Development of an in vitro model to study the biological effects of blinking.

PURPOSE: To develop a mechanical model in which a contact lens is swept over ocular surface cells under conditions that mimic the force and speed of the blink, and to investigate the resulting biological changes. METHODS: A computer controlled mechanical instrument was developed to hold a dish containing 3D cultured stratified human ocular surface epithelial cells, across which an arm bearing a contact lens was swept back and forth repeatedly at a speed and force mimicking the human blink. Cells were subjected to repeated sweep cycles for up to 1 h at a speed of 120 mm/s with or without an applied force of 19.6 mN (to mimic pressure exerted by upper eyelid), after which the cell layer thickness was measured, the cell layer integrity was investigated using fluorescent quantum dots (6 and 13 nm) and the phosphorylation levels of various protein kinases were analyzed by human phospho-kinase arrays. Data for selected kinases were further quantitated by enzyme immunoassays. RESULTS: The thickness of the cell layers did not change after exposure to sweep cycles with or without applied force. Quantum dots (6 and 13 nm) were able to penetrate the layers of cells exposed to sweep cycles but not layers of untreated control cells. The phosphorylation levels of HSP27 and JNK1/2/3 increased for cells exposed to sweep cycles with applied force compared to untreated control cells. CONCLUSIONS: The in vitro mechanical instrument is a useful tool to investigate the effects of blinking on the ocular surface.

Predicting Accommodative Response Using Paraxial Schematic Eye Models.

PURPOSE: Previous ultrasound biomicroscopy (UBM) studies showed that accommodative optical response (AOR) can be predicted from accommodative biometric changes in a young and a pre-presbyopic population from linear relationships between accommodative optical and biometric changes, with a standard deviation of less than 0.55D. Here, paraxial schematic eyes (SE) were constructed from measured accommodative ocular biometry parameters to see if predictions are improved. METHODS: Measured ocular biometry (OCT, A-scan, and UBM) parameters from 24 young and 24 pre-presbyopic subjects were used to construct paraxial SEs for each individual subject (individual SEs) for three different lens equivalent refractive index methods. Refraction and AOR calculated from the individual SEs were compared with Grand Seiko (GS) autorefractor measured refraction and AOR. Refraction and AOR were also calculated from individual SEs constructed using the average population accommodative change in UBM measured parameters (average SEs). RESULTS: Schematic eye calculated and GS measured AOR were linearly related (young subjects: slope = 0.77, r = 0.86; pre-presbyopic subjects: slope = 0.64, r = 0.55). The mean difference in AOR (GS - individual SEs) for the young subjects was -0.27D and for the pre-presbyopic subjects was 0.33D. For individual SEs, the mean ± SD of the absolute differences in AOR between the GS and SEs was 0.50 ± 0.39D for the young subjects and 0.50 ± 0.37D for the pre-presbyopic subjects. For average SEs, the mean ± SD of the absolute differences in AOR between the GS and the SEs was 0.77 ± 0.88D for the young subjects and 0.51 ± 0.49D for the pre-presbyopic subjects. CONCLUSIONS: Individual paraxial SEs predict AOR, on average, with a standard deviation of 0.50D in young and pre-presbyopic subject populations. Although this prediction is only marginally better than from individual linear regressions, it does consider all the ocular biometric parameters.

Distortion Correction of Visante Optical Coherence Tomography Cornea Images.

PURPOSE: Quantitative biometry measurements from uncorrected anterior segment optical coherence tomography (AS-OCT) images are inaccurate because of spatial and optical distortions. Prior reported distortion correction equations for the Visante AS-OCT were not reproducible. The goal was to calculate the distortions and provide equations to correct corneal parameters for the Visante AS-OCT to get a central corneal radius of curvature from young and older subjects. METHODS: Five contact lenses (CLs) of known front and back radii of curvature and central thickness were imaged using the Visante AS-OCT (Carl Zeiss, Dublin, CA). Contact lens surface coordinates from Visante images were identified and fitted with a circle using custom Matlab image analysis software. Spatial and optical distortions of the Visante image of the CL radii of curvature and thickness were calculated and corrected. Visante images were also captured from 24 younger (aged 21 to 36 years) and 30 older (aged 36 to 48 years) human subjects. Corneal radii of curvature and thickness measurements from these subjects were corrected, and intrasession and intersession repeatabilities of the corneal parameters were calculated. RESULTS: Root mean square error of radius and power of the CL surfaces after distortion correction were 0.02 mm and 0.18D for the front and 0.011 mm and 0.11D for the back, respectively. Intraclass correlation coefficient for intrasession and intersession repeatability for all the corneal parameters from the human subjects was greater than 0.88 in both age groups. CONCLUSIONS: A distortion correction algorithm was developed for the Visante AS-OCT and applied to extract human corneal radius of curvature measurements.

