Tissue Shear Deformation Stimulates Proteoglycan and Protein Biosynthesis in Bovine Cartilage Explants

2001 ◽  
Vol 395 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Moonsoo Jin ◽  
Eliot H. Frank ◽  
Thomas M. Quinn ◽  
Ernst B. Hunziker ◽  
Alan J. Grodzinsky
Keyword(s):  
Shear Deformation ◽  
Protein Biosynthesis ◽  
Cartilage Explants ◽  
Bovine Cartilage

scholarly journals A potential key role for alpha-haemolysin ofStaphylococcus aureusin mediating chondrocyte death in septic arthritis

2018 ◽  
Vol 7 (7) ◽  
pp. 457-467 ◽  
Author(s):  
I. D. M. Smith ◽  
K. M. Milto ◽  
C. J. Doherty ◽  
S. G. B. Amyes ◽  
A. H. R. W. Simpson ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Septic Arthritis ◽  
Cartilage Damage ◽  
Laser Scanning Microscopy ◽  
Cartilage Explants ◽  
Confocal Laser ◽  
Dead Cell ◽  
Chondrocyte Viability ◽  
Chondrocyte Death ◽  
Bovine Cartilage

ObjectivesStaphylococcus aureus (S. aureus) is the most commonly implicated organism in septic arthritis, a condition that may be highly destructive to articular cartilage. Previous studies investigating laboratory and clinical strains of S. aureus have demonstrated that potent toxins induced significant chondrocyte death, although the precise toxin or toxins that were involved was unknown. In this study, we used isogenic S. aureus mutants to assess the influence of alpha (Hla)-, beta (Hlb)-, and gamma (Hlg)-haemolysins, toxins considered important for the destruction of host tissue, on in situ bovine chondrocyte viability.MethodsBovine cartilage explants were cultured with isogenic S. aureus mutants and/or their culture supernatants. Chondrocyte viability was then assessed within defined regions of interest in the axial and coronal plane following live- and dead-cell imaging using the fluorescent probes 5-chloromethylfluorescein diacetate and propidium iodide, respectively, and confocal laser-scanning microscopy.ResultsHla-producing mutants caused substantial chondrocyte death compared with the toxin-deficient control (Hla-Hlb-Hlg-), whilst mutants producing Hlb and Hlg in the absence of Hla induced minimal chondrocyte death. Coronal studies established that Hla-induced chondrocyte death started in the superficial zone of cartilage and spread to deeper layers, whereas Hlb and Hlg toxins were without significant effect.ConclusionThis study identified Hla as a highly potent S. aureus toxin that caused rapid chondrocyte death in bovine cartilage, with other toxins or metabolic products produced by the bacteria playing a minor role. The identification of Hla in mediating chondrocyte death may assist in the development of therapeutic strategies aimed at reducing the extent of cartilage damage during and after an episode of septic arthritis. Cite this article: I. D. M. Smith, K. M. Milto, C. J. Doherty, S. G. B. Amyes, A. H. R. W. Simpson, A. C. Hall. A potential key role for alpha-haemolysin of Staphylococcus aureus in mediating chondrocyte death in septic arthritis. Bone Joint Res 2018;7:457–467. DOI: 10.1302/2046-3758.77.BJR-2017-0165.R1.

scholarly journals Chondrocyte-mediated catabolism of aggrecan: aggrecanase-dependent cleavage induced by interleukin-1 or retinoic acid can be inhibited by glucosamine

1998 ◽  
Vol 335 (1) ◽  
pp. 59-66 ◽  
Author(s):  
John D. SANDY ◽  
Dan GAMETT ◽  
Vivian THOMPSON ◽  
Christie VERSCHAREN
Keyword(s):  
Retinoic Acid ◽  
Core Protein ◽  
Interleukin 1 ◽  
Cartilage Explants ◽  
Biosynthetic Activity ◽  
Fragment Analysis ◽  
Analysis System ◽  
Chondrosarcoma Cell Line ◽  
Bovine Cartilage ◽  
Aggrecan Core Protein

A rat chondrosarcoma cell line and bovine cartilage explants have been used to study the control of aggrecan degradation by chondrocytes treated with interleukin-1 (IL-1) or retinoic acid (RA). Aggrecan fragment analysis with anti-neo-epitope antibodies suggests that aggrecanase (an as yet unidentified enzyme) is the only aggrecan-degrading proteinase active in these cultures. With rat cells, aggrecanase converts the aggrecan core protein into two major G1-domain-bearing products (60 kDa with a C-terminal Glu-373, and 220 kDa with a C-terminal Glu-1459). Both products were quantified on a standardized Western analysis system with a G1-specific antibody. Immunoblots were analysed by scanning densitometry and the sensitivity, linearity and reproducibility of the assay were established. With rat cells the aggrecanase response to IL-1 was optimal at about 2 mM glutamine, but was progressively inhibited at higher concentrations, with about 90% inhibition at 10 mM glutamine. Such inhibition by glutamine was not, however, observed with bovine explants. On the other hand, marked inhibition of aggrecanase-dependent cleavage was observed with both rat cells and bovine explants when d(+)-glucosamine was included at concentrations above 2 mM. Inhibition was apparently not due to cytotoxicity or interference with IL-1 signalling, since biosynthetic activity was not inhibited and inhibition of the aggrecanase response was also obtained when RA was used as the catabolic stimulator. Possible mechanisms for the inhibition of the aggrecanase response by glucosamine in chondrocytes treated with IL-1 or RA are discussed.

