In brief, the tissue samples being placed within eppendorf tubes after which the volume was made up to 100 μl with distilled water. Then 1 ml of Blyscan dye reagent was added to each specimen. Glycosaminoglycan standards were then prepared by employing aliquots with the following concentration of GAG: 1.0, 2.0, 3.0, 4.0 & 5.0 μg. The specimens were subsequently placed on a mechanical agitator at room temperature for 30 minutes to allow binding of the blyscan dye to the sulphated glycosaminoglycans. The next step involved centrifuging the specimens at 10000 rpm for 10 minutes. The specimens were then inverted and drained removing the unbound dye solution. The next step involved releasing of the bound dye, which, was achieved by adding 1 ml of dissociation reagent to each one of the cartilage samples. The specimens were then mixed by way of a vortex. The contents were transferred to a well plate which was clearly labelled. This was inserted within a photospectrometer to calculate absorption ratios of the samples. The absorption ratios for the standards were plotted on a graph and these were used to calculate the content of GAG (n = 3).