Glycosaminoglycans in the pericellular matrix of chondrons and chondrocytes.
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 (gal beta 1,4glcNAc6S, gal6S beta 1,4glcNAc6S, glcNAc beta 1,3gal and glcNAc6S beta 1,3gal) 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.
Wang
Qi Guang
QG
Institute for Science & Technology in Medicine, University of Keele, Huxley Building, Staffordshire, ST5 5BG, UK.
El Haj
Alicia J
AJ
Kuiper
Nicola J
NJ
D000818:Animals; D002358:Cartilage, Articular / Q000737:chemistry* / Q000166:cytology; D002417:Cattle; D002452:Cell Count; D002469:Cell Separation / Q000379:methods; D019902:Chondrocytes / Q000737:chemistry*; D004586:Electrophoresis / Q000379:methods; D005109:Extracellular Matrix / Q000737:chemistry*; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans / Q000032:analysis*; D007150:Immunohistochemistry; D007632:Keratan Sulfate / Q000032:analysis; D023822:Tissue Engineering
D016326:Extracellular Matrix Proteins; D005456:Fluorescent Dyes; D006025:Glycosaminoglycans; D007632:Keratan Sulfate