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Hari A Reddi - One of the best experts on this subject based on the ideXlab platform.
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modulation of Superficial Zone protein lubricin prg4 by kartogenin and transforming growth factor β1 in surface Zone chondrocytes in bovine articular cartilage
Cartilage, 2016Co-Authors: Kazumasa Miyatake, Kenjiro Iwasa, Sean M. Mcnary, Gordon Peng, Hari A ReddiAbstract:ObjectiveSuperficial Zone protein (SZP)/lubricin/PRG4 functions as a boundary lubricant in articular cartilage to decrease friction and wear. As articular cartilage lubrication is critical for normal joint function, the accumulation of SZP at the surface of cartilage is important for joint homeostasis. Recently, a heterocyclic compound called kartogenin (KGN) was found to induce chondrogenic differentiation and enhance mRNA expression of lubricin. The objective of this study was to determine whether KGN can stimulate synthesis of SZP in Superficial Zone, articular chondrocytes.DesignWe investigated the effects of KGN and transforming growth factor-β1 (TGF-β1) on articular cartilage and synovium of the bovine knee joint by evaluating SZP secretion by enzyme-linked immunosorbent assay analysis. Monolayer, micromass, and explant cultures of articular cartilage, and monolayer culture of synoviocytes, were treated with KGN. SZP accumulation in the medium was evaluated and mRNA expression was measured through q...
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Superficial Zone extracellular matrix extracts enhance boundary lubrication of self assembled articular cartilage
Cartilage, 2016Co-Authors: Gordon Peng, Sean M. Mcnary, Kyriacos A Athanasiou, Hari A ReddiAbstract:Previous work has shown that increasing the production of boundary lubricant, Superficial Zone protein (SZP), did not reduce the friction coefficient of self-assembled articular cartilage constructs and was possibly due to poor retention of the lubricant. The aim of this investigation was to reduce the friction coefficient of self-assembled articular cartilage constructs through enhancing SZP retention by the exogenous addition of extracellular matrix (ECM) extracted from the Superficial Zone of native articular cartilage.Superficial Zone cartilage was shaved from juvenile bovine femoral condyles using a dermatome, minced finely with razor blades, extracted with 4 M guanidine-hydrochloride, buffer exchanged with culture medium, and added directly to the culture medium of self-assembled articular cartilage constructs at low (10 µg/mL) and high (100 µg/mL) concentrations for 4 weeks. Biochemical and biomechanical properties were determined at the conclusion of 4 weeks culture.ECM treatment increased compressive and tensile stiffness of self-assembled articular cartilage constructs and decreased the friction coefficient. Glycosaminoglycan content decreased and collagen content increased significantly in self-assembled constructs by the ECM treatment.Friction coefficients of self-assembled articular cartilage constructs were reduced by adding extracted Superficial Zone ECM into the culture medium of self-assembled articular cartilage constructs.
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regulation of lubricin Superficial Zone protein by wnt signalling in bovine synoviocytes
Journal of Tissue Engineering and Regenerative Medicine, 2016Co-Authors: Atsuyuki Inui, Takashi Iwakura, Hari A ReddiAbstract:Lubricin, homologous to Superficial Zone protein (SZP), functions as a boundary lubricant in articular cartilage and plays an essential role in the maintenance of joint function and homeostasis. Wnt signalling plays a key role in joint development, including synovial joint formation, and several Wnt proteins are expressed in the synovium and articular cartilage in arthritis. The aim of this study was to determine the role of Wnt signalling on SZP accumulation in synoviocytes. Isolated synoviocytes from bovine knee joints were cultured with Wnt proteins (Wnt-3a and Wnt-5a) and antagonists or agonists of the Wnt–β-catenin pathway or Wnt–Ca2+ pathway in serum-free chemically defined medium. SZP accumulation in the culture medium was determined by enzyme-linked immunosorbent assay. Wnt-3a suppressed SZP accumulation via a Wnt–β-catenin-dependent pathway. In contrast, Wnt-5a stimulated SZP accumulation via a β-catenin independent pathway. The present investigation provides novel insights into the role of the Wnt signalling pathways in SZP accumulation in synoviocytes and their roles in the homeostasis of normal joints. Copyright © 2013 John Wiley & Sons, Ltd.
