Finite element analysis of zonular forces.

To determine the effect of zonular forces on lens capsule topography, a finite element (FE) analyses of lens capsules with no lens stroma and constant and variable thickness with anterior capsulotomies of 1.5 mm-6.5 mm were evaluated when subjected to equatorial (Ez), anterior (Az) and posterior (Pz) zonular forces. The lens capsule was considered in the unaccommodated state when the total initial zonular force was 0.00075 N or 0.3 N. From the total 0.00075 N zonular force, the Ez force was increased in 0.000125 N steps for a maximum force of 0.03 N and simultaneously the Az plus Pz force was reduced in 0.000125 N steps to zero. In addition, the force of all the zonules was reduced from 0.00075 N and separately from 0.3 N in 0.000125 N steps to zero. Only when Ez force was increased as Az and Pz force was reduced did the capsule topography simulate in vivo observations with the posterior capsule pole bowing posteriorly. The posterior bowing was directly related to Ez force and capsulotomy size. Whether the total force of all the zonules in the unaccommodated state was 0.00075 N or 0.3 N and reduced in steps to zero, the lens capsule topography did not emulate the in vivo observations. The FE analysis demonstrated that Ez tension increases while the Az and Pz tension decreases and that all the zonules do not relax during ciliary muscle contraction.

Mechanism of accommodation assessed by change in precisely registered ocular images associated with concurrent change in auto-refraction.

PURPOSE: Our purpose was to determine the changes in anterior chamber depth (ACD) and central lens thickness (CLT) during pharmacologically induced accommodation. METHODS: Following pupillary dilation with phenylephrine 10%, baseline auto-refractions and swept-source optical coherence tomographic biometric images (Zeiss IOLMaster 700) were obtained from the right eyes of 25 subjects aged 19 to 24 years. Pilocarpine 4% and phenylephrine 10% were then instilled into these right eyes. One hour later, auto-refractions and biometric imaging were repeated. Only data from eight of 25 subjects met the following stringent criteria to be included in the study analysis: pre and post-pilocarpine biometric foveal images were registerable, the images of the corneal centers were shifted by ≤100 µm, pupils >5 mm and the pharmacologically induced refractive change was ≥ -7 diopters. RESULTS: The mean auto-refractive accommodative change for the eight included subjects was -12.45 diopters (± 3.45 diopters). The mean change in CLT was 81 µm (± 54 µm) and the mean change in ACD was -145 µm (± 86 µm). Superimposition of the registered pre and post-pilocarpine biometric images of the sagittal sections of the whole eye from each subject demonstrated that the position of the whole lens did not shift either anteriorly, posteriorly or vertically during pharmacologically induced accommodation. CONCLUSIONS: A small increase in lens thickness was associated with a large change in accommodative amplitude and no significant change in lens position as predicted by the Schachar theory.

Axitinib inhibits retinal and choroidal neovascularization in in vitro and in vivo models.

Age-related Macular Degeneration (AMD) is the leading cause of visual impairment and blindness in the elderly in developed countries. Neovascular/exudative (wet) AMD is the aggressive form of AMD and can involve choroidal neovascularization and vascular leakage. Anti-vascular endothelial growth factor (anti-VEGF) medications have significantly improved treatment of wet-AMD. However, only approximately 40% of patients obtain full benefit from anti-VEGF therapy and the medications are given by intravitreal injection. Axitinib, a small molecule multi-receptor tyrosine kinase inhibitor used for the treatment of advanced renal cell carcinoma, is taken orally and inhibits VEGF activity by blocking VEGF receptors. Axitinib also has the advantage of blocking platelet derived growth factor (PDGF) receptors which play a role in neovascularization. Using in vitro human retinal microvascular endothelial cells (HRMVECs), human brain vascular pericytes (HBVRs), 3D co-culture vessel sprout assay, and in vivo laser induced rat choroidal neovascularization (CNV) models, the effect of axitinib on neovascularization was evaluated. Axitinib inhibited neovascularization better than anti-VEGF and/or anti-hPDGF-B mAb in the in vitro models demonstrating that combined inhibition of both VEGF and PDGF pathways may be synergistic in treating wet-AMD. Additionally, axitinib showed good efficacy at a low dose (0.875 mg/day) in laser-induced CNV model in rats. In conclusion our data shows that axitinib, an inhibitor of VEGF and PDGF-B pathways may be useful in ameliorating wet-AMD therapy.

