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      Contenido de glicosaminoglicanos, aldehídos y proteínas en el líquido sinovial de la articulación metacarpofalángica equina normal y alterada Translated title: Concentration of glycosaminoglycan, aldehydes and protein in synovial fluid from normal and damaged equine metacarpophalangeal joints

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          Abstract

          El objetivo del trabajo fue relacionar la severidad del cuadro osteoartrítico con la concentración de moléculas de la matriz extracelular del cartílago en el líquido sinovial. Para ello se midieron las concentraciones de proteína total y de grupos aldehídos como productos de degradación del colágeno. Además, se midió la concentración de glicosaminoglicanos (GAGs) del líquido sinovial en su valor total (GAGs T) y de una fracción de éstos, que corresponde a los GAGs diferentes del ácido hialurónico o GAGs sulfatados (GAGs S), entre los que se describen esencialmente condroitín sulfato y queratán sulfato. El contenido de ácido hialurónico se calculó por diferencia entre las concentraciones de GAGs T y de GAGs S. Las muestras de líquido sinovial se obtuvieron por artrocéntesis aséptica desde articulaciones metacarpofalángicas de equinos mestizos en matadero, inmediatamente después del beneficio de los animales y luego separadas en cuatro grupos, después del examen visual post mortem de las articulaciones: uno normal como control (n=17) y tres grupos alterados provenientes de articulaciones con diferentes grados de alteración articular (+ = 15; ++ = 12 y +++ = 18). La concentración de grupos aldehidos se determinó a través de una reacción colorimétrica entre aldehídos y N-Metil Benzotiazolidon Hidrazona. Además, para cuantificar los GAGs T y los GAGs S se utilizó un método colorimétrico con Alcian Blue usando diferentes concentraciones de MgCl2. El grupo con daño articular más severo (+++) mostró incremento de la concentración de proteínas y disminución de productos de degradación de colágeno y de la proteína colagenosa (P <0,05), mientras que la concentración de GAGs T, de GAGs S y de ácido hialurónico no mostró diferencias significativas entre los grupos analizados (P 0,05), detectándose sólo tendencias a disminuir en la concentración ácido hialurónico y a aumentar en la concentración de GAGs S en el grupo con daño articular más severo (+++). Estos resultados indican que la concentración de productos de degradación del colágeno en el líquido sinovial podría ser utilizada como marcador precoz de los cambios iniciales del proceso osteoartrítico.

          Translated abstract

          The purpose of this study was to evaluate the influence of severity of osteoarthritis on the concentration of degradation products from the cartilage extracellular matrix on synovial fluid. Total protein and aldehyde group concentrations were determined as indicators of collagen degradation. The concentration of glycosaminoglycans (GAGs) in sinovial liquid was also determined, both the total amount (GAGsT) and the fractions corresponding to the GAGs other than hyaluronic acid or sulfated GAGs (GAGsG), that consist mostly of chondroitin and keratin sulfate. The hyaluronic acid content of the synovial fluid was calculated as the difference between the concentrations of GAGsT and GAGsS. Samples of synovial fluid were collected from metacarpophalangeal joints of crossbred equines immediatelly after slaughter, by aseptic needle aspiration and then, following post mortem joint examination, were divided into four groups: a normal group as control (n = 17 ) and three alterated groups, obtained from joints with different degrees damage (+ = 15 ; ++ = 12 and +++ = 18 samples). The synovial fluid aldehyde concentrations were determined by a colorimetric reaction between aliphatic aldehydes and N-Methyl Benzothiazolidon Hydrazone. A colorimetric method with Alcian Blue using different MgCl2 concentrations, was used to quantify GAGsT and GAGsS. These results showed an increase of total protein concentration in the more alterated group (+++) (P <0,05). The collagen degradation product concentration increased in the group less altered (+) compared whith the more alterated group (+++) and collagen protein concentration showed an increase in both less alterated groups (+ and ++) compared with the more alterated group (+++) (P <0,05). The GAGsT, GAGsS and hyaluronic acid concentrations did not show statisticant differences between groups. GAGsT and hyaluronic acid concentrations showed a tendency to decrease in the more damaged samples. These results indicate that collagen degradation product concentration in synovial fluid could be used as a marker of initial changes in the osteoarthritis process.

