Viability and volume of in situ bovine articular chondrocytes—changes following a single impact and effects of medium osmolarity  Dr Peter G. Bush, Ph.D.,

Slides:



Advertisements
Similar presentations
Y. Tochigi, P. Zhang, M. J. Rudert, T. E. Baer, J. A. Martin, S. L
Advertisements

Oxidized low-density lipoprotein (ox-LDL) binding to lectin-like ox-LDL receptor-1 (LOX- 1) in cultured bovine articular chondrocytes increases production.
Moderate dynamic compression inhibits pro-catabolic response of cartilage to mechanical injury, tumor necrosis factor-α and interleukin-6, but accentuates.
C.P. Neu, T. Novak, K.F. Gilliland, P. Marshall, S. Calve 
Expression of superficial zone protein in mandibular condyle cartilage
IL-10 reduces apoptosis and extracellular matrix degradation after injurious compression of mature articular cartilage  P. Behrendt, A. Preusse-Prange,
L. Ding, E. Heying, N. Nicholson, N. J. Stroud, G. A. Homandberg, J. A
Interleukin-1 inhibits osmotically induced calcium signaling and volume regulation in articular chondrocytes  S. Pritchard, Ph.D., B.J. Votta, M.S., S.
Infrapatellar fat pad aggravates degeneration of acute traumatized cartilage: a possible role for interleukin-6  J. He, Y. Jiang, P.G. Alexander, V. Ulici,
D.A. Houston, A.K. Amin, T.O. White, I.D.M. Smith, A.C. Hall 
M. Z. Ilic, Ph. D. , B. Martinac, Ph. D. , T. Samiric, Ph. D. , C. J
Differential proteome analysis of normal and osteoarthritic chondrocytes reveals distortion of vimentin network in osteoarthritis  S. Lambrecht, M.Pharm.,
Articular chondrocytes derived from distinct tissue zones differentially respond to in vitro oscillatory tensile loading  E.J. Vanderploeg, Ph.D., C.G.
Analysis of radial variations in material properties and matrix composition of chondrocyte-seeded agarose hydrogel constructs  T.-A.N. Kelly, Ph.D., K.W.
Clathrin-Mediated Endocytosis Persists during Unperturbed Mitosis
The role of the PCM in reducing oxidative stress induced by radical initiated photoencapsulation of chondrocytes in poly(ethylene glycol) hydrogels  N.
Increased stromelysin-1 (MMP-3), proteoglycan degradation (3B3- and 7D4) and collagen damage in cyclically load-injured articular cartilage  Peggy M.
Endoplasmic reticulum stress-induced apoptosis contributes to articular cartilage degeneration via C/EBP homologous protein  Y. Uehara, J. Hirose, S.
S. E. Cisewski, L. Zhang, J. Kuo, G. J. Wright, Y. Wu, M. J. Kern, H
K. Murata, N. Kanemura, T. Kokubun, T. Fujino, Y. Morishita, K
A.J. McGregor, B.G. Amsden, S.D. Waldman  Osteoarthritis and Cartilage 
H.T. Kokkonen, J.S. Jurvelin, V. Tiitu, J. Töyräs 
The volume and morphology of chondrocytes within non-degenerate and degenerate human articular cartilage  P.G Bush, Ph.D., A.C Hall, Ph.D.  Osteoarthritis.
Assessment of strategies to increase chondrocyte viability in cryopreserved human osteochondral allografts: evaluation of the glycosylated hydroquinone,
Osmolarity effects on bovine articular chondrocytes during three-dimensional culture in alginate beads  X. Xu, J.P.G. Urban, U.K. Tirlapur, Z. Cui  Osteoarthritis.
Promotion of the intrinsic damage–repair response in articular cartilage by fibroblastic growth factor-2  F.M.D. Henson, Ph.D., E.A. Bowe, Ph.D., M.E.
M.L. Delco  Osteoarthritis and Cartilage 
The use of hyperosmotic saline for chondroprotection: implications for orthopaedic surgery and cartilage repair  N.M. Eltawil, S.E.M. Howie, A.H.R.W.
Oxidized low-density lipoprotein (ox-LDL) binding to lectin-like ox-LDL receptor-1 (LOX- 1) in cultured bovine articular chondrocytes increases production.
Glutamine protects articular chondrocytes from heat stress and NO-induced apoptosis with HSP70 expression  H. Tonomura, M.D., K.A. Takahashi, M.D., Ph.D.,
Chondrocyte pro-proliferative compounds may utilize extracellular matrix remodeling processes to regenerate cartilage  D. Reker, C. Kjelgaard-Petersen,
Oxidative stress induces expression of osteoarthritis markers procollagen IIA and 3B3(−) in adult bovine articular cartilage  I.M. Khan, Ph.D., S.J. Gilbert,
Inhibition of caspase-9 reduces chondrocyte apoptosis and proteoglycan loss following mechanical trauma  C.A.M. Huser, M.Sc., M. Peacock, M.E. Davies,