Biological Properties of Allogenic Articular Chondrocytes on the Surface of Bovine Cartilage Explants in vitro

2003 ◽  
Vol 50 (8) ◽  
pp. 418-423 ◽  
Author(s):  
G. Kim ◽  
M. Okumura ◽  
D. Bosnakovski ◽  
T. Ishiguro ◽  
T. Kadosawa ◽  
...  
Keyword(s):  
Articular Chondrocytes ◽  
Biological Properties ◽  
Cartilage Explants ◽  
Bovine Cartilage

scholarly journals Raman Needle Arthroscopy for In Vivo Molecular Assessment of Cartilage

2021 ◽  
Author(s):  
Kimberly Kroupa ◽  
Man I Wu ◽  
Juncheng Zhang ◽  
Magnus Jensen ◽  
Wei Wong ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Low Cost ◽  
Femoral Condyle ◽  
Cartilage Explants ◽  
Anatomical Site ◽  
Superficial Zone ◽  
Human Cartilage ◽  
Polarized Raman ◽  
Bovine Cartilage

The development of treatments for osteoarthritis (OA) is burdened by the lack of standardized biomarkers of cartilage health that can be applied in clinical trials. We present a novel arthroscopic Raman probe that can optically biopsy cartilage and quantify key ECM biomarkers for determining cartilage composition, structure, and material properties in health and disease. Technological and analytical innovations to optimize Raman analysis include: 1) multivariate decomposition of cartilage Raman spectra into ECM-constituent-specific biomarkers (glycosaminoglycan [GAG], collagen [COL], water [H2O] scores), and 2) multiplexed polarized Raman spectroscopy to quantify superficial zone collagen anisotropy via a PLS-DA-derived Raman collagen alignment factor (RCAF). Raman measurements were performed on a series of ex vivo cartilage models: 1) chemically GAG-depleted bovine cartilage explants (n=40), 2) mechanically abraded bovine cartilage explants (n=30), 3) aging human cartilage explants (n=14), and 4) anatomical-site-varied ovine osteochondral explants (n=6). Derived Raman GAG score biomarkers predicted 95%, 66%, and 96% of the variation in GAG content of GAG-depleted bovine explants, human explants, and ovine explants, respectively (p<0.001). RCAF values were significantly different for explants with abrasion-induced superficial zone collagen loss (p<0.001). The multivariate linear regression of Raman-derived ECM biomarkers (GAG and H2O scores) predicted 94% of the variation in elastic modulus of ovine explants (p<0.001). Finally, we demonstrated the first in vivo Raman arthroscopy assessment of an ovine femoral condyle through intraarticular entry into the synovial capsule. This work advances Raman arthroscopy towards a transformative low cost, minimally invasive diagnostic platform for objective monitoring of treatment outcomes from emerging OA therapies.

Hypo-Osmolarity Decreases the Ability of Cells in Articular Cartilage to Survive a Load-Induced Injury

2003 ◽  
Author(s):  
Adam Levin ◽  
Peter A. Torzilli ◽  
C. T. Christopher Chen
Keyword(s):  
Cell Death ◽  
Articular Cartilage ◽  
Cell Viability ◽  
Cartilage Explants ◽  
Hypotonic Solution ◽  
Pericellular Matrix ◽  
Isotonic Solution ◽  
Confined Compression ◽  
Superficial Zone ◽  
Bovine Cartilage

A recent study showed that exposure to hypotonic conditions increased chondrocyte surface area up to 234% by stretching the folded membrane, which may reduce the chondrocyte’s ability to deform under load. The goal of this study was to determine the effect of hypo-osmolarity on chondrocyte survival from load-induced injury. Bovine cartilage explants were incubated in either isotonic or hypotonic pH-buffered solution for 20 minutes, loaded with cyclic confined compression at 5MPa for 1 hour, and then assessed for cell viability using cell vital dyes and for pericellular matrix (PCM) using an type VI collagen antibody. Cell death in loaded explants was significantly greater than that of non-loaded controls (p&lt;0.001). However, explants loaded in the hypotonic solution showed significantly greater cell death than those loaded in the isotonic solution. An increase of dead cells with flatten PCM were located in the superficial zone. Our findings suggest that hypo-osmolarity decreases the ability of chondrocytes in articular cartilage to survive from load-induced injury.

