In order to be able to use second-generation ACT techniques for the repair of cartilage defects in patients with OA, it is highly important to investigate whether OA chondrocytes have an irreversibly altered phenotype or if these cells can differentiate towards a hyaline cartilage phenotype after in vitro expansion. Today, there are conflicting data whether OA chondrocytes fulfill the prerequisites for ACT treatment or not [12, 13, 15, 21]. This encouraged us to investigate more thoroughly the chondrogenic differentiation potential of human OA chondrocytes using microarray technology in order to determine whether OA chondrocytes might possibly be used in second-generation ACT.. Microarray analysis of human OA and ND chondrocytes cultured in ML indicated that the OA chondrocytes were in a less differentiated state compared with the ND chondrocytes. This is thus in accordance with the differences detected in vivo between OA and ND cartilage [10, 22]. Re-differentiation in scaffold cultures ...
Aigner, Thomas, Gebhard, Pia Margarethe, Schmid, Erik, Bau, Brigitte, Harley, Vincent and Poschl, Ernst (2003) SOX9 expression does not correlate with type II collagen expression in adult articular chondrocytes. Matrix Biology, 22 (4). pp. 363-372. ISSN 1569-1802 Full text not available from this repository. (Request a copy ...
Background: Recent studies have provided evidence that integrins play roles in recognition of mechanical stimuli and its translation into a cellular response. Integrin signaling may be regulated by a number of mechanisms including accessory proteins such as CD98 (4F2 antigen). Objectives: To determine CD98 expression by human articular chondrocytes and its involvement in human articular mechanotransduction. Methods: CD98 expression was assessed by immunostaining of cryostat sections of snap frozen articular cartilage and in cultured cells by western blotting. Chondrocytes enzymatically isolated from macroscopically normal and osteoarthritic (OA) articular cartilage were grown in short term, primary monolayer culture and used in a resting state or following mechanical stimulation at 0.33Hz. Results: Human articular chondrocytes express CD98 and immunoreactivity revealed a similar heterogeneous pattern of CD98 in both normal and osteoarthritic (OA) human articular cartilage. No role of CD98 was detected
TY - JOUR. T1 - Expression and regulation of Toll-like receptor 2 by IL-1β and fibronectin fragments in human articular chondrocytes. AU - Su, S. L.. AU - Tsai, C. D.. AU - Lee, C. H.. AU - Salter, D. M.. AU - Lee, Herng Sheng. PY - 2005/10. Y1 - 2005/10. N2 - Objective: The objective of this study was to examine expression and regulation of Toll-like receptor 2 (TLR2) in human articular chondrocytes. Methods: Human articular chondrocytes were enzymatically isolated from normal and osteoarthritic knee cartilage. Immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to assess the expression of toll-like receptors. Following stimulation of chondrocytes in vitro by IL-1β and fibronectin proteolytic fragments, the relative levels of mRNA for TLR2 were determined by quantitative real-time PCR. MyD88 activation and nuclear factor-κB (NF-κB) translocation were evaluated by immunoprecipitation and electrophoretic mobility shift assay, ...
TY - JOUR. T1 - Evaluation of the post-treatment anti-inflammatory capacity of osteoarthritic chondrocytes. T2 - An in vitro study using baicalein. AU - Wu, Chang Chin. AU - Chen, Yi Ru. AU - Lu, Dai Hua. AU - Hsu, Li Ho. AU - Yang, Kai Chiang. AU - Sumi, Shoichiro. PY - 2020/6. Y1 - 2020/6. N2 - Introduction: Targeting inflammatory cascades is considered a promising way to prevent knee osteoarthritis (OA) progression. In terms of down-regulating the expression of inducible nitric oxide synthase (iNOS), interleukin (IL)-6, and matrix metalloproteinases (MMPs), pre-treatment with the flavonoid baicalein reportedly protects articular chondrocytes against the cytotoxicity of IL-1β. However, the benefits of post-treatment baicalein on osteoarthritic chondrocytes are not fully elucidated. Methods: In this study, primary human chondrocytes were stimulated with IL-1β prior to baicalein application to evaluate the therapeutic effect of post-treatment. Results: Post-treatment baicalein alleviated cell ...
