondrocytes. Perturbed [Ca2+]i of chondrocytes may be associated with joint disease. The objective of the study was to compare large animal models for investigating Ca2+ homeostasis in chondrocytes. Materials and Methods: The gross anatomy of the metacarpophalangeal joint (MCP) of cattle and sheep was compared, along with the effect of various manoeuvres used to study the mechanisms of Ca2+ homeostasis in chondrocytes from load-bearing areas. The gross anatomy was observed before and after dissection, and internal architecture was examined after sectioning. Cartilage thickness was measured with a digital micrometer. Chondrocyte yield was determined after isolation. Chondrocytes were incubated with Fura-2 and Ca2+i followed in different extracellular conditions. A hypotonic shock (HTS) was used to mimic removal of a load. Results: The results showed that ovids and bovids were skeletally immature and aspects of Ca2+ homeostasis were similar. Ovine chondrocytes had higher resting fluorescence, consistent with elevated resting Ca2+ levels. Results from ion substitution experiments were consistent with a role for Na+/Ca2+ exchange, and swelling-induced Ca2+ enters into the cytoplasm via the plasma membrane and intracellular stores. Conclusions: Ca2+ homeostasis in chondrocytes from both species behaved in a similar manner to HTS and ion substitutions. Differences in resting [Ca2+]i could be associated with species, stage of maturation, or Fura-2 itself and require further investigation. These findings contribute to our understanding of the physiology of articular cartilage in different species, and their potential use as models for studying joint disease in humans.
Structure and secretory activity of cultured chondrocytes from patients with osteoarthritis
Montrull,Hilda L.; Brizuela,Nilda Y.; Demurtas,Silvia L.; Spitale,Luis; Meirovich,Carlos I.;
Biocell , 2005,
Abstract: cartilage samples were taken from oa patients in order to describe and quantify pro-inflammatory mediators. samples were cultured under aseptic conditions in dulbecco's modified eagle medium at 37°c for 10 days. control samples, taken from non-inflammatory cartilage, were cultured under the same conditions. the levels of no-2 and no-3 were measured in the supernatant using a spectrophotometric assay. the activity of mmp-1 was quantified by elisa. the concentration of no-x was 47.3 ± 4.1 μm in the oa cartilague and 10.7 ± 1.8 μm in the controls. the average mmp-1 activity was 3,650 ± 387 ng/ml in the oa cartilage and 2,150 ± 190 ng/ml in the control samples. these increased values of mmp-1 and no-x observed in the oa cartilage suggest a higher catabolic activity. a morphological analysis of oa chondral tissue using light microscopy shows that the surface of the tissue is characterized by the presence of aggregated chondrocytes or &"clones&" but in the deeper areas isolated cells are found. these results could be a significant contribution towards the identification of biological markers indicating the presence of oa activity.
The effects of transforming growth factor β1, insulin-like growth factor 1 and leptin on the proliferation of fetal chondrocytes
I. Fenichel,Z. Evron,Z. Nevo
Journal of Orthopaedics and Traumatology , 2007, DOI: 10.1007/s10195-007-0152-2
Abstract: One method to exogenously enhance the repair of articular cartilage is the local application of growth factors. This method is based on the chondrogenic effects of some agents and their potential ability to enhance cell migration. Human fetal chondrocytes were isolated and cultured. Their proliferation under the influence of different agents was microscopically evaluated. Fetal calf serum at 5% and 10% concentrations induced microscopically visible cell proliferation. Transforming growth factor beta one (5 and 10 ng/ml), insulin-like growth factor 1 (5 and 10 nmol/l) and leptin (1 and 2 ng/ml) accelerated proliferation of the cells towards the increasing gradient of the agents. Fibroblast growth factor beta (5 and 10 ng/ml), bone morphogenic protein two (10 ng/ml) and laminin (1 mg/ml) did not affect cell proliferation. This study suggests that different agents can play a role in the proliferation of fetal chondrocytes.
Structure and secretory activity of cultured chondrocytes from patients with osteoarthritis
Hilda L. Montrull,Nilda Y. Brizuela,Silvia L. Demurtas,Luis Spitale
Biocell , 2005,
Abstract: Cartilage samples were taken from OA patients in order to describe and quantify pro-inflammatory mediators. Samples were cultured under aseptic conditions in Dulbecco's modified Eagle medium at 37°C for 10 days. Control samples, taken from non-inflammatory cartilage, were cultured under the same conditions. The levels of NO-2 and NO-3 were measured in the supernatant using a spectrophotometric assay. The activity of MMP-1 was quantified by ELISA. The concentration of NO-x was 47.3 ± 4.1 μM in the OA cartilague and 10.7 ± 1.8 μM in the controls. The average MMP-1 activity was 3,650 ± 387 ng/ml in the OA cartilage and 2,150 ± 190 ng/ml in the control samples. These increased values of MMP-1 and NO-x observed in the OA cartilage suggest a higher catabolic activity. A morphological analysis of OA chondral tissue using light microscopy shows that the surface of the tissue is characterized by the presence of aggregated chondrocytes or &"clones&" but in the deeper areas isolated cells are found. These results could be a significant contribution towards the identification of biological markers indicating the presence of OA activity.
