Regional differences in electroactive response of the sclera.

The sclera exhibits mechanical response when subjected to an external electric stimulation. The scleral electroactive response is a function of its charge density, mechanical properties, thickness, and strength of the applied electric voltage. The primary objective of the present work was to investigate the regional differences in the electroactive response of porcine sclera. To this end, we cut scleral strips in meridional directions from superior-temporal, superior-nasal, inferior-temporal, and inferior-nasal quadrants. In addition, we excised samples circumferentially from the posterior, equatorial, and anterior regions. The electroactive bending response of these samples was measured under 10 and 15 V in 0.15 M NaCl solution. The meridional samples were tested under two different configurations by clamping them either from their anterior or posterior end. It was observed that the scleral electroactive deformation increased with increasing the the electric voltage. Furthermore, regardless of the region from which meridional strips were excised, their electroactive response was considerably larger when they were clamped from their anterior end. Unlike meridional strips, the electroactive response of circumferential samples was significantly dependent on the location, that is, the average maximum bending angle of posterior samples was significantly larger than that of equatorial and anterior strips. The regionally different electroactive bending response of the sclera was discussed in terms of the variation in its biochemical and biomechanical properties throughout the eyeball.

Finite Deformation of Scleral Tissue under Electrical Stimulation: An Arbitrary Lagrangian-Eulerian Finite Element Method.

The sclera is considered as the principal load-bearing tissue within the eye. The sclera is negatively charged; thus, it exhibits mechanical response to electrical stimulation. We recently demonstrated the electroactive behavior of sclera by performing experimental measurements that captured the deformation of the tip of scleral strips subjected to electric voltage. We also numerically analyzed the electromechanical response of the tissue using a chemo-electro-mechanical model. In the pre-sent study, we extended our previous work by experimentally characterizing the deformation profile of scleral strips along their length under electrical stimulation. In addition, we improved our previous mathematical model such that it could numerically capture the large deformation of samples. For this purpose, we considered the transient variability of the fixed charge density and the coupling between mechanical and chemo-electrical phenomena. These improvements in-creased the accuracy of the computational model, resulting in a better numerical representation of experimentally measured bending angles.

On the Mechanical Roles of Glycosaminoglycans in the Tensile Properties of Porcine Corneal Stroma.

Purpose: The cornea is primarily composed of collagen fibrils that are embedded in a ground substance rich in proteoglycans and other glycoproteins. It is known that glycosaminoglycan (GAG) side chains of proteoglycans form anti-parallel duplexes between collagen fibrils. The present work was done in order to investigate the mechanical function of GAGs in defining the tensile properties of porcine corneal stroma. Methods: Porcine corneal stromal strips dissected from the nasal-temporal direction were divided into control, buffer-treated, and enzyme-treated groups. The samples in the control group were used immediately after dissection. However, the buffer-treated and enzyme-treated samples were, respectively, incubated for 18 hours at 37°C in a buffer solution made up of 100-mM sodium acetate at pH 6.0 or in an enzyme solution containing keratanase II. The Blyscan assay was used to quantify the total GAG content and assess GAG depletion in the samples treated with the enzyme and buffer solutions. Uniaxial tensile tests were also performed to determine the effect of GAG removal on mechanical properties of the cornea. Results: The GAG content in enzyme-treated samples was significantly lower than that of the normal and buffer-treated specimens (P < 0.05). Moreover, GAG-depleted strips showed significantly softer mechanical responses in comparison with the control and buffer samples (P < 0.05). Conclusions: Removing GAGs from the corneal extracellular matrix led to significant tensile property reduction; supporting the hypothesis that there exists a strong correlation between the GAG content and mechanical properties of the corneal stroma.

Modeling and experimental investigation of electromechanical properties of scleral tissue; a CEM model using an anisotropic hyperelastic constitutive relation.