Osteoarthritis (OA), a non-inflammatory, degenerative disease of articular cartilages, is a common cause of poor performance and early retirement in equine athletes. Pathologically, OA is characterized by matrix degradation and decreased chondrocyte numbers. A mechanical stress is believed to be the major etiologic factor of OA development. Recent studies have indicated that apoptosis is responsible for hypocellularity in OA cartilage and that chondrocyte death by apoptosis could directly contribute to matrix degradation. Increased nitric oxide (NO), a free radical, has been implicated as a cause of chondrocyte apoptosis. No studies, however, have been performed on chondrocyte apoptosis in equine OA. We investigated chondrocyte apoptosis in equine OA cartilage and its relationship to matrix degradation and NO production. Furthermore, we studied whether mechanical stress could induce chondrocyte apoptosis and how NO production and Bcl-2 and caspase-3 proteins contribute to chondrocyte apoptosis by using
Background/Purpose: Osteoarthritis (OA) is the most common form of arthritis, affecting nearly 10% of the US population. With age and injury, chondrocytes have diminished mitochondrial content and mitochondrial production of ATP contributing to OA pathogenesis. We have previously reported that chondrocytes release ATP, which is converted extracellularly to adenosine and maintains chondrocyte homeostasis via endogenous stimulation of the A2AR. Injured/inflamed chondrocytes have lower ATP levels and release less ATP resulting in diminished extracellular adenosine and A2AR stimulation. Mice and humans lacking the capacity to convert extracellular ATP to adenosine (ecto-5nucleotidase deficient) develop spontaneous OA as do mice lacking A2AR (A2ARKO). We therefore studied the effect of A2AR stimulation on mitochondrial health and function in chondrocytes from WT and A2ARKO mice and in a human chondrocytic cell line. Methods: A human chondrocyte cell line, T/C28-a2, or neonatal chondrocytes isolated ...
The PI3K pathway has been shown to affect numerous cellular processes in a tissue-specific fashion; for example, it is required for survival in different cell types such as cardiomyocytes [34], cellular differentiation in the case of osteoclasts and keratinocytes [35, 36], and proliferation and differentiation of osteoblasts [35]. It also stimulates differentiation of CD4+ T-cells [37] and development and proliferation of B cells [38, 39]. We hypothesized that the PI3K pathway has similar effects in the growth plate, promoting endochondral bone growth by increasing proliferation and differentiation of chondrocytes and by suppressing apoptosis.. We found that inhibition of PI3K with LY294002 results in decreased differentiation, in both primary chondrocytes (micromass cultures) and organ cultures. Markers of both early chondrocyte differentiation such as collagen II and glycosaminoglycans and of late hypertrophic differentiation such as collagen X, p57, Alkaline phosphatase activity and calcium ...
Intercalation movement of proliferative chondrocytes is crucial for their columnar organization which is essential for proper function of growth plate cartilage. The conventional motor protein kinesin‐1 directionally transporting various cargos along microtubules might be involved in this polarized cell movement. Kinesin‐1 is suggested to transport unknown cargo(s) modulating focal adhesion (FA) turnover which is a key step in cell movement. To investigate kinesin‐1s role in chondrocytes intercalation, we generate kinesin‐1 heavy chain (Kif5b) knockout mouse. In the growth plate of KIF5B deficient mouse, we observed abnormal cell morphology and disrupted columnar structure. Isolated mutant chondrocytes show reduced motility and adhesion ability to ECM proteins. Vinculin, the key regulator of focal adhesions, is found as a potential protein associated with KIF5B in mouse chondrocytes. Further study will investigate whether KIF5B affects chondrocytes motility and adhesion via FAs ...
Osteoarthritis (OA) is a debilitating disease of the joints characterized by cartilage degradation but to date there is no available pharmacological treatment to inhibit disease progression neither is there any available biomarker to predict its development. In the present study, we examined the expression level and possible involvement of novel cell-ECM adhesion-related molecules such as Iintegrin Linked Kinase (ILK), PINCH, parvin, Mig-2 and Migfilin in OA pathogenesis using primary human articular chondrocytes from healthy individuals and OA patients. Our findings show that only ILK and Migfilin were upregulated in OA compared to the normal chondrocytes. Interestingly, Migfilin silencing in OA chondrocytes rather exacerbated than ameliorated the osteoarthritic phenotype, as it resulted in even higher levels of catabolic and hypertrophic markers while at the same time induced reduction in ECM molecules such as aggrecan. Furthermore, we also provide a link between Migfilin and beta-catenin ...
Hypertrophic chondrocytes contained the highest numerical abundance of peroxisomes compared to proliferative chondrocytes as examples for endochondral ossification.
