J Winkler1, H Lünsdorf. 1. Department of Ophthalmology, Medical University of Lübeck, Germany. jorgw@yahoo.com
Abstract
PURPOSE: Asteroid hyalosis is a disease of the vitreous, characterized by brilliant reflecting particles, termed asteroid bodies, which are surrounded by a tightly adhering network of fibrils. The composition and mode of formation of asteroid bodies is not yet understood in detail. The purpose of this study was to investigate the ultrastructure of asteroid bodies and to identify the intrinsic inorganic and organic components that contribute to the nature and development of asteroid bodies. METHODS: Electron energy loss spectroscopy and energy-filtered transmission electron microscopy were used for the elemental analysis of asteroid bodies. The ultrastructural localization of glycosaminoglycans was investigated, using lectin and antibody conjugates in conjunction with transmission electron microscopy and epifluorescence microscopy. Anionic sites of glycosaminoglycans were detected with 15 nm cationic colloidal gold at low pH, applied as a postembedding technique. Ultrastructural details of asteroid bodies were documented using fast Fourier transform analysis of zero-loss filtered images. RESULTS: Element mapping of asteroid bodies by electron spectroscopic imaging revealed a homogeneous distribution of calcium, phosphorus, and oxygen. The electron energy loss spectra of these elements showed details similar to those found for hydroxyapatite. Additionally, high contrast and sensitivity against a calcium-specific chelator highlighted the crystalline, apatite-like nature of asteroid bodies. Immunofluorescence microscopy revealed the presence of chondroitin-6-sulfate at the periphery of asteroid bodies, which is in agreement with the ultrastructural colocalization of anionic sites. Fast Fourier transform analysis revealed that each 7-nm periodicity of asteroid lamellar stacks is divided by a fine, parallel-oriented line, separating each 7-nm layer into two halves of 3.5-nm thickness. Carbohydrates specific for hyaluronic acid were observed by lectin-gold labeling to be part of the inner matrix of asteroid bodies. CONCLUSIONS: The results of this study demonstrate the structural and elemental similarity of asteroid bodies with hydroxyapatite. Proteoglycans and their glycosaminoglycan side chains are implicated in playing a role in regulating the biomineralization process.
PURPOSE:Asteroid hyalosis is a disease of the vitreous, characterized by brilliant reflecting particles, termed asteroid bodies, which are surrounded by a tightly adhering network of fibrils. The composition and mode of formation of asteroid bodies is not yet understood in detail. The purpose of this study was to investigate the ultrastructure of asteroid bodies and to identify the intrinsic inorganic and organic components that contribute to the nature and development of asteroid bodies. METHODS: Electron energy loss spectroscopy and energy-filtered transmission electron microscopy were used for the elemental analysis of asteroid bodies. The ultrastructural localization of glycosaminoglycans was investigated, using lectin and antibody conjugates in conjunction with transmission electron microscopy and epifluorescence microscopy. Anionic sites of glycosaminoglycans were detected with 15 nm cationic colloidal gold at low pH, applied as a postembedding technique. Ultrastructural details of asteroid bodies were documented using fast Fourier transform analysis of zero-loss filtered images. RESULTS: Element mapping of asteroid bodies by electron spectroscopic imaging revealed a homogeneous distribution of calcium, phosphorus, and oxygen. The electron energy loss spectra of these elements showed details similar to those found for hydroxyapatite. Additionally, high contrast and sensitivity against a calcium-specific chelator highlighted the crystalline, apatite-like nature of asteroid bodies. Immunofluorescence microscopy revealed the presence of chondroitin-6-sulfate at the periphery of asteroid bodies, which is in agreement with the ultrastructural colocalization of anionic sites. Fast Fourier transform analysis revealed that each 7-nm periodicity of asteroid lamellar stacks is divided by a fine, parallel-oriented line, separating each 7-nm layer into two halves of 3.5-nm thickness. Carbohydrates specific for hyaluronic acid were observed by lectin-gold labeling to be part of the inner matrix of asteroid bodies. CONCLUSIONS: The results of this study demonstrate the structural and elemental similarity of asteroid bodies with hydroxyapatite. Proteoglycans and their glycosaminoglycan side chains are implicated in playing a role in regulating the biomineralization process.