A.W. Palmer, C.G. Wilson, E.J. Baum, M.E. Levenston 
Effects of insulin-like growth factor-1 and dexamethasone on cytokine-challenged cartilage: relevance to post-traumatic osteoarthritis  Y. Li, Y. Wang,
Y. Tochigi, P. Zhang, M. J. Rudert, T. E. Baer, J. A. Martin, S. L
Y. Zhou, L. Resutek, L. Wang, X. Lu  Osteoarthritis and Cartilage 
Primary cilia disassembly down-regulates mechanosensitive hedgehog signalling: a feedback mechanism controlling ADAMTS-5 expression in chondrocytes  C.L.
A.C. Dang, M.D., A.P. Warren, M.D., H.T. Kim, M.D., Ph.D. 
Structural adaptations in compressed articular cartilage measured by diffusion tensor imaging  S.K. de Visser, B.Eng. (Med.), R.W. Crawford, D.Phil.,
A stable isotope method for the simultaneous measurement of matrix synthesis and cell proliferation in articular cartilage in vivo  K.W. Li, S.A. Siraj,
Dr J. Deschner, D. M. D. , Ph. D. , Dr B. Rath-Deschner, D. M. D. , Ph
Rapid in situ chondrocyte death induced by Staphylococcus aureus toxins in a bovine cartilage explant model of septic arthritis  I.D.M. Smith, J.P. Winstanley,
K. D. Novakofski, R. M. Williams, L. A. Fortier, H. O. Mohammed, W. R
L. C. Davies, B. Sc. , Ph. D. , E. J. Blain, B. Sc. , Ph. D. , B
D.R. Rich, A.L. Clark  Osteoarthritis and Cartilage 
Pharmaceutical nanocarrier association with chondrocytes and cartilage explants: influence of surface modification and extracellular matrix depletion 
H. Sadeghi, D.E.T. Shepherd, D.M. Espino  Osteoarthritis and Cartilage 
The rate of hypo-osmotic challenge influences regulatory volume decrease (RVD) and mechanical properties of articular chondrocytes  Z. Wang, J. Irianto,
V. Morel, Ph.D., A. Merçay, M.Sc., T.M. Quinn, Ph.D. 
P. I. Milner, Ph. D. , B. V. Sc. , B. Sc. (Hons. ), M. R. C. V. S. , R
S.I. Paterson, A.K. Amin, A.C. Hall  Osteoarthritis and Cartilage 
Characterizing osteochondrosis in the dog: potential roles for matrix metalloproteinases and mechanical load in pathogenesis and disease progression 
Mevastatin reduces cartilage degradation in rabbit experimental osteoarthritis through inhibition of synovial inflammation  Y. Akasaki, M.D., S. Matsuda,
In vitro glycation of articular cartilage alters the biomechanical response of chondrocytes in a depth-dependent manner  J.M. Fick, M.R.J. Huttu, M.J.
Scaffold degradation elevates the collagen content and dynamic compressive modulus in engineered articular cartilage  K.W. Ng, Ph.D., L.E. Kugler, B.S.,
K.P. Arkill, Ph.D., C.P. Winlove, D.Phil.  Osteoarthritis and Cartilage 
Comparison between chondroprotective effects of glucosamine, curcumin, and diacerein in IL-1β-stimulated C-28/I2 chondrocytes  S. Toegel, M.Pharm.S.,
W.C. Bae, Ph.D., B.L. Schumacher, B.S., R.L. Sah, M.D., Sc.D. 
V.K. Shekhawat, M.P. Laurent, C. Muehleman, M.A. Wimmer 
Effect of a glucosamine derivative on impact-induced chondrocyte apoptosis in vitro. A preliminary report  C.A.M. Huser, Ph.D., M.E. Davies, Ph.D.  Osteoarthritis.
Intracellular calcium oscillations in articular chondrocytes induced by basic calcium phosphate crystals lead to cartilage degradation  C. Nguyen, M.
Mechanical injury of bovine cartilage explants induces depth-dependent, transient changes in MAP kinase activity associated with apoptosis  D.H. Rosenzweig,
PTHrP overexpression partially inhibits a mechanical strain-induced arthritic phenotype in chondrocytes  D. Wang, J.M. Taboas, R.S. Tuan  Osteoarthritis.
Cellular origin of neocartilage formed at wound edges of articular cartilage in a tissue culture experiment  P.K. Bos, M.D., Ph.D., N. Kops, B.Sc., J.A.N.
Estrogen reduces mechanical injury-related cell death and proteoglycan degradation in mature articular cartilage independent of the presence of the superficial.
C. Pascual Garrido, A. A. Hakimiyan, L. Rappoport, T. R. Oegema, M. A
Enhanced phagocytic capacity endows chondrogenic progenitor cells with a novel scavenger function within injured cartilage  C. Zhou, H. Zheng, J.A. Buckwalter,
Expression of superficial zone protein in mandibular condyle cartilage
Effects of physical stimulation with electromagnetic field and insulin growth factor-I treatment on proteoglycan synthesis of bovine articular cartilage 
Presentation transcript:

Viability and volume of in situ bovine articular chondrocytes—changes following a single impact and effects of medium osmolarity  Dr Peter G. Bush, Ph.D., Peter D. Hodkinson, M.Sc., Georgina L. Hamilton, B.Sc., Dr Andrew C. Hall, Ph.D.  Osteoarthritis and Cartilage  Volume 13, Issue 1, Pages 54-65 (January 2005) DOI: 10.1016/j.joca.2004.10.007 Copyright © 2004 OsteoArthritis Research Society International Terms and Conditions

Fig. 1 An overview of the pattern of chondrocyte death arising from the application of a single injurious impact to bovine articular cartilage. Explants of cartilage were incubated with calcein-AM and PI as described (see Materials and Methods) and then subjected to (a) no impact, or (b) a single impact of 100g from 10cm. The cartilage was then imaged by CLSM after 60min, and the projected image viewed perpendicular to the cartilage surface is shown (see Materials and Methods). Intracellular calcein or PI fluorescence appears green or red, respectively. Confocal images were acquired at 10μm z-step intervals using a 5× dry objective lens to a depth of ∼60μm. Bar=100μm. Osteoarthritis and Cartilage 2005 13, 54-65DOI: (10.1016/j.joca.2004.10.007) Copyright © 2004 OsteoArthritis Research Society International Terms and Conditions

Fig. 2 Chondrocyte death following a single impact. Explants of bovine articular cartilage were subjected to (a) no impact, (b) 100g from 10cm, or (c) 200g from 10cm and then maintained in culture. Maximum projected confocal images of calcein (live cells; green) and PI (dead cell nuclei; red) were taken of transverse sections 24h after impact. Confocal images were acquired at 10μm z-step intervals using a 10× dry objective lens. For the experimental protocols for this and other figures, see Materials and Methods. Bar=100μm. Osteoarthritis and Cartilage 2005 13, 54-65DOI: (10.1016/j.joca.2004.10.007) Copyright © 2004 OsteoArthritis Research Society International Terms and Conditions