Prediction of accommodative optical response in prepresbyopic subjects using ultrasound biomicroscopy.

PURPOSE: To determine whether relatively low-resolution ultrasound biomicroscopy (UBM) can be used to predict the accommodative optical response in prepresbyopic eyes as well as in a previous study of young phakic subjects, despite lower accommodative amplitudes. SETTING: College of Optometry, University of Houston, Houston, USA. DESIGN: Observational cross-sectional study. METHODS: Static accommodative optical response was measured with infrared photorefraction and an autorefractor (WR-5100K) in subjects aged 36 to 46 years. A 35 MHz UBM device (Vumax, Sonomed Escalon) was used to image the left eye, while the right eye viewed accommodative stimuli. Custom-developed Matlab image-analysis software was used to perform automated analysis of UBM images to measure the ocular biometry parameters. The accommodative optical response was predicted from biometry parameters using linear regression, 95% confidence intervals (CIs), and 95% prediction intervals. RESULTS: The study evaluated 25 subjects. Per-diopter (D) accommodative changes in anterior chamber depth (ACD), lens thickness, anterior and posterior lens radii of curvature, and anterior segment length were similar to previous values from young subjects. The standard deviations (SDs) of accommodative optical response predicted from linear regressions for UBM-measured biometry parameters were ACD, 0.15 D; lens thickness, 0.25 D; anterior lens radii of curvature, 0.09 D; posterior lens radii of curvature, 0.37 D; and anterior segment length, 0.42 D. CONCLUSIONS: Ultrasound biomicroscopy parameters can, on average, predict accommodative optical responses with SDs of less than 0.55 D using linear regressions and 95% CIs. Ultrasound biomicroscopy can be used to visualize and quantify accommodative biometric changes and predict accommodative optical response in prepresbyopic eyes.

Can ultrasound biomicroscopy be used to predict accommodation accurately?

PURPOSE: Clinical accommodation testing involves measuring either accommodative optical changes or accommodative biometric changes. Quantifying both optical and biometric changes during accommodation might be helpful in the design and evaluation of accommodation restoration concepts. This study aims to establish the accuracy of ultrasound biomicroscopy (UBM) in predicting the accommodative optical response (AOR) from biometric changes. METHODS: Static AOR from 0 to 6 diopters (D) stimuli in 1-D steps were measured with infrared photorefraction and a Grand Seiko autorefractor (WR-5100 K; Shigiya Machinery Works Ltd., Hiroshima, Japan) in 26 human subjects aged 21 to 36 years. Objective measurements of accommodative biometric changes to the same stimulus demands were measured from UBM (Vu-MAX; Sonomed Escalon, Lake Success, NY) images in the same group of subjects. AOR was predicted from biometry using linear regressions, 95% confidence intervals, and 95% prediction intervals. RESULTS: Bland-Altman analysis showed 0.52 D greater AOR with photorefraction than with the Grand Seiko autorefractor. Per-diopter changes in accommodative biometry were: anterior chamber depth (ACD): -0.055 mm/D, lens thickness (LT): +0.076 mm/D, anterior lens radii of curvature (ALRC): -0.854 mm/D, posterior lens radii of curvature (PLRC): -0.222 mm/D, and anterior segment length (ASL): +0.030 mm/D. The standard deviation of AOR predicted from linear regressions for various biometry parameters were: ACD: 0.24 D, LT: 0.30 D, ALRC: 0.24 D, PLRC: 0.43 D, ASL: 0.50 D. CONCLUSIONS: UBM measured parameters can, on average, predict AOR with a standard deviation of 0.50 D or less using linear regression. UBM is a useful and accurate objective technique for measuring accommodation in young phakic eyes.

Defining the ideal femtosecond laser capsulotomy.

