Consequences of chondrocyte hypertrophy on osteoarthritic cartilage: potential effect on angiogenesis L. Pesesse, C. Sanchez, J.-P. Delcour, A. Bellahcène,

Slides:

Advertisements

Similar presentations

High molecular weight hyaluronic acid regulates osteoclast formation by inhibiting receptor activator of NF-κB ligand through Rho kinase W. Ariyoshi,

Advertisements

M. M. J. Caron, P. J. Emans, M. M. E. Coolsen, L. Voss, D. A. M

CXC chemokine ligand 12a enhances chondrocyte proliferation and maturation during endochondral bone formation G.-W. Kim, M.-S. Han, H.-R. Park, E.-J.

Pro-inflammatory stimulation of meniscus cells increases production of matrix metalloproteinases and additional catabolic factors involved in osteoarthritis.

High molecular weight hyaluronic acid regulates osteoclast formation by inhibiting receptor activator of NF-κB ligand through Rho kinase W. Ariyoshi,

Muscle cell-derived factors inhibit inflammatory stimuli-induced damage in hMSC- derived chondrocytes R.S. Rainbow, H. Kwon, A.T. Foote, R.C. Preda, D.L.

GDF5 reduces MMP13 expression in human chondrocytes via DKK1 mediated canonical Wnt signaling inhibition L. Enochson, J. Stenberg, M. Brittberg, A. Lindahl

Detecting new microRNAs in human osteoarthritic chondrocytes identifies miR-3085 as a human, chondrocyte-selective, microRNA N. Crowe, T.E. Swingler,

Connexin43 hemichannels mediate small molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage

Effective reduction of the interleukin-1β transcript in osteoarthritis-prone guinea pig chondrocytes via short hairpin RNA mediated RNA interference influences.

Insulin-like growth factor-1 boosts the developing process of condylar hyperplasia by stimulating chondrocytes proliferation Y. Chen, J. Ke, X. Long,

Upregulated ank expression in osteoarthritis can promote both chondrocyte MMP-13 expression and calcification via chondrocyte extracellular PPi excess

L. Pesesse, C. Sanchez, D. A. Walsh, J. -P. Delcour, C. Baudouin, P

Histone deacetylase inhibitors suppress mechanical stress-induced expression of RUNX-2 and ADAMTS-5 through the inhibition of the MAPK signaling pathway.

M. -H. Moon, J. -K. Jeong, Y. -J. Lee, J. -W. Seol, C. J. Jackson, S

Combination of ADMSCs and chondrocytes reduces hypertrophy and improves the functional properties of osteoarthritic cartilage M.R. Ahmed, A. Mehmood,

M. M. J. Caron, P. J. Emans, M. M. E. Coolsen, L. Voss, D. A. M

Impaired glycolytic metabolism causes chondrocyte hypertrophy-like changes via promotion of phospho-Smad1/5/8 translocation into nucleus T. Nishida,

CXC chemokine ligand 12a enhances chondrocyte proliferation and maturation during endochondral bone formation G.-W. Kim, M.-S. Han, H.-R. Park, E.-J.

Fibroblast Growth Factor 23 drives MMP13 expression in human osteoarthritic chondrocytes in a Klotho-independent manner A. Bianchi, M. Guibert, F. Cailotto,

TGFβ inhibition during expansion phase increases the chondrogenic re-differentiation capacity of human articular chondrocytes R. Narcisi, L. Signorile,

M. Wang, H. Jin, D. Tang, S. Huang, M.J. Zuscik, D. Chen

X. Zhang, I. Prasadam, W. Fang, R. Crawford, Y. Xiao

NF-κBp65-specific siRNA inhibits expression of genes of COX-2, NOS-2 and MMP-9 in rat IL-1β-induced and TNF-α-induced chondrocytes Dr C. Lianxu, Ph.D.,

Long-term NSAID treatment directly decreases COX-2 and mPGES-1 production in the articular cartilage of patients with osteoarthritis M.A. Álvarez-Soria,

Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like.

Subtractive gene expression profiling of articular cartilage and mesenchymal stem cells: serpins as cartilage-relevant differentiation markers S. Boeuf,

M. Wang, H. Jin, D. Tang, S. Huang, M.J. Zuscik, D. Chen

Fibroblast growth factor-2 induced chondrocyte cluster formation in experimentally wounded articular cartilage is blocked by soluble Jagged-1 I.M. Khan,

Restriction of spontaneous and prednisolone-induced leptin production to dedifferentiated state in human hip OA chondrocytes: role of Smad1 and β-catenin.

