Hyperpolarisation of cultured human chondrocytes following cyclical pressure‐induced strain: Evidence of a role for α5β1 integrin as a chondrocyte mechanoreceptor

Wright, M O; Nishida, Kozue; Bavington, Charles D.; Godolphin, J L; Dunne, Eimear; Walmsley, Sarah R.; Jobanputra, Paresh; Nuki, G; Salter, Donald

doi:10.1002/jor.1100150517

Cited by 153 publications

(138 citation statements)

References 19 publications

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“…Among the integrins expressed on chondrocytes, ␣5␤1 integrin is the one most studied to date. Previous studies have shown that this integrin regulates various aspects of chondrocyte biology (21)(22)(23)(24)(25). However, the involvement of this integrin in dedifferentiation was not supported by the results of the present study.…”

Section: Discussioncontrasting

confidence: 99%

“…Chondrocytes express several types of integrins (17)(18)(19)(20), which are known to regulate various aspects of cell behavior. In addition to cell attachment, integrins on the chondrocytes are involved in proliferation, survival, differentiation, matrix remodeling, and response to mechanical stimuli (21)(22)(23)(24)(25). Integrins are also believed to play a role in the dedifferentiation of chondrocytes, but their involvement in the process has not yet been demonstrated.…”

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confidence: 99%

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αvβ5 Integrin promotes dedifferentiation of monolayer-cultured articular chondrocytes

Fukui¹,

Ikeda²,

Tanaka³

et al. 2011

Arthritis & Rheumatism

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Objective. When cultured in monolayers, articular chondrocytes undergo an obvious phenotypic change. Although the involvement of integrins has been suggested, the exact mechanisms of the change have not been determined. This study was undertaken to clarify the mechanisms underlying the loss of chondrocyte phenotype early after plating.Methods. Primary cultured human articular chondrocytes were used for the experiments. Involvement of respective integrins in the phenotypic change was investigated in RNA interference (RNAi) experiments. A signaling pathway involved in the change was identified in experiments using specific inhibitors and adenoviruses encoding mutated genes involved in the pathway. Adenoviruses carrying mutated GTPases were used to determine the involvement of small GTPases in the process.Results. In monolayer-cultured chondrocytes, suppression of ␣v or ␤5 integrin expression by RNAi inhibited morphologic changes in the cells and increased (or prevented a reduction in) the expression of various cartilage matrix genes. Consistent results were obtained in experiments using a blocking antibody and a synthetic inhibitor of ␣v␤5 integrin. The decrease in cartilage matrix gene expression in chondrocytes after plating was mediated by ERK signaling, which was promoted primarily by ␣v␤5 integrin. In articular chondrocytes, the affinity of ␣v␤5 integrin for ligands was regulated by the small GTPase R-Ras. R-Ras was gradually activated in monolayer-cultured chondrocytes after plating, which caused a gradual decline in cartilage matrix gene expression through enhanced ␣v␤5 integrin activation and the subsequent increase in ERK signaling.Conclusion. Our findings indicate that ␣v␤5 integrin may be involved in the change that occurs in monolayer-cultured chondrocytes after plating.

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Section: Discussioncontrasting

confidence: 99%

mentioning

confidence: 99%

αvβ5 Integrin promotes dedifferentiation of monolayer-cultured articular chondrocytes

Fukui¹,

Ikeda²,

Tanaka³

et al. 2011

Arthritis & Rheumatism

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“…Alternatively, cells could respond directly to pressur through specific mechanoreceptors. The presence of potential mechanoreceptor molecules, like integrins [35] and stretch activated ion channels [8] strengthen this line of thinking.…”

Section: Discussionmentioning

confidence: 90%

The enhancement of periosteal chondrogenesis in organ culture by dynamic fluid pressure

Mukherjee

Saris

Schultz

et al. 2001

Journal Orthopaedic Research

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Cartilage repair by autologous periosteal arthroplasty is enhanced by continuous passive motion (CPM) of the joint after transplantation of the periosteal graft. However, the mechanisms by which CPM stimulate chondrogenesis are unknown. Based on the observation that an oscillating intra-synovial pressure fluctuation has been reported to occur during CPM (0.&10 kPa), it was hypothesized that the oscillating pressure experienced by the periosteal graft as a result of CPM has a beneficial effect on the chondrogenic response of the graft. We have developed an in vitro model with which dynamic fluid pressures (DFP) that mimic those during CPM can be applied to periosteal explants while they are cultured in agarose gel suspension. In this study periosteal explants were treated with or without DFP during suspension culture in agarose, which is conducive to chondrogenesis. Different DFP application times (30 min, 4 h, 24 Wday) and pressure magnitudes (13, 103 kPa or stepwise 13 to 54 to 103 kPa) were compared for their effects on periosteal chondrogenesis. Low levels of DFP (13 kPa at 0.3 Hz) significantly enhanced chondrogenesis over controls (34 f 7% vs 14 f 5%; P < 0.05), while higher pressures (103 kPa at 0.3 Hz) completely inhibited chondrogenesis, as determined from the percentage of tissue that was determined to be cartilage by histomorphometry. Application of low levels of DFP to periosteal explants also resulted in significantly increased concentrations of Collagen Type I1 protein (43 f 8% vs 10 f 5%; P < 0.05). New proteoglycan synthesis, as measured by 35S-sulphate uptake was increased by 30% in periosteal explants stimulated with DFP (350 f 50 DPM vs 250 f 75 DPM of 35S-sulphate uptakdpg total protein), when compared to controls though this difference was not statistically significant. The DFP effect at low levels was dose-dependant for time of application as well, with 4 hlday stimulation causing significantly higher chondrogenesis than just 30 midday (34 f 7 vs 12 f 4% cartilage; P < 0.05) and not significantly less than that obtained with 24 hlday of DFP (48 f 9% cartilage, P > 0.05). These observations may partially explain the beneficial effect on cartilage repair by CPM. They also validate an in vitro model permitting studies aimed at elucidating the mechanisms of action of mechanical factors regulating chondrogenesis. The fact that these tissues were successfully cultured in a mechanical environment for six weeks makes it possible to study the actions of mechanical factors on the entire chondrogenic pathway, from induction to maturation. Finally, these data support the theoretical predictions regarding the role of hydrostatic compression in fracture healing.