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the distribution of Superficial Zone protein szp lubricin prg4 and boundary mode frictional properties of the bovine diarthrodial joint
Journal of Biomechanics, 2015Co-Authors: Gordon Peng, Sean M. Mcnary, Kyriacos A Athanasiou, Hari A ReddiAbstract:The diarthrodial, knee joint is a remarkably efficient bearing system; articulating cartilage surfaces provide nearly frictionless performance with minimal wear. The low friction properties of the cartilage surfaces are due in part to the boundary lubricant, Superficial Zone protein (SZP); also known as lubricin or proteoglycan 4 (PRG4). In previous work, SZP localization and cartilage friction were examined across the femoral condyles. Studies in the literature have also individually investigated the other tissues that comprise the human knee and four-legged animal stifle joint, such as the meniscus or patella. However, comparisons between individual studies are limited due to the variable testing conditions employed. Friction is a system property that is dependent on the opposing articulating surface, entraining speed, and loading. A cross-comparison of the frictional properties and SZP localization across the knee/stifle joint tissues utilizing a common testing configuration is therefore needed. The objective of this investigation was to determine the friction coefficient and SZP localization of the tissues comprising the three compartments of the bovine stifle joint: patella, patellofemoral groove, femoral condyles, meniscus, tibial plateau, and anterior cruciate ligament. The boundary mode coefficient of friction was greater in tissues of the patellofemoral compartment than the lateral and medial tibiofemoral compartments. SZP immunolocalization followed this trend with reduced depth of staining and intensity in the patella and patellofemoral groove compared to the femoral condyles and tibial plateau. These results illustrate the important role of SZP in reducing friction in the tissues and compartments of the knee/stifle joint.
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stimulation of Superficial Zone protein lubricin prg4 by transforming growth factor β in Superficial Zone articular chondrocytes and modulation by glycosaminoglycans
Tissue Engineering Part A, 2015Co-Authors: Araceli Cuellar, Hari A ReddiAbstract:Superficial Zone protein (SZP), also known as lubricin and proteoglycan 4 (PRG4), plays an important role in the boundary lubrication of articular cartilage and is regulated by transforming growth factor (TGF)-β. Here, we evaluate the role of cell surface glycosaminoglycans (GAGs) during TGF-β1 stimulation of SZP/lubricin/PRG4 in Superficial Zone articular chondrocytes. We utilized primary monolayer Superficial Zone articular chondrocyte cultures and treated them with various concentrations of TGF-β1, in the presence or absence of heparan sulfate (HS), heparin, and chondroitin sulfate (CS). The cell surface GAGs were removed by pretreatment with either heparinase I or chondroitinase-ABC before TGF-β1 stimulation. Accumulation of SZP/lubricin/PRG4 in the culture medium in response to stimulation with TGF-β1 and various exogenous GAGs was demonstrated by immunoblotting and quantitated by enzyme-linked immunosorbent assay. We show that TGF-β1 and exogenous HS enhanced SZP accumulation of Superficial Zone cho...
A. Hari Reddi - One of the best experts on this subject based on the ideXlab platform.
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Optimization of Methods for Articular Cartilage Surface Tissue Engineering: Cell Density and Transforming Growth Factor Beta Are Critical for Self-Assembly and Lubricin Secretion
Tissue Engineering Part C-methods, 2017Co-Authors: Kenjiro Iwasa, A. Hari ReddiAbstract:Objective: Lubricin/Superficial Zone protein (SZP)/proteoglycan4 (PRG4) plays an important role in boundary lubrication in articular cartilage. Lubricin is secreted by Superficial Zone chondrocytes and synoviocytes of the synovium. The specific objective of this investigation is to optimize the methods for tissue engineering of articular cartilage surface. The aim of this study is to investigate the effect of cell density on the self-assembly of Superficial Zone chondrocytes and lubricin secretion as a functional assessment. Design: Superficial Zone chondrocytes were cultivated as a monolayer at low, medium, and high densities. Chondrocytes at the three different densities were treated with transforming growth factor beta (TGF-β)1 twice a week or daily, and the accumulated lubricin in the culture medium was analyzed by immunoblots and quantitated by enzyme-linked immunosorbent assay (ELISA). Results: Cell numbers in low and medium densities were increased by TGF-β1; whereas cell numbers in high-density ce...
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Modulation of Superficial Zone Protein/Lubricin/PRG4 by Kartogenin and Transforming Growth Factor-β1 in Surface Zone Chondrocytes in Bovine Articular Cartilage.
