In vitro inhibition of aggrecanase activity by tetracyclines and proteoglycan loss from osteoarthritic human articular cartilage

2010 ◽  
Vol 28 (6) ◽  
pp. 828-833 ◽  
Author(s):  
Jürgen Steinmeyer ◽  
Jens Kordelle ◽  
Henning Stürz
Keyword(s):  
Articular Cartilage ◽  
Human Articular Cartilage ◽  
In Vitro Inhibition

scholarly journals Biosynthesis of proteoglycan in vitro by cartilage from human osteochondrophytic spurs

1982 ◽  
Vol 206 (2) ◽  
pp. 329-341 ◽  
Author(s):  
Charles J. Malemud ◽  
Victor M. Goldberg ◽  
Roland W. Moskowitz ◽  
Lee L. Getzy ◽  
Robert S. Papay ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Deae Cellulose ◽  
Culture Conditions ◽  
Human Articular Cartilage ◽  
Guanidinium Chloride ◽  
Tissue Slices ◽  
Keratan Sulphate ◽  
Defined Culture

Proteoglycan biosynthesis by human osteochondrophytic spurs (osteophytes) obtained from osteoarthritic femoral heads at the time of surgical joint replacement was studied under defined culture conditions in vitro. Osteophytes were primarily present in two anatomic locations, marginal and epi-articular. Minced tissue slices were incubated in the presence of [35S]sulphate or [14C]glucosamine. Osteophytes incorporated both labelled precursors into proteoglycan, which was subsequently characterized by CsCl-isopycnic-density-gradient ultracentrifugation and chromatography on Sepharose CL-2B. The material extracted with 0.5m-guanidinium chloride showed 78.1% of [35S]sulphate in the A1 fraction after centrifugation. Only 23.0% of the [35S]sulphate in this A1 fraction was eluted in the void volume of Sepharose CL-2B under associative conditions. About 60–80% of the [35S]sulphate in the tissue 4m-guanidinium chloride extract was associated with monomeric proteoglycan (fraction D1). The average partition coefficient (Kav.) of the proteoglycan monomer on Sepharose CL-2B was 0.28–0.33. Approx. 12.4% of this monomer formed stable aggregates with high-molecular-weight hyaluronic acid in vitro. Sepharose CL-2B chromatography of fractions with lower buoyant densities (fractions D2–D4) demonstrated elution profiles on Sepharose CL-2B substantially different than that of fraction D1, indicative of the polydisperse nature of the newly synthesized proteoglycan. Analysis of the composition and chain size of the glycosaminoglycans showed the following: (1) preferential elution of both [35S]sulphate and [14C]glucosamine in the 0.5m-LiCl fraction on DEAE-cellulose; (2) the predominant sulphated glycosaminoglycan was chondroitin 6-sulphate (60–70%), with 9–11% keratan sulphate in the monomer proteoglycan; (3) Kav. values of 0.38 on Sephadex G-200 and 0.48 on Sepharose CL-6B were obtained with papain-digested and NaBH4-treated D1 monomer respectively. A comparison of the synthetic with endogenous glycosaminoglycans indicated similar types. These studies indicated that human osteophytes synthesized in vitro sulphated proteoglycans with some characteristics similar to those of mature human articular cartilage, notably in the size of their proteoglycan monomer and predominance of chondroitin 6-sulphate. They differed from articular cartilage primarily in the lack of substantial quantities of keratan sulphate and aggregation properties associated with monomer interaction with hyaluronic acid.

scholarly journals Recombinant human gelsolin promotes the migration of human articular cartilage chondrocytes by regulating gene expression in vitro

2020 ◽  
Vol 2 (4) ◽  
pp. 100124
Author(s):  
Jessica Feldt ◽  
Jessica Welss ◽  
Verena Schropp ◽  
Kolja Gelse ◽  
Michael Tsokos ◽  
...  
Keyword(s):  
Gene Expression ◽  
Articular Cartilage ◽  
Human Articular Cartilage

scholarly journals Characterization of proteins from human synovium and mononuclear leucocytes that induce resorption of cartilage proteoglycan in vitro

