Glycosaminoglycans in the pericellular matrix of chondrons and chondrocytes

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Blackwell Science Inc

RESUMO

This is the first study to quantitate and profile the glycosaminoglycan (GAG) composition of the pericellular matrix (PCM) of chondrons and chondrocytes using the highly sensitive technique; fluorophore-assisted carbohydrate electrophoresis (FACE). Bovine articular chondrocytes and chondrons were isolated enzymatically. High cell yield and viability were obtained for both preparations. Chondrons had strong immunofluorescent labeling for keratan sulphate and chondroitin-6 sulphate but no labeling for hyaluronan. We compared the immunofluorescent data with FACE. The quantities of total keratan sulphate were determined to be 0.013±0.002 pg cell−1 and 0.032 ±0.003 pg cell−1 in the chondrocyte and chondron preparations, respectively. Four internal keratan sulphate sugars were detected (galβ1, 4glcNAc6S, gal6Sβ1, 4glcNAc6S, glcNAcβ1, 3gal and glcNAc6Sβ1, 3gal) for both preparations but they were present at significantly higher concentrations in chondron preparations (P<0.01). Total chondroitin sulphate (CS) was determined to be 0.054±0.004 pg cell−1 and 0.077±0.005 pg cell−1 for chondrocyte and chondron preparations, respectively. Unsulphated CS disaccharide levels were similar but chondrons had significantly more chondroitin-4 sulphated disaccharides and chondroitin-6 sulphated disaccharides (P<0.05). Hyaluronan acid was present at low concentrations (0.010±0.001 pg cell−1) in both chondrocytes and chondrons. In this study, enzyme digestion coupled with FACE separation revealed new information about the differences in GAGs from isolated chondrocyte and chondron preparations. Further investigation of the differences in GAGs from chondrocytes and chondrons from different zones of articular cartilage may be useful for tissue engineering approaches.

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