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“…The investigation of isolated chondrocytes provides one way to study intracellular signals, without the need for extensive processing to remove the extracellular matrix present in native cartilage. Such an approach has yielded information on the ability of mechanical stimulation of chondrocytes to affect signaling pathways involving intracellular Ca2' (27,76), transmembrane potential (74,75), and nitric oxide (16). The current study used rabbit chondrocytes, which were expanded by as many as two passages of monolayer culture to obtain adequate cell numbers.…”

Section: Discussionmentioning

confidence: 99%

Biomechanical regulation of matrix metalloproteinase‐9 in cultured chondrocytes

Jin

Sah

et al. 2000

Journal Orthopaedic Research

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Summary:Abnormal mechanical loading of joints may induce degeneration of articular cartilage. Shear stress is one mode of mechanical loading that may regulate chondrocyte metabolism. We investigated the mechanism by which shear stress induccs the gene encoding matrix metalloproteinase-9, a mediator of the progressive degradation of articular cartilage in ostcoarthritis. In vifro experiments using passaged rabbit chondrocytes in monolayer culture subjected to a shear stress of 16 dynicn? (1.6 Pa) in a flow channel showed increased expression of the matrix mctalloproteinase-9 gcne. The induction of matrix metalloproteinase-9 appeared to depend on a region in the 5' promoter of the gcnc that contains a 12-0-tetradecanoylphorbol 13-acetateresponsive element. Transfection experiments using a construct containing a luciferase reporter driven by a 12-0-tetradecanoylphorbol 13-acetate-responsive clement indicated that shear stress activated a 12-0-tetradecanoylphorbol 13-acetate-responsive element-mediated transcription in chondrocytes. Similar experiments showed that shear stress induced a matrix metalloproteinase-9 promoter construct (matrix metalloproteinase-9-luciferase). Shear stress activated c-Jun NH1-terniinal kinasc, cxtraccllular signal-regulated kinase. and p3X. 'Transfection of matrix inetalloproteinase-9-lucifcrase together with thc dominant negative mutant of c-.Tun NH,-terminal kinase. hut not with that of cxtraccllular signal-regulated kinase or p38. attenuated the shearinduced matrix metalloproteinasc-9 promoter activity. In addition. transfection of constructs encoding dominant negative mutants o C Ras. Rac. and Cdc42 attenuated the induction of c-Jun transcriptional activity by shear stress. Thus. shear stimulation of chondrocytes stimulates Kas, Rac, and Cdc42, which subsequcntly activate c-Jun NH?-terminal kinasc to induce a 12-0-tetradecanoylphorhol 13-acetate-responsive clementmcdiatcd expression of matrix metalloproteinase-9. ~The metabolic activity of chondrocytes is regulated by mechanical factors (28). Such biomechanical regulation map affect the balance between anabolic and catabolic processes in articular cartilage. In osteoarthritis, a metabolic imbalance results in lie1 matrix degradation and the degeneration of articular cartilage (59). Many in vivo and in vitro studies have examined the effect of biomechanical stimuli on the synthesis of matrix proteins by chondrocytes (28). However. relatively few studies have analyzed the extracellular or cellular mechanisms by which mechanical stimuli may alter cartilage matrix degradationincluding the modulation of matrix protcinascs and their inhibitors (68)-and thereby contribute to the initiation or progression of osteoarthritis.In osteoarthritis, the synthesis of matrix metalloprotcinascs (MMPs) by chondrocytes is incrcased (20) and the balancc betwcen proteases and protease

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Hyperpolarisation of cultured human chondrocytes following cyclical pressure‐induced strain: Evidence of a role for α5β1 integrin as a chondrocyte mechanoreceptor

Cited by 153 publications

References 19 publications

αvβ5 Integrin promotes dedifferentiation of monolayer-cultured articular chondrocytes

αvβ5 Integrin promotes dedifferentiation of monolayer-cultured articular chondrocytes

The enhancement of periosteal chondrogenesis in organ culture by dynamic fluid pressure

Biomechanical regulation of matrix metalloproteinase‐9 in cultured chondrocytes

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