December 1998
Volume 39, Issue 13
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Articles  |   December 1998
Advanced glycation end products in vitreous: Structural and functional implications for diabetic vitreopathy.
Author Affiliations
  • A W Stitt
    Department of Ophthalmology, The Queen's University of Belfast, Northern Ireland, United Kingdom.
  • J E Moore
    Department of Ophthalmology, The Queen's University of Belfast, Northern Ireland, United Kingdom.
  • J A Sharkey
    Department of Ophthalmology, The Queen's University of Belfast, Northern Ireland, United Kingdom.
  • G Murphy
    Department of Ophthalmology, The Queen's University of Belfast, Northern Ireland, United Kingdom.
  • D A Simpson
    Department of Ophthalmology, The Queen's University of Belfast, Northern Ireland, United Kingdom.
  • R Bucala
    Department of Ophthalmology, The Queen's University of Belfast, Northern Ireland, United Kingdom.
  • H Vlassara
    Department of Ophthalmology, The Queen's University of Belfast, Northern Ireland, United Kingdom.
  • D B Archer
    Department of Ophthalmology, The Queen's University of Belfast, Northern Ireland, United Kingdom.
Investigative Ophthalmology & Visual Science December 1998, Vol.39, 2517-2523. doi:
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      A W Stitt, J E Moore, J A Sharkey, G Murphy, D A Simpson, R Bucala, H Vlassara, D B Archer; Advanced glycation end products in vitreous: Structural and functional implications for diabetic vitreopathy.. Invest. Ophthalmol. Vis. Sci. 1998;39(13):2517-2523.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: Advanced glycation end products (AGEs) form irreversible cross-links with many macromolecules and have been shown to accumulate in tissues at an accelerated rate in diabetes. In the present study, AGE formation in vitreous was examined in patients of various ages and in patients with diabetes. Ex vivo investigations were performed on bovine vitreous incubated in glucose to determine AGE formation and cross-linking of vitreous collagen. METHODS: By means of an AGE-specific enzyme-linked immunosorbent assay (ELISA), AGE formation was investigated in vitreous samples obtained after pars plana vitrectomy in patients with and without diabetes. In addition, vitreous AGEs were investigated in bovine vitreous collagen after incubation in high glucose, high glucose with aminoguanidine, or normal saline for as long as 8 weeks. AGEs and AGE cross-linking was subsequently determined by quantitative and qualitative assays. RESULTS: There was a significant correlation between AGEs and increasing age in patients without diabetes (r = 0.74). Furthermore, a comparison between age-matched diabetic and nondiabetic vitreous showed a significantly higher level of AGEs in the patients with diabetes (P < 0.005). Collagen purified from bovine vitreous incubated in 0.5 M glucose showed an increase in AGE formation when observed in dot blot analysis, immunogold labeling, and AGE ELISA. Furthermore, there was increased cross-linking of collagen in the glucose-incubated vitreous, when observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and protein separation. This cross-linking was effectively inhibited by coincubation with 10 mM aminoguanidine. CONCLUSIONS: This study suggests that AGEs may form in vitreous with increasing age. This process seems to be accelerated in the presence of diabetes and as a consequence of exposure to high glucose. Advanced glycation and AGE cross-linking of the vitreous collagen network may help to explain the vitreous abnormalities characteristic of diabetes.

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