PURPOSE: We define the ideal anterior capsulotomy through consideration of capsular histology and biomechanics. Desirable qualities include preventing posterior capsular opacification (PCO), maintaining effective lens position (ELP) and optimising capsular strength. METHODS: Laboratory study of capsular biomechanics and literature review of histology and published clinical results. RESULTS: Parameters of ideal capsulotomy construction include complete overlap of the intraocular lens to prevent PCO, centration on the clinical approximation of the optical axis of the lens to ensure concentricity with the capsule equator, and maximal capsular thickness at the capsulotomy edge to maintain integrity. CONCLUSIONS: Constructing the capsulotomy centred on the clinical approximation of the optical axis of the lens with diameter 5.25 mm optimises prevention of PCO, consistency of ELP and capsular strength.

Objective measurement of accommodative biometric changes using ultrasound biomicroscopy.

PURPOSE: To demonstrate that ultrasound biomicroscopy (UBM) can be used for objective quantitative measurements of anterior segment accommodative changes. SETTING: College of Optometry, University of Houston, Houston, Texas, USA. DESIGN: Prospective cross-sectional study. METHODS: Anterior segment biometric changes in response to 0 to 6.0 diopters (D) of accommodative stimuli in 1.0 D steps were measured in eyes of human subjects aged 21 to 36 years. Imaging was performed in the left eye using a 35 MHz UBM (Vumax) and an A-scan ultrasound (A-5500) while the right eye viewed the accommodative stimuli. An automated Matlab image-analysis program was developed to measure the biometry parameters from the UBM images. RESULTS: The UBM-measured accommodative changes in anterior chamber depth (ACD), lens thickness, anterior lens radius of curvature, posterior lens radius of curvature, and anterior segment length were statistically significantly linearly correlated with accommodative stimulus demands. Standard deviations of the UBM-measured parameters were independent of the accommodative stimulus demands (ACD: 0.0176 mm; lens thickness: 0.0294 mm; anterior lens radius of curvature: 0.3350 mm; posterior lens radius of curvature: 0.1580 mm; and anterior segment length: 0.0340 mm). The mean difference between the A-scan and UBM measurements was -0.070 mm for ACD and 0.166 mm for lens thickness. CONCLUSIONS: Accommodating phakic eyes imaged using UBM allowed visualization of the accommodative response, and automated image analysis of the UBM images allowed reliable, objective, quantitative measurements of the accommodative intraocular biometric changes. FINANCIAL DISCLOSURE: Neither author has a financial or proprietary interest in any material or method mentioned.

Pharmacologically and Edinger-Westphal stimulated accommodation in rhesus monkeys does not rely on changes in anterior chamber pressure.

This study was undertaken to understand the role of anterior chamber pressure (ACP) during pharmacological and Edinger-Westphal (EW) stimulated accommodation in anesthetized monkeys. Experiments were performed on one iridectomized eye each of 7 anesthetized adolescent rhesus monkeys. Accommodation was induced by EW stimulation (n = 2) and intravenous administration of 0.25-4.0 mg/kg pilocarpine (n = 6). Accommodative refractive and biometric changes were measured with continuous 60 Hz infrared photorefraction (n = 6) and 100 Hz A-scan ultrasound biometry (n = 1). An ocular perfusion system was used to measure and manipulate ACP. Pressure was recorded via a 27-gauge needle in the anterior chamber connected to a pressure transducer (n = 7). The needle was also connected to a fluid reservoir to allow ACP to be manipulated and clamped (n = 4) by raising or lowering the fluid reservoir. In all six pharmacologically stimulated monkeys ACP increased during accommodation, from 0.70 to 2.38 mmHg, four of which showed pressure decreases preceding the pressure increases. Two eyes also showed increases in ACP during EW-stimulated accommodation of 2.8 and 7.2 mmHg. ACP increased with increasing EW stimulus amplitudes (n = 2). Clamping or externally manipulating ACP had no effect on resting refraction or on EW and pharmacologically stimulated accommodation in four eyes. The results show that EW stimulated and pharmacologically stimulated accommodation do not rely on ACP in rhesus monkeys.

Comparison between carbachol iontophoresis and intravenous pilocarpine stimulated accommodation in anesthetized rhesus monkeys.