The Use of Human Amniotic Membrane for Cartilage Repair: A Sheep Study  [PDF]
Samuel K. Tabet, David M. Conner, Davis A. Guebert
Stem Cell Discovery (SCD) , 2015, DOI: 10.4236/scd.2015.54005
Abstract: Studies show that human amniotic cells’ pluripotentency can be influenced to produce chondrocytes and osteocytes through adding demineralized bone (DMB). Objective: This study evaluates the human amniotic membrane (HAM) mixed with DMB to fill defects in sheep models. We hypothesized this membrane would fill these defects with hyaline-like cartilage with chondrocytes populating the matrix. Design: Six adult sheep were used in this study. One hindquarter knee of each sheep was utilized to make two cartilage defects: one on the femoral condyle and one in the trochlear grove. Three control sheep had unfilled defects. Three sheep received HAM/DMB from a placenta to fill the defects. The membrane was folded so the cellular layer faced the defect and the joint while demineralized bone was placed between the layers. The membranes were fixed to the femur and to the trochlear grove. At six months, the sheep were sacrificed for evaluation. Results: Of the controls, defects did not fill with hyaline or fibrocartilage. In HAM/DMB sheep, 50% of the defects retained the membrane, consistent with other animal models. Membrane defects were examined histologically by a validated scoring system. A strong correlation of little statistical difference between the test and the normal cartilages was observed. The defects that retained membranes had evidence of diffuse chondrocyte-like cell proliferation of stromal matrix similar to hyaline cartilage. Conclusions: HAM/DMB is a potential source of pluripotent cells that can influence chondrogenesis in defects in sheep models. The implications for application in a human model are promising and warrant further study.
The Use of Hypothermically Stored Amniotic Membrane for Cartilage Repair: A Sheep Study  [PDF]
Samuel K. Tabet, Alisa L. Clark, Eric B. Chapman, Doug Thal
Stem Cell Discovery (SCD) , 2015, DOI: 10.4236/scd.2015.54007
Abstract: Objective: To evaluate the use of hypothermically stored human amniotic membrane for cartilage repair in adult sheep. Studies show that human amniotic membrane contains pluripotent mesenchymal stem cells that can be influenced to produce chondrocytes. It is unknown if human amniotic cells can produce hyaline-like cartilage. This study evaluates the use of hypothermically stored amniotic membrane (HSAM) to fill chondral defects in a sheep model. We hypothesized HSAM would fill defects with hyaline-like cartilage with chondrocytes populating the matrix. One sheep was used as a control, and four sheep received amniotic membrane. Two of these sheep were used as a normal control comparison. A 1 cm2 defect was created on the trochlear grove in all specimens. Each membrane was sized and laid over with the stromal layer facing the subchondral bone and covered with Fibrin sealant. The knees were harvested at five months and underwent morphological, histological, and immunohistological evaluation based on the original validated scoring system by O’Driscoll. The control defect didn’t fill in with hyaline cartilage or fibrocartilage. The defects that successfully retained the graft had evidence of diffuse chondrocyte cell proliferation and showed a stromal matrix similar to hyaline cartilage. The graft samples showed a near 100% morphological fill in the HSAM defect contrasting to <10% fill in the control defect. The retained HSAM grafts scored 2.5 on a 0 - 3 cartilage appearance scale compared with 0.5 for the control defects. HSAM is a potential source of pluripotent cells that can influence chondrogenesis in a sheep model. The implications for application in a human model are promising and warrant further study.
Chondrocyte Production of Pro-Inflammatory Chemokine MCP-1 (CCL-2) and Prostaglandin E-2 Is Inhibited by Avocado/Soybean Unsaponifiables, Glucosamine, Chondroitin Sulfate Combination  [PDF]
Erica J. Secor, Mark W. Grzanna, Ann M. Rashmir-Raven, Carmelita G. Frondoza
Pharmacology & Pharmacy (PP) , 2018, DOI: 10.4236/pp.2018.91002
Abstract: Osteoarthritis (OA) is a chronic, painful disease affecting articulating joints in man and animals. It is characterized by cartilage breakdown, bone remodeling, osteophyte formation and joint inflammation. Currently used non-steroidal anti-inflammatory drugs for the management of OA are known to have deleterious side effects. To address the need for alternatives, we evaluated the anti-inflammatory effects of a combination of avocado/soybean unsaponifiables (ASU), glucosamine (GLU) and chondroitin sulfate (CS) by measuring chemokine MCP-1 (monocyte chemoattractant protein 1, CCL2) and prostaglandin E-2 (PGE2) in stimulated chondrocytes. As the only cellular constituents of cartilage, chondrocytes are the source of pro-inflammatory mediators that play critical roles in the pathogenesis of OA. Chondrocytes were incubated: with: 1) control media, 2) [ASU + GLU + CS] combination, or 3) Phenylbutazone (PBZ) for 24 hours. Cells were next stimulated with IL-1β or LPS for another 24 hrs. MCP-1 and PGE2 from supernatants were quantitated by immunoassay. Another set of chondrocytes seeded in chamber slides were stimulated with IL-1β for 1 hour and then immunostained for NF-κB. Chondrocytes stimulated with IL-1β or LPS significantly increased MCP-1 and PGE2 production which were significantly reduced after treatment with [ASU + GLU + CS]. In contrast, PBZ significantly reduced PGE2 but not MCP-1 production. IL-1β stimulation induced nuclear translocation of NF-κB, which was inhibited by pre-treatment with either [ASU + GLU + CS] or PBZ. The present study provides evidence that the production of MCP-1 by chondrocytes can be inhibited by the combination of [ASU + GLU + CS] but not by PBZ. In contrast, PGE2 production was inhibited by either treatment suggesting that the production of MCP-1 and PGE2 could be independently regulated. The finding of distinct effects of [ASU + GLU + CS] on MCP-1 and PGE2 synthesis supports a scientific rationale for a multimodal treatment approach in the management of OA.
