An ultrastructural and systemic analysis of glycosaminoglycans in thyroid-associated ophthalmopathy

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Abstract
Purpose To determine the ultrastructural localisation of glycosaminoglycans (GAGs) in the extraocular muscles (EOMs) of patients with thyroid-associated ophthalmopathy (TAO) and to see whether the quantity and type of GAGs present in blood and urine are markers of the disease. Methods Biopsies of affected EOMs were taken and studied by transmission electron microscopy (TEM). These were either fixed conventionally for TEM, or in 0.5% tannic acid and others for immunogold staining. Serum hyaluronan (HA) was measured using a radioimmunoassay in patients with TAO as well as control subjects, and urinary GAG levels assessed by photometric quantitation of hexuronic acid after reaction with carbazole. The excretion pattern of the urinary GAGs was determined by discontinuous electrophoresis. Results TEM showed that there is a marked ficpansion of the endomysial space in TAO EOM biopsies as compared with non-TAO strabismus specimens. This is caused by an increased number of collagen fibres. interspersed with a granular amorphous material surrounding striated collagen fibres shown to be hyaluronan by immunogold ftaining. In contrast, serum hyaluronan concentrations were similar in TAO and control patients, although there was a statistically significant difference in the ttrinary GAG excretion between the two groups of patients examined. By discontinuous electrophoresis, chondroitin ivlphate and heparan sulphate were present in both patients and controls. Conclusion GAGs and in particularly HA are present at the EOM level in patients with jecently inactive TAO. However, serum levels of HA and urinary GAGs are not sensitive indicators for their presence within the EOMs.