1983 ◽  
Vol 209 (2) ◽  
pp. 337-344 ◽  
Author(s):  
J Saklatvala ◽  
S J Sarsfield ◽  
L M C Pilsworth
Keyword(s):  
Articular Cartilage ◽  
Synovial Tissue ◽  
Gel Filtration ◽  
Human Articular Cartilage ◽  
Nasal Cartilage ◽  
Cartilage Proteoglycan ◽  
Mononuclear Leucocytes ◽  
Culture Supernatants

Both human synovial tissue in culture and lectin-stimulated mononuclear leucocytes produced a protein that induced proteoglycan resorption in explants of bovine nasal cartilage and human articular cartilage. On gel filtration the protein had Mr 16000-20000 and on isoelectric focusing its pI was 5.2-5.3. The protein corresponded to catabolin, which has previously been identified as a product of cultured porcine synovial tissue and mononuclear leucocytes. The action of partially purified human catabolin was not inhibited by cortisol, although the activity of the leucocyte supernatants from which it had been isolated was inhibited. For this reason it is not possible to be sure that the active factor detected in the bioassay of the crude leucocyte culture supernatants is in fact catabolin.

scholarly journals In vitro method for 3D morphometry of human articular cartilage chondrons based on micro-computed tomography

2018 ◽  
Vol 26 (8) ◽  
pp. 1118-1126 ◽  
Author(s):  
I. Kestilä ◽  
J. Thevenot ◽  
M.A. Finnilä ◽  
S.S. Karhula ◽  
I. Hadjab ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Articular Cartilage ◽  
Human Articular Cartilage ◽  
Micro Computed Tomography ◽  
In Vitro Method

scholarly journals Recapitulation of physiological spatiotemporal signals promotes in vitro formation of phenotypically stable human articular cartilage

2017 ◽  
Vol 114 (10) ◽  
pp. 2556-2561 ◽  
Author(s):  
Johnathan J. Ng ◽  
Yiyong Wei ◽  
Bin Zhou ◽  
Jonathan Bernhard ◽  
Samuel Robinson ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Endochondral Ossification ◽  
Human Articular Cartilage ◽  
Control Groups ◽  
Human Cartilage ◽  
Chondrocyte Maturation ◽  
Self Assembling ◽  
Spatiotemporal Regulation

Standard isotropic culture fails to recapitulate the spatiotemporal gradients present during native development. Cartilage grown from human mesenchymal stem cells (hMSCs) is poorly organized and unstable in vivo. We report that human cartilage with physiologic organization and in vivo stability can be grown in vitro from self-assembling hMSCs by implementing spatiotemporal regulation during induction. Self-assembling hMSCs formed cartilage discs in Transwell inserts following isotropic chondrogenic induction with transforming growth factor β to set up a dual-compartment culture. Following a switch in the basal compartment to a hypertrophic regimen with thyroxine, the cartilage discs underwent progressive deep-zone hypertrophy and mineralization. Concurrent chondrogenic induction in the apical compartment enabled the maintenance of functional and hyaline cartilage. Cartilage homeostasis, chondrocyte maturation, and terminal differentiation markers were all up-regulated versus isotropic control groups. We assessed the in vivo stability of the cartilage formed under different induction regimens. Cartilage formed under spatiotemporal regulation in vitro resisted endochondral ossification, retained the expression of cartilage markers, and remained organized following s.c. implantation in immunocompromised mice. In contrast, the isotropic control groups underwent endochondral ossification. Cartilage formed from hMSCs remained stable and organized in vivo. Spatiotemporal regulation during induction in vitro recapitulated some aspects of native cartilage development, and potentiated the maturation of self-assembling hMSCs into stable and organized cartilage resembling the native articular cartilage.

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