Aggrecanases and cartilage matrix degradation
Hideaki Nagase, Masahide Kashiwagi
Arthritis Research & Therapy , 2003, DOI: 10.1186/ar630
Abstract: Cartilage consists of a relatively small number of chondrocytes and abundant extracellular matrix (ECM) components. While numerous macromolecules have been identified in cartilage, the major constituents are collagen fibrils and aggrecan, a large aggregating proteoglycan [1]. Collagen fibrils consisting mainly of type II collagen and, to a lesser extent, of collagen type IX and type XI form an oriented meshwork that provides the cartilage with tensile strength. Aggrecans fill the interstices of the collagen meshwork by forming large aggregated complexes interacting with hyaluronan and link proteins. Aggrecan monomers are approximately 2.5 million Da and consist of a 250-kDa core protein to which chondroitin sulfate and keratan sulfate glycosaminoglycan (GAG) chains are covalently attached. Aggrecans are highly hydrated because of their negatively charged long polysaccharide chains, and thus provide the cartilage with its ability to resist compressive loads.Chondrocytes synthesize and catabolize ECM macromolecules, while the matrix in turn functions to maintain the homeostasis of the cellular environment and the structure of cartilage. In diseases such as osteoarthritis (OA) and rheumatoid arthritis (RA), degradation of the ECM exceeds its synthesis, resulting in a net decrease in the amount of cartilage matrix or even in the complete erosion of the cartilage overlying the bone at the joint surface. Although many possible causes of cartilage destruction have been suggested, such as hypoxic conditions and oxygen-derived free radicals [2,3], the primary cause of this process is thought to be an elevation in the activities of proteolytic enzymes. The loss of aggrecan is considered a critical early event of arthritis, occurring initially at the joint surface and progressing to the deeper zones. This is followed by degradation of collagen fibrils and mechanical failure of the tissue.The matrix metalloproteinases (MMPs) have been considered the main enzymes responsible for
Insulin impairs the maturation of chondrocytes in vitro
Torres, E.S.;Andrade, C.V.;Fonseca, E.C.;Mello, M.A.;Duarte, M.E.L.;
Brazilian Journal of Medical and Biological Research , 2003, DOI: 10.1590/S0100-879X2003000900007
Abstract: the precise nature of hormones and growth factors directly responsible for cartilage maturation is still largely unclear. since longitudinal bone growth occurs through endochondral bone formation, excess or deficiency of most hormones and growth factors strongly influences final adult height. the structure and composition of the cartilaginous extracellular matrix have a critical role in regulating the behavior of growth plate chondrocytes. therefore, the maintenance of the three-dimensional cell-matrix interaction is necessary to study the influence of individual signaling molecules on chondrogenesis, cartilage maturation and calcification. to investigate the effects of insulin on both proliferation and induction of hypertrophy in chondrocytes in vitro we used high-density micromass cultures of chick embryonic limb mesenchymal cells. culture medium was supplemented with 1% fcs + 60 ng/ml (0.01 μm) insulin and cultures were harvested at regular time points for later analysis. proliferating cell nuclear antigen immunoreactivity was widely detected in insulin-treated cultures and persisted until day 21 and [3h]-thymidine uptake was highest on day 14. while apoptosis increased in control cultures as a function of culture time, terminal deoxynucleotidyl transferase-mediated dutp nick end labeling (tunel)-labeled cells were markedly reduced in the presence of insulin. type ii collagen production, alkaline phosphatase activity and cell size were also lower in insulin-treated cultures. our results indicate that under the influence of 60 ng/ml insulin, chick chondrocytes maintain their proliferative potential but do not become hypertrophic, suggesting that insulin can affect the regulation of chondrocyte maturation and hypertrophy, possibly through an antiapoptotic effect.
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