Optical development in the murine eye lens of accelerated senescence-prone SAMP8 and senescence-resistant SAMR1 strains.

The eye lens is responsible for focusing objects at various distances onto the retina and its refractive power is determined by its surface curvature as well as its internal gradient refractive index (GRIN). The lens continues to grow with age resulting in changes to the shape and to the GRIN profile. The present study aims to investigate how the ageing process may influence lens optical development. Murine lenses of accelerated senescence-prone strain (SAMP8) aged from 4 to 50 weeks; senescence-resistant strain (SAMR1) aged from 5 to 52 weeks as well as AKR strain (served as control) aged from 6 to 70 weeks were measured using the X-ray interferometer at the SPring-8 synchrotron Japan within three consecutive years from 2020 to 2022. Three dimensional distributions of the lens GRIN were reconstructed using the measured data and the lens shapes were determined using image segmentation in MatLab. Variations in the parameters describing the lens shape and the GRIN profile with age were compared amongst three mouse strains. With advancing age, both the lens anterior and posterior surface flattens and the lens sagittal thickness increase in all three mouse strains (Anterior radius of curvature increase at 0.008 mm/week, 0.007 mm/week and 0.002 mm/week while posterior radius of curvature increase at 0.002 mm/week, 0.007 mm/week and 0.003 mm/week respectively in AKR, SAMP8 and SAMR1 lenses). Compared with the AKR strain, the SAMP8 samples demonstrate a higher rate of increase in the posterior curvature radius (0.007 mm/week) and the thickness (0.015 mm/week), whilst the SAMR1 samples show slower increases in the anterior curvature radius (0.002 mm/week) and its thickness (0.013 mm/week). There are similar age-related trends in GRIN shape in the radial direction (in all three types of murine lenses nr2 and nr6 increase with age while nr4 decrease with age consistently) but not in the axial direction amongst three mouse strains (nz1 of AKR lens decrease while of SAMP8 and SAMR1 increase with age; nz2 of all three models increase with age; nz3 of AKR lens increase while of SAMP8 and SAMR1 decrease with age). The ageing process can influence the speed of lens shape change and affect the GRIN profile mainly in the axial direction, contributing to an accelerated decline rate of the optical power in the senescence-prone strain (3.5 D/week compared to 2.3 D/week in the AKR control model) but a retardatory decrease in the senescence-resistant strain (2.1 D/week compared to the 2.3D/week in the AKR control model).

A comparative analysis of the influence of refractive error on image acuity using three eye models.

PURPOSE: To analyse and compare image acuity for different refractive errors generated by either altering axial length or corneal curvature and using three human eye models with two pupil sizes. METHODS: Three different eye models, Liou-Brennan, Goncharov and Navarro, were used. Simulations were made (using Ansys Zemax OpticStudio 22.3) for real pupil sizes of 3 and 6 mm with refractive errors ranging from -2 to +2 D in 0.25 D increments. Refractive errors were simulated by varying axial length or corneal curvature. Root mean square (RMS) values were used to determine image acuity. RESULTS: For the 3-mm pupil, all models gave similar results, with the Navarro model having slightly higher RMS values for the emmetropic eye. For the 6-mm pupil, the Liou-Brennan and Goncharov eye models gave similar results, with RMS values lower than for the Navarro eye model. The highest RMS value was visible in the axial length-induced refractive errors. Refractive errors generated by altering corneal curvature give smaller RMS values than those generated by altering axial length. The axial length and corneal radius simulations indicate a wide spread of results for myopic, hyperopic and emmetropic eyes. There are multiple outcomes that give the same refractive error, even within a single-eye model. The axial length/corneal curvature ratio showed a higher ratio for myopes than hyperopes for every model. CONCLUSIONS: The influence of refractive error on image acuity varied depending on the simulation method of refractive error and the model used. The origins of refractive error and the influence it has on image acuity need further investigation. As models become more sophisticated, personalised and biologically relevant, they will better represent the image acuity of the eye for varying refractive errors, ethnicities, ages and pupil sizes.

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.

Optical development in the zebrafish eye lens.