Cartilage, 2016Co-Authors: Kazumasa Miyatake, Kenjiro Iwasa, Sean M. Mcnary, Gordon Peng, A. Hari ReddiAbstract:ObjectiveSuperficial Zone protein (SZP)/lubricin/PRG4 functions as a boundary lubricant in articular cartilage to decrease friction and wear. As articular cartilage lubrication is critical for normal joint function, the accumulation of SZP at the surface of cartilage is important for joint homeostasis. Recently, a heterocyclic compound called kartogenin (KGN) was found to induce chondrogenic differentiation and enhance mRNA expression of lubricin. The objective of this study was to determine whether KGN can stimulate synthesis of SZP in Superficial Zone, articular chondrocytes.DesignWe investigated the effects of KGN and transforming growth factor-β1 (TGF-β1) on articular cartilage and synovium of the bovine knee joint by evaluating SZP secretion by enzyme-linked immunosorbent assay analysis. Monolayer, micromass, and explant cultures of articular cartilage, and monolayer culture of synoviocytes, were treated with KGN. SZP accumulation in the medium was evaluated and mRNA expression was measured through q...
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The distribution of Superficial Zone protein (SZP)/lubricin/PRG4 and boundary mode frictional properties of the bovine diarthrodial joint.
Journal of Biomechanics, 2015Co-Authors: Gordon Peng, Sean M. Mcnary, Kyriacos A Athanasiou, A. Hari ReddiAbstract:The diarthrodial, knee joint is a remarkably efficient bearing system; articulating cartilage surfaces provide nearly frictionless performance with minimal wear. The low friction properties of the cartilage surfaces are due in part to the boundary lubricant, Superficial Zone protein (SZP); also known as lubricin or proteoglycan 4 (PRG4). In previous work, SZP localization and cartilage friction were examined across the femoral condyles. Studies in the literature have also individually investigated the other tissues that comprise the human knee and four-legged animal stifle joint, such as the meniscus or patella. However, comparisons between individual studies are limited due to the variable testing conditions employed. Friction is a system property that is dependent on the opposing articulating surface, entraining speed, and loading. A cross-comparison of the frictional properties and SZP localization across the knee/stifle joint tissues utilizing a common testing configuration is therefore needed. The objective of this investigation was to determine the friction coefficient and SZP localization of the tissues comprising the three compartments of the bovine stifle joint: patella, patellofemoral groove, femoral condyles, meniscus, tibial plateau, and anterior cruciate ligament. The boundary mode coefficient of friction was greater in tissues of the patellofemoral compartment than the lateral and medial tibiofemoral compartments. SZP immunolocalization followed this trend with reduced depth of staining and intensity in the patella and patellofemoral groove compared to the femoral condyles and tibial plateau. These results illustrate the important role of SZP in reducing friction in the tissues and compartments of the knee/stifle joint.
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Stimulation of Superficial Zone Protein/Lubricin/PRG4 by Transforming Growth Factor-β in Superficial Zone Articular Chondrocytes and Modulation by Glycosaminoglycans
Tissue Engineering Part A, 2014Co-Authors: Araceli Cuellar, A. Hari ReddiAbstract:Superficial Zone protein (SZP), also known as lubricin and proteoglycan 4 (PRG4), plays an important role in the boundary lubrication of articular cartilage and is regulated by transforming growth factor (TGF)-β. Here, we evaluate the role of cell surface glycosaminoglycans (GAGs) during TGF-β1 stimulation of SZP/lubricin/PRG4 in Superficial Zone articular chondrocytes. We utilized primary monolayer Superficial Zone articular chondrocyte cultures and treated them with various concentrations of TGF-β1, in the presence or absence of heparan sulfate (HS), heparin, and chondroitin sulfate (CS). The cell surface GAGs were removed by pretreatment with either heparinase I or chondroitinase-ABC before TGF-β1 stimulation. Accumulation of SZP/lubricin/PRG4 in the culture medium in response to stimulation with TGF-β1 and various exogenous GAGs was demonstrated by immunoblotting and quantitated by enzyme-linked immunosorbent assay. We show that TGF-β1 and exogenous HS enhanced SZP accumulation of Superficial Zone cho...
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Induction of chondrogenesis and expression of Superficial Zone protein in synovial explants with TGF-β1 and BMP-7.
