Glycosaminoglycan loss from cartilage after anterior cruciate ligament rupture: influence of time since rupture and chondral injury

OBJECTIVE: To quantify the concentration of sulfated glycosaminoglycans (GAGs) concentration in the synovial fluid (SF) of knees with chronic anterior cruciate ligament (ACL) rupture and to identify possible associations between GAG concentration in SF and the time elapsed since rupture and degree of chondral injury. METHOD: Fourteen adult male subjects with total unilateral ACL rupture, which had occurred between 5 and 144 months earlier, were assessed. All subjects underwent joint aspiration; it was possible to collect SF from ten individuals. The samples were quantified to determine the GAG concentration using dimethylmethylene blue (DMMB) staining. The degree of chondral injury was macroscopically evaluated using the modified Mankin histological scale. Spearman correlation test (< 0.05) was used to evaluate the association between GAG concentration and chondral injury, and Pearson correlation test (< 0.05) was used to evaluate the association between GAG concentration and the time elapsed since rupture. RESULTS: The GAG concentration in SF showed a mean variation of 73.84 ± 40.75 µg/ml, with a mean time of 40.4 ± 40.3 months since the rupture. There was no correlation between GAG concentration and time since the rupture (r= -0.09, p= 0.81). The chondral injury grades found were 0, 1, 4 and 5. There was no correlation between chondral injury grade and GAG concentration in SF (r= -0.41, p= 0.24). CONCLUSION: After at least 5 months, the GAG concentration in SF from knees with ACL rupture is independent of the time elapsed since rupture and/or the severity of chondral injury.

OBJETIVO: Quantificar a concentração de glicosaminoglicanas sulfatadas (GAGs) no líquido sinovial (LS) de joelhos com ruptura crônica do ligamento cruzado anterior (LCA) e identificar uma possível correlação entre a concentração de GAGs no LS e o tempo pós-ruptura e grau de lesão condral. MÉTODOS: Foram avaliados 14 indivíduos adultos do sexo masculino com ruptura total unilateral do LCA, ocorrida entre cinco a 144 meses. Todos os sujeitos foram puncionados, sendo possível a coleta de LS em dez indivíduos. As amostras foram quantificadas para determinar a concentração de GAGs usando a coloração azul de dimetilmetileno, método descrito por Farndale21. O grau de lesão condral foi macroscopicamente avaliado pela escala histológica de Mankin modificada por Messner14. As correlações entre concentração de GAGs e lesão condral foram feitas pelo teste de correlação de Sperman (p< 0,05) e a concentração de GAGs e tempo pós-ruptura pelo teste de correlação de Pearson (p< 0,05). RESULTADOS: Concentração de GAGs no LS apresentou variação média de 73,84 ± 40,75µg/mL, sendo o tempo médio pós-ruptura de 40,4 + 40,3 meses. Não houve correlação entre concentração de GAGs e o tempo pós-ruptura (r= -0,09, p= 0,81). Os graus de lesão condral encontrados foram de 0, 1, 4 e 5. Não houve correlação entre grau de lesão condral e a concentração de GAGs no LS (r= -0,41, p= 0,24). CONCLUSÕES: Após no mínimo cinco meses, a concentração de GAGs no LS de joelhos com ruptura do LCA independe do tempo pós-ruptura e/ou do grau de lesão condral.

Eye pigments of the blood-sucking insect, Triatoma infestans klug (Hemiptera, Reduviidae)

A pigmentação de olhos pretos (forma selvagem) e vermelhos (forma mutante) de Triatoma infestans foi estudada por espectrofotometria e comparada à de olhos vermelhos (selvagem) e brancos (mutante) de Drosophila melanogaster. Os perfis do espectro de absorção dos pigmentos de olho preto e vermelho de T. infestans foram semelhantes entre si e ao dos olhos de tipo selvagem de D. melanogaster. A similaridade com a forma selvagem de D. melanogaster indicou que ambos os tipos de olho de T. infestans continham omocromos do tipo xantomatina, o que foi confirmado por cromatografia ascendente em papel. Não foram detectadas pteridinas, melaninas e ominas como pigmentos de olho em T. infestans. A diferença na cor de olho em T. infestans foi considerada uma função da concentração de xantomatina, sendo menor o conteúdo de omocromo nos olhos vermelhos, embora isso provavelmente não afete a acuidade visual do inseto. Esses resultados estão de acordo com dados de outros autores quanto a semelhanças envolvendo outras características entre espécimes de olho preto e vermelho de T. infestans.