The sclera is a soft tissue primarily consisting of collagen fibers, elastin, and proteoglycans. The proteoglycans are composed of a core protein and negatively charged glycosaminoglycan side chains. The fixed electric charges inside the scleral extracellular matrix play a key role in its swelling and are expected to cause the tissue to deform in response to an electric field. However, the electroactive response of the sclera has not yet been investigated. The present work experimentally demonstrates that sclera behaves similar to an anionic electrosensitive hydrogel and develops a chemo-electro-mechanical (CEM) mathematical framework for its electromechanical response. In the numerical model, a hyperelastic constitutive law with distributed collagen fibers is used to capture the nonlinear mechanical properties of the sclera, and the coupled Poisson-Nernst-Planck equations represent the distribution of mobile ions throughout the domain. After calibrating the proposed numerical CEM model against the experimental measurements, we employ it to investigate the effects of different parameters on the scleral electromechanical response including the voltage and fixed charge density. The experimental and numerical findings of the present study confirm that sclera behaves as an electroactive hydrogel and provide new insight into the mechanical response of this ocular tissue.

Effect of corneal collagen crosslinking on viscoelastic shear properties of the cornea.

The cornea is responsible for most of the refractive power in the eye and acts as a protective layer for internal contents of the eye. The cornea requires mechanical strength for maintaining its precise shape and for withstanding external and internal forces. Corneal collagen crosslinking (CXL) is a treatment option to improve corneal mechanical properties. The primary objective of this study was to characterize CXL effects on viscoelastic shear properties of the porcine cornea as a function of compressive strain. For this purpose, corneal buttons were prepared and divided into three groups: control group (n = 5), pseudo-crosslinked group (n = 5), and crosslinked group (n = 5). A rheometer was used to perform dynamics torsional shear experiments on corneal disks at different levels of compressive strain (0%-40%). Specifically, strain sweep experiments and frequency sweep tests were done in order to determine the range of linear viscoelasticity and frequency dependent shear properties, respectively. It was found that the shear properties of all samples were dependent on the shear strain magnitude, loading frequency, and compressive strain. With increasing the applied shear strain, all samples showed a nonlinear viscoelastic response. Furthermore, the shear modulus of samples increased with increasing the frequency of the applied shear strain and/or increasing the compressive strain. Finally, the CXL treatment significantly increased the shear storage and loss moduli when the compressive strain was varied from 0% to 30% (p < 0.05); larger shear moduli were observed at compressive 40% strain but the difference was not significant (P = 0.12).

Experimental and numerical analysis of electroactive characteristics of scleral tissue.

The sclera provides mechanical support to retina and protects internal contents of the eye against external injuries. The scleral extracellular matrix is mainly composed of collagen fibers and proteoglycans (PGs). At physiological pH, collagen molecules are neutral but PGs contain negatively charged glycosaminoglycan chains. Thus, the sclera can be considered as a polyelectrolyte hydrogel and is expected to exhibit mechanical response when subjected to electrical stimulations. In this study, we mounted scleral strips, dissected from the posterior part of porcine eyes, at the center of a custom-designed container between two electrodes. The container was filled with NaCl solution and the bending deformation of scleral strips as a function of the applied electric voltage was measured experimentally. It was found that scleral strips reached to an average bending angle of 3°, 10° and 23° when subjected to 5V, 10V, and 15V, respectively. We also created a chemo-electro-mechanical finite element model for simulating the experimental measurements by solving coupled Poisson-Nernst-Plank and equilibrium mechanical field equations. The scleral fixed charge density and modulus of elasticity were found by fitting the experimental data. The ion concentration distribution inside the domain was found numerically and was used to explain the underlying mechanisms for the scleral electroactive response. The numerical simulations were also used to investigate the effects of various parameters such as the electric voltage and fixed charge density on the scleral deformation under an electric field. STATEMENT OF SIGNIFICANCE: This manuscript investigates the electroactive response of scleral tissue. It demonstrates that the sclera deforms mechanically when subjected to electrical stimulations. A chemo-electro-mechanical model is also presented in order to numerically capture the electromechanical response of the sclera. This numerical model is used to explain the experimental observations by finding the ion distribution inside the tissue under an electric field. This work is significant because it shows that the sclera is an electroactive polyanionic hydrogel and it provides new information about the underlying mechanisms governing its mechanical and electrical properties.

Tensile Viscoelastic Properties of the Sclera after Glycosaminoglycan Depletion.

