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      Analysis of the Viscoelastic Properties of the Human Cornea Using Scheimpflug Imaging in Inflation Experiment of Eye Globes

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          Abstract

          Purpose

          To demonstrate a Scheimpflug-based imaging procedure for investigating the depth- and time-dependent strain response of the human cornea to inflation testing of whole eye globes.

          Methods

          Six specimens, three of which with intact corneal epithelium, were mounted in a customized apparatus within a humidity and temperature-monitored wet chamber. Each specimen was subjected to two mechanical tests in order to measure corneal strain resulting from application of cyclic ( cyclic regimen) and constant ( creep regimen) stress by changing the intra-ocular pressure (IOP) within physiological ranges (18–42 mmHg). Corneal shape changes were analyzed as a function of IOP and both corneal stress-strain curves and creep curves were generated.

          Results

          The procedure was highly accurate and repeatable. Upon cyclic stress application, a biomechanical corneal elasticity gradient was found in the front-back direction. The average Young's modulus of the anterior cornea ranged between 2.28±0.87 MPa and 3.30±0.90 MPa in specimens with and without intact epithelium (P = 0.05) respectively. The Young's modulus of the posterior cornea was on average 0.21±0.09 MPa and 0.17±0.06 MPa (P>0.05) respectively. The time-dependent strain response of the cornea to creep testing was quantified by fitting data to a modified Zener model for extracting both the relaxation time and compliance function.

          Conclusion

          C yclic and creep mechanical tests are valuable for investigating the strain response of the intact human cornea within physiological IOP ranges, providing meaningful results that can be translated to clinic. The presence of epithelium influences the results of anterior corneal shape changes when monitoring deformation via Scheimpflug imaging in inflation experiments of whole eye globes.

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          Most cited references43

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          The three-dimensional organization of collagen fibrils in the human cornea and sclera.

          The organization of collagen fibrils in the human cornea and sclera was studied by scanning electron microscopy, after digestion of cellular elements by sodium hydroxide, and by conventional transmission electron microscopy. The collagen fibrils in the cornea had a uniform diameter of about 25 nm. In Bowman's layer, individual collagen fibrils were interwoven densely to form a felt-like sheet. In the stroma, most of the collagen fibrils ran abreast in lamellae, with varying widths and thickness. These lamellae were arranged basically parallel to the corneal surface but often communicated with those of adjacent layers by interchanging their fibrils. In the innermost stromal region adjacent to Descemet's membrane, collagen fibrils were oriented in various directions and interlaced, forming loose fibrillar networks. The sclera, however, was composed of collagen fibrils with various diameters ranging from 25-230 nm. Although these collagen fibrils formed bundles, they were not parallel but were entangled in individual bundles. These collagen bundles varied in width and thickness, often gave off branches, and intertwined with each other.
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            Brillouin optical microscopy for corneal biomechanics.

            The mechanical properties of corneal tissue are linked to prevalent ocular diseases and therapeutic procedures. Brillouin microscopy is a novel optical technology that enables three-dimensional mechanical imaging. In this study, the feasibility of this noncontact technique was tested for in situ quantitative assessment of the biomechanical properties of the cornea. Brillouin light-scattering involves a spectral shift proportional to the longitudinal modulus of elasticity of the tissue. A 532-nm single-frequency laser and a custom-developed ultrahigh-resolution spectrometer were used to measure the Brillouin frequency. Confocal scanning was used to perform Brillouin elasticity imaging of the corneas of whole bovine eyes. The longitudinal modulus of the bovine corneas was compared before and after riboflavin corneal collagen photo-cross-linking. The Brillouin measurements were then compared with conventional stress-strain mechanical test results. High-resolution Brillouin images of the cornea were obtained, revealing a striking depth-dependent variation of the elastic modulus across the cornea. Along the central axis, the Brillouin frequency shift varied gradually from 8.2 GHz in the epithelium to 7.5 GHz near the endothelium. The coefficients of the down slope were measured to be approximately 1.09, 0.32, and 2.94 GHz/mm in the anterior, posterior, and innermost stroma, respectively. On riboflavin collagen cross-linking, marked changes in the axial Brillouin profiles (P < 0.001) were noted before and after cross-linking. Brillouin imaging can assess the biomechanical properties of cornea in situ with high spatial resolution. This novel technique has the potential for use in clinical diagnostics and treatment monitoring.
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              Comparative study of corneal strip extensometry and inflation tests.

              Strip extensometry tests are usually considered less reliable than trephinate inflation tests in studying corneal biomechanics. In spite of the evident simplicity of strip extensometry tests, several earlier studies preferred inflation tests in determining the constitutive relationship of the cornea and its other material properties, such as Young's modulus and the hysteresis behaviour. In this research, the deficiencies of the strip tests are discussed and a mathematical procedure presented to take account of these deficiencies when obtaining the corneal material properties. The study also involves testing 10 pairs of porcine corneas using both strip extensometry and trephinate inflation techniques and the results are subjected to mathematical back analysis in order to determine the stress-strain behaviour. The behaviour obtained from the strip extensometry tests and using the new mathematical analysis procedure is shown to match closely the inflation test results.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                14 November 2014
                : 9
                : 11
                : e112169
                Affiliations
                [1 ]CNR-IPCF, Unit of Support of Cosenza, Ponte P. Bucci, 87036 Rende, Italy
                [2 ]Vision Engineering Italy S.r.l., Via Adda 7, 00198 Rome, Italy
                [3 ]Fondazione G.B. Bietti IRCCS, Via Livenza 3, 00198 Rome, Italy
                University of Missouri-Columbia, United States of America
                Author notes

                Competing Interests: The authors declare that no competing interests exist.

                Conceived and designed the experiments: GL ML. Performed the experiments: MR ML. Analyzed the data: ML GL SS. Contributed reagents/materials/analysis tools: GL. Wrote the paper: GL SS ML. Revised the manuscript: GL SS ML.

                Article
                PONE-D-14-18124
                10.1371/journal.pone.0112169
                4232387
                25397674
                c6e60478-0511-43a1-bb0c-596ab3fd64a0
                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 25 April 2014
                : 13 October 2014
                Page count
                Pages: 10
                Funding
                This work was supported by a grant from the National Framework Program for Research and Innovation PON (research program n. 01_00110). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Ocular System
                Ocular Anatomy
                Cornea
                Biomechanics
                Physical Sciences
                Chemistry
                Chemical Properties
                Viscosity
                Custom metadata
                The authors confirm that all data underlying the findings are fully available without restriction. All relevant data are within the paper.

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                Uncategorized

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