Joel R. Palko

Dynamic testing of regional viscoelastic behavior of canine sclera.

Intraocular pressure (IOP) fluctuations have gained recent clinical interest and thus warrant an understanding of how the sclera responds to dynamic mechanical insults. The objective of this study was to characterize the regional dynamic viscoelastic properties of canine sclera under physiological cyclic loadings. Scleral strips were excised from the anterior, equatorial, and posterior sclera in ten canine eyes. The dimensions of each strip were measured using a high resolution ultrasound imaging system. The strips were tested in a humidity chamber at approximately 37 °C using a Rheometrics Systems Analyzer. A cyclic strain input (0.25%, 1 Hz) was applied to the strips, superimposed upon pre-stresses corresponding to an IOP of 15, 25, and 45 mmHg. The cyclic stress output was recorded and the dynamic properties were calculated based on linear viscoelasticity. Uni-axial tensile tests were also performed on the same samples and the results were compared to those reported for human eyes. The results showed that the scleras resistance to dynamic loading increased significantly while the damping capability decreased significantly with increasing pre-stresses for all regions of sclera (P < 0.001). Anterior sclera appeared to have a significantly higher damping capability than equatorial and posterior sclera (P = 0.003 and 0.018, respectively). The secant modulus from uni-axial tensile tests showed a decreasing trend from anterior to posterior sclera, displaying a similar pattern as in the human eye. In conclusion, all scleral regions in the canine eyes exhibited an increased ability to resist and a decreased ability to dampen cyclic stress insults at increasing pre-stress (i.e., increasing steady-state IOP). The regional variation of the dynamic properties differed from those of uni-axial tensile tests. Dynamic testing may provide useful information to better understand the mechanical behavior of the sclera in response to dynamic IOP.

Biaxial mechanical testing of posterior sclera using high-resolution ultrasound speckle tracking for strain measurements.

This study aimed to characterize the mechanical responses of the sclera, the white outer coat of the eye, under equal-biaxial loading with unrestricted shear. An ultrasound speckle tracking technique was used to measure tissue deformation through sample thickness, expanding the capabilities of surface strain techniques. Eight porcine scleral samples were tested within 72 h postmortem. High resolution ultrasound scans of scleral cross-sections along the two loading axes were acquired at 25 consecutive biaxial load levels. An additional repeat of the biaxial loading cycle was performed to measure a third normal strain emulating a strain gage rosette for calculating the in-plane shear. The repeatability of the strain measurements during identical biaxial ramps was evaluated. A correlation-based ultrasound speckle tracking algorithm was used to compute the displacement field and determine the distributive strains in the sample cross-sections. A Fung type constitutive model including a shear term was used to determine the material constants of each individual specimen by fitting the model parameters to the experimental stress-strain data. A non-linear stress-strain response was observed in all samples. The meridian direction had significantly larger strains than that of the circumferential direction during equal-biaxial loadings (Ps<0.05). The stiffness along the two directions was also significantly different (P=0.02) but highly correlated (R(2)=0.8). These results showed that the mechanical properties of the porcine sclera were nonlinear and anisotropic under biaxial loading. This work has also demonstrated the feasibility of using ultrasound speckle tracking for strain measurements during mechanical testing.

Spatially heterogeneous corneal mechanical responses before and after riboflavin-ultraviolet-A crosslinking.

Purpose To determine the heterogeneous through‐thickness strains in the cornea at physiologic intraocular pressures before and after corneal collagen crosslinking (CXL) using noninvasive ultrasound. Setting Department of Biomedical Engineering, Ohio State University, Columbus, Ohio, USA. Design Experimental study. Methods Sixteen paired canine corneoscleral shells were divided into 2 groups. The CXL group completed a standard CXL protocol using riboflavin–ultraviolet‐A (UVA) irradiation. The control group was given an identical treatment except UVA irradiation. Ultrasound scans (at 55 MHz) of the cornea were obtained before and after treatment as the corneoscleral shell was inflated from 5 mm Hg to 45 mm Hg to calculate the distributive through‐thickness strains in the cornea. The mean radial and tangential strains of the whole cornea layer, as well as those of the anterior, middle, and posterior thirds of the cornea, were compared before and after treatment in the control group and CXL group using linear mixed models with repeated measures. Results Significant reductions in tangential and radial strains occurred in the CXL group (P=.003 and P=.0025, respectively) but not the control group (P=.08 and P=.63, respectively). The anterior third had the smallest strains in all pretreated corneas (P<.001) and posttreated corneas (CXL group, P=.023; control group, P=.01). Conclusions Ultrasound speckle tracking showed heterogeneous strain distributions through the cornea and confirmed that CXL results in a stiffer corneal response (ie, smaller strains during physiologic loadings). This technique may provide a clinical tool to quantify the biomechanical effects of CXL. Financial Disclosure No author has a financial or proprietary interest in any material or method mentioned.

The effects of decorin and HGF-primed vocal fold fibroblasts in vitro and ex vivo in a porcine model of vocal fold scarring.

Vocal fold injury can be irreversible, leading to vocal fold scarring, with permanent functional effects and no optimal treatment. A porcine model of vocal fold scarring was used to test effects of decorin and primed vocal fold fibroblasts in vitro using a cell migration assay and immunoblotting, and by using functional measurements of porcine larynges and excised porcine vocal folds.

Biomechanical properties and correlation with collagen solubility profile in the posterior sclera of canine eyes with an ADAMTS10 mutation.

PURPOSE We examined the biomechanical properties and correlation with the collagen solubility profile of the posterior sclera in a canine model of primary open-angle glaucoma caused by the G661R missense mutation in the ADAMTS10 gene. METHODS Scleral strips from ADAMTS10-mutant (affected) dogs and age-matched controls were collected. Viscoelastic properties (i.e., complex modulus and tan[δ]) were measured using dynamic mechanical analysis (DMA) with a 0.15% sinusoidal strain at different frequencies superimposed upon different preloads. A tensile ramp was performed following DMA. The collagen solubility profile was examined using a colorimetric hydroxyproline assay to determine the amount of soluble and insoluble collagen. The viscoelastic properties were compared between groups using linear mixed models for repeated measures at different preloads and frequencies. The correlation between the biomechanical properties and collagen content were evaluated using Pearson correlations. RESULTS Complex modulus and tan(δ) were significantly lower in the affected group (P < 0.001), and the differences were consistent at different preloads and frequencies. The B value from the tensile ramp test also was significantly lower in the affected group (P = 0.02). The insoluble collagen was significantly lower in the affected group (P < 0.05) and correlated positively with the complex modulus (R = 0.88, P < 0.005). CONCLUSIONS An inherently weaker and biochemically distinct posterior sclera was observed in dogs with the G661R missense mutation in ADAMTS10 before clinical indications of optic nerve damage. It remains to be shown whether and how the altered scleral biomechanics may affect the rate of glaucoma progression following intraocular pressure elevation.

Influence of Age on Ocular Biomechanical Properties in a Canine Glaucoma Model with ADAMTS10 Mutation.

Soft tissue often displays marked age-associated stiffening. This study aims to investigate how age affects scleral biomechanical properties in a canine glaucoma model with ADAMTS10 mutation, whose extracellular matrix is concomitantly influenced by the mutation and an increased mechanical load from an early age. Biomechanical data was acquired from ADAMTS10-mutant dogs (n = 10, 21 to 131 months) and normal dogs (n = 5, 69 to 113 months). Infusion testing was first performed in the whole globes to measure ocular rigidity. After infusion experiments, the corneas were immediately trephined to prepare scleral shells that were mounted on a pressurization chamber to measure strains in the posterior sclera using an inflation testing protocol. Dynamic viscoelastic mechanical testing was then performed on dissected posterior scleral strips and the data were combined with those reported earlier by our group from the same animal model (Palko et al, IOVS 2013). The association between age and scleral biomechanical properties was evaluated using multivariate linear regression. The relationships between scleral properties and the mean and last measured intraocular pressure (IOP) were also evaluated. Our results showed that age was positively associated with complex modulus (p<0.001) and negatively associated with loss tangent (p<0.001) in both the affected and the normal groups, suggesting an increased stiffness and decreased mechanical damping with age. The regression slopes were not different between the groups, although the complex modulus was significantly lower in the affected group (p = 0.041). The posterior circumferential tangential strain was negatively correlated with complex modulus (R = -0.744, p = 0.006) showing consistent mechanical evaluation between the testing methods. Normalized ocular rigidity was negatively correlated with the last IOP in the affected group (p = 0.003). Despite a mutation that affects the extracellular matrix and a chronic IOP elevation in the affected dogs, age-associated scleral stiffening and loss of mechanical damping were still prominent and had a similar rate of change as in the normal dogs.

Three-Dimensional Strains in Human Posterior Sclera Using Ultrasound Speckle Tracking

Intraocular pressure (IOP) induced strains in the peripapillary sclera may play a role in glaucoma progression. Using inflation testing and ultrasound speckle tracking, the 3D strains in the peripapillary sclera were measured in nine human donor globes. Our results showed that the peripapillary sclera experienced through-thickness compression and meridional stretch during inflation, while minimal circumferential dilation was observed when IOP was increased from 10 to 19 mmHg. The maximum shear was primarily oriented in the through-thickness, meridional cross sections and had a magnitude slightly larger than the first principal strain. The tissue volume had minimal overall change, confirming near-incompressibility of the sclera. Substantial strain heterogeneity was present in the peripapillary region, with local high strain areas likely corresponding to structural heterogeneity caused by traversing blood vessels. These 3D strain characteristics provide new insights into the biomechanical responses of the peripapillary sclera during physiological increases of IOP. Future studies are needed to confirm these findings and investigate the role of these biomechanical characteristics in ocular diseases.

Skew Deviation and Partial Ocular Tilt Reaction Due to Intratympanic Gentamicin Injection, with Review of the Literature

ABSTRACT Skew deviation is a rare side effect of intratympanic gentamicin injection for intractable Meniere’s disease. When the skew deviation is accompanied by pathologic head tilt and ocular torsion, the result is an ocular tilt reaction (OTR). The authors report the case of a 56-year-old man with refractory Meniere’s disease who developed binocular vertical diplopia following intratympanic gentamicin injection and was found to have skew deviation and a partial ocular tilt reaction. The authors also review the reported cases of skew deviation following intratympanic gentamicin and confirm this phenomenon, which has only rarely been reported in the literature.