The optics of the eye is the key to a functioning visual system. The exact nature of the correlation between ocular optics and eye development is not known because of the paucity of knowledge about the growth of a key optical element, the eye lens. The sophisticated optics of the lens and its gradient of refractive index provide the superior optical quality that the eye needs and which, it is thought, has a major influence on the development of proper visual function. The nature of a gradient refractive index lens, however, renders accurate measurements of its development difficult to make and has been the reason why the influence of lens growth on visual function remains largely unknown. Novel imaging techniques have made it possible to investigate growth of the eye lens in the zebrafish. This study shows measurements using X-ray Talbot interferometry of three-dimensional gradient index profiles in eye lenses of zebrafish from late larval to adult stages. The zebrafish lens shows evidence of a gradient of refractive index from the earliest stages measured and its growth suggests an apparent coincidence between periods of rapid increase in refractive index in the lens nucleus and increased expression of a particular crystallin protein group.

Comparative analysis of the corneal birefringence pattern in healthy children and adults.

PURPOSE: To undertake a comparative analysis of the corneal shape, thickness and isochromatics in the eyes of children and adults in order to determine the extent of similarities and differences between the cohorts. METHODS: The study involved 24 children (aged 8 years) and 37 young White adults (aged between 22-24 years) with no apparent or known health or ocular conditions. Measurements were made of corneal radius of curvature, both central (CCT) and paracentral (PCT) corneal thickness and intraocular pressure (IOP). Images of the isochromatics were captured using a slit lamp and a circular polarizer. The geometry of fringe I and II of the isochromatics was analysed. RESULTS: Statistically significant differences were found between CCT and PCT in nasal and temporal regions for both the children and adult cohorts. The same trends were observed in the radii of the cornea. Statistically significant differences between side lengths and angles of isochromatic fringes were found. No differences in asymmetry of shape for fringe I between adults and children were detected; greater symmetry was seen in fringe II in children than for adults. CONCLUSIONS: The asymmetry in corneal shape and curvature contributes to the shape of the isochromatic fringes. This is likely linked to the orientation and parameters of the collagen fibres and to the muscles' forces, and be relevant for surgical procedures such as corneal transplantation.

How a dynamic optical system maintains image quality: Self-adjustment of the human eye.

The eyeball is continually subjected to forces that cause alterations to its shape and dimensions, as well as to its optical components. Forces that induce accommodation result in an intentional change in focus; others, such as the effect of intraocular pressure fluctuations, are more subtle. Although the mechanical properties of the eyeball and its components permit mediation of such subtle forces, the concomitant optical changes are not detected by the visual system. Optical self-adjustment is postulated as the mechanism that maintains image quality. The purpose of this study was to investigate how self-adjustment occurs by using an optical model of the eyeball and to test the requisite optical and biometric conditions.

Cell compaction is not required for the development of gradient refractive index profiles in the embryonic chick lens.

The development of the eye requires the co-ordinated integration of optical and neural elements to create a system with requisite optics for the given animal. The eye lens has a lamellar structure with gradually varying protein concentrations that increase towards the centre, creating a gradient refractive index or GRIN. This provides enhanced image quality compared to a homogeneous refractive index lens. The development of the GRIN during ocular embryogenesis has not been investigated previously. This study presents measurements using synchrotron X-ray Talbot interferometry and scanning electron microscopy of chick eyes from embryonic day 10: midway through embryonic development to E18: a few days before hatching. The lens GRIN profile is evident from the youngest age measured and increases in magnitude of refractive index at all points as the lens grows. The profile is parabolic along the optic axis and has two distinct regions in the equatorial plane. We postulate that these may be fundamental for the independent central and peripheral processes that contribute to the optimisation of image quality and the development of an eye that is emmetropic. The spatial distributions of the distinct GRIN profile regions match with previous measurements on different fibre cell groups in chick lenses of similar developmental stages. Results suggest that tissue compaction may not be necessary for development of the GRIN in the chick eye lens.

Treatment of Human Lens Epithelium with High Levels of Nanoceria Leads to Reactive Oxygen Species Mediated Apoptosis.