Osteoarthritis (OA) is characterized by the degradation of articular cartilage, marked by the breakdown of matrix proteins. Studies demonstrated the involvement of chemokines in this process, and some may potentially serve as diagnostic markers and therapeutic targets; however, the underlying signal transductions are not well understood. We investigated the effects of the CC chemokine eotaxin-1 (CCL11) on the matrix metalloproteinase (MMP) expression and secretion in the human chondrocyte cell line SW1353 and primary chondrocytes. Eotaxin-1 significantly induced MMP-3 mRNA expression in a dose-dependent manner. Inhibitors of extracellular signal-regulated kinase (ERK) and p38 kinase were able to repress eotaxin-1-induced MMP-3 expression. On the contrary, Rp-adenosine-3,5-cyclic monophosphorothioate (Rp-cAMPs), a competitive cAMP antagonist for cAMP receptors, and H-89, a protein kinase A (PKA) inhibitor, markedly enhanced eotaxin-1-induced MMP-3 expression. These results suggest that MMP-3 expression
15-Lipoxygenases and their metabolites have been shown to exhibit anti-inflammatory and immunomodulatory properties, but little is known regarding their expression and function in chondrocytes. The objective of this study was to evaluate the expression of 15-lipoxygenase-1 and -2 in human articular chondrocytes, and to investigate the effects of their metabolites 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids on IL-1β-induced matrix metalloproteinase (MMP)-1 and MMP-13 expression. The expression levels of 15-lipoxygenase-1 and -2 were analyzed by reverse transcription PCR and Western blotting in chondrocytes, and by immunohistochemistry in cartilage. Chondrocytes or cartilage explants were stimulated with IL-1β in the absence or presence of 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids, and the levels of MMP-1 and MMP-13 protein production and type II collagen cleavage were evaluated using immunoassays. The role of peroxisome proliferator-activated
FILGUEIRAS, R.R et al. Effect of freezing on rabbit cultured chondrocytes. Arq. Bras. Med. Vet. Zootec. [online]. 2011, vol.63, n.1, pp.46-55. ISSN 1678-4162. http://dx.doi.org/10.1590/S0102-09352011000100008.. This work evaluated the effect of freezing on chondrocytes maintained in culture, aiming the establishment of a cell bank for future application as heterologous implant. Chondrocytes extracted from joint cartilage of nine healthy New Zealand White rabbits were cultivated and frozen with the cryoprotector 5% dimethylsulfoxide for six months. Phenotypic and scanning electron microscopy analyses were carried out to identify morphological and functional differences between fresh and thawed cells. After enzymatic digestion, a total of 4.8x105cells per rabbit were obtained. Fresh chondrocytes showed a high mitotic rate and abundant matrix was present up to 60 days of culture. Loss of phenotypic stability was notable in the thawed chondrocytes, with a low labeling of proteoglycans and weak ...
During longitudinal development of the long bone cartilage, periarticular chondrocyte differentiation, which adds cells to the columnar region, is followed by chondrocyte hypertrophy, which reduces cells in the columnar region. Therefore, the length of the columnar chondrocyte region is determined by three parameters: the pace of periarticular chondrocyte differentiation, the pace of chondrocyte hypertrophy and the rate of columnar chondrocyte proliferation (Fig. 7). As upregulated Ihh signaling promotes periarticular chondrocyte differentiation and increases the rate of columnar chondrocyte proliferation (Kobayashi et al., 2005b), the proliferating columnar chondrocyte region would be increased if chondrocyte hypertrophy were not altered. Our observation that the columnar chondrocyte region was shorter in the PTHrP-/-;Ptch1c/-; Col2a1-Cre double mutant than in the PTHrP-/- single mutant (Fig. 2B, Fig. 3A) demonstrates that Hh signaling also acts to promote chondrocyte hypertrophy in the absence ...
INTRODUCTION: Currently available treatments for osteoarthritis (OA) are restricted to nonsteroidal anti-inflammatory drugs, which exhibit numerous side effects and are only temporarily effective. Thus novel, safe and more efficacious anti-inflammatory agents are needed for OA. Naturally occurring polyphenolic compounds, such as curcumin and resveratrol, are potent agents for modulating inflammation. Both compounds mediate their effects by targeting the NF-kappaB signalling pathway. METHODS: We have recently demonstrated that in chondrocytes resveratrol modulates the NF-kappaB pathway by inhibiting the proteasome, while curcumin modulates the activation of NF-kappaB by inhibiting upstream kinases (Akt). However, the combinational effects of these compounds in chondrocytes has not been studied and/or compared with their individual effects. The aim of this study was to investigate the potential synergistic effects of curcumin and resveratrol on IL-1beta-stimulated human chondrocytes in vitro using ...