Model of zonular forces on the lens capsule during accommodation.

How the human eye focuses for near; i.e. accommodates, is still being evaluated after more than 165 years. The mechanism of accommodation is essential for understanding the etiology and potential treatments for myopia, glaucoma and presbyopia. Presbyopia affects 100% of the population in the fifth decade of life. The lens is encased in a semi-elastic capsule with attached ligaments called zonules that mediate ciliary muscle forces to alter lens shape. The zonules are attached at the lens capsule equator. The fundamental issue is whether during accommodation all the zonules relax causing the central and peripheral lens surfaces to steepen, or the equatorial zonules are under increased tension while the anterior and posterior zonules relax causing the lens surface to peripherally flatten and centrally steepen while maintaining lens stability. Here we show with a balloon capsule zonular force model that increased equatorial zonular tension with relaxation of the anterior and posterior zonules replicates the topographical changes observed during in vivo rhesus and human accommodation of the lens capsule without lens stroma. The zonular forces required to simulate lens capsule configuration during in vivo accommodation are inconsistent with the general belief that all the zonules relax during accommodation.

Evaluation of the siRNA PF-04523655 versus ranibizumab for the treatment of neovascular age-related macular degeneration (MONET Study).

OBJECTIVE: To evaluate the efficacy of different dosing paradigms of PF-04523655 (PF) versus ranibizumab (comparator) in subjects with neovascular age-related macular degeneration (AMD). DESIGN: Multicenter, open-label, prospective, randomized, comparator-controlled exploratory study. PARTICIPANTS: A total of 151 patients with subfoveal choroidal neovascularization (CNV) secondary to neovascular AMD who were naive to AMD therapy. METHODS: In this phase 2 study, patients were randomized to 1 of 5 treatment groups with equal ratio. All groups received ranibizumab 0.5 mg at baseline and (a) PF 1 mg every 4 weeks (Q4W) from week 4 to week 12; (b) PF 3 mg Q4W from week 4 to week 12; (c) PF 3 mg every 2 weeks (Q2W) from week 4 to week 12; (d) PF 1 mg + ranibizumab (combination) Q4W from baseline to week 12; and (e) ranibizumab Q4W to week 12. All study treatments were given as intravitreal injections. MAIN OUTCOME MEASURES: The primary end point was the mean change in best-corrected visual acuity (BCVA) from baseline at week 16; secondary end points included the percentage of patients gaining ≥ 10 and ≥ 15 letters in BCVA and mean change in retinal central subfield thickness, lesion thickness, and CNV area. RESULTS: At week 16, the PF 1 mg + ranibizumab combination group achieved numerically greater improvement in mean BCVA from baseline (9.5 letters) than the ranibizumab group (6.8 letters). The difference was not statistically significant. The BCVA improvement in the PF monotherapy groups was less than in the ranibizumab group. Similar trends were observed in the percentage of patients who gained ≥ 10 and ≥ 15 letters. From baseline to week 16 (last observed carried forward), the combination and ranibizumab groups had similar mean reductions in central subfield retinal thickness and total CNV area, which were greater than in all PF monotherapy groups. There were no clinically meaningful differences in reduction of lesion thickness among treatment groups. CONCLUSIONS: In this early, underpowered study evaluating treatments for neovascular AMD, the combination of PF with ranibizumab led to an average gain in BCVA that was more than with ranibizumab monotherapy. No safety concerns were identified.

The effect of high concentration and exposure duration of nanoceria on human lens epithelial cells.