Fig. 3 The time course of chondrocyte death resulting from a single impact load. Bovine articular cartilage incubated with calcein-AM and PI, was positioned with the synovial surface uppermost, and subjected to a single impact load of 100g from 10cm, and then viewed by CLSM. Upper panels show the maximum projected confocal images of calcein (live cells; green) and PI (dead cell nuclei; red) (a) 5min, (b) 15min, (c) 25min, (d) 35min, and (e) 45min after impact. To visualise the time course of cell death more easily, the lower panels indicate the additional cell death observed in sequential images over the following periods (0–5min, 5–15min, 15–25min, 25–35min, and 35–45min). Confocal images were acquired at 10μm z-step intervals using a 5× dry objective lens. Bar=200μm. Osteoarthritis and Cartilage 2005 13, 54-65DOI: (10.1016/j.joca.2004.10.007) Copyright © 2004 OsteoArthritis Research Society International Terms and Conditions

Fig. 4 Changes to the viability of SZ chondrocytes near the injured edge resulting from a single impact load. Bovine articular cartilage was subjected to a single impact load (100g from 10cm) and viewed from the synovial surface. Panels show maximum projected confocal images of calcein (live cells; green) and PI (dead cell nuclei; red): (a) 3min, (b) 6min, (c) 12min, and (d) 20min after impact. The chondrocytes used for volume determinations in this experiment are identified individually using a white spot [panel (d)]. Confocal images were acquired at 1μm z-step intervals using a 63× water-immersion objective lens. Bar=25μm; image approximately 150μm×150μm. Osteoarthritis and Cartilage 2005 13, 54-65DOI: (10.1016/j.joca.2004.10.007) Copyright © 2004 OsteoArthritis Research Society International Terms and Conditions

Fig. 5 The decrease in chondrocyte viability after two single impacts of different magnitude. The proportion of viable cells as a percentage of the total cell number studied was determined over 20min for (1) control cartilage explants (i.e., not exposed to impact), and for explants subjected to (2) 100g from 5cm and (3) 100g from 10cm. The data are shown at the time points corresponding to initiation of the scan, although note that the total scanning time required for each measurement was ∼90s. At the first time point studied, there was a significant (P<0.001, indicated by †) decrease in the proportion of viable cells. Although the subsequent decrease in viability appeared greater for 100g from 10cm compared to the same load from 5cm, this difference was only significant at 20min (P<0.01, indicated by *). Over the entire time course, the rate of cell death was not significantly different (P=0.718; two-way ANOVA; see Results). Data (means±s.e.m.) are from at least five experiments under each condition. Osteoarthritis and Cartilage 2005 13, 54-65DOI: (10.1016/j.joca.2004.10.007) Copyright © 2004 OsteoArthritis Research Society International Terms and Conditions

Fig. 6 The change in cell volume of in situ chondrocytes following a single impact. Cartilage explants were positioned with the synovial surface uppermost, and then impacted. The edge of the injury was located quickly as described and the volume of viable chondrocytes near the edge determined over 20min (see Materials and Methods, and Figs. 1 and 4). The control cartilage was not subjected to impact and the volume of in situ chondrocytes determined at the time points indicated, with the mean initial value taken as 100%. For both impact loads, chondrocyte shrinkage was significantly different (P<0.006, indicated by *) but the rate of shrinkage was not different between the two loads (P=0.089; two-way ANOVA; see Results). Data (n[N]) are given as means±s.e.m. for (4[18]), (3[26]) and (5[17]), respectively. Osteoarthritis and Cartilage 2005 13, 54-65DOI: (10.1016/j.joca.2004.10.007) Copyright © 2004 OsteoArthritis Research Society International Terms and Conditions