Rhesus monkeys are an animal model for human accommodation and presbyopia and consistent and repeatable methods are needed to stimulate and measure accommodation in anesthetized rhesus monkeys. Accommodation has typically been pharmacologically stimulated with topical pilocarpine or carbachol iontophoresis. Intravenous (i.v.) pilocarpine has recently been shown to produce more natural, rapid and reproducible accommodative responses compared to topical pilocarpine. Here, i.v. pilocarpine was compared to carbachol iontophoresis stimulated accommodation. Experiments were performed under anaesthesia on five previously iridectomized monkeys aged 10-16 years. In three monkeys, accommodation was stimulated with carbachol iontophoresis in five successive experiments and refraction measured with a Hartinger coincidence refractometer. In separate experiments, accommodation was stimulated using a 5 mg/kg bolus of i.v. pilocarpine given over 30 s followed by a continuous infusion of 20 mg/kg/hr for 5.5 min in three successive experiments with the same monkeys as well as in single experiments with two additional monkeys. Refraction was measured continuously using photorefraction with baseline and accommodated refraction also measured with the Hartinger. In subsequent i.v. pilocarpine experiments with each monkey, accommodative changes in lens equatorial diameter were measured in real-time with video-image analysis. Maximum accommodation of three monkeys with carbachol iontophoresis (five repeats) was (mean ± SD; range) 14.0 ± 3.5; 9.9-20.3 D and with i.v. pilocarpine stimulation (three repeats) was 11.1 ± 1.1; 9.9-13.0 D. The average of the standard deviations of maximum accommodation from each monkey was 0.8 ± 0.3 D from carbachol iontophoresis and 0.3 ± 0.2 from i.v. pilocarpine. The average latency to the start of the response after carbachol iontophoresis was 2.5 ± 3.9; 0.0-12.0 min with a time constant of 12.7 ± 9.5; 2.3-29.2 min. The average latency after i.v. pilocarpine was 0.31 ± 0.03; 0.25-0.34 min with a time constant of 0.19 ± 0.07; 0.11-0.31 s. During i.v. pilocarpine stimulated accommodation in five monkeys, lens diameters decreased by 0.54 ± 0.09; 0.42-0.64 mm with a rate of change of 0.052 ± 0.002; 0.050-0.055 mm/D. Accommodative responses with i.v. pilocarpine were more rapid, consistent and stable than those with carbachol iontophoresis. The accommodative decrease in lens diameter with i.v. pilocarpine as a function of age was consistent with previous results using constant topical pilocarpine. Intravenous pilocarpine stimulated accommodation is safe, more consistent and more rapid than carbachol iontophoresis and it requires no contact with or obstruction of the eye thus allowing continuous and uninterrupted refraction and ocular biometry measurements.

Long-term reproducibility of Edinger-Westphal stimulated accommodation in rhesus monkeys.

If longitudinal studies of accommodation or accommodation restoration procedures are undertaken in rhesus monkeys, the methods used to induce and measure accommodation must remain reproducible over the study period. Stimulation of the Edinger-Westphal (EW) nucleus in anesthetized rhesus monkeys is a valuable method to understand various aspects of accommodation. A prior study showed reproducibility of EW-stimulated accommodation over 14 months after chronic electrode implantation. However, reproducibility over a period longer than this has not been investigated and therefore remains unknown. To address this, accommodation stimulation experiments in four eyes of two rhesus monkeys (13.7 and 13.8 years old) were evaluated over a period of 68 months. Carbachol iontophoresis stimulated accommodation was first measured with a Hartinger coincidence refractometer (HCR) two weeks before electrode implantation to determine maximum accommodative amplitudes. EW stimulus-response curves were initially measured with the HCR one month after electrode implantation and then repeated at least six times for each eye in the following 60 months. At 64 months, carbachol iontophoresis induced accommodation was measured again. At 68 months, EW stimulus-response curves were measured with an HCR and photorefraction every week over four consecutive weeks to evaluate the short-term reproducibility over one month. In the four eyes studied, long-term EW-stimulated accommodation decreased by 7.00 D, 3.33 D, 4.63 D, and 2.03 D, whereas carbachol stimulated accommodation increased by 0.18 D-0.49 D over the same time period. The short-term reproducibility of maximum EW-stimulated accommodation (standard deviations) over a period of four weeks at 68 months after electrode implantation was 0.48 D, 0.79 D, 0.55 D and 0.39 D in the four eyes. Since the long-term decrease in EW-stimulated accommodation is not matched by similar decreases in carbachol iontophoresis stimulated accommodation, the decline in accommodation cannot be due to the progression of presbyopia but is likely to result from variability in EW electrode position. Therefore, EW-stimulated accommodation in anesthetized monkeys is not appropriate for long-term longitudinal studies of age-related loss of accommodation or accommodation restoration procedures.

Advances in lens implant technology.