Nanotechnology has the potential for treating diseases and conditions of ageing. The eye is particularly vulnerable, because chronic pathologies can lead to sight loss. Human lens epithelial cells were exposed to 10, 20, and 100 µg/mL of negatively charged nanoceria for 48 and 72 hours; DNA damage and cell growth were assessed. Concentrations up to 100 µg/mL for 48 hours did not cause measurable genotoxic effects. For exposures of 72 hours, concentrations above 10 µg/mL showed small but statistically significant differences in DNA damage from negative controls. All treated samples were less damaged than positive controls. Cell growth, monitored for up to 7 days, did not show deviations in cell morphology or growth between treated and untreated samples. Whereas time of exposure may have greater effect than dosage, indicating potential for genotoxicity at higher exposures, human lens epithelial cells can sustain normal growth when exposed to concentrations of nanoceria of up to 100 µg/mL. From the Clinical Editor: Human lens epithelial cells were exposed to various concentrations of negatively charged nanoceria for 48 and 72 hours to assess DNA damage and cell growth. The authors demonstrate that epithelial cells can sustain normal growth when exposed to concentrations of up to 100 µg/mL, with time of exposure having a greater effect than dosage, indicating potential genotoxicity at higher exposures.

Nanoceria have no genotoxic effect on human lens epithelial cells.

There are no treatments for reversing or halting cataract, a disease of the structural proteins in the eye lens, that has associations with other age-related degenerative conditions such as Alzheimer's disease. The incidence of cataract and associated conditions is increasing as the average age of the population rises. Protein folding diseases are difficult to assess in vivo as proteins and their age-related changes are assessed after extraction. Nanotechnology can be used to investigate protein changes in the intact lens as well as for a potential means of drug delivery. Nanoparticles, such as cerium oxide (CeO(2)) which have antioxidant properties, may even be used as a means of treating cataract directly. Prior to use in treatments, nanoparticle genotoxicity must be tested to assess the extent of any DNA or chromosomal damage. Sister chromatid exchanges were measured and DNA damage investigated using the alkaline COMET assay on cultured human lens epithelial cells, exposed to 5 and 10 microg ml(-1) of CeO(2) nanoparticles (nanoceria). Nanoceria at these dosages did not cause any DNA damage or significant increases in the number of sister chromatid exchanges. The absence of genotoxic effects on lens cells suggests that nanoceria, in the doses and exposures tested in this study, are not deleterious to the eye lens and have the potential for use in studying structural alterations, in developing non-surgical cataract treatments and in investigating other protein folding diseases.

Image registration of the human accommodating eye demonstrates equivalent increases in lens equatorial radius and central thickness.

AIM: To compare the results of in vivo human high resolution image registration studies of the eye during accommodation to the predictions of mathematical and finite element models of accommodation. METHODS: Data from published high quality image registration studies of pilocarpine induced accommodative changes of equatorial lens radius (ELR) and central lens thickness (CLT) were statistically analyzed. RESULTS: The mean changes in ELR and CLT were 6.76 µm/diopter and 6.51 µm/diopter, respectively. The linear regressions, reflecting the association between ELR and accommodative amplitude (AAELR) was: slope=6.58 µm/diopter, r2 =0.98, P<0.0001 and between CLT and AACLT was: slope=6.75 µm/diopter, r2 =0.83, P<0.001. On the basis of these relationships, the CLT slope and the AAELR were used to predict the measured change in ELR (ELRpredicted). There was no statistical difference between ELRpredicted and the measured ELR as demonstrated by a Student's paired t-test: P=0.96 and linear regression analysis: slope=0.97, r2 =0.98, P<0.00001. CONCLUSION: Image registration with invariant positional references demonstrates that ELR and CLT equivalently minimally increase ∼7.0 µm/diopter during accommodation. The small equivalent increases in ELR and CLT are associated with a large accommodative amplitude. These findings are consistent with the predictions of mathematical and finite element models that specified the stiffness of the lens nucleus is the same or greater than the lens cortex and that accommodation involves a small force (<5 g).

Optical coherence tomography measurements of the fresh porcine eye and response of the outer coats of the eye to volume increase.