scholarly journals Monoclonal antibodies that specifically recognize neoepitope sequences generated by ‘aggrecanase’ and matrix metalloproteinase cleavage of aggrecan: application to catabolism in situ and in vitro

1995 ◽  
Vol 305 (3) ◽  
pp. 799-804 ◽  
Author(s):  
C E Hughes ◽  
B Caterson ◽  
A J Fosang ◽  
P J Roughley ◽  
J S Mort
Keyword(s):  
Monoclonal Antibodies ◽  
Retinoic Acid ◽  
Degradation Products ◽  
Interleukin 1 ◽  
Proteolytic Cleavage ◽  
Cartilage Explants ◽  
Reactive Material ◽  
Culture Systems ◽  
Bovine Cartilage

Monoclonal antibodies have been prepared that react specifically with the neoepitopes present on proteoglycan degradation products generated from the proteolytic cleavage of aggrecan in the interglobular domain. Antibody BC-3 recognizes the new N-terminus (ARGSV...) on aggrecan degradation products produced by the action of the as yet uncharacterized proteolytic activity, ‘aggrecanase’, and antibody BC-4 recognizes the new C-terminus (...DIPEN) generated by the proteolytic action of matrix metalloproteinases. Specificity for these neoepitope sequences was determined in competitive e.l.i.s.a. using synthetic peptide antigens as inhibitors. Antibody BC-3 was used in the detection of aggrecan degradation products in the culture medium obtained from two different in vitro culture systems: bovine cartilage explants treated with either retinoic acid or interleukin-1, and secondly, rat chondrosarcoma cells treated with retinoic acid. Both interleukin-1 and retinoic acid treatment caused an increase in aggrecan catabolism resulting in an increased release to the medium of specific aggrecan degradation products containing the BC-3 neoepitope generated by the action of ‘aggrecanase'. However, several additional aggrecan catabolites were present that were not immunoreactive with antibody BC-3. In addition, under control conditions, in the bovine cartilage cultures the BC-3 epitope was found on some of these aggrecan catabolites. In contrast, no immune-reactive material was found in the aggrecan degradation products present in control media of rat chondrosarcoma cells cultured in the absence of retinoic acid. Collectively, these results demonstrate that ‘aggrecanase’ activity is not a constitutive event in all cartilage culture systems and also suggest that proteolytic agents other than ‘aggrecanase’ are involved in aggrecan catabolism in normal turnover compared with pathological conditions. Antibody BC-4 was used to demonstrate the identity of the G1 domain of aggrecan following proteolytic cleavage of a purified G1-G2 preparation with collagenase, gelatinase A or stromelysin. The G2 product of this cleavage did not react with antibody BC-3, indicating that, under the experimental conditions used, none of these enzymes exhibited ‘aggrecanase’ activity. It is expected that both of these antibodies will play a pivotal role in detailed studies elucidating molecular mechanisms of aggrecan degradation and they will be particularly useful for the sensitive monitoring of aggrecan degradation products in tissue extracts and body fluids.

scholarly journals Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments

2009 ◽  
Vol 2 (1) ◽  
pp. 259 ◽  
Author(s):  
Bijue Wang ◽  
Pingping Chen ◽  
Anne-Christine Jensen ◽  
Morten A Karsdal ◽  
Suzi H Madsen ◽  
...  
Keyword(s):  
Cartilage Explants ◽  
Bovine Cartilage

Effect of Impact Load on Articular Cartilage: Cell Metabolism and Viability, and Matrix Water Content

1999 ◽  
Vol 121 (5) ◽  
pp. 433-441 ◽  
Author(s):  
P. A. Torzilli ◽  
R. Grigiene ◽  
J. Borrelli ◽  
D. L. Helfet
Keyword(s):  
Cell Death ◽  
Water Content ◽  
Impact Load ◽  
Cell Metabolism ◽  
Cartilage Explants ◽  
Critical Threshold ◽  
Limited Information ◽  
Impact Stress ◽  
Stress Dependent ◽  
Bovine Cartilage

Significant evidence exists that trauma to a joint produced by a single impact load below that which causes subchondral bone fracture can result in permanent damage to the cartilage matrix, including surface fissures, loss of proteoglycan, and cell death. Limited information exists, however, on the effect of a varying impact stress on chondrocyte biophysiology and matrix integrity. Based on our previous work, we hypothesized that a stress-dependent response exists for both the chondrocyte’s metabolic activity and viability and the matrix’s hydration. This hypothesis was tested by impacting bovine cartilage explants with nominal stresses ranging from 0.5 to 65 MPa and measuring proteoglycan biosynthesis, cell viability, and water content immediately after impaction and 24 hours later. We found that proteoglycan biosynthesis decreased and water content increased with increasing impact stress. However, there appeared to be a critical threshold stress (15–20 MPa) that caused cell death and apparent rupture of the collagen fiber matrix at the time of impaction. We concluded that the cell death and collagen rupture are responsible for the observed alterations in the tissue’s metabolism and water content, respectively, although the exact mechanism causing this damage could not be determined.

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