May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Regeration of the Inner Collagenous Layer (ICL) of Human Bruch's Membrane by Cleaning and Extracellular Matrix Protein Coating: Morphometric Evidence
Author Affiliations & Notes
  • T.H. Tezel
    Department of Ophthalmology, Columbia Univ College of Physicians and Surgeons, New York, NY, United States
  • O. Ishida
    Department of Ophthalmology, Columbia Univ College of Physicians and Surgeons, New York, NY, United States
  • H.J. Kaplan
    Department of Ophthalmology, University of Louisville, Kentucky Lions Eye Center, Louisville, KY, United States
  • L.V. Del Priore
    Department of Ophthalmology, University of Louisville, Kentucky Lions Eye Center, Louisville, KY, United States
  • Footnotes
    Commercial Relationships  T.H. Tezel, None; O. Ishida, None; H.J. Kaplan, None; L.V. Del Priore, None.
  • Footnotes
    Support  Foundation Fighting Blindness, the Macula Foundation, and unrestricted funds from RPB, Inc.
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 1712. doi:
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      T.H. Tezel, O. Ishida, H.J. Kaplan, L.V. Del Priore; Regeration of the Inner Collagenous Layer (ICL) of Human Bruch's Membrane by Cleaning and Extracellular Matrix Protein Coating: Morphometric Evidence . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1712.

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

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Abstract

Abstract: : Purpose: We have previously shown that increased cross-linking of thickened collagen fibers and accumulation of large extracellular debris characterize aging of the ICL of human Bruch's membrane. Here we demonstrate that cleaning and/or extracellular protein recoating of the native collagen framework of aged (>60 years old) human Bruch's membrane restores the architecture of this layer. Methods: Explants of peripheral ICL were prepared from 4 aged donor human cadaver eyes (63-89 years old). Triplicate explants were cleaned by treatment with 0.1% Triton-X / 0.1% sodium citrate, and/or resurfaced with an extracellular matrix protein mixture (ECM-P) containing laminin (330µ/ml), fibronectin (250µ/ml), and vitronectin (33µ/ml). Random SEM images were analyzed morphometrically using NIH image analysis software to determine the following morphometrical parameters in each group: total pore area (nm2 per µ2), number of pores (per µ2), average diameter of collagen fibers (nm), fractal dimension of collagen fibers, area of deposits (nm2 per µ2), number of deposits (per µ2) and percent deposit coverage of the surface (%). Morphometrical data compared with untreated samples from the same donors. Results: The number of pores within the ICL increases slightly (5.9%) by cleaning but decreases by cleaning and extracellular protein coating (21.7%). Detergent treatment increases the diameter of individual collagen fibers (26.9%) and results in more individual fibers (11.3 % increase) by breaking collagen cross-links. 41.9% of the deposits are removed with cleaning. Cleaning followed by extracellular protein coating doubles the amount of matrix proteins on the collagen framework without increasing the occupied area (66.9%). Conclusions: Cleaning and extracellular protein coating of the Bruch's membrane can reestablish its native structure. Regeneration of Bruch's membrane may constitute an important step in reconstructive approaches for AMD and other aged-matrix diseases.

Keywords: Bruch's membrane • age-related macular degeneration • extracellular matrix 
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