Nanoceria (cerium oxide nanoparticles) have been shown to protect human lens epithelial cells (HLECs) from oxidative stress when used at low concentrations. However, there is a lack of understanding about the mechanism of the cytotoxic and genotoxic effects of nanoceria when used at higher concentrations. Here, we investigated the impact of 24-hour exposure to nanoceria in HLECs. Nanoceria's effects on basal reactive oxygen species (ROS), mitochondrial morphology, membrane potential, ATP, genotoxicity, caspase activation and apoptotic hallmarks were investigated. Scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM-EDX) studies on isolated mitochondria revealed significant uptake and localization of nanoceria in the mitochondria. At high nanoceria concentrations (400 µg mL-1), intracellular levels of ROS were increased and the HLECs exhibited classical hallmarks of apoptosis. These findings concur with the cells maintaining normal ATP levels necessary to execute the apoptotic process. These results highlight the need for nanoceria dose-effect studies on a range of cells and tissues to identify therapeutic concentrations in vitro or in vivo.

Contributions of shape and stiffness to accommodative loss in the ageing human lens: a finite element model assessment.

Ageing changes to the various components of the accommodative system of the eye lens contribute to the loss of focusing power. The relative contributions of each ageing component, however, are not well defined. This study investigates the contribution of geometric parameters and material properties on accommodation, simulated using models based on human lenses aged 16, 35, and 48 years. Each model was tested using two different sets of material properties and a range of zonular fiber angles and was compared to results from in vivo measurements. The geometries and material parameters of older and younger lens models were interchanged to investigate the role of shape and material on accommodative capacity. Results indicate that geometry has the greater role in accommodation.

Data analysis of the ocular response analyzer for improved distinction and detection of glaucoma.

The purpose of this study was to evaluate the clinical utility of the output parameters of the ocular response analyzer (ORA) and those calculated from the raw ORA in subjects with healthy eyes and those with suspected glaucoma, and in patients with two types of glaucoma. The raw ORA data were analyzed using a custom software that included the Gaussian filtering of applanation curves for three different window sizes. To the best of our knowledge, these findings present a novel means of optimizing the use of measurements from the ORA, which can refine the characteristics of corneal biomechanics, enabling a distinction between the types of glaucoma and leading to an improvement in diagnosing and early detection.

Coil conversion to ß-strand induced by dimerization.

Most molecular processes in living organisms rely on protein-protein interactions, many of which are mediated by ß-sheet interfaces; this study investigates the formation of ß-sheet interfaces through the conversion of coils into ß-strands. Following an exhaustive search in the Protein Data Bank, the corresponding structural dimorphic fragments were extracted, characterized, and analyzed. Their short strand lengths and specific amino acid profiles indicate that dimorphic ß-strand interfaces are likely to be less stable than standard ones and could even convert to coil interfaces if their environment changes. Moreover, the construction of a simple classifier able to discriminate between the sequences of dimorphic and standard ß-strand interfaces suggests that the nature of those dimorphic sequences could be predicted, providing a novel means of identifying proteins capable of forming dimers.

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.

A critique of the regulation of data science in healthcare research in the European Union.

The EU offers a suitable milieu for the comparison and harmonisation of healthcare across different languages, cultures, and jurisdictions (albeit with a supranational legal framework), which could provide improvements in healthcare standards across the bloc. There are specific ethico-legal issues with the use of data in healthcare research that mandate a different approach from other forms of research. The use of healthcare data over a long period of time is similar to the use of tissue in biobanks. There is a low risk to subjects but it is impossible to gain specific informed consent given the future possibilities for research. Large amounts of data on a subject present a finite risk of re-identification. Consequently, there is a balancing act between this risk and retaining sufficient utility of the data. Anonymising methods need to take into account the circumstances of data sharing to enable an appropriate balance in all cases. There are ethical and policy advantages to exceeding the legal requirements and thereby securing the social licence for research. This process would require the examination and comparison of data protection laws across the trading bloc to produce an ethico-legal framework compatible with the requirements of all member states. Seven EU jurisdictions are given consideration in this critique.

