Hyaluronidase treatment, collagen fibril packing, and normal transparency in rabbit corneas.

PURPOSE: Hyaluronidase treatment is the initial step of corneaplasty, a treatment under development that induces stromal softening and involves the application of a custom designed forming lens to achieve modification of refractive error. The purpose of this investigation was to examine changes in the arrangement of stromal collagen fibrils after hyaluronidase treatment. METHODS: Rabbit corneas were evaluated by slit-lamp microscopy at 0, 2 and 7 days after treatment and haze was assessed by subjective observation. Molecular and interfibrillar Bragg spacing of corneal collagen were measured from synchrotron x-ray scattering patterns. Transmission electron microscopy and digital image analysis were used to calculate radial distribution functions from the positions of collagen fibrils. The calculated fibril sizes and positions were also used to predict the transmission of visible light through these corneas. RESULTS: Hyaluronidase-treated corneas were shown to have a decreased interfibrillar Bragg spacing of 15% to 21%. Fibril hydration did not change. Transparency of these corneas remained unaltered. CONCLUSIONS: Hyaluronidase reduced the hydration of the corneal stroma, which led to a more compacted collagen fibril arrangement. This compression was predicted to cause a small reduction in the transmission of visible light through the cornea but not to a point likely to cause visual impairment.

Organization of collagen fibrils in the corneal stroma in relation to mechanical properties and surgical practice.

We have used synchrotron x-ray diffraction to obtain quantitative information about the precise orientation of the collagen fibrils in the human cornea and sclera, and how these fuse at the limbus. We have shown that the human corneal stroma has preferred collagen orientation in the inferior-superior and nasal-temporal directions. At the limbus, however, the preferred orientation is tangential to the cornea. We demonstrated that these limbal fibrils are in the form of a circumcorneal annulus and quantified how this annulus varies with position. We have also started to unravel how the corneal collagen fibrils fuse with the collagen in the annulus. The results have both mechanical and surgical implications. Keratoplasty is performed without considering the preferred directions of the collagen fibrils. Surgery is increasingly used to correct astigmatism and myopia. The site and direction of an incision during, for example, cataract surgery, can influence the eventual level of astigmatism. X-ray diffraction helps our understanding of the underlying structural and hence mechanical reasons for refractive problems following surgery.

The integration of the corneal and limbal fibrils in the human eye.

The precise orientation of the collagen fibrils in human cornea and sclera and the method by which these two areas fuse together at the limbus have never been determined, despite the importance of this information. From a consideration of the mechanics of the system, fibril orientation in the tissue has the potential to affect the curvature of the cornea so, by inference, refractive problems such as astigmatism involving an incorrect curvature of the cornea may be related to fibril orientation. The high intensity and small beam size of a synchrotron x-ray source has enabled us to study fibril orientation in post-mortem human cornea and sclera. Previously we have revealed two preferred directions of orientation in the cornea (Meek, K. M., T. Blamires, G. F. Elliot, T. Y. Gyi, and C. J. Nave. 1987. Curr. Eye Res. 6:841-846) and a circumcorneal annulus in the limbus (Newton, R. H., and K. M. Meek. 1998. Invest. Ophthalmol. & Visual Sci. 39: 1125-1134). Here we present the results of our investigation into the relationship between these two features. Our measurements indicate that the corneal fibrils oriented in the two preferred directions bend at the limbus to run circumferentially. On the basis of these results we propose a model as to how the human cornea and sclera fuse together.

Circumcorneal annulus of collagen fibrils in the human limbus.

PURPOSE: To quantify the orientation of the collagen fibrils in the human limbus and to compare it with the orientation in the cornea and the sclera. METHODS: Fibril orientation was measured from 100 synchrotron x-ray diffraction patterns collected at intervals along lines across the cornea, limbus, and sclera. RESULTS: A distinct circumcorneal annulus of collagen fibrils was revealed in the limbus. For the individual cornea investigated, the annulus was not uniform around the corneal circumference; its width, fibril angular spread, and fibril density all varied with position. The average width of the annulus was narrower in the superior sector (1.5 mm) than in the inferior sector (2.0 mm). The results from the edge of the cornea suggested that the preferentially aligned corneal fibrils bend sharply at the limbus to run circumferentially or that, together with this bending or alone, extra fibrils arising in the sclera run across this zone. In the cornea the biaxial preferred directions became more pronounced from the center of the cornea toward the limbus along the superior-inferior axis. CONCLUSIONS: The excess of circumferential fibrils in the limbus, predicted from the sharp change of curvature in the surface of the eye at the limbus, appeared to take the form of a well-defined annulus. From a consideration of the mechanics of the system it seemed probable that the purpose of this annulus was to help maintain the correct curvature of the cornea. Before further research, it was hypothesized that some refractive problems associated with an incorrect curvature of the cornea may be related in part to abnormalities of this circumcorneal annulus.

Regional cerebral blood flow imaging: a quantitative comparison of technetium-99m-HMPAO SPECT with C15O2 PET.

The aim of this study was to compare technetium-99m-hexamethylpropyleneamineoxime (99mTc-HMPAO) single-photon emission computed tomography (SPECT) with regional cerebral blood flow (rCBF) imaging using positron emission tomography (PET). As investigation of dementia is likely to be one of the main uses of routine rCBF imaging, 18 demented patients were imaged with both techniques. The PET data were compared quantitatively with three versions of the SPECT data. These were, first, data normalized to the SPECT cerebellar uptake, second, data linearly corrected using the PET cerebellar value and, finally, data Lassen corrected for washout from the high flow areas. Both the linearly-corrected (r = 0.81) and the Lassen-corrected (r = 0.79) HMPAO SPECT data showed good correlation with the PET rCBF data. The relationship between the normalized HMPAO SPECT data and the PET data was nonlinear. It is not yet possible to obtain rCBF values in absolute units from HMPAO SPECT without knowledge of the true rCBF in one reference region for each patient.