Tissue Engineering Part A, 2013Co-Authors: Takashi Iwakura, Ryosuke Sakata, A. Hari ReddiAbstract:Superficial Zone protein (SZP) functions as a boundary lubricant in articular cartilage and decreases the coefficient of friction. As lubrication of articular cartilage is critical for normal joint function, the ability to secrete SZP at the surface of tissue-engineered cartilage is a prerequisite for optimal lubrication. Synovium-derived mesenchymal stem cells (MSCs) are thought to be an attractive cell source for cartilage regeneration. However, optimization of a three-dimensional environment is necessary for tissue engineering. In this study, we investigated whether synovial explants, which would preserve the physiologic microenvironment for MSCs therein, have the potential of SZP secretion after chondrogenic differentiation by treatment with transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7). Immunostaining and enzyme-linked immunosorbent assay analysis demonstrated that synovial explants can synthesize and secrete SZP following chondrogenic differentiation in response to ...
Thomas M Schmid - One of the best experts on this subject based on the ideXlab platform.
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Induction of Superficial Zone protein (SZP)/lubricin/PRG 4 in muscle-derived mesenchymal stem/progenitor cells by transforming growth factor-β1 and bone morphogenetic protein-7.
Arthritis Research & Therapy, 2012Co-Authors: Jose A Andrades, Thomas M Schmid, S C K M Motaung, Silvia Claros, Jose Becerra, Pedro Jiménez-palomo, José M. López-puerta, A. Hari ReddiAbstract:Introduction Articular cartilage (AC) is an avascular tissue with precise polarity and organization. The three distinct Zones are: surface, middle and deep. The production and accumulation of the Superficial Zone protein (SZP), also known as lubricin, by the surface Zone is a characteristic feature of AC. To date, there is a wealth of evidence showing differentiation of AC from mesenchymal stem cells. Most studies that described chondrogenic differentiation did not focus on AC with characteristic surface marker SZP/lubricin. The present investigation was initiated to determine the induction of SZP/lubricin in skeletal muscle-derived mesenchymal stem/progenitor cells (MDMSCs) by transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7).
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induction of Superficial Zone protein szp lubricin prg 4 in muscle derived mesenchymal stem progenitor cells by transforming growth factor β1 and bone morphogenetic protein 7
Arthritis Research & Therapy, 2012Co-Authors: Jose A Andrades, Hari A Reddi, Thomas M Schmid, S C K M Motaung, Pedro Jimenezpalomo, Silvia Claros, Jose M Lopezpuerta, Jose BecerraAbstract:Introduction Articular cartilage (AC) is an avascular tissue with precise polarity and organization. The three distinct Zones are: surface, middle and deep. The production and accumulation of the Superficial Zone protein (SZP), also known as lubricin, by the surface Zone is a characteristic feature of AC. To date, there is a wealth of evidence showing differentiation of AC from mesenchymal stem cells. Most studies that described chondrogenic differentiation did not focus on AC with characteristic surface marker SZP/lubricin. The present investigation was initiated to determine the induction of SZP/lubricin in skeletal muscle-derived mesenchymal stem/progenitor cells (MDMSCs) by transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7).
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increased friction coefficient and Superficial Zone protein expression in patients with advanced osteoarthritis
Arthritis & Rheumatism, 2010Co-Authors: A. Hari Reddi, Thomas M Schmid, K Komvopoulos, P E Di CesareAbstract:Objective To quantify the concentration of Superficial Zone protein (SZP) in the articular cartilage and synovial fluid of patients with advanced osteoarthritis (OA) and to further correlate the SZP content with the friction coefficient, OA severity, and levels of proinflammatory cytokines. Methods Samples of articular cartilage and synovial fluid were obtained from patients undergoing elective total knee replacement surgery. Additional normal samples were obtained from donated body program and tissue bank sources. Regional SZP expression in cartilage obtained from the femoral condyles was quantified by enzyme-linked immunosorbent assay (ELISA) and visualized by immunohistochemistry. Friction coefficient measurements of cartilage plugs slid in the boundary lubrication system were obtained. OA severity was graded using histochemical analyses. The concentrations of SZP and proinflammatory cytokines in synovial fluid were determined by ELISA. Results A pattern of SZP localization in knee cartilage was identified, with load-bearing regions exhibiting high SZP expression. SZP expression patterns were correlated with friction coefficient and OA severity; however, SZP expression was observed in all samples at the articular surface, regardless of OA severity. SZP expression and aspirate volume of synovial fluid were higher in OA patients than in normal controls. Expression of cytokines was elevated in the synovial fluid of some patients. Conclusion Our findings indicate a mechanochemical coupling in which physical forces regulate OA severity and joint lubrication. The findings of this study also suggest that SZP may be ineffective in reducing joint friction in the boundary lubrication mode at an advanced stage of OA, where other mechanisms may dominate the observed tribological behavior.