The Effect of the General Data Protection Regulation on Medical Research.

BACKGROUND: The enactment of the General Data Protection Regulation (GDPR) will impact on European data science. Particular concerns relating to consent requirements that would severely restrict medical data research have been raised. OBJECTIVE: Our objective is to explain the changes in data protection laws that apply to medical research and to discuss their potential impact. METHODS: Analysis of ethicolegal requirements imposed by the GDPR. RESULTS: The GDPR makes the classification of pseudonymised data as personal data clearer, although it has not been entirely resolved. Biomedical research on personal data where consent has not been obtained must be of substantial public interest. CONCLUSIONS: The GDPR introduces protections for data subjects that aim for consistency across the EU. The proposed changes will make little impact on biomedical data research.

Critical appraisal of alternative irritation models: three decades of testing ophthalmic pharmaceuticals.

BACKGROUND: Testing ocular tolerability of ocular pharmaceuticals is an essential regulatory requirement. The current approved reference model (gold standard) for ocular irritation testing is the Draize test. However this method is subjective and involves using live animals, hence the need to develop alternative in vitro and ex vivo testing strategies. SOURCE OF DATA: Pubmed, Science Direct, Scopus, Google Scholar, Medline, Current Content, Web of Science and validation reports from international regulatory bodies; The Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and European Centre for the Validation of Alternative Methods (ECVAM) were searched for in vitro alternatives. AREA OF AGREEMENT: Whilst no single in vitro test can effectively replace the Draize eye irritation test, regulatory bodies and cosmetic/pharmaceutical industries agree that there is a need for in vitro alternatives with validated endpoints to evaluate pharmaceutical ingredients and finished eye products. AREA OF CONTROVERSY: There is no single in vitro test / assay that can predict the ocular irritation potential of mild to moderate test substances. AREA TIMELY FOR DEVELOPING RESEARCH: This review provides a critical appraisal of the selected in vitro and ex vivo ocular toxicity models recommended by international regulatory bodies. These include cytotoxicity methods, biochemical systems and ex vivo assays. The latter are approved by ECVAM as in vitro alternatives for the well-known Draize test. Hen's egg test-chorioallantoic membrane and the isolated rabbit eye test are also accepted by regulatory agencies in France, Germany, the Netherlands and the UK. A combination of ex vivo assays along with histological examination of excised bovine cornea can predict the conjunctival and corneal tolerability and cover a wider range of ocular pharmaceutical substances.

Refractive index degeneration in older lenses: A potential functional correlate to structural changes that underlie cataract formation.

A major structure/function relationship in the eye lens is that between the constituent proteins, the crystallins and the optical property of refractive index. Structural breakdown that leads to cataract has been investigated in a number of studies; the concomitant changes in the optics, namely increases in light attenuation have also been well documented. Specific changes in the refractive index gradient that cause such attenuation, however, are not well studied because previous methods of measuring refractive index require transparent samples. The X-ray Talbot interferometric method using synchrotron radiation allows for measurement of fine changes in refractive index through lenses with opacities. The findings of this study on older human lenses show disruptions to the refractive index gradient and in the refractive index contours. These disruptions are linked to location in the lens and occur in polar regions, along or close to the equatorial plane or in lamellar-like formations. The disruptions that are seen in the polar regions manifest branching formations that alter with progression through the lens with some similarity to lens sutures. This study shows how the refractive index gradient, which is needed to maintain image quality of the eye, may be disturbed and that this can occur in a number of distinct ways. These findings offer insight into functional changes to a major optical parameter in older lenses. Further studies are needed to elicit how these may be related to structural degenerations reported in the literature.

Analysis of 2-oxothiazolidine-4-carboxylic acid by hydrophilic interaction liquid chromatography: application for ocular delivery using chitosan nanoparticles.