Chondrogenesis occurs via three steps: (a) commitment to chondrocyte differentiation by mesenchymal cells, seen as mesenchymal condensation; (b) chondrocyte proliferation in the growth plate to facilitate longitudinal development; and (c) differentiation of proliferating chondrocytes to hypertrophic chondrocytes (for review see references 1, 2). During endochondral ossification, chondrocyte hypertrophy is critical, because cells alter the extracellular matrix and induce vascular invasion. Extrinsic factors such as bone morphogenetic proteins, Indian hedgehog, and modulators such as Sox9, Runx2, Smads, and histone deacetylase 4 are reportedly essential for chondrogenesis (1, 2). Loss of histone deacetylase 4 causes early onset of hypertrophy, followed by growth retardation (6). Overexpression of Runx2 under the control of a type II collagen promoter/enhancer induces dwarfism because of precocious endochondral ossification and accelerated chondrocyte differentiation (4, 5). Thus, regulation of ...
Mechanical forces can stimulate the production of extracellular matrix molecules. We tested the efficacy of ultrasound to increase proteoglycan synthesis in bovine primary chondrocytes. The ultrasound-induced temperature rise was measured and its contribution to the synthesis was investigated using bare heat stimulus. Chondrocytes from five cellular isolations were exposed in triplicate to ultrasound (1 MHz, duty cycle 20%, pulse repetition frequency 1 kHz) at average intensity of 580 mW/cm2 for 10 minutes daily for 1-5 days. Temperature evolution was recorded during the sonication and corresponding temperature history was created using a controllable water bath. This exposure profile was used in 10-minute-long heat treatments of chondrocytes. Heat shock protein 70 (Hsp70) levels after one-time treatment to ultrasound and heat was analyzed by Western blotting, and proteoglycan synthesis was evaluated by 35S-sulfate incorporation. Ultrasound treatment did not induce Hsp70, while heat treatment ...
TY - JOUR. T1 - Can microcarrier-expanded chondrocytes synthesize cartilaginous tissue in vitro?. AU - Surrao, Denver C. AU - Khan, Aasma A. AU - McGregor, Aaron J. AU - Amsden, Brian G. AU - Waldman, Stephen D. PY - 2011/8. Y1 - 2011/8. N2 - Tissue engineering is a promising approach for articular cartilage repair; however, it is challenging to produce adequate amounts of tissue in vitro from the limited number of cells that can be extracted from an individual. Relatively few cell expansion methods exist without the problems of de-differentiation and/or loss of potency. Recently, however, several studies have noted the benefits of three-dimensional (3D) over monolayer expansion, but the ability of 3D expanded chondrocytes to synthesize cartilaginous tissue constructs has not been demonstrated. Thus, the purpose of this study was to compare the properties of engineered cartilage constructs from expanded cells (monolayer and 3D microcarriers) to those developed from primary chondrocytes. Isolated ...
In cartilage tissue engineering, hydrogel is widely used as the scaffold for hosting and culturing chondrocyte suspension during neo-tissue formation. In order to develop cultured chondrocytes into a functional cartilage equivalent, the hydrogel must provide an ideal microenvironment for the rapidly proliferating chondrocytes. At the same time, the essential functions of chondrocytes, such as the secretion of type II collagen and glycosaminoglycans, must be maintained. In these studies, we quantitatively characterize the mechanobiology underlying a newly discovered "edge flourish" phenomenon of cultured chondrocytes within a three-dimensional agarose hydrogel, which may ultimately nurture scaffold-free cartilaginous tissue regeneration. First, real-time microscopy was used to track the spatiotemporal distributions of chondrocytes at different focal planes. The chondrocytes were observed to exhibit abundant neo-tissue outgrowth and significant cartilaginous phenotype at the edge of the hydrogel ...
Transformed chondrocyte cell lines and primary OA human articular chondrocytes (HAC) were grown in monolayer culture. Cells were exposed to cyclical mechanical stimulation (MS) using an apparatus that produces strain on the base of the culture dish, causing the deformation of attached cells. Chondrocytes were also incubated with a cytokine cocktail containing IL-1β, TNFα, IL-6 and IFNγ (CYT). The transformed chondrocyte cell lines C20A4 and C2812 did not express detectable levels of NOS protein or nitrite activity. However increased inducible NOS (iNOS) mRNA following CYT stimulation suggested that the cells were able to sense the CYT stimulation. Primary OA HAC showed increased production of iNOS mRNA, protein and nitrite following CYT or IL-1β stimulation. The simultaneous application of CYT and MS also showed elevated iNOS mRNA, protein and nitrite levels, although these were significantly lower than following CYT alone. An IL-4 neutralising antibody and a β1 integrin function blocking ...