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mechanotransduction of bovine articular cartilage Superficial Zone protein by transforming growth factor β signaling
Arthritis & Rheumatism, 2007Co-Authors: Afshin Khalafi, Thomas M Schmid, K Komvopoulos, Hari A ReddiAbstract:Objective Mechanical signals are key determinants in tissue morphogenesis, maintenance, and restoration strategies in regenerative medicine, although molecular mechanisms of mechanotransduction remain to be elucidated. This study was undertaken to investigate the mechanotransduction process of expression of Superficial Zone protein (SZP), a critical joint lubricant. Methods Regional expression of SZP was first quantified in cartilage obtained from the femoral condyles of immature bovines, using immunoblotting, and visualized by immunohistochemistry. Contact pressure mapping in whole joints was accomplished using pressure-sensitive film and a load application system for joint testing. Friction measurements on cartilage plugs were acquired under boundary lubrication conditions using a pin-on-disk tribometer modified for reciprocating sliding. Direct mechanical stimulation by shear loading of articular cartilage explants was performed with and without inhibition of transforming growth factor β (TGFβ) signaling, and SZP content in media was quantified by enzyme-linked immunosorbent assay. Results An unexpected pattern of SZP localization in knee cartilage was initially identified, with anterior regions exhibiting high levels of SZP expression. Regional SZP patterns were regulated by mechanical signals and correlated with tribological behavior. Direct relationships were demonstrated between high levels of SZP expression, maximum contact pressures, and low friction coefficients. Levels of SZP expression and accumulation were increased by applying shear stress, depending on location within the knee, and were decreased to control levels with the use of a specific inhibitor of TGFβ receptor type I kinase and subsequent phospho-Smad2/3 activity. Conclusion These findings indicate a new role for TGFβ signaling in the mechanism of cellular mechanotransduction that is especially significant for joint lubrication.
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increased accumulation of Superficial Zone protein szp in articular cartilage in response to bone morphogenetic protein 7 and growth factors
Journal of Orthopaedic Research, 2007Co-Authors: Afshin Khalafi, Thomas M Schmid, Hari A ReddiAbstract:The purpose of this study was to investigate the role of bone morphogenetic proteins (BMPs), such as BMP-7, growth factors, and cytokines, in the accumulation of Superficial Zone protein (SZP) in bovine articular cartilage. Calf Superficial articular cartilage discs and chondrocytes were obtained for explant and monolayer culture systems, respectively. Dose- and time-dependent actions of BMP-7 on SZP accumulation were investigated in both explant and monolayer culture systems. In addition, actions of various morphogens and growth factors [BMP-2, BMP-4, fibroblast growth factor 2 (FGF-2), insulin-like growth factor 1 (IGF-1), platelet-derived growth factor (PDGF), and transforming growth factor β (TGF-β1)], and cytokines [interleukin (IL)-1α, IL-1β, and tumor necrosis factor (TNF-α)] alone, and in combination with BMP-7, on SZP accumulation were investigated in monolayer culture systems. SZP accumulation was quantified in both the cartilage and the medium using SDS-PAGE and subsequent immunoblotting. In both explant and monolayer cultures, BMP-7 increased SZP accumulation in a dose- and time-dependent fashion (p < 0.05). Furthermore, SZP accumulation was significantly increased in monolayer cultures by FGF-2, IGF-1, PDGF, and TGF-β1 (p < 0.05). Both IL-1α and TNF-α significantly reduced SZP accumulation (p < 0.05). The inhibition of SZP accumulation by TNF-α was partially alleviated by concurrent treatment with BMP-7. The results of this investigation provide novel insights into the role of morphogens, especially BMP-7, growth factors, and cytokines in the accumulation of SZP in articular cartilage. This information has clinical implications because stimulation of SZP may ameliorate the pathology of joint function in arthritis. Furthermore, tissue engineering approaches to articular cartilage may depend on the optimal synthesis and assembly of SZP in the Superficial Zone to ensure functional tissue architecture. © 2006 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 2007
Sean M. Mcnary - One of the best experts on this subject based on the ideXlab platform.