Distribution of small proteoglycans and glycosaminoglycans in humerus-related articular cartilage of chickens

The expression of components present in the cartilaginous extracellular matrix is related to development, gender, and genotype, as well as to the biomechanical properties of each type of cartilage. In the present study, we analyzed small proteoglycans and glycosaminoglycans present in different cartilages of the chicken wing after extraction with guanidine hydrochloride or papain. Quantitative analysis of glycosaminoglycans showed a larger amount in humeral cartilage (around 200 mg/g tissue) than in articular cartilage of the radius and ulna, with 138 and 80 mg/g tissue, respectively. Non-collagenous proteins isolated were predominantly from cartilage in the proximal regions of the humerus and radius. D4 fractions obtained by ultracentrifugation were separated by DEAE-Sephacel and Octyl-Sepharose chromatography and analyzed by SDS-PAGE. Two bands of 57 and 70-90 kDa were observed for all samples treated with ß-mercaptoethanol. Immunoblotting of these proteins was positive for the small proteoglycans fibromodulin and decorin, respectively. Apparently, the 57-kDa protein is present in macromolecular complexes of 160 and 200 kDa. Chondroitin sulfate was detected in all regions. HPLC analysis of the products formed by chondroitinase AC and ABC digestion mainly revealed ß-D-glucuronic acid and N-acetyl ß-D-galactosamine residues. The 4-sulfation/6-sulfation ratio was close to 3, except for the proximal cartilage of the radius (2.5). These results suggest functional differences between the scapula-humerus, humerus-ulna, and humerus-radius joints of the chicken wing. This study contributes to the understanding of the physiology of cartilage and joints of birds under different types of mechanical stress.

Is there a relationship between the state of aggregation os small proteoglycans and the biomechanical properties of tissues?

The small proteoglycans fibromodulin and decorin may play an important role in regulating collagen fibrillogenesis and interactions with growth factors. Here, we describe the presence of these proteoglycans in cartilage submitted to different biomechanical forces. Fibromodulin from chicken and bovine articular cartilage was shown to self-associate. The different states of fibromodulin aggregation due to disulfide bonding demonstrable in different regions of the same joint suggest that the presence of different biomechanical forces results in the differential expression of small proteoglycans. A 250-kDa complex found in chicken tibiotarsal cartilage, which migrates as a 59-kDa component in SDS-PAGE under reducing conditions, and which was recognized by anti-fibromodulin antibodies, was not demonstrable in tarsometatarsal cartilage where a different fibromodulin complex has been recently demonstrated. Biglycan and decorin were not expressed in the same way in different regions of the bovine knee joint, suggesting that there is a relationship between the expression of small proteoglycans and the different biomechanical properties of a tissue.

Detection of a small proteoglycan present in xiphoid cartilage regions submitted to different biomechanical forces

1. The effect of biomechanical forces on larges proteglycans and collagen of cartilage has deserved intensive study, enhacing the importance of these molecules to support a better distribution of compressive forces especially in articular cartilage. In the present study, other extracellular matrix components, non-collagenous proteins and small proteglycans, have been evaluated in terms of biomechanical tension. 2. Different parts of chicken xiphoid cartilage, lateral (R and L) and central (C) portions, which bear different biomechanical tensions, were analyzed. DEAE-cellulose chromatography profiles were similar for R and L portions. SDS-PAGE analyses revealed proteins of 29, 60 and 70 KDa for R and L. The 20-and 70-KDa proteins were not detected in the C portion while the 60-KDaprotein was presented at a high level. 3. The differences found between lateral (R and L) and central portions of the xiphoid cartilage may be related to the struycture of the cartilage which bears higher tension forces than the lateral parts