The mechanical environment of the chondrocyte is an important factor that influences the maintenance of the articular cartilage extracellular matrix. Previous studies have utilized theoretical models of chondrocytes within articular cartilage to predict the stress-strain and fluid flow environments around the cell, but little is currently known regarding the cellular properties which are required for implementation of these models. The objectives of this study were to characterize the mechanical behavior of primary human chondrocytes and to determine the Youngs modulus of chondrocytes from non-osteoarthritic (`normal) and osteoarthritic cartilage. A second goal was to quantify changes in the volume of isolated chondrocytes in response to mechanical deformation. The micropigette aspiration technique was used to measure the deformation of a single chondrocyte into a glass micropipette in response to a prescribed pressure. The results of this study indicate that the human chondrocyte behaves as a ...
Low density lipoprotein receptor (LDLR)-related protein 1 (LRP1/CD91) is one of the receptors of CCN2 that conducts endochondral ossification and cartilage repair. LRP1 is a well-known endocytic receptor, but its distribution among chondrocytes remains to be elucidated. We herein demonstrate for the first time that the distribution of LRP1 in chondrocytes except for hypertrophic chondrocytes in vivo and in vitro. Interestingly, the LRP1 levels were higher in mature chondrocytic HCS-2/8 and osteoblastic SaOS-2 than in other cells, whereas the other LDLR family members involved in ossification were detected at lower levels in HCS-2/8. It was interesting to note that in HCS-2/8, LRP1 was observed not only on the cell surface and in the cytoplasm, but also in the nucleus. Exogenously added CCN2 was incorporated into HCS-2/8, which was partially co-localized with LRP1, and targeted to the recycling endosomes and nucleus as well as the lysosomes. These findings suggest specific roles of LRP1 in ...
An early event in skeletal joint development is the specification of articular chondrocytes at the joint surface. Articular chondrocytes are distinct in producing lower levels of cartilage matrix and not being replaced by bone, yet how they acquire these properties remains poorly understood. Here, we show that two members of the Iroquois transcriptional repressor family, Irx7 and Irx5a, function to block chondrocyte maturation at the developing hyoid joint of zebrafish. These Irx factors suppress the production of cartilage matrix at the joint in part by preventing the activation of a col2a1a enhancer by Sox9a. Further, both zebrafish Irx7 and mouse IRX1 are able to repress cartilage matrix production in a murine chondrogenic cell line. Iroquois proteins may therefore have a conserved role in keeping chondrocytes in an immature state, with the lower levels of cartilage matrix produced by these immature cells contributing to joint flexibility ...
... during extension, and age-related drop in chondrogenic activity present critical road blocks to the usage of autologous chondrocyte implantation for cartilage fix. neocartilage made by juvenile chondrocytes was 100-flip greater than WHI-P180 in neocartilage made by adult cells. Collagen type type and II IX mRNAs in clean juvenile chondrocytes had been 100- and 700-collapse higher, respectively, than in adult chondrocytes. The distributions of collagens II and IX had been similar in indigenous juvenile cartilage and in neocartilage created by juvenile cells. Juvenile cells grew considerably quicker in monolayer civilizations than adult cells (p = 0.002) and proteoglycan amounts stated in agarose lifestyle was significantly higher in juvenile cells than in adult cells after multiple passages (p < 0.001). Juvenile chondrocytes didnt stimulate lymphocyte proliferation. Conclusions These outcomes record a dramatic age group ...
Both mechanical load and elevated levels of proinflammatory cytokines have been associated with the risk for developing osteoarthritis (OA), yet the potential interaction of these mechanical and biological factors is not well understood. The purpose of this study was to evaluate the response of chondrocytes to the effects of dynamic unconfined compression, TNF-α, and the simultaneous effects of dynamic unconfined compression and TNF-α. The response to these three treatments was markedly different and, taken together, the response in the gene expression of chondrocytes to the different treatment conditions suggest a complex interaction between structure, biology, and mechanical loading.. ...
