| Changes in the Refractive Index of the Stroma and Its Extrafibrillar Matrix When the Cornea Swells Biophysical Journal, Volume 85, Issue 4, 1 October 2003, Pages 2205-2212 Keith M. Meek, Sally Dennis and Shukria Khan Abstract The transparency of the corneal stroma is critically dependent on the hydration of the tissue; if the cornea swells, light scattering increases. Although this scattering has been ascribed to the disruption caused to the arrangement of the collagen fibrils, theory predicts that light scattering could increase if there is an increased mismatch in the refractive indices of the collagen fibrils and the material between them. The purpose of this article is to use Gladstone and Dale's law of mixtures to calculate volume fractions for a number of different constituents in the stroma, and use these to show how the refractive indices of the stroma and its constituent extrafibrillar material would be expected to change as more solvent enters the tissue. Our calculations predict that solvent entering the extrafibrillar space causes a reduction in its refractive index, and hence a reduction in the overall refractive index of the bovine stroma according to the equation =1.335+0.04/(0.22+0.24 ′), where is the refractive index and ′ is the hydration of the swollen stroma. This expression is in reasonable agreement with our experimental measurements of refractive index versus hydration in bovine corneas. When the hydration of the stroma increases from =3.2 to =8.0, we predict that the ratio of the refractive index of the collagen fibrils to that of the material between them increases from 1.041 to 1.052. This change would be expected to make only a small contribution to the large increase in light scattering observed when the cornea swells to =8. Abstract | Full Text | PDF (126 kb) |
| Light Transmission in the Human Cornea as a Function of Position across the Ocular Surface: Theoretical and Experimental Aspects Biophysical Journal, Volume 95, Issue 11, 1 December 2008, Pages 5092-5099 James Doutch, Andrew J. Quantock, Valerie A. Smith and Keith M. Meek Abstract This article investigates the theoretical basis for differences in visible light transmission through the human cornea as a function of distance from the center. Experimentally, transmission decreases approximately linearly up to 3mm from the central axis, then quadratically beyond this. It is known that collagen fibril number density and collagen fibril radii change from the central region to the corneal periphery. We modeled, using the direct-summation-of-scattered-fields method, the effects these ultrastructural changes would be expected to have on light transmission, accounting for the increase in corneal thickness from center to edge. Fibril positions for the modeling were obtained from electron micrographs of human cornea. Theoretically, transmission remains fairly constant across the central cornea; then, as the fibril diameter increases, the predicted scattering increases. Interfibrillar spacing changes alter the refractive index ratio between matrix and fibril; this was modeled in our theoretical deductions. Fibril number density had a minimal effect on light propagation. Our theoretical deductions were in broad agreement with our experimental data. It is concluded that the reduced transparency in the peripheral stroma is primarily caused by changes in fibril radius and an increase in refractive index ratio between the fibril and the interfibrillar substance. Abstract | Full Text | PDF (587 kb) |
| Swelling Studies on the Cornea and Sclera: The Effects of pH and Ionic Strength Biophysical Journal, Volume 77, Issue 3, 1 September 1999, Pages 1655-1665 Yifei Huang and Keith M. Meek Abstract The biophysical properties of the cornea and sclera depend on the precise maintenance of tissue hydration. We have studied the swelling of the tissues as a function of pH and ionic strength of the bathing medium, using an equilibration technique that prevents the loss of proteoglycans during swelling. Synchrotron x-ray diffraction was used to measure the average intermolecular and interfibrillar spacings, the fibril diameters, and the collagen D-periodicity. We found that both tissues swelled least near pH 4, that higher hydrations were achieved at lower ionic strengths, and that sclera swelled about one-third as much as cornea under most conditions. In the corneal stroma, the interfibrillar spacing increased most with hydration at pH values near 7. Fibril diameters and D-periodicity were independent of tissue hydration and pH at hydrations above 1. Intermolecular spacings in both tissues decreased as the ionic strength was increased, and there was a significant difference between cornea and sclera. Finally, we observed that corneas swollen near pH 7 transmitted significantly more light than those swollen at lower pH levels. The results indicate that the isoelectric points of both tissues are close to pH 4. The effects of ionic strength can be explained in terms of chloride binding within the tissues. The higher light transmission achieved in corneas swollen at neutral pH may be related to the fact that the interfibrillar fluid is more evenly distributed under these conditions. Abstract | Full Text | PDF (319 kb) |
Copyright © 1997 The Biophysical Society. All rights reserved.
Biophysical Journal, Volume 72, Issue 3, 1382-1387, 1 March 1997
doi:10.1016/S0006-3495(97)78784-8
Research Article
D.W. Leonard and K.M. Meek
Open University, Oxford Research Unit, United Kingdom.
Ultrastructural data from x-ray diffraction studies of the cornea were used to estimate the refractive indices of the collagen fibrils and extrafibrillar material of human, ox, trout, and rabbit corneas. X-ray diffraction measurements of the size and spacing of the collagen fibrils and the separation between the constituent molecules of the fibrils were taken from a previous species study. The tissue volume fractions occupied by the stromal components were estimated and their refractive indices were calculated using the Gladstone-Dale law of mixtures. For the fibrils and extrafibrillar material, the refractive indices in the human cornea were 1.411 and 1.365; for the ox 1.413 and 1.357; for the rabbit 1.416 and 1.357; and for the trout 1.418 and 1.364, respectively. An alternative estimate based on the physical properties and chemical composition of bovine cornea, accounting for interfibrillar type VI collagen and cellular water, produced similar estimates of 1.416 and 1.356 for the fibrils and extrafibrillar material, respectively.