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.

VEGF-A-induced changes in distal outflow tract structure and function.

PURPOSE: To investigate changes in distal outflow tract vessels caused by VEGF-A and their impact on outflow. METHODS: We compared VEGF-A perfused porcine anterior segments with and without trabecular meshwork (TM) to control eyes. In the first experiment (n=48), we analyzed live changes of the outflow tract with spectral-domain optical coherence tomography (SD-OCT) over 3 h and reconstructed them in 3D. In a second experiment (n=32), we measured the intraocular pressure (IOP) variation in response to VEGF-A over 48 h and computed the outflow facility. RESULTS: VEGF-A increased the vessel volume of the distal outflow tract by 16.8±10.6% while control eyes remained unchanged (0.5±6.8%). Volume changes occurred within the first 100 min before plateauing at 140 min. VEGF-A enhanced the outflow facility in eyes without TM by 38.6±25.5% at 24 h as compared to controls (p<0.05). CONCLUSION: VEGF-A dilated vessels of the distal outflow tract and increased the outflow facility even after TM removal, pointing to a regulatory mechanism independent of proximal structures.

Suitability of Slaughterhouse-Acquired Pig Eyes as Model Systems for Refractive Ultraviolet and Infrared Femtosecond Laser Research.

PURPOSE: To elucidate whether it is feasible to use porcine eyes from scalded, abattoir-acquired animals for refractive femtosecond laser research. METHODS: An infrared laser (FS 200) and an ultraviolet laser (prototype version) were tested for their applicability on scalded pig eyes. Fifty porcine eyes were divided into two equally-sized groups and assigned to either the infrared or the ultraviolet laser. Both laser groups were comprised of five subgroups of n = 5 eyes each. Group A: non-scalded eyes (negative control); group B: eyes taken from tunnel-scalded animals; group C1: eyes taken from tank-scalded animals without opaque corneal lesion; group C2: eyes taken from animals with opaque corneal lesion; group D: eyes scalded in toto in the laboratory (positive control). In each group the lasers were employed to create a stromal flap. The quality of the laser cuts and the resulting flap beds, as well as of the porcine corneas themselves, was examined by anterior segment optical coherence tomography and scanning electron microscopy. RESULTS: All scalded specimens exhibited substantial corneal swelling, most pronounced in group C2. After ultraviolet laser application, the tank- and tunnel-scalded samples displayed marked irregularities and an increased degree of surface roughness in the flap beds. After infrared laser application, this was only the case in the tank-scalded specimens. CONCLUSION: It is not recommended to use eyes taken from scalded pigs for ultraviolet femtosecond laser experiments. For infrared femtosecond lasers, eyes taken from tunnel-scalded animals may represent an acceptable alternative, if non-scalded eyes are not available.

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).

Employing Nanostructured Scaffolds to Investigate the Mechanical Properties of Adult Mammalian Retinae Under Tension.

Numerous eye diseases are linked to biomechanical dysfunction of the retina. However, the underlying forces are almost impossible to quantify experimentally. Here, we show how biomechanical properties of adult neuronal tissues such as porcine retinae can be investigated under tension in a home-built tissue stretcher composed of nanostructured TiO2 scaffolds coupled to a self-designed force sensor. The employed TiO2 nanotube scaffolds allow for organotypic long-term preservation of adult tissues ex vivo and support strong tissue adhesion without the application of glues, a prerequisite for tissue investigations under tension. In combination with finite element calculations we found that the deformation behavior is highly dependent on the displacement rate which results in Young's moduli of (760-1270) Pa. Image analysis revealed that the elastic regime is characterized by a reversible shear deformation of retinal layers. For larger deformations, tissue destruction and sliding of retinal layers occurred with an equilibration between slip and stick at the interface of ruptured layers, resulting in a constant force during stretching. Since our study demonstrates how porcine eyes collected from slaughterhouses can be employed for ex vivo experiments, our study also offers new perspectives to investigate tissue biomechanics without excessive animal experiments.

Dose-dependent effects of netarsudil, a Rho-kinase inhibitor, on the distal outflow tract.