Effect of glucosamine and its peptidyl-derivative on the production of extracellular matrix components by human primary chondrocytes D. Stoppoloni, L.

Endoplasmic reticulum stress-induced apoptosis contributes to articular cartilage degeneration via C/EBP homologous protein Y. Uehara, J. Hirose, S.

Chondrocyte calcium-sensing receptor expression is up-regulated in early guinea pig knee osteoarthritis and modulates PTHrP, MMP-13, and TIMP-3 expression

Interaction between zonal populations of articular chondrocytes suppresses chondrocyte mineralization and this process is mediated by PTHrP J. Jiang,

Expression and function of the insulin receptor in normal and osteoarthritic human chondrocytes: modulation of anabolic gene expression, glucose transport.

Connexin43 hemichannels mediate small molecule exchange between chondrocytes and matrix in biomechanically-stimulated temporomandibular joint cartilage

Human osteoarthritic chondrocytes are impaired in matrix metalloproteinase-13 inhibition by IFN-γ due to reduced IFN-γ receptor levels R. Ahmad, M. El.

MicroRNA-320 regulates matrix metalloproteinase-13 expression in chondrogenesis and interleukin-1β-induced chondrocyte responses F. Meng, Z. Zhang, W.

Differential expression of leptin and leptin's receptor isoform (Ob-Rb) mRNA between advanced and minimally affected osteoarthritic cartilage; effect.

Depletion of primary cilia in articular chondrocytes results in reduced Gli3 repressor to activator ratio, increased Hedgehog signaling, and symptoms.

Pro-inflammatory stimulation of meniscus cells increases production of matrix metalloproteinases and additional catabolic factors involved in osteoarthritis.

CaMKII inhibition in human primary and pluripotent stem cell-derived chondrocytes modulates effects of TGFβ and BMP through SMAD signaling B. Saitta,

Glucosamine promotes chondrogenic phenotype in both chondrocytes and mesenchymal stem cells and inhibits MMP-13 expression and matrix degradation A.

Differentiation potential of human muscle-derived cells towards chondrogenic phenotype in alginate beads culture R. Andriamanalijaona, Ph.D., E. Duval,

H.H. Lee, M.J. O'Malley, N.A. Friel, C.R. Chu

Cytochrome c oxidase levels in chondrocytes during monolayer expansion and after return to three dimensional culture O.A. Boubriak, J.T.S. Brooks, J.P.G.

C. Sanchez, Ph. D. , O. Gabay, Ph. D. , C. Salvat, B. Sc. , Y. E

Expression and regulation of Toll-like receptor 2 by IL-1β and fibronectin fragments in human articular chondrocytes S.-L. Su, M.S., C.-D. Tsai, Ph.D.,

V. Queirolo, D. Galli, E. Masselli, R. M. Borzì, S. Martini, F

Osteoblasts from the sclerotic subchondral bone downregulate aggrecan but upregulate metalloproteinases expression by chondrocytes. This effect is mimicked.

M.M.-G. Sun, F. Beier Osteoarthritis and Cartilage

Expression of the semicarbazide-sensitive amine oxidase in articular cartilage: its role in terminal differentiation of chondrocytes in rat and human

DIO2 modifies inflammatory responses in chondrocytes

H Jo, M. D. , J. S Park, Ph. D. , E. M Kim, M. S. , M. Y Jung, Ph. D

Selenomethionine inhibits IL-1β inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX2) expression in primary human chondrocytes A.W.M. Cheng,

Enhancing and maintaining chondrogenesis of synovial fibroblasts by cartilage extracellular matrix protein matrilins M. Pei, M.D., Ph.D., J. Luo, M.D.,

M. M. J. Caron, P. J. Emans, M. M. E. Coolsen, L. Voss, D. A. M

Dr J. Deschner, D. M. D. , Ph. D. , Dr B. Rath-Deschner, D. M. D. , Ph

Autologous chondrocyte implantation (ACI) for aged patients: development of the proper cell expansion conditions for possible therapeutic applications

Z. Lin, M. B. , N. J. Pavlos, B. Sc. (Hons. ), Ph. D. , M. A. Cake, B

Activation by IL-1 of bovine articular chondrocytes in culture within a 3D collagen-based scaffold. An in vitro model to address the effect of compounds.