Cell-to-cell diffusion of second messengers across intercellular channels allows tissues to co-ordinate responses to extracellular stimuli. Intercellular diffusion of inositol 1,4,5-trisphosphate, locally produced by focal stimulations, sustains the propagation of intercellular Ca2+ waves, by stimulating the release of intracellular Ca2+ in neighbouring cells. We previously demonstrated that in cultured articular chondrocytes and HIG-82 synovial cells, studied with digitial fluorescence video imaging, mechanical stimulation of a single cell induced intercellular Ca2+ waves dependent on the presence of gap junctions. In the absence of extracellular Ca2+ the propagating distance of the wave decreased significantly in HIG-82 cells, but appeared unaffected in chondrocytes. We now show that both cells types express connexin 43 and a similar functional coupling, thus suggesting that the different Ca2+ sensitivity of intercellular waves is not due to major differences in gap junction constituent ...
The various forms of joint loading have been found to differentially influence anatomical and molecular responses, which together are aimed to maintain the joint homeostasis. To elucidate these mechanisms of mechanotransduction, we review the roles of the distinct joint components, as well as numerous in vitro studies that have been performed to ... read more unravel chondrocyte responses in changing environments. The main signalling pathways in transduction of load signals are initiated by integrins sensing matrix deformation and by altered interstitial pH, influencing ion fluxes through membrane channels. Downstream signalling after static compression occurs mainly via the MAPK pathways of ERK1/2, SAPK (Jnk) and p38, which directly influence transcription factors of genes involved in cartilage breakdown. In contrast, cyclic compression and mild shear forces lead to membrane hyperpolarisation and subsequently stimulates cartilage matrix synthesis. These findings add further comprehension with ...
The development of biologically and mechanically competent hydrogels is a prerequisite in cartilage engineering. We recently demonstrated that a marine exopolysaccharide, GY785, stimulates the in vitro chondrogenesis of adipose stromal cells. In the present study, we thus hypothesized that enriching our silated hydroxypropyl methylcellulose hydrogel (Si-HPMC) with GY785 might offer new prospects in the development of scaffolds for cartilage regeneration. The interaction properties of GY785 with growth factors was tested by surface plasmon resonance (SPR). The biocompatibility of Si-HPMC/GY785 towards rabbit articular chondrocytes (RACs) and its ability to maintain and recover a chondrocytic phenotype were then evaluated in vitro by MTS assay, cell counting ...
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Authors: Filip, Anna , Bianchi, Arnaud , Mainard, Didier , Lacolley, Patrick , Magdalou, Jacques , Mercier, Nathalie Article Type: Research Article Abstract: OBJECTIVES: Chondrocytes hypertrophy is a physiological process observed in endochondral ossification during development until adolescence in human. It can also be observed during pathophysiological conditions such as osteoarthritis. Hypertrophic chondrocytes synthesise collagen X and express matrix metalloproteinase 13 and alkaline phosphatase. The cellular models available to study this process are either not convenient, they might lead to a rapid dedifferentiation of chondrocytes, or they are far from the physiological conditions. The objective of this study was to design an user-friendly 2D-primary cell culture of young articular chondrocytes of rat able to follow the terminal differentiation process. EXPERIMENTAL DESIGN: After …confluence, chondrocytes were cultured according to 4 differentiation protocols. Protocol 1 contained ...
TY - JOUR. T1 - Expression of runtB is modulated during chondrocyte differentiation. AU - Castagnola, Patrizio. AU - Gennari, Massimo. AU - Gaggero, Alessia. AU - Rossi, Fabio. AU - Daga, Antonio. AU - Corsetti, Maria Teresa. AU - Calabi, Franco. AU - Cancedda, Ranieri. PY - 1996/3/15. Y1 - 1996/3/15. N2 - The runt locus in Drosophila encodes a nuclear protein involved in embryo segmentation, sex determination/X dosage compensation, and neurogenesis. runt homologues have been identified in higher vertebrates. The encoded proteins share a domain of 128 amino acids called the runt domain. It has been reported that this domain mediates DNA binding and heterodimerization. Here, we analyze runtB expression during chondrocyte differentiation in vitro and in vivo. We have first isolated, from a chondrocyte library, a cDNA clone coding for a runtB chicken homologue and containing a complete open reading frame. The predicted protein product is 84% identical to the mouse PEBP2αB2 isoform. By RT-PCR ...
TY - JOUR. T1 - Increased TGF-β and BMP Levels and Improved Chondrocyte-Specific Marker Expression In Vitro under Cartilage-Specific Physiological Osmolarity. AU - Timur, Ufuk Tan. AU - Caron, Marjolein. AU - van den Akker, Guus. AU - van der Windt, Anna. AU - Visser, Jenny. AU - van Rhijn, Lodewijk. AU - Weinans, Harrie. AU - Welting, Tim. AU - Emans, Pieter. AU - Jahr, Holger. PY - 2019/2/2. Y1 - 2019/2/2. KW - chondrocyte. KW - osmolarity. KW - TGF- superfamily. KW - signalling. KW - collagen type II. KW - bone morphogenetic proteins. KW - HUMAN ARTICULAR CHONDROCYTES. KW - EXTRACELLULAR-MATRIX. KW - CELL-MEMBRANE. KW - BETA. KW - PHENOTYPE. KW - OSTEOARTHRITIS. KW - STIMULATION. KW - HYPERTROPHY. KW - ELEMENTS. KW - ENDOGLIN. U2 - 10.3390/ijms20040795. DO - 10.3390/ijms20040795. M3 - Article. VL - 20. JO - International Journal of Molecular Sciences. JF - International Journal of Molecular Sciences. SN - 1422-0067. IS - 4. M1 - 795. ER - ...
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In the following sections, we primarily review our own work on the expression, tissue distribution and function of integrins in selected in vitro models of articular chondrocytes and limb bud mesenchymal cells. The expression pattern of α1-, α3-, αv- and α5β1-integrins and their specific ligand binding were investigated in monolayer cultures of chondrocytes from 17-day-old mouse embryos using morphological and immunomorphological methods. After a 3-h culture period of chondrocytes from 17-day-old mouse embryos, numerous cells had already adhered in the form of a monolayer. After a 1-day culture period αv-, α3- and α5β1-integrins were observed on the chondrocytes. During the first 4 days, the number of the cells had increased, a matrix became perceptible. After a 5-day culture period, the flat fibroblast-like cells, often of bipolar shape, increased in number at the expense of the chondrocytes. © 2008 Springer-Verlag Berlin Heidelberg.. ...
Systems and methods for modifying the environment of target cell using genetically altered chondrocytes are provided. The genetically engineered chondrocytes can be used to express a therapeutic agent in a subject, including in an environment typically associated with chondrocytes and in an environment not typically associated with chondrocytes.
The first step in growing new cartilage is initiating chondrogenesis, or convincing the mesenchymal stem cells to differentiate into chondrocytes, which in turn generate the spongy matrix of collagen and sugars that cushions joints. One challenge in prompting this differentiation is that, despite the low density of adult chondrocytes in tissues, the actual formation of cartilage begins with cells in close proximity. "In typical hydrogels used in cartilage tissue engineering," Burdick said, "were spacing cells apart, so theyre losing that initial signal and interaction. Thats when we started thinking about cadherins, which are molecules that these cells use to interact with each other, particularly at the point they first become chondrocytes.". To simulate that environment, the researchers used a peptide sequence that mimics these cadherin interactions, which they bound to the hydrogels used to encapsulate the mesenchymal stem cells.. "While the direct link between cadherins and chondrogenesis ...
Conceição N, Viegas M, Fidalgo J, M. Cancela L. Development and characterization of Xl1, a Xenopus laevis chondrocyte-like cell culture. Mol Cell Biochem. 2013;373(1-2):41-51. doi:10.1007/s11010-012-1473-x ...
Osteoarthritis is a pain-associated progressive disease and pain mediators, such as opioid receptors, expressed in articular cartilage could represent novel therapeutic targets. Acute and chronic stages of OA indicate different metabolic abilities of the chondrocytes depending on inflammatory state. This study aimed to investigate the response of healthy and osteoarthritic chondrocytes and their expression and release of pain mediators in response to acute inflammation. Interleukin-1 beta (IL-1β) and lipopolysaccharide (LPS) were used to induce an acute inflammatory response in cultured equine chondrocytes harvested from healthy joints (HC) and osteoarthritic joints (OAC), the latter representing acute exacerbation of a chronic inflammatory state. Intracellular Ca2+ release was determined after exposure to serotonin (5-hydroxytryptamine (5-HT), glutamate or ATP. Protein expression levels of F- and G-actin, representing actin rearrangement, and opioid receptors were investigated. Glutamate ...