PURPOSE: To characterize the effects of netarsudil on the aqueous humor outflow tract distal to the trabecular meshwork (TM). We hypothesized that netarsudil increases outflow facility in eyes with and without circumferential ab interno trabeculectomy (AIT) that removes the TM. METHODS: Sixty-four porcine anterior segment cultures were randomly assigned to groups with (n = 32) and without circumferential AIT (n = 32). Cultures were exposed to 0.1, 1, and 10 µM netarsudil (N = 8 eyes per concentration). For each concentration, IOP and vessel diameters were compared with their respective pretreatment baselines. Outflow tract vessel diameters were assessed by spectral-domain optical coherence tomography (SDOCT) and rendered in 4D (XYZ time series). RESULTS: Netarsudil at 1 µM reduced IOP both in eyes with TM (- 0.60 ± 0.24 mmHg, p = 0.01) and in eyes without TM (- 1.79 ± 0.42 mmHg, p < 0.01). At this concentration, vessels of the distal outflow tract dilated by 72%. However, at 0.1 µM netarsudil elevated IOP in eyes with TM (1.59 ± 0.36 mmHg, p < 0.001) as well as in eyes without TM (0.23 ± 0.32 mmHg, p < 0.001). Vessels of the distal outflow tract constricted by 31%. Similarly, netarsudil at a concentration of 10 µM elevated IOP both in eyes with TM (1.91 ± 0.193, p < 0.001) and in eyes without TM (3.65 ± 0.86 mmHg, p < 0.001). At this concentration, outflow tract vessels constricted by 27%. CONCLUSION: In the porcine anterior segment culture, the dose-dependent IOP changes caused by netarsudil matched the diameter changes of distal outflow tract vessels. Hyper- and hypotensive properties of netarsudil persisted after TM removal.

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.

Characterization of Corneal Biomechanical Properties and Determination of Natural Intraocular Pressure Using CID-GAT.

PURPOSE: The intraocular pressure (IOP) measured using Goldmann Applanation Tonometry (GAT) is confounded by individual corneal properties. We investigated a modified method that removes the confoundment by incorporating corneal properties into the Imbert-Fick's law is investigated. METHOD: Porcine eyes were pressurized between 10 and 40 mm Hg using a manometer. The eyes were indented using a flat cylindrical indenter. A modified corneal indentation device (CID) procedure was used to obtain the corneal moduli Eqs . The calculated IOPNC from the Imbert-Fick's Law using the corneal moduli Eqs was compared to the natural IOPN, measured using pressure sensor inserted into the eye. RESULTS: Test results showed that IOP-dependent corneal modulus Eqs is a primary confounding factor in IOP calculation. The average elastic modulus Eqs is 0.173 ± 0.018 MPa at 20 mm Hg, and increases with IOP at a linear rate of 0.0066 MPa per mm Hg (r = 0.997, P < 0.001). Incorporation of individual Eqs into IOPNC calculation showed that IOPNC are in good agreement with reference IOPN (slope = 0.999, r = 0.939, P < 0.001). CONCLUSIONS: The IOP-dependent corneal modulus Eqs is a primary confounding factor in IOP calculation. A modified CID-GAT procedure to obtain natural cornea-independent IOPNC is developed and verified in this study. The CID-GAT IOP modification may be used in place of conventional GAT when the confounding effects in eyes with atypical cornea (e.g., laser-assisted in situ keratomileusis [LASIK] thinned) are significant. TRANSLATIONAL RELEVANCE: Confoundment from corneal properties results in IOP measurement errors. The study showed that the CID-GAT method can significantly reduce the confounding corneal errors.

Evaluation of the effect of fasudil hydrochloride on outflow facility in enucleated eyes of different animals / 眼科新进展

Objective To evaluate the influence of a selective Rho-associated protein kinase inhibitor (fasudil hydrochloride) on outflow facility in enucleated porcine,rabbit and bovine eyes.Methods At the constant perfusion pressure of 15 mm-Hg (1 kPa =7.5 mmHg),the baseline coefficient of outflow facility (C0) of the isolated porcine,rabbit and bovine eyes was recorded respectively.The enucleated porcine eyes were divided into two groups randomly (n =6),and they were control group and experimental group.The same grouping method was also used-C0 the ribbit and bovine eyes.The control group was subjected to GPBS perfusion,while the experimental group was treated with 100 μmol · L-1 fasudil solution,followed by recording the experimental coefficient of outflow facility (C1),as well as calculating ΔC (ΔC =C1-C0) and ΔC％ (ΔC％ =ΔC/C0).Finally,the paired t test and one-way analysis of variance were performed using SPSS 17.0.Results As for porcine eyes,the ΔC％ of the control group was (17.83 ± 3.84) ％ while the experimental group was (44.00 ± 6.44) ％;as for rabbit eyes,the ΔC％ of the control group was (15.50 ± 2.93) ％,while the experimental group was (31.67 ±6.54)％;as for bovine eyes,the ΔC％ of the control group was (11.67 ± 1.17)％,while the experimental group was (37.17 ± 4.48)％.The ΔC％ in the experimental group was significantly increased when compared with the control group in three animals,with significant difference (all P ＜ 0.05).There was no statistical difference in ΔC％ of three experimental groups among different kinds of animals (all P ＜ 0.05).Conclusion Fasudil can improve outflow facility in enucleated eyes of animals,and it can redistribute aqueous humor drainage to a wider area through directly regulating the cytoskeleton of cells and matrix,resulting in increased coefficient of outflow facility.