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modulation of Superficial Zone protein lubricin prg4 by kartogenin and transforming growth factor β1 in surface Zone chondrocytes in bovine articular cartilage
Cartilage, 2016Co-Authors: Kazumasa Miyatake, Kenjiro Iwasa, Sean M. Mcnary, Gordon Peng, Hari A ReddiAbstract:ObjectiveSuperficial Zone protein (SZP)/lubricin/PRG4 functions as a boundary lubricant in articular cartilage to decrease friction and wear. As articular cartilage lubrication is critical for normal joint function, the accumulation of SZP at the surface of cartilage is important for joint homeostasis. Recently, a heterocyclic compound called kartogenin (KGN) was found to induce chondrogenic differentiation and enhance mRNA expression of lubricin. The objective of this study was to determine whether KGN can stimulate synthesis of SZP in Superficial Zone, articular chondrocytes.DesignWe investigated the effects of KGN and transforming growth factor-β1 (TGF-β1) on articular cartilage and synovium of the bovine knee joint by evaluating SZP secretion by enzyme-linked immunosorbent assay analysis. Monolayer, micromass, and explant cultures of articular cartilage, and monolayer culture of synoviocytes, were treated with KGN. SZP accumulation in the medium was evaluated and mRNA expression was measured through q...
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Superficial Zone extracellular matrix extracts enhance boundary lubrication of self assembled articular cartilage
Cartilage, 2016Co-Authors: Gordon Peng, Sean M. Mcnary, Kyriacos A Athanasiou, Hari A ReddiAbstract:Previous work has shown that increasing the production of boundary lubricant, Superficial Zone protein (SZP), did not reduce the friction coefficient of self-assembled articular cartilage constructs and was possibly due to poor retention of the lubricant. The aim of this investigation was to reduce the friction coefficient of self-assembled articular cartilage constructs through enhancing SZP retention by the exogenous addition of extracellular matrix (ECM) extracted from the Superficial Zone of native articular cartilage.Superficial Zone cartilage was shaved from juvenile bovine femoral condyles using a dermatome, minced finely with razor blades, extracted with 4 M guanidine-hydrochloride, buffer exchanged with culture medium, and added directly to the culture medium of self-assembled articular cartilage constructs at low (10 µg/mL) and high (100 µg/mL) concentrations for 4 weeks. Biochemical and biomechanical properties were determined at the conclusion of 4 weeks culture.ECM treatment increased compressive and tensile stiffness of self-assembled articular cartilage constructs and decreased the friction coefficient. Glycosaminoglycan content decreased and collagen content increased significantly in self-assembled constructs by the ECM treatment.Friction coefficients of self-assembled articular cartilage constructs were reduced by adding extracted Superficial Zone ECM into the culture medium of self-assembled articular cartilage constructs.
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Modulation of Superficial Zone Protein/Lubricin/PRG4 by Kartogenin and Transforming Growth Factor-β1 in Surface Zone Chondrocytes in Bovine Articular Cartilage.
Cartilage, 2016Co-Authors: Kazumasa Miyatake, Kenjiro Iwasa, Sean M. Mcnary, Gordon Peng, A. Hari ReddiAbstract:ObjectiveSuperficial Zone protein (SZP)/lubricin/PRG4 functions as a boundary lubricant in articular cartilage to decrease friction and wear. As articular cartilage lubrication is critical for normal joint function, the accumulation of SZP at the surface of cartilage is important for joint homeostasis. Recently, a heterocyclic compound called kartogenin (KGN) was found to induce chondrogenic differentiation and enhance mRNA expression of lubricin. The objective of this study was to determine whether KGN can stimulate synthesis of SZP in Superficial Zone, articular chondrocytes.DesignWe investigated the effects of KGN and transforming growth factor-β1 (TGF-β1) on articular cartilage and synovium of the bovine knee joint by evaluating SZP secretion by enzyme-linked immunosorbent assay analysis. Monolayer, micromass, and explant cultures of articular cartilage, and monolayer culture of synoviocytes, were treated with KGN. SZP accumulation in the medium was evaluated and mRNA expression was measured through q...
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the distribution of Superficial Zone protein szp lubricin prg4 and boundary mode frictional properties of the bovine diarthrodial joint
Journal of Biomechanics, 2015Co-Authors: Gordon Peng, Sean M. Mcnary, Kyriacos A Athanasiou, Hari A ReddiAbstract:The diarthrodial, knee joint is a remarkably efficient bearing system; articulating cartilage surfaces provide nearly frictionless performance with minimal wear. The low friction properties of the cartilage surfaces are due in part to the boundary lubricant, Superficial Zone protein (SZP); also known as lubricin or proteoglycan 4 (PRG4). In previous work, SZP localization and cartilage friction were examined across the femoral condyles. Studies in the literature have also individually investigated the other tissues that comprise the human knee and four-legged animal stifle joint, such as the meniscus or patella. However, comparisons between individual studies are limited due to the variable testing conditions employed. Friction is a system property that is dependent on the opposing articulating surface, entraining speed, and loading. A cross-comparison of the frictional properties and SZP localization across the knee/stifle joint tissues utilizing a common testing configuration is therefore needed. The objective of this investigation was to determine the friction coefficient and SZP localization of the tissues comprising the three compartments of the bovine stifle joint: patella, patellofemoral groove, femoral condyles, meniscus, tibial plateau, and anterior cruciate ligament. The boundary mode coefficient of friction was greater in tissues of the patellofemoral compartment than the lateral and medial tibiofemoral compartments. SZP immunolocalization followed this trend with reduced depth of staining and intensity in the patella and patellofemoral groove compared to the femoral condyles and tibial plateau. These results illustrate the important role of SZP in reducing friction in the tissues and compartments of the knee/stifle joint.
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The distribution of Superficial Zone protein (SZP)/lubricin/PRG4 and boundary mode frictional properties of the bovine diarthrodial joint.
Journal of Biomechanics, 2015Co-Authors: Gordon Peng, Sean M. Mcnary, Kyriacos A Athanasiou, A. Hari ReddiAbstract:The diarthrodial, knee joint is a remarkably efficient bearing system; articulating cartilage surfaces provide nearly frictionless performance with minimal wear. The low friction properties of the cartilage surfaces are due in part to the boundary lubricant, Superficial Zone protein (SZP); also known as lubricin or proteoglycan 4 (PRG4). In previous work, SZP localization and cartilage friction were examined across the femoral condyles. Studies in the literature have also individually investigated the other tissues that comprise the human knee and four-legged animal stifle joint, such as the meniscus or patella. However, comparisons between individual studies are limited due to the variable testing conditions employed. Friction is a system property that is dependent on the opposing articulating surface, entraining speed, and loading. A cross-comparison of the frictional properties and SZP localization across the knee/stifle joint tissues utilizing a common testing configuration is therefore needed. The objective of this investigation was to determine the friction coefficient and SZP localization of the tissues comprising the three compartments of the bovine stifle joint: patella, patellofemoral groove, femoral condyles, meniscus, tibial plateau, and anterior cruciate ligament. The boundary mode coefficient of friction was greater in tissues of the patellofemoral compartment than the lateral and medial tibiofemoral compartments. SZP immunolocalization followed this trend with reduced depth of staining and intensity in the patella and patellofemoral groove compared to the femoral condyles and tibial plateau. These results illustrate the important role of SZP in reducing friction in the tissues and compartments of the knee/stifle joint.
R A Kandel - One of the best experts on this subject based on the ideXlab platform.
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yap taz regulates the expression of proteoglycan 4 and tenascin c in Superficial Zone chondrocytes
European Cells & Materials, 2020Co-Authors: Elizabeth Delve, Vivian Co, Suresh C Regmi, J Parreno, Tannin A Schmidt, R A KandelAbstract:: The roles of cell division control protein 42 homologue (CDC42) and actin polymerisation in regulating the phenotype of Superficial-Zone chondrocytes (SZCs) have been demonstrated in vitro; however, the signalling pathway(s) downstream have yet to be fully elucidated. The study hypothesis was that Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) act downstream to regulate proteoglycan 4 (PRG4) and tenascin C (TNC). Bovine SZCs grown in monolayer were treated with ML141 (CDC42 inhibitor) or the actin depolymerising agents, latrunculin B and cytochalasin D, to determine the effect on YAP/TAZ. Verteporfin (YAP/TAZ inhibitor) and YAP/TAZ siRNA-mediated knockdown were used to determine their role in regulating PRG4 and TNC. ML141 treatment reduced total YAP/TAZ protein, nuclear TAZ levels and the YAP/TAZ target gene, connective tissue growth factor (CTGF) mRNA levels. Latrunculin B decreased nuclear TAZ, while cytochalasin D treatment trended towards increased nuclear TAZ (p = 0.06), correlating with decreased and increased CTGF mRNA levels, respectively. Verteporfin treatment decreased PRG4 and TNC expression, with no effect on actin polymerisation. siRNA-mediated knockdown of YAP/TAZ revealed that PRG4 was regulated by YAP/TAZ while TNC was regulated by TAZ only. As cytochalasin D can activate myocardin-related transcription factor-A (MRTF-A), siRNA-mediated knockdown was performed to determine the role of MRTF-A in regulating YAP/TAZ. Although nuclear TAZ decreased, no significant changes in total protein levels were observed. Findings suggested that CDC42 and actin polymerisation regulated SZCs through multiple actin-regulated pathways. Understanding the regulation of these chondroprotective molecules may have important implications for prevention/treatment of osteoarthritis.
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YAP/TAZ regulates the expression of proteoglycan 4 and tenascin C in Superficial-Zone chondrocytes.
European Cells & Materials, 2020Co-Authors: Elizabeth Delve, Vivian Co, Suresh C Regmi, J Parreno, Tannin A Schmidt, R A KandelAbstract:: The roles of cell division control protein 42 homologue (CDC42) and actin polymerisation in regulating the phenotype of Superficial-Zone chondrocytes (SZCs) have been demonstrated in vitro; however, the signalling pathway(s) downstream have yet to be fully elucidated. The study hypothesis was that Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) act downstream to regulate proteoglycan 4 (PRG4) and tenascin C (TNC). Bovine SZCs grown in monolayer were treated with ML141 (CDC42 inhibitor) or the actin depolymerising agents, latrunculin B and cytochalasin D, to determine the effect on YAP/TAZ. Verteporfin (YAP/TAZ inhibitor) and YAP/TAZ siRNA-mediated knockdown were used to determine their role in regulating PRG4 and TNC. ML141 treatment reduced total YAP/TAZ protein, nuclear TAZ levels and the YAP/TAZ target gene, connective tissue growth factor (CTGF) mRNA levels. Latrunculin B decreased nuclear TAZ, while cytochalasin D treatment trended towards increased nuclear TAZ (p = 0.06), correlating with decreased and increased CTGF mRNA levels, respectively. Verteporfin treatment decreased PRG4 and TNC expression, with no effect on actin polymerisation. siRNA-mediated knockdown of YAP/TAZ revealed that PRG4 was regulated by YAP/TAZ while TNC was regulated by TAZ only. As cytochalasin D can activate myocardin-related transcription factor-A (MRTF-A), siRNA-mediated knockdown was performed to determine the role of MRTF-A in regulating YAP/TAZ. Although nuclear TAZ decreased, no significant changes in total protein levels were observed. Findings suggested that CDC42 and actin polymerisation regulated SZCs through multiple actin-regulated pathways. Understanding the regulation of these chondroprotective molecules may have important implications for prevention/treatment of osteoarthritis.
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cdc42 regulates the expression of Superficial Zone molecules in part through the actin cytoskeleton and myocardin related transcription factor a
Journal of Orthopaedic Research, 2018Co-Authors: Elizabeth Delve, Vivian Co, J Parreno, Pohan Wu, Jasmine Chong, Matteo Di Scipio, R A KandelAbstract:: Osteoarthritis (OA) is a degenerative disease that initially manifests as loss of the Superficial Zone (SZ) of articular cartilage. SZ chondrocytes (SZC) differ in morphology from other chondrocytes as they are elongated and oriented parallel to the tissue surface. Proteoglycan 4 (PRG4) and tenascin C (TNC) are molecules expressed by SZC, which have been shown to be chondroprotective. Identification of the signalling pathway(s) regulating expression of SZ molecules may lead to a therapeutic target that can be used to delay or prevent the onset of OA. The hypothesis of this study is that expression of SZ molecules are regulated in part, by the CDC42-actin-myocardin-related transcription factor-A (MRTF-A) signaling pathway. SZC from bovine metacarpal-phalangeal joints were isolated and grown in monolayer culture. Each target in the CDC42-actin-MRTF-A pathway was inhibited and the effect on cell shape, actin cytoskeleton status, and expression of PRG4 and TNC were determined. Treatment with the CDC42 inhibitor ML141 decreased PRG4 and TNC expression, and correlated with increased cell circularity and G-/F-actin ratio. PRG4 and TNC expression were differentially regulated by actin depolymerizing agents, latrunculin B and cytochalasin D. Chemical inhibition of MRTF-A resulted in decreased expression of both PRG4 and TNC; however, specific knockdown by small interfering RNA only decreased expression of TNC indicating that TNC, but not PRG4, is regulated by MRTF-A. Although PRG4 and TNC expression are both regulated by CDC42 and actin, it appears to occur through different downstream signaling pathways. Further study is required to elucidate the pathway regulating PRG4. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:2421-2430, 2018.