Cataract surgery is one of the oldest and the most frequent outpatient clinic operations in medicine performed worldwide. The clouded human crystalline lens is replaced by an artificial intraocular lens implanted into the capsular bag. During the last six decades, cataract surgery has undergone rapid development from a traumatic, manual surgical procedure with implantation of a simple lens to a minimally invasive intervention increasingly assisted by high technology and a broad variety of implants customized for each patient's individual requirements. This review discusses the major advances in this field and focuses on the main challenge remaining - the treatment of presbyopia. The demand for correction of presbyopia is increasing, reflecting the global growth of the ageing population. Pearls and pitfalls of currently applied methods to correct presbyopia and different approaches under investigation, both in lens implant technology and in surgical technology, are discussed.

Quantification of age-related and per diopter accommodative changes of the lens and ciliary muscle in the emmetropic human eye.

PURPOSE: To calculate age-related and per diopter (D) accommodative changes in crystalline lens and ciliary muscle dimensions in vivo in a single cohort of emmetropic human adults ages 30 to 50 years. METHODS: The right eyes of 26 emmetropic adults were examined using ultrasonography, phakometry, anterior segment optical coherence tomography, and high resolution magnetic resonance imaging. Accommodation was measured both subjectively and objectively. RESULTS: In agreement with previous research, older age was linearly correlated with a thicker lens, steeper anterior lens curvature, shallower anterior chamber, and lower lens equivalent refractive index (all P < 0.01). Age was not related to ciliary muscle ring diameter (CMRD) or lens equatorial diameter (LED). With accommodation, lens thickness increased (+0.064 mm/D, P < 0.001), LED decreased (-0.075 mm/D, P < 0.001), CMRD decreased (-0.105 mm/D, P < 0.001), and the ciliary muscle thickened anteriorly (+0.013 to +0.026 mm/D, P < 0.001) and thinned posteriorly (-0.011 to -0.015, P < 0.01). The changes per diopter of accommodation in LED, CMRD, and ciliary muscle thickness were not related to subject age. CONCLUSIONS: The per diopter ciliary muscle contraction is age independent, even as total accommodative amplitude declines. Quantifying normal biometric dimensions of the accommodative structures and changes with age and accommodative effort will further the development of new IOLs designed to harness ciliary muscle forces.

Manipulation of intraocular pressure for studying the effects on accommodation.

A reliable experimental system in which IOP can be manipulated or a rapid IOP change can be induced while simultaneously and continuously measuring IOP and the ocular accommodative changes would be useful for understanding the physiological effect of intraocular pressure (IOP) on the accommodative mechanism. In this study, an IOP perfusion and recording system was developed and tested using 13 enucleated pig eyes. The vitreous chamber of the pig eyes was cannulated with a needle connected to two fluid reservoirs at different heights. One reservoir was set to achieve one of three baseline pressures of 5.5 mmHg, 13.0 mmHg and 20.5 mmHg. The other reservoir was moved to achieve pressures of 1.5 mmHg, 3.0 mmHg, 4.5 mmHg and 6.0 mmHg higher than the baseline pressure. The height differential between the reservoirs determined the amplitude of IOP changes. Rapid IOP changes were induced by switching the reservoirs with a solenoid pinch-valve. Two needles, one each attached to a pressure transducer were inserted into the anterior chamber and vitreous chamber respectively. Custom developed software was used to measure the anterior chamber pressure and vitreous chamber pressure at 80 Hz. A high-resolution continuous A-scan ultrasound biometer (CUB) was used to dynamically measure changes in ocular biometry including anterior chamber depth (ACD), lens thickness (LT) and vitreous chamber depth (VCD) while the vitreous chamber pressure was manipulated. The changes in ACD, LT and VCD were analyzed as a function of the pressure change. Perfusion-induced axial biometric changes were quantified by the slopes of linear regression relationships. Both anterior chamber pressure and vitreous chamber pressure changed relatively systematically with the induced vitreous chamber pressure changes (anterior chamber: y = 0.863x + 0.030, r(2) = 0.983; vitreous chamber: y = 0.883x + 0.009, r(2) = 0.981). At perfusion pressures of 5.5, 13.0 and 20.5 mmHg, the slopes for ACD were -5.72, -2.75 and -2.36 µm/mmHg, for LT were -3.31, -1.59 and -1.03 µm/mmHg and for VCD were 19.05, 8.63 and 5.18 µm/mmHg. The system was able to manipulate and monitor IOP while axial biometry changes were recorded. This system will allow the relationship between IOP and accommodation to be studied in non-human primate eyes.