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Research Papers

J Biomech Eng. 2011;133(9):091001-091001-5. doi:10.1115/1.4004947.

Numerical simulation is conducted to evaluate the wind and posture effects on the aerodynamic performance of a skier during the flight stage. Both steady and unsteady models are applied on a 2D geometry. Using the Fluent code, the fundamental equations of fluid flow are solved simultaneously. In particular we focus on the influence of wind speed and direction on aerodynamic forces with several different postures of the skier in steady modeling. For a chosen case, the unsteady models are used to predict the transient characteristics of streamline distributions and aerodynamic forces. It is found that the skier’s postures, wind speed, and direction play a significant role. The wind condition affects the pressure force (the form drag) on the skier and makes it a resistance or thrust regarding wind directions. The optimized posture with a minimization of resistance under a facing wind is determined as a moving-forward body of the skier. The unsteady modeling reveals that the wake around the skier and aerodynamic forces are strongly dependent on time. This initial study not only provides a qualitative and theoretical basis for the athletes to understand the effects of wind and postures, and then to optimize their postures according to the wind condition during the flight stage of skiing, but also builds the foundation for the systematic study of skiing process with more advanced CFD models in the future.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091002-091002-6. doi:10.1115/1.4004948.

Previous studies by our laboratory have demonstrated that implanting a stiffer tissue engineered construct at surgery is positively correlated with repair tissue stiffness at 12 weeks. The objective of this study was to test this correlation by implanting a construct that matches normal tissue biomechanical properties. To do this, we utilized a soft tissue patellar tendon autograft to repair a central-third patellar tendon defect. Patellar tendon autograft repairs were contrasted against an unfilled defect repaired by natural healing (NH). We hypothesized that after 12 weeks, patellar tendon autograft repairs would have biomechanical properties superior to NH. Bilateral defects were established in the central-third patellar tendon of skeletally mature (one year old), female New Zealand White rabbits (n = 10). In one limb, the excised tissue, the patellar tendon autograft, was sutured into the defect site. In the contralateral limb, the defect was left empty (natural healing). After 12 weeks of recovery, the animals were euthanized and their limbs were dedicated to biomechanical (n = 7) or histological (n = 3) evaluations. Only stiffness was improved by treatment with patellar tendon autograft relative to natural healing (p = 0.009). Additionally, neither the patellar tendon autograft nor natural healing repairs regenerated a normal zonal insertion site between the tendon and bone. Immunohistochemical staining for collagen type II demonstrated that fibrocartilage-like tissue was regenerated at the tendon-bone interface for both repairs. However, the tissue was disorganized. Insufficient tissue integration at the tendon-to-bone junction led to repair tissue failure at the insertion site during testing. It is important to re-establish the tendon-to-bone insertion site because it provides joint stability and enables force transmission from muscle to tendon and subsequent loading of the tendon. Without loading, tendon mechanical properties deteriorate. Future studies by our laboratory will investigate potential strategies to improve patellar tendon autograft integration into bone using this model.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091003-091003-8. doi:10.1115/1.4004994.

We derive a cellular solids approach to the design of bone scaffolds for stiffness and pore size. Specifically, we focus on scaffolds made of stacked, alternating, orthogonal layers of hydroxyapatite rods, such as those obtained via micro-robotic deposition, and aim to determine the rod diameter, spacing and overlap required to obtain specified elastic moduli and pore size. To validate and calibrate the cellular solids model, we employ a finite element model and determine the effective scaffold moduli via numerical homogenization. In order to perform an efficient, automated execution of the numerical studies, we employ a geometry projection method so that analyses corresponding to different scaffold dimensions can be performed on a fixed, non-conforming mesh. Based on the developed model, we provide design charts to aid in the selection of rod diameter, spacing and overlap to be used in the robotic deposition to attain desired elastic moduli and pore size.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091004-091004-8. doi:10.1115/1.4004533.

The study provides a pathway to design a mechanics-matching vascular graft for an end-to-end anastomosis to a host artery. For functional equivalence, we submit that the graft and a host artery should have equal inner deformed diameters, equal pressure-radius module, and experience equal axial forces when subjected to mean arterial pressure. These criteria for mechanical equivalence are valid for a large class of materials that can be considered as elastic incompressible and orthotropic solids. As an example, specific known artery properties were used to design or select a graft made from a new synthetic biomaterial to demonstrate that reliable and robust technology for graft fabrication is possible.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091005-091005-6. doi:10.1115/1.4004919.