PURPOSE: Fibrillar collagen network and glycosaminoglycans (GAGs) are the primary components of extracellular matrix (ECM) of the sclera. The main goal of this study was to investigate the possible structural roles of GAGs in the scleral tensile properties as a function of preconditioning and displacement rate. METHODS: Four-step uniaxial stress-relaxation tests were used for characterizing the viscoelastic tensile response of the posterior porcine sclera with and without enzymatic GAG removal. The scleral strips were divided into different groups based on the displacement rate and the presence or absence of a preconditioning step in the loading protocol. The groups were (1) displacement rate of 0.2 mm/min without preconditioning, (2) displacement rate of 1 mm/min without preconditioning, (3) displacement rate of 0.2 mm/min with preconditioning, and (4) displacement rate of 1 mm/min with preconditioning. The peak stress, equilibrium stress, and the equilibrium elastic modulus were calculated for all specimens and compared against each other. RESULTS: Increasing the displacement rate from 0.2 mm/min to 1.0 mm/min was found to cause an insignificant change in the equilibrium stress and equilibrium elastic modulus of porcine scleral strips. Removal of GAGs resulted in an overall stiffer tensile behavior independent of the displacement rate in samples that were not preconditioned (P < .05). The behavior of preconditioned samples with and without GAG removal was not significantly different from each other. CONCLUSIONS: The experimental measurements of the present study showed that GAGs play an important role in the mechanical properties of the posterior porcine sclera. Furthermore, using a preconditioning step in the uniaxial testing protocol resulted in not being able to identify any significant difference in the tensile behavior of GAG depleted and normal scleral strips.

Regional Differences in the Glycosaminoglycan Role in Porcine Scleral Hydration and Mechanical Behavior.

Purpose: This study characterized the role of glycosaminoglycans (GAGs) in the hydration, thickness, and biomechanical properties of posterior and anterior porcine sclera. Methods: The scleral discs and strips were obtained from the anterior and posterior parts of porcine eyes, and their initial hydration and thickness were measured. The anterior and posterior scleral discs were used to show the efficacy of the GAG removal protocol by quantifying their GAG content. The strips were divided into three groups of PBS treatment, buffer treatment, and enzyme treatment in order to assess the effects of different treatment procedures on the thickness, hydration, and viscoelastic properties of the samples. The mechanical properties of the strips were determined by performing uniaxial tensile stress relaxation experiments. Results: It was found that the control and buffer groups had insignificant differences in all measured quantities. The samples from the posterior region had a significantly larger GAG content and thickness in comparison with those from anterior region; however, there was an insignificant difference in their hydration. The GAG depletion process decreased the hydration of both anterior and posterior samples significantly (P < 0.05). Furthermore, the mechanical tests showed that the removal of GAGs resulted in stiffer mechanical behavior in both anterior and posterior samples (P < 0.05). In particular, the peak stress and equilibrium stress were significantly larger for the strips in the enzyme treatment group. Conclusions: GAGs and their interaction with the collagen network are important in defining the hydration and mechanical properties of both posterior and anterior sclera.

Nonlinear Mechanical Properties of Prestressed Branched Fibrous Networks.

Random fiber networks constitute the solid skeleton of many biological materials such as the cytoskeleton of cells and extracellular matrix of soft tissues. These random networks show unique mechanical properties such as nonlinear shear strain-stiffening and strain softening when subjected to preextension and precompression, respectively. In this study, we perform numerical simulations to characterize the influence of axial prestress on the nonlinear mechanical response of random network structures as a function of their micromechanical and geometrical properties. We build our numerical network models using the microstructure of disordered hexagonal lattices and quantify their nonlinear shear response as a function of uniaxial prestress strain. We consider three different material models for individual fibers and fully characterize their influence on the mechanical response of prestressed networks. Moreover, we investigate both the influence of geometric disorder keeping the network connectivity constant and the influence of the randomness in the stiffness of individual fibers keeping their mean stiffness constant. The effects of network connectivity and bending rigidity of fibers are also determined. Several important conclusions are made, including that the tensile and compressive prestress strains, respectively, increase and decrease the initial network shear stiffness but have no effect on the maximal shear modulus. We discuss the findings in terms of microstructural properties such as the local strain energy distribution.

Constitutive Modeling of Corneal Tissue: Influence of Three-Dimensional Collagen Fiber Microstructure.