Corneal and scleral thickness and anterior chamber dimensions are required for understanding developmental and pathological processes. Parameters of the eyeball are also required to calculate optical and material properties. As the eyeball resembles a pressure vessel, it has been suggested that elasticity of the cornea and sclera could be calculated from the measurements of thickness. Baseline corneal and scleral thicknesses and anterior chamber dimensions and how these change with incremental increases of intraocular fluid are measured in fresh porcine eyes using the Visante OCT (optical coherence tomography). At baseline, corneal thickness is almost constant. Anterior scleral thickness is variable, decreasing from 0.91+/-0.07 mm near the limbus to a minimum of 0.58+/-0.13. Posterior scleral thickness is more constant with an average of 0.78+/-0.09 mm. Near the optic nerve the thickness increases to 1.00+/-0.09 mm. Average baseline anterior chamber angle, diameter, and depth were found to be 33.15+/-4.91 deg, 13.60+/-0.38 mm, and 2.13+/-0.22 mm, respectively. After fluid injections, maximum changes in corneal and scleral thicknesses were 9 to 10 and 1 to 3%, respectively. Anterior chamber angle and depth decreased slightly but significantly. Changes in the eyeball coats with fluid injections, indicate that the pressure vessel model can be applied to the eye to calculate corneal and scleral elasticities.

Maximum human objectively measured pharmacologically stimulated accommodative amplitude.

PURPOSE: To measure the maximum, objectively measured, accommodative amplitude, produced by pharmacologic stimulation. METHODS: Thirty-seven healthy subjects were enrolled, with a mean age of 20.2±1.1 years, corrected visual acuity of 20/20, and mean spherical equivalent refraction (SER) =-0.83±1.60 diopters. For each subject, the right pupil was dilated with phenylephrine 10%. After 30 minutes, the pupil was measured, the left eye was patched, and the right eye was autorefracted. Pilocarpine 4% was then instilled in the right eye, followed by phenylephrine. At 45 minutes after the pilocarpine, autorefraction and pupil size were again measured. RESULTS: Mean pupil size pre- and postpilocarpine was 8.0±0.8 mm and 4.4±1.9 mm, respectively. Pre- and postpilocarpine, the mean SER was -0.83±1.60 and -10.55±4.26 diopters, respectively. The mean pilocarpine-induced accommodative amplitude was 9.73±3.64 diopters. Five subjects had accommodative amplitudes ≥14.00 diopters. Accommodative amplitude was not significantly related to baseline SER (p-value =0.24), pre- or postpilocarpine pupil size (p-values =0.13 and 0.74), or change in pupil size (p-value =0.37). Iris color did not statistically significantly affect accommodative amplitude (p-value =0.83). CONCLUSION: Following topically applied pilocarpine, the induced objectively measured accommodation in the young eye is greater than or equal to the reported subjectively measured voluntary maximum accommodative amplitude.

A Long-term Safety Study of Latanoprost in Pediatric Patients With Glaucoma and Ocular Hypertension: A Prospective Cohort Study.

PURPOSE: The objective was to assess the long-term effect of treatment with latanoprost on ocular development and safety in pediatric patients with glaucoma and ocular hypertension. DESIGN: Prospective cohort study. METHODS: This was a prospective 3-year cohort study conducted in 14 countries in Europe and South America. Patients aged < 18 years with glaucoma or ocular hypertension were enrolled into either the latanoprost or non-prostaglandin (non-PG) group in this observational study. The primary endpoint was change in best-corrected visual acuity (BCVA) from baseline to 3 years. Several secondary endpoints were evaluated, including corneal thickness and ocular hyperpigmentation. For treatment comparison, analysis of covariance (ANCOVA) was used for continuous endpoints and Fisher exact test was applied for proportion of participants with clinically significant deterioration events. RESULTS: A total of 175 patients were enrolled: 102 in the latanoprost group (median follow-up: 36.7 months) and 73 in the non-PG group (median follow-up: 36.1 months). There was no statistically significant difference between the latanoprost and the non-PG groups (aged 5 to <18 years) in BCVA change from baseline (least square mean logMAR difference -0.03 [95% confidence interval: -0.12, 0.06]), corneal thickness, or ocular hyperpigmentation. CONCLUSIONS: Latanoprost had an acceptable safety profile with no evidence of inducing clinically meaningful or statistically significant changes in ocular development or ocular hyperpigmentation in pediatric patients with glaucoma and ocular hypertension.

A CCR2/5 Inhibitor, PF-04634817, Is Inferior to Monthly Ranibizumab in the Treatment of Diabetic Macular Edema.