Perfusion bioreactors are a promising in vitro strategy to engineer bone tissue because they supply needed oxygen and nutrients and apply an osteoinductive mechanical stimulus to osteoblasts within large porous three-dimensional scaffolds. Model two-dimensional studies have shown that dynamic flow conditions (e.g., pulsatile oscillatory waveforms) elicit an enhanced mechanotransductive response and elevated expression of osteoblastic proteins relative to steady flow. However, dynamic perfusion of three-dimensional scaffolds has been primarily examined in short term cultures to probe for early markers of mechanotransduction. Therefore, the objective of this study was to investigate the effect of extended dynamic perfusion culture on osteoblastic differentiation of primary mesenchymal stem cells (MSCs). To accomplish this, rat bone marrow-derived MSCs were seeded into porous foam scaffolds and cultured for 15 days in osteogenic medium under pulsatile regimens of 0.083, 0.050, and 0.017 Hz. Concurrently, MSCs seeded in scaffolds were also maintained under static conditions or cultured under steady perfusion. Analysis of the cells after 15 days of culture indicated that alkaline phosphatase (ALP) activity, mRNA expression of osteopontin (OPN), and accumulation of OPN and prostaglandin E2 were enhanced for all four perfusion conditions relative to static culture. ALP activity, OPN and OC mRNA, and OPN protein accumulation were slightly higher for the intermediate frequency (0.05 Hz) as compared with the other flow conditions, but the differences were not statistically significant. Nevertheless, these results demonstrate that dynamic perfusion of MSCs may be a useful strategy for stimulating osteoblastic differentiation in vitro.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091006-091006-7. doi:10.1115/1.4004944.

The intervertebral disc (IVD) receives important nutrients, such as glucose, from surrounding blood vessels. Poor nutritional supply is believed to play a key role in disc degeneration. Several investigators have presented finite element models of the IVD to investigate disc nutrition; however, none has predicted nutrient levels and cell viability in the disc with a realistic 3D geometry and tissue properties coupled to mechanical deformation. Understanding how degeneration and loading affect nutrition and cell viability is necessary for elucidating the mechanisms of disc degeneration and low back pain. The objective of this study was to analyze the effects of disc degeneration and static deformation on glucose distributions and cell viability in the IVD using finite element analysis. A realistic 3D finite element model of the IVD was developed based on mechano-electrochemical mixture theory. In the model, the cellular metabolic activities and viability were related to nutrient concentrations, and transport properties of nutrients were dependent on tissue deformation. The effects of disc degeneration and mechanical compression on glucose concentrations and cell density distributions in the IVD were investigated. To examine effects of disc degeneration, tissue properties were altered to reflect those of degenerated tissue, including reduced water content, fixed charge density, height, and endplate permeability. Two mechanical loading conditions were also investigated: a reference (undeformed) case and a 10% static deformation case. In general, nutrient levels decreased moving away from the nutritional supply at the disc periphery. Minimum glucose levels were at the interface between the nucleus and annulus regions of the disc. Deformation caused a 6.2% decrease in the minimum glucose concentration in the normal IVD, while degeneration resulted in an 80% decrease. Although cell density was not affected in the undeformed normal disc, there was a decrease in cell viability in the degenerated case, in which averaged cell density fell 11% compared with the normal case. This effect was further exacerbated by deformation of the degenerated IVD. Both deformation and disc degeneration altered the glucose distribution in the IVD. For the degenerated case, glucose levels fell below levels necessary for maintaining cell viability, and cell density decreased. This study provides important insight into nutrition-related mechanisms of disc degeneration. Moreover, our model may serve as a powerful tool in the development of new treatments for low back pain.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091007-091007-9. doi:10.1115/1.4004993.

The material properties of passive skeletal muscle are critical to proper function and are frequently a target for therapeutic and interventional strategies. Investigations into the passive viscoelasticity of muscle have primarily focused on characterizing the elastic behavior, largely neglecting the viscous component. However, viscosity is a sizeable contributor to muscle stress and extensibility during passive stretch and thus there is a need for characterization of the viscous as well as the elastic components of muscle viscoelasticity. Single mouse muscle fibers were subjected to incremental stress relaxation tests to characterize the dependence of passive muscle stress on time, strain and strain rate. A model was then developed to describe fiber viscoelasticity incorporating the observed nonlinearities. The results of this model were compared with two commonly used linear viscoelastic models in their ability to represent fiber stress relaxation and strain rate sensitivity. The viscous component of mouse muscle fiber stress was not linear as is typically assumed, but rather a more complex function of time, strain and strain rate. The model developed here, which incorporates these nonlinearities, was better able to represent the stress relaxation behavior of fibers under the conditions tested than commonly used models with linear viscosity. It presents a new tool to investigate the changes in muscle viscous stresses with age, injury and disuse.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091008-091008-9. doi:10.1115/1.4004996.

Treatments for coarctation of the aorta (CoA) can alleviate blood pressure (BP) gradients (Δ), but long-term morbidity still exists that can be explained by altered indices of hemodynamics and biomechanics. We introduce a technique to increase our understanding of these indices for CoA under resting and nonresting conditions, quantify their contribution to morbidity, and evaluate treatment options. Patient-specific computational fluid dynamics (CFD) models were created from imaging and BP data for one normal and four CoA patients (moderate native CoA: Δ12 mmHg, severe native CoA: Δ25 mmHg and postoperative end-to-end and end-to-side patients: Δ0 mmHg). Simulations incorporated vessel deformation, downstream vascular resistance and compliance. Indices including cyclic strain, time-averaged wall shear stress (TAWSS), and oscillatory shear index (OSI) were quantified. Simulations replicated resting BP and blood flow data. BP during simulated exercise for the normal patient matched reported values. Greatest exercise-induced increases in systolic BP and mean and peak ΔBP occurred for the moderate native CoA patient (SBP: 115 to 154 mmHg; mean and peak ΔBP: 31 and 73 mmHg). Cyclic strain was elevated proximal to the coarctation for native CoA patients, but reduced throughout the aorta after treatment. A greater percentage of vessels was exposed to subnormal TAWSS or elevated OSI for CoA patients. Local patterns of these indices reported to correlate with atherosclerosis in normal patients were accentuated by CoA. These results apply CFD to a range of CoA patients for the first time and provide the foundation for future progress in this area.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091009-091009-11. doi:10.1115/1.4005166.

The vibration characteristics of shell structures such as eyes have been shown to vary with intraocular pressure (IOP). Therefore, vibration characteristics of the eye have the potential to provide improved correlation to IOP over traditional IOP measurements. As background to examine an improved IOP correlation, this paper develops a finite element model of an eye subject to vibration. The eye is modeled as a shell structure filled with inviscid pressurized fluid in which there is no mean flow. This model solves a problem of a fluid with coupled structural interactions of a generally spherically shaped shell system. The model is verified by comparing its vibrational characteristics with an experimental modal analysis of an elastic spherical shell filled with water. The structural dynamic effects due to change in pressure of the fluid are examined. It is shown that the frequency response of this fluid-solid coupled system has a clear increase in natural frequency as the fluid pressure rises. The fluid and structure interaction is important for accurate prediction of system dynamics. This model is then extended to improve its accuracy in modeling the eye by including the effect of the lens to study corneal vibration. The effect of biomechanical parameters such as the thicknesses of different parts of the eye and eye dimensions in altering measured natural frequencies is investigated and compared to the influence of biomechanical parameters in Goldmann applanation tonometry models. The dynamic response of the eye is found to be less sensitive to biomechanical parameters than the applanation tonometry model.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091010-091010-7. doi:10.1115/1.4005168.

In this work we present test methods, devices, and preliminary results for the mechanical characterization of the small bowel for intra luminal robotic mobility. Both active and passive forces that affect mobility are investigated. Four investigative devices and testing methods to characterize the active and passive forces are presented in this work: (1) a novel manometer and a force sensor array that measure force per cm of axial length generated by the migrating motor complex, (2) a biaxial test apparatus and method for characterizing the biomechanical properties of the duodenum, jejunum, and ileum, (3) a novel in vitro device and protocol designed to measure the energy required to overcome the self-adhesivity of the mucosa, and (4) a novel tribometer that measures the in vivo coefficient of friction between the mucus membrane and the robot surface. The four devices are tested on a single porcine model to validate the approach and protocols. Mean force readings per cm of axial length of intestine that occurred over a 15 min interval in vivo were 1.34 ± 0.14 and 1.18 ± 0.22 N cm−1 in the middle and distal regions, respectively. Based on the biaxial stress/stretch tests, the tissue behaves anisotropically with the circumferential direction being more compliant than the axial direction. The mean work per unit area for mucoseparation of the small bowel is 0.08 ± 0.03 mJ cm−2 . The total energy to overcome mucoadhesion over the entire length of the porcine small bowel is approximately 0.55 J. The mean in vivo coefficient of friction (COF) of a curved 6.97 cm2 polycarbonate sled on live mucosa traveling at 1 mm s−1 is 0.016 ± 0.002. This is slightly lower than the COF on excised tissue, given the same input parameters. We have initiated a comprehensive program and suite of test devices and protocols for mechanically characterizing the small bowel for in vivo mobility. Results show that each of the four protocols and associated test devices has successfully gathered preliminary data to confirm the validity of our test approach.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):091011-091011-7. doi:10.1115/1.4005170.

In a previous work (Raghupathy and Barocas, 2010, “Generalized Anisotropic Inverse Mechanics for Soft Tissues,”J. Biomech. Eng., 132 (8), pp. 081006), a generalized anisotropic inverse mechanics method applicable to soft tissues was presented and tested against simulated data. Here we demonstrate the ability of the method to identify regional differences in anisotropy from full-field displacements and boundary forces obtained from biaxial extension tests on soft tissue analogs. Tissue heterogeneity was evaluated by partitioning the domain into homogeneous subdomains. Tests on elastomer samples demonstrated the performance of the method on isotropic materials with uniform and nonuniform properties. Tests on fibroblast-remodeled collagen cruciforms indicated a strong correlation between local structural anisotropy (measured by polarized light microscopy) and the evaluated local mechanical anisotropy. The results demonstrate the potential to quantify regional anisotropic material behavior on an intact tissue sample.

Commentary by Dr. Valentin Fuster

Technical Briefs

J Biomech Eng. 2011;133(9):094501-094501-4. doi:10.1115/1.4004821.

Posterior wall fracture is one of the most common fracture types of the acetabulum and a conventional approach is to perform open reduction and internal fixation with a plate and screws. Percutaneous screw fixations, on the other hand, have recently gained attention due to their benefits such as less exposure and minimization of blood loss. However their biomechanical stability, especially in terms interfragmentary movement, has not been investigated thoroughly. The aims of this study are twofold: (1) to measure the interfragmentary movements in the conventional open approach with plate fixations and the percutaneous screw fixations in the acetabular fractures and compare them; and (2) to develop and validate a fast and efficient way of predicting the interfragmentary movement in percutaneous fixation of posterior wall fractures of the acetabulum using a 3D finite element (FE) model of the pelvis. Our results indicate that in single fragment fractures of the posterior wall of the acetabulum, plate fixations give superior stability to screw fixations. However screw fixations also give reasonable stability as the average gap between fragment and the bone remained less than 1 mm when the maximum load was applied. Our finite element model predicted the stability of screw fixation with good accuracy. Moreover, when the screw positions were optimized, the stability predicted by our FE model was comparable to the stability obtained by plate fixations. Our study has shown that FE modeling can be useful in examining biomechanical stability of osteosynthesis and can potentially be used in surgical planning of osteosynthesis.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):094502-094502-5. doi:10.1115/1.4004916.

Transforming decades’ old methodology, electromechanical reshaping (EMR) may someday replace traditionally destructive surgical techniques with a less invasive means of cartilage reshaping for reconstructive and esthetic facial surgery. Electromechanical reshaping is essentially accomplished through the application of voltage to a mechanically deformed cartilage specimen. While the capacity of the method for effective reshaping has been consistently shown, its associated effects on cartilage mechanical properties are not fully comprehended. To begin to explore the mechanical effect of EMR on cartilage, the tangent moduli of EMR-treated rabbit septal and auricular cartilage were calculated and compared to matched control values. Between the two main EMR parameters, voltage and application time, the former was varied from 2–8 V and the latter held constant at 2 min for septal cartilage, 3 min for auricular cartilage. Flat platinum electrodes were used to apply voltage, maintaining the flatness of the specimens for more precise mechanical testing through a uniaxial tension test of constant strain rate 0.01 mm/s. Above 2 V, both septal and auricular cartilage demonstrated a slight reduction in stiffness, quantified by the tangent modulus. A thermal effect was observed above 5 V, a newly identified EMR application threshold to avoid the dangers associated with thermoforming cartilage. Optimizing EMR application parameters and understanding various side effects bridge the gap between EMR laboratory research and clinical use, and the knowledge acquired through this mechanical study may be one additional support for that bridge.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):094503-094503-4. doi:10.1115/1.4004917.

Vertebral burst fractures are commonly studied with experimental animal models. There is however a lack of consensus as to what parameters are important to create an unstable burst fracture with a significant canal encroachment on such model. This study aims to assess the effect of the loading rate, flexion angle, spinal level, and their interactions on the production of a vertebral thoracolumbar burst fracture on a porcine model. Sixteen functional spinal units composed of three vertebrae were harvested from mature Yucatan minipigs. Two loading rates (0.01 and 500 mm/s), two flexion angles (0° and 15°), and two spinal levels (T11-T13 and T14-L2) were studied, following a full factorial experimental plan with one repetition. Compression was applied to each functional unit to create a vertebral fracture. The load-to-failure, loss of compressive stiffness, final canal encroachment, and fracture type were used as criteria to evaluate the resulting fracture. All specimens compressed without flexion resulted in burst fractures. Half of the specimens compressed with the 15° flexion angle resulted in compression fractures. Specimens positioned without flexion lost more of their compressive stiffness and had more significant canal encroachment. Fractured units compressed with a higher loading rate resulted in a greater loss of compressive stiffness. The spinal level had no significant effect on the resulting fractures. The main parameters which affect the resulting fracture are the loading rate and the flexion angle. A higher loading rate and the absence of flexion favors the production of burst fractures with a greater canal encroachment.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):094504-094504-4. doi:10.1115/1.4004920.

The goal of tissue engineering is to use substitutes to repair and restore organ function. Bioreactors are an indispensable tool for monitoring and controlling the unique environment for engineered constructs to grow. However, in order to determine the biochemical properties of engineered constructs, samples need to be destroyed. In this study, we developed a novel technique to nondestructively online-characterize the water content and fixed charge density of cartilaginous tissues. A new technique was developed to determine the tissue mechano-electrochemical properties nondestructively. Bovine knee articular cartilage and lumbar annulus fibrosus were used in this study to demonstrate that this technique could be used on different types of tissue. The results show that our newly developed method is capable of precisely predicting the water volume fraction (less than 3% disparity) and fixed charge density (less than 16.7% disparity) within cartilaginous tissues. This novel technique will help to design a new generation of bioreactors which are able to actively determine the essential properties of the engineered constructs, as well as regulate the local environment to achieve the optimal conditions for cultivating constructs.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):094505-094505-5. doi:10.1115/1.4004949.

Numerical simulation of soft tissue mechanical properties is a critical step in developing valuable biomechanical models of live organisms. A cubic Hermitian spline optimization routine is proposed in this paper to model nonlinear experimental force-elongation curves of soft tissues, in particular when modeled as lumped elements. Boundary conditions are introduced to account for the positive definiteness and the particular curvature of the experimental curve to be fitted. The constrained least-square routine minimizes user intervention and optimizes fitting of the experimental data across the whole fitting range. The routine provides coefficients of a Hermitian spline or corresponding knots that are compatible with a number of constraints that are suitable for modeling soft tissue tensile curves. These coefficients or knots may become inputs to user-defined component properties of various modeling software. Splines are particularly advantageous over the well-known exponential model to account for the traction curve flatness at low elongations and to allow for more flexibility in the fitting process. This is desirable as soft tissue models begin to include more complex physical phenomena.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):094506-094506-5. doi:10.1115/1.4004995.

Homogeneous contractility is usually assigned to the remote region, border zone (BZ), and the infarct in existing infarcted left ventricle (LV) mathematical models. Within the LV, the contractile function is therefore discontinuous. Here, we hypothesize that the BZ may in fact define a smooth linear transition in contractility between the remote region and the infarct. To test this hypothesis, we developed a mathematical model of a sheep LV having an anteroapical infarct with linearly–varying BZ contractility. Using an existing optimization method (Sun , 2009, “A Computationally Efficient Formal Optimization of Regional Myocardial Contractility in a Sheep With Left Ventricular Aneurysm,” J. Biomech. Eng., 131 (11), pp. 111001), we use that model to extract active material parameter Tmax and BZ width dn that “best” predict in–vivo systolic strain fields measured from tagged magnetic resonance images (MRI). We confirm our hypothesis by showing that our model, compared to one that has homogeneous contractility assigned in each region, reduces the mean square errors between the predicted and the measured strain fields. Because the peak fiber stress differs significantly (∼15%) between these two models, our result suggests that future mathematical LV models, particularly those used to analyze myocardial infarction treatment, should account for a smooth linear transition in contractility within the BZ.

Commentary by Dr. Valentin Fuster
J Biomech Eng. 2011;133(9):094507-094507-7. doi:10.1115/1.4005167.

Blood damage and thrombosis are major complications that are commonly seen in patients with implanted mechanical heart valves. For this in vitro study, we isolated the closing phase of a bileaflet mechanical heart valve to study near valve fluid velocities and stresses. By manipulating the valve housing, we gained optical access to a previously inaccessible region of the flow. Laser Doppler velocimetry and particle image velocimetry were used to characterize the flow regime and help to identify the key design characteristics responsible for high shear and rotational flow. Impact of the closing mechanical leaflet with its rigid housing produced the highest fluid stresses observed during the cardiac cycle. Mean velocities as high as 2.4 m/s were observed at the initial valve impact. The velocities measured at the leaflet tip resulted in sustained shear rates in the range of 1500–3500 s−1 , with peak values on the order of 11,000–23,000 s−1 . Using velocity maps, we identified regurgitation zones near the valve tip and through the central orifice of the valve. Entrained flow from the transvalvular jets and flow shed off the leaflet tip during closure combined to generate a dominant vortex posterior to both leaflets after each valve closing cycle. The strength of the peripheral vortex peaked within 2 ms of the initial impact of the leaflet with the housing and rapidly dissipated thereafter, whereas the vortex near the central orifice continued to grow during the rebound phase of the valve. Rebound of the leaflets played a secondary role in sustaining closure-induced vortices.

Commentary by Dr. Valentin Fuster

Design Innovation

J Biomech Eng. 2011;133(9):095001-095001-7. doi:10.1115/1.4004921.

The biomechanical model of glaucoma considers intraocular pressure-related stress and resultant strain on load bearing connective tissues of the optic nerve and surrounding peripapillary sclera as one major causative influence that effects cellular, vascular, and axonal components of the optic nerve. By this reasoning, the quantification of variations in the microstructural architecture and macromechanical response of scleral shells in glaucomatous compared to healthy populations provides an insight into any variations that exist between patient populations. While scleral shells have been tested mechanically in planar and pressure-inflation scenarios the link between the macroscopic biomechanical response and the underlying microstructure has not been determined to date. A potential roadblock to determining how the microstructure changes based on pressure is the ability to mount the spherical scleral shells in a method that does not induce unwanted stresses to the samples (for instance, in the flattening of the spherical specimens), and then capturing macroscopic and microscopic changes under pressure. Often what is done is a macroscopic test followed by sample fixation and then imaging to determine microstructural organization. We introduce a novel device and method, which allows spherical samples to be pressurized and macroscopic and microstructural behavior quantified on fully hydrated ocular specimens. The samples are pressurized and a series of markers on the surface of the sclera imaged from several different perspectives and reconstructed between pressure points to allow for mapping of nonhomogenous strain. Pictures are taken from different perspectives through the use of mounting the pressurization scheme in a gimbal that allows for positioning the sample in several different spherical coordinate system configurations. This ability to move the sclera in space about the center of the globe, coupled with an upright multiphoton microscope, allows for collecting collagen, and elastin signal in a rapid automated fashion so the entire globe can be imaged.

Commentary by Dr. Valentin Fuster

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