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      Experimental evaluation of stiffening effect induced by UVA/Riboflavin corneal cross-linking using intact porcine eye globes

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          Abstract

          UVA/riboflavin corneal cross-linking (CXL) is a common used approach to treat progressive keratoconus. This study aims to investigate the alteration of corneal stiffness following CXL by mimicking the inflation of the eye under the in vivo loading conditions. Seven paired porcine eye globes were involved in the inflation test to examine the corneal behaviour. Cornea-only model was constructed using the finite element method, without considering the deformation contribution from sclera and limbus. Inverse analysis was conducted to calibrate the non-linear material behaviours in order to reproduce the inflation test. The corneal stress and strain values were then extracted from the finite element models and tangent modulus was calculated under stress level at 0.03 MPa. UVA/riboflavin cross-linked corneas displayed a significant increase in the material stiffness. At the IOP of 27.25 mmHg, the average displacements of corneal apex were 307 ± 65 μm and 437 ± 63 μm (p = 0.02) in CXL and PBS corneas, respectively. Comparisons performed on tangent modulus ratios at a stress of 0.03 MPa, the tangent modulus measured in the corneas treated with the CXL was 2.48 ± 0.69, with a 43±24% increase comparing to its PBS control. The data supported that corneal material properties can be well-described using this inflation methods following CXL. The inflation test is valuable for investigating the mechanical response of the intact human cornea within physiological IOP ranges, providing benchmarks against which the numerical developments can be translated to clinic.

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          Stress-strain measurements of human and porcine corneas after riboflavin-ultraviolet-A-induced cross-linking.

          To evaluate the biomechanical effect of combined riboflavin-ultraviolet A (UVA) treatment on porcine and human corneas. Department of Ophthalmology, Technical University of Dresden, Dresden, Germany. Corneal strips from 5 human enucleated eyes and 20 porcine cadaver corneas were treated with the photosensitizer riboflavin and irradiated with 2 double UVA diodes (370 nm, irradiance = 3 mW/cm2) for 30 minutes. After cross-linking, static stress-strain measurements of the treated and untreated corneas were performed using a microcomputer-controlled biomaterial tester with a prestress of 5 x 10(3) Pa. There was a significant increase in corneal rigidity after cross-linking, indicated by a rise in stress in treated porcine corneas (by 71.9%) and human corneas (by 328.9%) and in Young's modulus by the factor 1.8 in porcine corneas and 4.5 in human corneas. The mean central corneal thickness was 850 microm +/- 70 (SD) in porcine corneas and 550 +/- 40 microm in human corneas. Riboflavin-UVA-induced collagen cross-linking led to an increase in mechanical rigidity in porcine corneas and an even greater increase in human corneas. As collagen cross-linking is maximal in the anterior 300 microm of the cornea, the greater stiffening effect in human corneas can be explained by the relatively larger portion of the cornea being cross-linked in the overall thinner human cornea.
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            Large Deformation Isotropic Elasticity - On the Correlation of Theory and Experiment for Incompressible Rubberlike Solids

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              Riboflavin/ultraviolet-a-induced collagen crosslinking for the treatment of keratoconus.

              In animal eyes, a significant increase in corneal biomechanical stiffness has been found after collagen crosslinking by combined riboflavin/ultraviolet-A (UVA) treatment. The aim of the present study was to evaluate the clinical usefulness of riboflavin/UVA-induced collagen crosslinking for bringing the progression of keratoconus to a halt. Prospective, nonrandomized clinical pilot study. Twenty-three eyes of 22 patients with moderate or advanced progressive keratoconus (maximum K value, 48-72 diopters) were included. After central corneal abrasion, photosensitizing riboflavin drops were applied and the eyes exposed to UVA (370 nm, 3 mW/cm(2)) in a 1-cm distance for 30 minutes. Postoperative examinations were performed in 6-month intervals, including visual acuity testing, corneal topography, slit-lamp examination, measurement of endothelial cell density, and photographic documentation. The follow-up time was between 3 months and 4 years. In all treated eyes, the progression of keratoconus was at least stopped. In 16 eyes (70%) regression with a reduction of the maximal keratometry readings by 2.01 diopters and of the refractive error by 1.14 diopters was found. Corneal and lens transparency, endothelial cell density, and intraocular pressure remained unchanged. Visual acuity improved slightly in 15 eyes (65%). Collagen crosslinking may be a new way for stopping the progression of keratectasia in patients with keratoconus. The need for penetrating keratoplasty might then be significantly reduced in keratoconus. Given the simplicity and minimal costs of the treatment, it might also be well-suited for developing countries. Long-term results are necessary to evaluate the duration of the stiffening effect and to exclude long term side-effects.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: Writing – review & editing
                Role: ConceptualizationRole: MethodologyRole: Writing – review & editing
                Role: Methodology
                Role: ConceptualizationRole: Formal analysisRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                4 November 2020
                2020
                : 15
                : 11
                : e0240724
                Affiliations
                [1 ] School of Engineering, University of Liverpool, Liverpool, United Kingdom
                [2 ] Department of Chemical Engineering, Feng Chia University, Taichung, Taiwan
                Cardiff University, UNITED KINGDOM
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                [¤a]

                Current address: School of Engineering, University of Liverpool, Liverpool, United Kingdom

                [¤b]

                Current address: Department of Chemical Engineering, Feng Chia University, Taichung, Taiwan

                Author information
                https://orcid.org/0000-0002-4473-1900
                https://orcid.org/0000-0001-5772-9474
                Article
                PONE-D-20-16381
                10.1371/journal.pone.0240724
                7641398
                33147249
                4ff32477-2415-49a0-bbb4-24017b633a1a
                © 2020 Chang et al

                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
                : 30 May 2020
                : 2 October 2020
                Page count
                Figures: 6, Tables: 2, Pages: 13
                Funding
                This work was supported by grants from the Tawian Ministry of Science and Technology (MOST 109-2221-E-035 -036 -MY3 and MOST 109-2221-E-035 -007 -MY3).
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Ocular System
                Ocular Anatomy
                Cornea
                Medicine and Health Sciences
                Anatomy
                Ocular System
                Ocular Anatomy
                Cornea
                Biology and Life Sciences
                Anatomy
                Head
                Eyes
                Medicine and Health Sciences
                Anatomy
                Head
                Eyes
                Biology and Life Sciences
                Anatomy
                Ocular System
                Eyes
                Medicine and Health Sciences
                Anatomy
                Ocular System
                Eyes
                Physical Sciences
                Materials Science
                Material Properties
                Mechanical Properties
                Stiffness
                Physical Sciences
                Mathematics
                Geometry
                Tangents
                Physical Sciences
                Physics
                Classical Mechanics
                Deformation
                Physical Sciences
                Physics
                Classical Mechanics
                Damage Mechanics
                Deformation
                Physical Sciences
                Chemistry
                Physical Chemistry
                Chemical Bonding
                Cross-Linking
                Biology and Life Sciences
                Biochemistry
                Proteins
                Collagens
                Biology and Life Sciences
                Biomechanics
                Custom metadata
                All relevant data are available from Dryad (doi: 10.5061/dryad.z8w9ghx9f).

                Uncategorized
                Uncategorized

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