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          Matrix metalloproteinases and tissue inhibitors of metalloproteinases in synovial fluids from patients with rheumatoid arthritis or osteoarthritis.

          Matrix metalloproteinases (MMPs) are expressed in joint tissues of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). The objective of this study was to define the steady state levels of seven different MMPs and two tissue inhibitors of metalloproteinases (TIMPs) as well as the potential metalloproteinase activity in the synovial fluid (SF) to provide more insight into the role of MMPs in cartilage destruction in RA and OA. Levels of MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-13, TIMP-1, and TIMP-2 in SF aspirated from knee joints of 97 patients with RA and 103 patients with OA were measured by the corresponding one step sandwich enzyme immunoassays. Proteolytic activity of MMPs in these SFs was examined in an assay using [(3)H]carboxymethylated transferrin substrate in the presence of inhibitors of serine and cysteine proteinases after activation with p-aminophenylmercuric acetate (APMA). Destruction of RA knee joints was radiographically evaluated. Levels of MMP-1, MMP-2, MMP-3, MMP-8, and MMP-9 were significantly higher in RA SF than in OA SF. MMP-7 and MMP-13 were detectable in more than 45% of RA SFs and in less than 20% of OA SFs, respectively. Among the MMPs examined, MMP-3 levels were extremely high compared with those of other MMPs. Direct correlations were seen between the levels of MMP-1 and MMP-3 and between those of MMP-8 and MMP-9 in RA SF. Although the levels of MMP-1 and MMP-3 increased even in the early stage of RA, those of MMP-8 and MMP-9 were low in the early stage and increased with the progression of RA. Molar ratios of the total amounts of the MMPs to those of the TIMPs were 5.2-fold higher in patients with RA than in OA, which was significant. APMA-activated metalloproteinase activity in SF showed a similar result, and a direct correlation was seen between the molar ratios and the activity in RA SF. Our results show that high levels of MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, and TIMP-1 are present in RA SF and suggest that once these MMPs are fully activated, they have an imbalance against TIMPs, which may contribute to the cartilage destruction in RA.
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            Biostatistical Analisis

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              Aggrecan turnover in human articular cartilage: use of aspartic acid racemization as a marker of molecular age.

              Aggrecan is a key component of the cartilage matrix. During aging, many changes occur in its composition and structure; in particular, there is an increase in the proportion of lower molecular weight monomers and of the "free" binding region. An important question has been whether these changes represent alterations in biosynthesis or whether they are due to the accumulation with age of the partially degraded fragments of the originally synthesized large monomer. In the present work we have used an independent tool, viz., the extent of racemization of aspartic acid to study the molecular "age" of different buoyant density fractions of the aggrecan of human articular cartilage, as well as of isolated free binding region and link protein. By measuring the D/LAsp ratio of the different aggrecan species, we were able to establish directly the relative residence times of these molecules in the cartilage matrix and, in combination with compositional and structural analyses, to define their "history" and calculate some of the kinetics constants characterizing their turnover. The value of the turnover constant for the large monomer in fraction A1D1 is 0.206 per year, which corresponds to a half-life of 3.4 years, while the turnover constant for the free binding region is 0.027 per year, which corresponds to a half-life of 25 years. It is thus clear that the rate of formation and turnover of the large monomer is much more rapid than the final degradation of the free binding region fragments, which explains the accumulation of the latter in cartilage during aging.
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                Author and article information

                Journal
                amv
                Archivos de medicina veterinaria
                Arch. med. vet.
                Facultad de Ciencias Veterinarias, Universidad Austral de Chile (Valdivia, , Chile )
                0301-732X
                2006
                : 38
                : 1
                : 47-52
                Affiliations
                [01] Santiago orgnameUniversidad de Chile orgdiv1Facultad de Ciencias Veterinarias y Pecuarias Chile hadarmes@ 123456uchile.cl
                Article
                S0301-732X2006000100007 S0301-732X(06)03800100007
                022688f0-9415-46ff-983a-b8e0278c6c07

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

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                Figures: 0, Tables: 0, Equations: 0, References: 37, Pages: 6
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                SciELO Chile

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                ARTICULOS ORIGINALES

                glicosaminoglicanos,glycosaminoglycans,synovial fluid,collagen,líquido sinovial,colágeno

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