[Fiberless miniature chandelier LED endoilluminator for pars plana vitrectomy]. / Faserloser Miniatur-Chandelier-LED-Endoilluminator für die Pars-plana-Vitrektomie.

BACKGROUND: Currently available chandelier endoilluminators for pars plana vitrectomy consist of conventional optical fibers coupled to a light source. The light probes of these fibers now provide wide emittance angles but it is still often not possible to illuminate the whole intraocular space via just one incision. Therefore, several light probes or additional handheld endoilluminators have to be used simultaneously or the lights have to be repositioned during surgery. OBJECTIVE: The presented prototype of a fiberless chandelier light-emitting diode (LED) endoilluminator aims at illuminating the whole intraocular space with just one incision while reducing the risk of harming the retina. MATERIAL AND METHODS: The light source is a white LED with a conical tip that allows stable fixation within an incision. The physical properties of these LEDs were determined and used for calculating the relevant irradiance to assess the risk of causing harm to the eye. The illumination of the intraocular space was investigated using porcine eyes. RESULTS: The illumination of porcine eyes with the modified LEDs was bright and homogeneous. Measurements and subsequent calculations proved that the expected thermal load and photochemical hazard were very low for human eyes. CONCLUSION: The hitherto existing experimental results on porcine eyes and the theoretical considerations on human eyes were found to be positive; therefore, it is expected that this new fiberless chandelier LED endoilluminator will prove to be advantageous for human patients. It promises a reduced number of incisions with a simplified handling and need to add handheld light sources only if minimal intensity oblique illumination is helpful together with the chandelier illumination. This contributes to a significant reduction of phototoxicity risks and additionally there is a chance for a cost reduction because expensive xenon or mercury lamps are no longer necessary. These expectations have to be verified by further studies on human eyes.

Outflow facility efficacy of five drugs in enucleated porcine eyes by a method of constant-pressure perfusion.

This study aimed to characterize a technique that assesses the outflow facility (C) efficacy of five kinds of IOP-lowering drugs commonly used clinically in enucleated porcine Eyes. Eyes were perfused at 15 mmHg with GPBS first to establish the baseline outflow facility (C0). Then the anterior chamber contents were exchanged for GPBS with corresponding concentration eye drops (4.9×10(3) nM Brimonidine, 41.1 nM Latanoprost, 3.4×10(3) nM Levobunolol, 3.0×10(3) nM Brinzolamide, 8.3×10(3) nM Pilocarpine) in five groups (n = 6 each), while 6 eyes received GPBS alone as control. The mean stable facility obtained after drug administration (C1) was continuously recorded. The changes between C0 and C1 (ΔC = C1-C0) were analyzed. Finally, for drugs among the five experiment groups with statistical significance, the concentration was reduced 3 times, otherwise the drugs' concentration was increased to 10 times to confirm its effectiveness further using the same methods (n = 6 each). We found that the average baseline outflow facility was 0.24±0.01 µl·min(-1)·mmHg(-1). C increased significantly in Brimonidine and Latanoprost groups, even the concentration of Brimonidine and Latanoprost was decreased 3 times (P < 0.05). However, there was no significantly increase in Levobunolol, Brinzolamide, Pilocarpine and control group (P > 0.05), but when drugs' concentration was increased to 10 times, the C value of Pilocarpine decreased significantly (P = 0.04). No significant washout effects in porcine eyes were observed. To conclude, outflow facility efficacy of five drugs in enucleated porcine eyes may provide a reference for clinical medicine. A constant-pressure perfusion technique should be useful to evaluate effect of pharmacologic agents or surgical manipulations on aqueous humor dynamics.