Genome editing is revolutionising biomedical research. It enables the introduction of targeted genomic sequence changes in isolated cells or whole organisms, and as such is an extremely powerful research tool with huge therapeutic potential. Cas9 is a nuclease guided by small RNAs (sgRNAs) to complementary sites in the genome. It induces double stranded breaks (DSBs) which repair through non-homologous end-joining (NHEJ), or in the presence of a suitable DNA template, by homology-directed repair (HDR). The CRISPR-Cas9 system thus enables highly specific gene editing giving it great potential for correction of genetic (particularly monogenic) diseases or for revolutionising current cell therapies, such as autologous chondrocyte implantation, where the cells could be edited in vitro before re-implantation back in the body. In the present proposal we will apply genome editing techniques recently developed with our collaborators (1) with the aim of creating modified populations of human chondrocytes ...
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Objective(s): Osteoarthritis (OA), characterized by degradation of articular cartilage, is a leading cause of disability. As the only cell type present in cartilage, chondrocytes play curial roles in the progression of OA. In our study, we aimed to explore the roles of miR-23b in the lipopolysaccharide (LPS)-induced inflammatory injury. Materials and Methods: LPS-induced cell injury of ATDC5 cells was evaluated by the loss of cell viability, enhancement of cell apoptosis, alteration of apoptosis-associated proteins, and release of inflammatory cytokines. Then, miR-23b level after LPS treatment was assessed by qRT-PCR. Next, the effects of aberrantly expressed miR-23b on the LPS-induced inflammatory injury were explored. The possible target genes of miR-23b were virtually screened by informatics and verified by luciferase assay. Subsequently, whether miR-23b functioned through regulating the target gene was validated. The involved signaling pathways were investigated finally.Results: Cell viability was
Articular cartilage is a tissue designed to withstand compression during joint movement and, in vivo, is subjected to a wide range of mechanical loading forces. Mechanosensitivity has been demonstrated to influence chondrocyte metabolism and cartilag
The past few months have brought unprecedented challenges and stress to all Americans and others around the globe. At Vericel Corporation, our thoughts are with those affected by COVID-19 and we are especially thankful to all healthcare workers for their critical efforts during this challenging time.. Our mission to help treat patients with serious knee cartilage injuries has remained steadfast throughout the pandemic. Vericel continues to manufacture MACI® (autologous cultured chondrocytes on porcine collagen membrane) for those in need of care. MACI is aseptically manufactured in an FDA licensed facility using sterile materials and reagents. Our clean rooms maintain the highest cleanliness standards through automated environmental controls and monitoring. A rotational cleaning program of the room and all work surfaces includes quality control testing and release of the rooms. All Vericel manufacturing personnel wear pre-sterilized gowning materials. Laminar flow biosafety cabinets located ...
Zamani, S. and Dehghani, Leila. and Drummen, G. and esfandiari, Ebrahim. and Abutorabi, Roshanak. and Rabbani, Hossein. and Tahani, Soheil. and Hashemi beni, Batool. (2016) Comparison of cartilage specific markers in articular and differentiated chondrocytes in pellet system. Biointerface Research in Applied Chemistry, 6 (6). Zamani, Saeed. and Hashemibeni, Batool. and esfandiari, Ebrahim. and Kabiri, Azadeh. and Rabbani, Hossein. and Abutorabi, Roshanak. (2014) Assessment of TGF-β3 on production of aggrecan by human articular chondrocytes in pellet culture system. Advanced Biomedical Research. ...
Differentiation of bmMPCs into chondrocytes.bmMPCs, pretreated with 5 mmol/L (control glucose) or 25 mmol/L (high glucose; HG) glucose for 7 days, were cultured
The potent aggrecanase ADAMTS-5 is constitutively secreted by chondrocytes, but it is rapidly endocytosed in normal cartilage via the cell surface endocytic receptor LRP1. Therefore it is difficult to detect the total ADAMTS-5 activity produced. In this study, we isolated a monoclonal anti-ADAMTS-5 antibody 1B7 that blocks LRP1-mediated internalization without affecting the aggrecanolytic activity. Addition of 1B7 to cultured human chondrocytes revealed the full aggrecanolytic activity of ADAMTS-5 generated by the cells. 1B7 is a useful tool to estimate the ADAMTS-5 activity and to identify its potential roles in the tissues.
Université de Liège - ULg , Département des sciences de la motricité , Unité de recherche sur los et le cartillage (U.R.O.C.) ,] ...
Principal Investigator:ITO Akira, Project Period (FY):1997 - 1998, Research Category:Grant-in-Aid for Scientific Research (C), Section:一般, Research Field:老化(加齢)
CKX-CCSW confluency checker software enables users to accurately estimate the proper timing for cell passage while minimizing human error.