Purpose: Ligands for the proinflammatory C-C chemokine receptor types 2 and 5 (CCR2 and CCR5) are elevated in the eyes of patients with diabetic macular edema (DME). We evaluated the efficacy and safety of PF-04634817, an oral CCR2/5 dual antagonist, versus intravitreal ranibizumab, in adult subjects with DME. Methods: In this phase II, randomized, placebo-controlled, double-masked study, eligible subjects (≥18 years of age) had type 1 or 2 diabetes and DME with best-corrected visual acuity (BCVA) of 20/32 or worse (letter score ≤ 78), and up to 20/320 or better (≥24 letter score), in the study eye. Subjects were assigned randomly 1:1 to once-daily (QD) oral PF-04634817 200 mg plus masked sham therapy as placebo or monthly intravitreal ranibizumab 0.3/0.5 mg plus QD oral placebo. The primary objective was to evaluate the efficacy of PF-04634817 compared with ranibizumab in change from baseline in BCVA after 12 weeks in a noninferiority design. Noninferiority was based on BCVA 80% confidence interval (CI): there had to be a less than three letter loss in the PF-04634817 arm compared with the ranibizumab arm. Results: A total of 199 subjects were randomized. Least squares mean difference in change in BCVA from baseline to week 12 in the study eye for the PF-04634817 arm was -2.41 letters (80% CI: -3.91, -0.91; P = 0.04) compared with ranibizumab. PF-04634817 was well tolerated. Conclusions: Treatment with oral CCR2/5 receptor dual antagonist PF-04634817 was associated with a modest improvement in BCVA, but did not meet the predefined noninferiority criteria compared with intravitreal ranibizumab.

Viscoelastic shear properties of the fresh porcine lens.

AIM: To determine the viscoelastic properties of the porcine lens METHODS: Linear viscoelastic shear properties of the stroma of four porcine lenses were measured within 5 hours post-mortem, using sinusoidal oscillatory shear deformation. The elastic shear modulus, viscous shear modulus, dynamic viscosity, damping ratio, and phase shift of the lenses were quantified by a controlled-strain, linear simple-shear rheometer at frequencies of 10-50 Hz. RESULTS: The mean viscoelastic properties and their standard deviations across the frequencies examined were: the elastic shear modulus, G' = 6.2+/-4.0 Pa, the viscous shear modulus, G'' = 19.2+/-2.5 Pa, the dynamic viscosity, eta' = 0.16+/-0.1 Pa x sec, the damping ratio zeta = 4.06+/-1.25, and the phase shift, delta = 76 degrees +/- 5.6 degrees. CONCLUSIONS: The measured viscoelastic shear properties of the porcine lens reflect a low dynamic viscosity with a high damping ratio. The porcine lens is viscoelastic and is more viscous than elastic. The magnitude of the complex shear modulus of the porcine lens, |G*|, is similar to the shear modulus of the young human lens. Understanding these viscoelastic properties of the natural lens may provide guidance in developing a lens substitute capable of accommodation in the post cataract patient.

The relationship between accommodative amplitude and the ratio of central lens thickness to its equatorial diameter in vertebrate eyes.

AIM: To determine the relationship between accommodative amplitude and central lens thickness/equatorial lens diameter (CLT/ELD) ratio in vertebrates. METHODS: Midsagittal sections of lenses from fixed, post mortem eyes from 125 different vertebrate species were photographed. Their CLT/ELD ratios were correlated with independently published measurements of their accommodative amplitudes. Using the non-linear finite element method (FEM), the efficiency of zonular traction (the absolute change in central radius of curvature per unit force [|DeltaCR|/F]) for model lenses with CLT/ELD ratios from 0.45 to 0.9 was determined. RESULTS: Vertebrates with CLT/ELD ratios < or =0.6 have high accommodative amplitudes. Zonular traction was found to be most efficient for those model lenses having CLT/ELD ratios < or =0.6. CONCLUSIONS: Vertebrates with lenses that have CLT/ELD ratios < or =0.6--i.e. "long oval" shapes--have the greatest accommodative amplitudes; e.g. primates, diving birds and diurnal birds of prey. Vertebrates that have oval or spherical shaped lenses, like owls and most mammals, have low accommodative amplitudes. Zonular traction was found to be most efficient when applied to model lenses with CLT/ELD ratios < or =0.6. The implications of these findings on the mechanism of accommodation are discussed.