The cornea, the transparent tissue in the front of the eye, along with the sclera, plays a vital role in protecting the inner structures of the eyeball. The precise shape and mechanical strength of this tissue are mostly determined by the unique microstructure of its extracellular matrix. A clear picture of the 3D arrangement of collagen fibrils within the corneal extracellular matrix has recently been obtained from the secondary harmonic generation images. However, this important information about the through-thickness distribution of collagen fibrils was seldom taken into account in the constitutive modeling of the corneal behavior. This work creates a generalized structure tensor (GST) model to investigate the mechanical influence of collagen fibril through-thickness distribution. It then uses numerical simulations of the corneal mechanical response in inflation experiments to assess the efficacy of the proposed model. A parametric study is also done to investigate the influence of model parameters on numerical predictions. Finally, a brief comparison between the performance of this new constitutive model and a recent angular integration (AI) model from the literature is given.

Numerical Investigation on the Role of Mechanical Factors Contributing to Globe Flattening in States of Elevated Intracranial Pressure.

Flattening of the posterior eye globe in the magnetic resonance (MR) images is a sign associated with elevated intracranial pressure (ICP), often seen in people with idiopathic intracranial hypertension (IIH). The exact underlying mechanisms of globe flattening (GF) are not fully known but mechanical factors are believed to play a role. In the present study, we investigated the effects of material properties and pressure loads on GF. For this purpose, we used a generic finite element model to investigate the deformation of the posterior eyeball. The degree of GF in numerical models and the significance of different mechanical factors on GF were characterized using an automated angle-slope technique and a statistical measure. From the numerical models, we found that ICP had the most important role in GF. We also showed that the angle-slope graphs pertaining to MR images from five people with high ICP can be represented numerically by manipulating the parameters of the finite element model. This numerical study suggests that GF observed in IIH patients can be accounted for by the forces caused by elevation of ICP from its normal level, while material properties of ocular tissues, such as sclera (SC), peripapillary sclera (PSC), and optic nerve (ON), would impact its severity.

Mechanical properties of subisostatic random networks composed of nonlinear fibers.

Fibrous protein networks provide structural integrity to different biological materials such as soft tissues. These networks display an unusual exponential strain-stiffening behavior when subjected to mechanical loads. This nonlinear strain-stiffening behavior has so far been explained in terms of the network microstructure and the flexibility of constituting fibers. Here, we conduct a comprehensive computational study to characterize the importance of material properties of individual fibers in the overall nonlinear mechanical response of random fiber networks. To this end, we consider three nonlinear material models, ranging from an almost linear form to a highly nonlinear one, for the fibers of subisostatic disordered networks. We characterize the amount of strain-stiffening as a function of bending rigidity of the fibers, the amount of nonlinearity of the fibers, and the connectivity of random networks. We find that networks composed of highly nonlinear fibers exhibit much more strain-stiffening than networks made up of linear fibers. Furthermore, the local strain distribution becomes more homogenous as the amount of nonlinearity in the material models increases. Increasing the network connectivity signifies the importance of the nonlinear material response of individual fibers in the overall mechanical behavior of networks. The constitutive behavior of fibers plays an important role in defining the failure response of networks particularly in the damage initiation and evolution. These important findings for how the mechanical response of individual fibers affects the overall mechanical properties of random networks could find applications in designing new biomimetic materials and could help scientists better understand the mechanical properties of biological materials.

A computational study of the behavior of colloidal gel networks at low volume fraction.

Colloidal gel networks appear in different scientific and industrial applications because of their unique properties. Molecular dynamics simulations could reveal the relation between molecular level and macroscopic properties of these systems. Nevertheless, the predictions of numerical simulations might depend on the specific form and parameters of interaction potentials. In this paper, a new effective interaction potential is used for characterizing the mechanical behavior of low volume fraction colloidal gels under large shear deformation. The findings are compared with those obtained from other available forms of interaction potentials in order to determine gel characteristics that are interaction potential independent. Furthermore, the macroscopic stress-strain behavior is discussed in terms of the behavior of different terms of the proposed interaction potential. The correlation between the stretch of interparticle bonds and their alignment in the direction of the maximum principal stress is also computed in order to provide microscopic explanations for the initial strain softening behavior. It is concluded that, in addition to topology, local mechanical interactions between colloidal particles are important in defining the mechanical response of soft gels.

Hydration related changes in tensile response of posterior porcine sclera.