Oxidative damage due to low levels of glutathione (GSH) is one of the main causes of cataract formation. It has been reported that 2-oxothiazolidine-4-carboxylic acid (OTZ), a cysteine prodrug, can increase the cellular level of GSH. Currently, there is no analytical method to separate and quantify OTZ from aqueous humour samples for cataract research. The present study aims to develop and validate a hydrophilic interaction liquid chromatography (HILIC) method for the quantification of OTZ in simulated aqueous humour (SAH). The developed method was validated according to FDA guidelines. Accuracy, precision, selectivity, sensitivity, linearity, lower limit of quantification (LLOQ), lower limit of detection (LLOD) and stability were the parameters assessed in the method validation. The developed method was found to be accurate and precise with LLOQ and LLOD of 200 and 100 ng/mL, respectively; method selectivity was confirmed by the absence of any matrix interference with the analyte peak. The constructed calibration curve was linear in the range of 0.2-10 µg/mL, with a regression coefficient of 0.999. In addition, the OTZ was found to be stable in SAH after three freeze/thaw cycles. Chitosan nanoparticles loaded with OTZ were formulated by the ionic gelation method. The nanoparticles were found to be uniform in shape and well dispersed with average size of 153 nm. The in vitro release of OTZ from the nanoparticles was quantified using the developed analytical method over 96 h. Permeation of OTZ through excised bovine cornea was measured using HILIC. The lag time and the flux were 0.2 h and 3.05 µg/cm(2) h, respectively.

Myopic Shift during Hyperbaric Oxygenation Attributed to Lens Index Changes.

PURPOSE: To examine ocular lens parameters and structural changes to elucidate mechanisms underlying the myopic shift and cataract-related changes that occur in some patients during hyperbaric oxygen (HBO) therapy. METHODS: Scheimpflug images (Nidek EAS-1000) of the crystalline lens, measurements of scattered light, objective refraction, keratometry, tonometry, and axial length of the eye were obtained after the first day of HBO therapy and repeated when patients had completed 19 days of the treatment. RESULTS: Significant reduction in mean (± SD) optical density was found in the lens nucleus, -2.8 (± 4.3) units (p = 0.009) and -2.2 (± 4.1) units (p = 0.027) within circular and oval areas, respectively. Significant decrease in mean (± SD) backward scattered light was measured, -0.4 (± 0.8) units (p = 0.022). Mean (± SD) myopic shift was -0.58 (± 0.39) diopters (p < 0.001), whereas cortical optical density, forward scattered light, lenticular parameters, keratometry, tonometry, anterior chamber depth, and axial length of the eye appeared unchanged. CONCLUSIONS: Transient myopic shift reported in patients during HBO therapy is attributed to changes in the refractive index of the lens. No changes in lens curvatures or thickness were found after treatment.

Adjustable internal structure for reconstructing gradient index profile of crystalline lens.

Employing advanced technologies in studying the crystalline lens of the eye has improved our understanding of the refractive index gradient of the lens. Reconstructing and studying such a complex structure requires models with adaptable internal geometry that can be altered to simulate geometrical and optical changes of the lens with aging. In this Letter, we introduce an optically well-defined, geometrical structure for modeling the gradient refractive index profile of the crystalline lens with the advantage of an adjustable internal structure that is not available with existing models. The refractive index profile assigned to this rotationally symmetric geometry is calculated numerically, yet it is shown that this does not limit the model. The study provides a basis for developing lens models with sophisticated external and internal structures without the need for analytical solutions to calculate refractive index profiles.

Optical properties of the lens: an explanation for the zones of discontinuity.

The structural basis of zones of discontinuity in the living human eye lens has not been elucidated, and there is no conclusive explanation for what relevance they may have to the structure and function of the lens. Newly developed synchrotron radiation based X-ray Talbot interferometry has enabled the detection of subtle fluctuations in the human eye lens which, when used in mathematical modelling to simulate reflected and scattered light, can recreate the image of the lens seen in the living human eye. The results of this study show that the zones of discontinuity may be caused by subtle fluctuations in the refractive index gradient as well as from random scattering in the central regions. As the refractive index contours are created by cell layers with progressively varying protein concentrations, the zones are linked to growth and will contain information about ageing and development. The index gradient is important for image quality and fluctuations in this gradient may add to quality optimisation and serve as models for designs of new generation implant lenses.