Regulation of mechanical stress-induced MMP-13 and ADAMTS-5 expression by RUNX-2 transcriptional factor in SW1353 chondrocyte-like cells T. Tetsunaga,

Activation of Indian hedgehog promotes chondrocyte hypertrophy and upregulation of MMP-13 in human osteoarthritic cartilage F. Wei, J. Zhou, X. Wei,

Synergistic effect of chondroitin sulfate and cyclic pressure on biochemical and morphological properties of chondrocytes from articular cartilage G.

Comparative effects of IL-1β and hydrogen peroxide (H2O2) on catabolic and anabolic gene expression in juvenile bovine chondrocytes G. Martin, Ph.D.,

Influence of rabbit notochordal cells on symptomatic intervertebral disc degeneration: anti-angiogenic capacity on human endothelial cell proliferation.

PTHrP overexpression partially inhibits a mechanical strain-induced arthritic phenotype in chondrocytes D. Wang, J.M. Taboas, R.S. Tuan Osteoarthritis.

John M. Lamar, Vandana Iyer, C. Michael DiPersio

Y. Akasaki, A. Hasegawa, M. Saito, H. Asahara, Y. Iwamoto, M.K. Lotz

Structured three-dimensional co-culture of mesenchymal stem cells with chondrocytes promotes chondrogenic differentiation without hypertrophy M.E. Cooke,

Presentation transcript:

Consequences of chondrocyte hypertrophy on osteoarthritic cartilage: potential effect on angiogenesis L. Pesesse, C. Sanchez, J.-P. Delcour, A. Bellahcène, C. Baudouin, P. Msika, Y. Henrotin Osteoarthritis and Cartilage Volume 21, Issue 12, Pages 1913-1923 (December 2013) DOI: 10.1016/j.joca.2013.08.018 Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Evolution of the expression of hypertrophy markers (A) col10a1 (ANOVA: P < 0.0001), (B) runx2 (ANOVA: P = 0.0002), (C) MMP13 (ANOVA: P < 0.0001), and the activity of mineralization enzymes (D) AP (ANOVA: P < 0.0001) and (E) NTPPPH (ANOVA: P = 0.0003) in OA chondrocytes cultivated in alginate beads for 28 days in medium supplemented with either 2% ULTROSER G or 10% FBS. The increase of expression/activity of hypertrophy markers was significant after 21 days (col10a1: P = 0.0363; runx2: P = 0.0379; MMP13: P = 0.0152; AP: P = 0.0047; NTPPPH: P = 0.0158), 24 days (col10a1: P = 0.0012; runx2: P = 0.0007; MMP13: P < 0.0001; AP: P = 0.0126; NTPPPH: P = 0.0103) and 28 days (col10a1: P = 0.0002; runx2: P = 0.0208; MMP13: P < 0.0001; AP: P = 0.0008; NTPPPH: P = 0.0054) when compared with expression/activity after 3 days of culture. Comparison of hypertrophy markers expression/activity by chondrocytes cultivated in FBS-enriched medium with the corresponding day of culture in 2% UG-supplemented medium revealed a significant increase after 21 days (runx2: P = 0.0473; MMP13: P = 0,0062; AP: P = 0.0052; NTPPPH: P = 0.0232) 24 days (col10a1: P = 0.0031; runx2: P = 0.0011; MMP13: P < 0.0001; AP: P = 0.0152; NTPPPH: P = 0.0320) and 28 days of culture (col10a1: P = 0.0015; runx2: P = 0.0237; MMP13: P < 0.0001; AP: P = 0.0011; NTPPPH: P = 0.0183). Data from three different chondrocytes cultures were collected every 3–4 days, normalized to the house-keeping gene HPRT or to the DNA content and presented in percentage of their respective control. Data were analyzed by Dunnett's test after one-way ANOVA and are presented with the mean ± 95% confidence interval. Osteoarthritis and Cartilage 2013 21, 1913-1923DOI: (10.1016/j.joca.2013.08.018) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Representative pictures of hematoxylin-eosin stained wax-embedded sections of OA chondrocytes cultivated in alginate beads during 3 days (A and B) or 21 days (C and D) in medium supplemented with either 2% UG (A and C) or 10% FBS (B and D) (magnification 20×). Osteoarthritis and Cartilage 2013 21, 1913-1923DOI: (10.1016/j.joca.2013.08.018) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Effect of non-hypertrophic (NH) and hypertrophic (H) OA chondrocyte conditioned media on wound healing by endothelial cells (HUVECS) (A and B), HUVECS real-time migration (C), real-time invasion (D) and adhesion (E). (A) Representative pictures (magnification 10×) of the wound healing experiment showing the gap in HUVECs at the beginning of the experiment (h0) and after 24 h of incubation (h24); (B) Wound healing (ANOVA: P = 0.0005) was quantified using image J SoftWare (www.rsbweb.nih.gov/ij/) and data are presented in percentage of non-hypertrophic chondrocyte conditioned media at h0. Hypertrophic chondrocyte conditioned medium significantly increased wound healing by endothelial cells at h24 compared to h0 (P = 0.0008) and compared to the effect of non-hypertrophic chondrocyte conditioned medium at h24 (P = 0.0250). (C and D) Real-time HUVECS migration (ANOVA: P < 0.0001) and invasion (ANOVA: P < 0.0001) were analyzed every 5 h on a period of 25 h. Hypertrophic chondrocyte conditioned medium significantly stimulated HUVECS migration after 25 h (P < 0.0001) and invasion was increased after 5 h (P = 0.0087), 10 h (P < 0.0001) and 15 h (P < 0.0001) compared to control condition and after 10 h (P < 0.0001) compared to non-hypertrophic chondrocyte conditioned medium (###). Non-hypertrophic chondrocyte conditioned medium stimulated HUVECS invasion after 15 h (P = 0.0041). In wound healing and real-time assays, control condition is a non-conditioned culture medium containing 10% FBS. (E) HUVECS adhesion was analyzed after 2 h incubation of HUVECS on different coating conditions (ANOVA: P = 0.0003). Hypertrophic chondrocyte conditioned medium significantly increased HUVECS adhesion (P = 0.0463) compared to non-hypertrophic chondrocyte conditioned medium. Positive and negative controls are respectively coatings of 10 μg/ml recombinant BSP or BSA. Data were analyzed by Tukey's test after one-way ANOVA and are presented with mean ± 95% confidence interval. Each data point is the mean of at least three measures and each experiment was performed on three different chondrocytes cultures. Osteoarthritis and Cartilage 2013 21, 1913-1923DOI: (10.1016/j.joca.2013.08.018) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 BSP detection in OA chondrocytes cultures by RT-PCR (A), western blot on cellular protein extracts (B) and western blot on chondrocytes conditioned media (C). (A) BSP gene expression by non-hypertrophic (NH) and hypertrophic (H) chondrocytes. Data from three different chondrocytes cultures were analyzed by F test to compare variances and are presented in percentage of control with the mean ± 95% confidence interval. The expression of BSP gene was significantly higher in hypertrophic than in non-hypertrophic chondrocytes (P = 0.0001). (B) BSP production in cellular extracts from NH and H chondrocytes. Equal protein loading was assessed by anti-tubulin-α immunoblotting. (C) BSP secretion in conditioned chondrocyte media from NH and H chondrocytes. Equal protein loading was assessed by Ponceau red staining. Molecular weight difference in control and chondrocytes conditioned medium is discussed in the discussion section. Western blot experiments were performed at least on three different cultures and representative data from one experiment are shown. Control (CTL) is constituted of Saos2 cellular protein extracts or conditioned medium. Osteoarthritis and Cartilage 2013 21, 1913-1923DOI: (10.1016/j.joca.2013.08.018) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Spearman's correlation and corresponding P-value were calculated to assess the association between BSP expression and specific markers of chondrocyte hypertrophy (A) collagen type X expression (P < 0.0001), (B) runx2 expression (P < 0.0001), (C) MMP13 expression (P = 0.0031), (D) AP activity (P = 0.007), (E) NTPPPH activity (P < 0.0001). Data were collected from non-hypertrophic and hypertrophic OA chondrocytes (n = 28). Osteoarthritis and Cartilage 2013 21, 1913-1923DOI: (10.1016/j.joca.2013.08.018) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Adhesion assay performed with a pre-incubation of HUVECS with the GRGDS peptide at 50nmoles/l (ANOVA: P = 0.0033). Blocking of HUVECS αvβ3 integrin receptors with the GRGDS reduced adhesion of HUVECs on hypertrophic chondrocyte conditioned media coating (H + GRGDS) with a significant decrease of 52% in HUVECs adhesion (P = 0.0039). The GRGDS pre-treatment had no effect on non-hypertrophic chondrocyte conditioned media coating (NH + GRGDS). Data from three different chondrocyte cultures were analyzed by Tukey's test after one-way ANOVA. Each experimental condition was performed in sextuple. Osteoarthritis and Cartilage 2013 21, 1913-1923DOI: (10.1016/j.joca.2013.08.018) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions