Adsorption of glycosaminoglycans onto hydroxyapatite using chromatography.
Proteoglycans are known to play an important role in the mineralization process, acting either as promoters or inhibitors. In this study the binding affinity of a variety of constituent glycosaminoglycan to hydroxyapatite was studied. Glycosaminoglycans (10-1000 microg ml(-1) in 0.02 M sodium acetate (pH 6.8) were constantly circulated through a hydroxyapatite column for 1 h. The total amount of glycosaminoglycan bound was determined by dimethylmethylene blue assay. The relative affinities of the different glycosaminoglycans remaining bound to hydroxyapatite was investigated by examining their release in a 0-1 M sodium phosphate gradient. Differences were noted between the desorption profiles of dermatan sulfate with two elution peaks and chondroitin 4-sulfate and chondroitin 6-sulfate each with a single peak. Dermatan sulfate and chondroitin 6-sulfate had a higher affinity for hydroxyapatite than chondroitin 4-sulfate possibly due to the presence of differing di-sulfated disaccharide ratios in the glycosaminoglycan chains. These findings suggest the presence of a variety of binding forms of each glycosaminoglycan or the differing orientation of these forms to yield different complexes with hydroxyapatite. The Ca2+ co-ordinates of the glycosaminoglycans are known to vary and may in part explain these findings.
Department of Basic Dental Science, Dental School, University of Wales College of Medicine, Heath Park, Cardiff, UK.
J
J
Okazaki
G
G
Embery
R C
RC
Hall
D T
DT
Hughes Wassell
R J
RJ
Waddington
A
A
Kamada
R J
RJ
Waddington
A
A
Kamada
D000327:Adsorption; D001665:Binding Sites; D001672:Biocompatible Materials / Q000737:chemistry*; D002853:Chromatography, Liquid / Q000379:methods; D017886:Durapatite / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008751:Methylene Blue / Q000031:analogs & derivatives; D015203:Reproducibility of Results
D001672:Biocompatible Materials; D006025:Glycosaminoglycans; D017886:Durapatite; D008751:Methylene Blue; C016401:1,9-dimethylmethylene blue