It has been shown that there exists significance dependence between hydration and biomechanical properties of hydrated tissues such as cornea. The primary purpose of this study was to determine hydration effects on mechanical properties of sclera. Scleral strips, dissected from the posterior part of pig eyes along the superior-inferior direction, were divided into four hydration groups by first drying them and then soaking them in PBS until their hydration reached to 75%, 100%, 150%, and 200%. The strips were subjected to ten consecutive cycles of loading and unloading up to 1 MPa. The response of samples at the tenth cycle was used to compute the tangent modulus, maximum strain, and hysteresis as a function of hydration. The experiments were done in oil in order to prevent hydration changes during the mechanical tests. The mechanical response of strips right after dissection, control group, was also measured. In general, significant softening of scleral strips was found with increasing hydration (p < 0.05). The stress-strain response of control group was between those of samples with hydration 150% and 200%. The experimental stress-strain data were successfully represented numerically with an exponential mathematical relation with R2 > 0.99. The present study showed that hydration would significantly alter the tensile response of scleral tissue. Thus, the hydration of scleral specimens during mechanical experimental measurements should be carefully controlled.

Colloidal particle gel models using many-body potential interactions.

Many-body effective interactions are commonly used in a molecular dynamics simulation study of gel networks formed by colloidal particles. Here we report an interaction potential that can be used to investigate the mechanical response of colloidal gel networks under shear deformation. We then investigate the dependence of the numerical simulation results on the form of mathematical expression used to define the interparticle interactions. This work reveals insight into particle gel models by discussing the physical origins of their mechanical response.

The contribution of sGAGs to stress-controlled tensile response of posterior porcine sclera.

Despite the significant progress in characterizing mechanical functions of individual scleral extracellular matrix (ECM) components, the biomechanical contribution of sulfated glycosaminoglycans (sGAGs) is still poorly understood. The primary purpose of this study was to determine the possible function of sGAGs in scleral mechanical response by characterizing the tensile behavior of normal and sGAG-depleted samples. We used chondroitinase ABC solution to remove sGAGs from scleral samples that were dissected from posterior porcine eyes. We performed biochemical analyses for assessing the efficacy of sGAG removal protocol. Furthermore, we conducted stress-controlled uniaxial tensile tests to characterize the influence of sGAG removal on mechanical properties of sclera. The tensile behavior of scleral strips right after dissection and after being soaked in buffer was also determined. Biochemical analyses confirmed that 18 hour incubation in 0.125 U/ml Chondroitinase ABC solution removed over 90% of chondroitin and dermatan sGAGs. No significant difference was observed in the thickness/hydration of samples because of enzyme- and buffer-treated samples. Furthermore, it was found that sGAG depletion did not significantly alter the tangent modulus, energy dissipation, and peak strain of posterior scleral strips. It was concluded that sGAGs did not influence the stress-controlled viscoelastic tensile response of sclera.

Influence of Microstructure on Stiffening Effects of Corneal Cross-linking Treatment.

PURPOSE: To investigate the stiffening effects of corneal cross-linking (CXL) on tensile properties of anterior and posterior corneal flaps. METHODS: A Descemet stripping automated endothelial keratoplasty system was used to prepare anterior and posterior flaps from porcine corneas. The flaps were subjected to CXL and their mechanical behavior was assessed by conducting uniaxial tensile experiments. Full-thickness corneas were also cross-linked from the posterior and anterior side and their tensile behavior was measured. RESULTS: The CXL procedure significantly improved biomechanical properties of anterior flaps (P < .05). CXL did not have any significant effect on tensile properties of posterior flaps. Cross-linking full-thickness porcine corneal stroma from the posterior side had no significant stiffening effect. CONCLUSIONS: The stiffening effect of CXL therapy depends significantly on the composition and microstructure of corneal extracellular matrix. [J Refract Surg. 2018;34(9):622-627.].

Relationship between initial corneal hydration and stiffening effects of corneal crosslinking treatment.

PURPOSE: To characterize the mechanical property improvement of riboflavin and ultraviolet light corneal crosslinking (CXL) procedure in artificially swollen human and porcine corneas. SETTING: Computational Biomechanics Research Laboratory, Mechanical and Industrial Engineering Department, University of Illinois at Chicago, Illinois, USA. DESIGN: Experimental study. METHODS: Porcine and human donor corneas were crosslinked at different hydration levels using riboflavin-dextran solutions of different osmolality. Four porcine groups (Hw [hydration in mg H2O/mg dry tissue] = 3.3 ± 0.2 [SD]; 4.0 ± 0.1; 5.1 ± 0.1; 5.6 ± 0.1) and 3 human groups (Hw = 3.2 ± 0.1; 3.9 ± 0.2; 5.3 ± 0.3) were considered. The mechanical properties were measured by uniaxial tensile experiments during which the hydration of samples was the same as the hydration at which they were crosslinked. Tensile properties of 2 porcine groups (Hw = 5.1 ± 0.1; 5.6 ± 0.1) were also measured when their average hydration was lowered to 4.0 mg H2O/mg dry tissue. RESULTS: The CXL procedure significantly increased tensile properties of both human and porcine samples in each hydration group (P < .05). The improvement in tensile properties was hydration-dependent, that is, samples crosslinked at higher hydration levels showed lower maximum tensile stress. The behavior of samples crosslinked at different initial hydration but tested mechanically at similar hydration showed insignificant difference (P = .7). CONCLUSION: Increasing the hydration of porcine and human corneal samples before the CXL treatment had insignificant influence on tensile property improvement, as measured by testing specimens at similar hydration.

UVA/riboflavin collagen crosslinking stiffening effects on anterior and posterior corneal flaps.

The UVA/riboflavin collagen crosslinking (CXL) is one of the treatment procedure for stopping the progression of keratoconus. The inclusion criterion for this procedure is a minimum corneal thickness of 400 µm, which is not often met in patients with advanced keratoconus. Preoperatively swelling thin corneas was shown to stabilize the keratectasia without any postoperative endothelial damage. Recently, we have shown that swelling porcine corneas prior to the CXL treatment had no significant effect on the resulting improvement in their tensile properties. In the present study, we extended this previous study and characterized the stiffening effects of CXL on anterior and posterior flaps as a function of their hydration. A DSAEK system was used to excise 10 mm corneal flaps from 80 porcine corneas. Individual flaps were crosslinked at different initial hydration levels by using riboflavin solutions composed of different dextran concentrations; the thickness was taken as a measure of flap hydration. A DMA machine was used to measure the tensile properties either immediately after the CXL treatment or after the thickness (hydration) of the crosslinked samples was brought down to a specific value. The average thickness of anterior groups was 670 µm, 540 µm, and 410 µm, and the average thickness of posterior groups was 845 µm, 650 µm, and 440 µm. It was found that although CXL significantly increased the tensile properties of all anterior groups, it had an insignificant effect on the stiffness of posterior flaps. Furthermore, except for the posterior flaps in 845 µm and 650 µm thickness groups, decreasing the hydration significantly increased the tensile modulus (p < 0.05). Finally, the anterior flaps that were crosslinked at higher hydration, i.e. swollen before CXL, showed significantly less amount of stiffening in comparison with those crosslinked at lower hydration when the tensile property measurement was done at similar hydration (p < 0.05).

Effect of UVA/Riboflavin Collagen Crosslinking on Biomechanics of Artificially Swollen Corneas.

Purpose: The purpose of this study was to characterize the relation between corneal hydration and stiffening effects of the UVA/riboflavin collagen crosslinking treatment and to investigate how artificially swelling the cornea prior to this treatment procedure affects tensile property improvement. Methods: Porcine corneas were collagen crosslinked in vitro at different hydration levels using a number of hypoosmolar and isoosmolar riboflavin solutions. Thickness of the specimens prior to crosslinking was taken as a proxy for their hydration and was used to divide them into different thickness groups. A Dynamic Mechanical Analysis (DMA) machine was used to perform mechanical tensile tests. The hydration of specimens during the mechanical tests was kept similar to the hydration at which they were crosslinked. The recorded force was used to calculate the maximum tensile stress and tangent modulus as a function of thickness (hydration) prior to collagen crosslinking treatment. Results: Collagen crosslinking with either a hypoosmolar or isoosmolar solution significantly increased corneal tensile modulus (P < 0.05). Corneas that were swollen prior to crosslinking showed significantly softer tensile properties compared with those that were crosslinked at lower hydration (P < 0.05). Although the degree of tensile property improvement was hydration dependent, the stiffness of samples crosslinked at higher hydration was not significantly different than the stiffness of those crosslinked at lower hydration when the hydration was kept similar in the mechanical experiments. Conclusions: Swelling porcine corneas to the different extents prior to collagen crosslinking treatment does not significantly change the amount of biomechanical improvement if tensile properties are measured at similar hydration.