The Impact of Photopigment Bleaching on the Human Rod Photoreceptor Subretinal Space Measured Via Optical Coherence Tomography.

Purpose: The purpose of this study was to investigate rod photopigment bleaching-driven intrinsic optical signals (IOS) in the human outer retina and its measurement repeatability based on a commercial optical coherence tomography (OCT) platform. Methods: The optical path length of the rod photoreceptor subretinal space (SRS), that is, the distance between signal bands of rod outer segment tips and retinal pigment epithelium, was measured in 15 healthy subjects in ambient light and during a long-duration bleaching white-light exposure. Results: On 2 identical study days (day 1 and day 2 [D1 and D2]), light stimulation resulted in a significant decrease in rod SRS by 21.3 ± 7.6% and 19.8 ± 8.5% (both P < 0.001), respectively. The test-retest reliability of the SRS maximum change of an individual subject was moderate for single measures (intraclass correlation coefficient [ICC] = 0.730, 95% confidence interval [CI] = 0.376, 0.900, P < 0.001) and good for average measures (ICC = 0.844, 95% CI = 0.546, 0.947, P < 0.001). The mean area under the stimulus response curve with values of 14.8 ± 9.4 and 15.5 ± 7.5 µm × minutes (P = 0.782) showed excellent agreement between the stimulus response on D1 and D2. Intermittent dark adaptation of the retina led to an initial increase of the SRS by 6.1% (P = 0.018) and thereafter showed a decrease toward baseline, despite continued dark adaptation. Conclusions: The data indicate the potential of commercial OCT in measuring slow IOS in the outer retina suggesting that the rod SRS could serve as a biomarker for photoreceptor function. The presented approach could provide an easily implementable clinical tool for the early detection of diseases affecting photoreceptor health.

Classification of Tear Film Lipid Layer En Face Maps Obtained Using Optical Coherence Tomography and Their Correlation With Clinical Parameters.

PURPOSE: The purpose of this study was to investigate the correlation between the pattern of optical coherence tomography (OCT) en face maps of the tear film lipid layer (TFLL) and lipid layer thickness (LLT), fluorescein breakup time (FBUT), and Schirmer I test values in healthy subjects. METHODS: Measurements from four clinical data sets were retrospectively analyzed, and TFLL patterns were classified into 3 categories: homogeneous (HOM), wavy (WAV), or dotted (DOT) appearance. Linear mixed model analyses were performed. Intraclass correlation coefficients and index of qualitative variation were computed to investigate interrater and intrasubject variabilities. RESULTS: For the LLT, a significant difference between HOM and DOT ( P < 0.001, ß HOMvsDOT = -6.42 nm) and WAV and DOT ( P = 0.002, ß WAVvsDOT = -4.04 nm) was found. Furthermore, the difference between WAV and DOT regarding FBUT ( P < 0.001, ß WAVvsDOT = -3.065 seconds) was significant, while no significant differences between any of the classes with respect to the Schirmer I test values were found. An intraclass correlation coefficient of 89.0% reveals a good interrater reliability, and an index of qualitative variation of 60.0% shows, on average, a considerable variability in TFLL pattern class for repeated measurements over 1 hour. CONCLUSIONS: A new classification method for OCT en face maps of the TFLL is presented. Significant differences between patterns were found with respect to LLT and FBUT. A dotted pattern on dark background appears to be the most stable type of TFLL. The analysis of OCT en face maps of the TFLL provides complimentary information to conventional imaging methods and might give new insights into the characteristics of the TFLL.

Quantification of intrinsic optical signals in the outer human retina using optical coherence tomography.

Intrinsic optical signals constitute a noninvasive biomarker promising the objective assessment of retinal photoreceptor function. We employed a commercial optical coherence tomography (OCT) system and an OCT signal model for evaluation of optical path length (OPL) changes in the temporal outer retina of five healthy subjects during light adaptation. Data were acquired at 30 time points, in ambient light and during long duration stimulation with white light, and analyzed, employing a signal model based on the sum of seven Gaussian curves corresponding to all relevant anatomical structures of the outer retina. During light stimulation, mean OPL between rod outer segment tips (ROST) and the retinal pigment epithelium (RPE) decreased by 21.4 ± 3.5%. Further, OPL between the external-limiting membrane (ELM) and the RPE decreased by 5.2 ± 0.9% versus baseline, while OPL between ELM and ROST showed an initial decrease by 2.1 ± 1.6% versus baseline and, thereafter, increased by 2.8 ± 2.1% versus baseline. Thus, the presented approach allowed for assess to dynamic changes in the outer retina in response to light. The change in the subretinal space occurring in the context of light adaptation could be measured using a standard OCT platform and a dedicated signal model.

Influence of Perfluorohexyloctane Eye Drops on Tear Film Thickness in Patients with Mild to Moderate Dry Eye Disease: A Randomized Controlled Clinical Trial.

Purpose: The aim of this mechanistic clinical study was to explore the effect of water-free perfluorohexyloctane eye drops on tear film thickness (TFT) in patients with dry eye disease (DED). Methods: Forty-eight patients with mild to moderate DED participated in this randomized, single-masked, observer-blinded parallel group study in a 1:1 ratio to receive either perfluorohexyloctane or unpreserved 0.9% saline solution (Hydrabak®, Thea, France) eye drops 4 times daily in both eyes for 4 weeks. A custom-built ultrahigh-resolution optical coherence tomography system was used to measure TFT. Furthermore, evaluation of lipid layer thickness (LLT) and noninvasive tear film breakup time, as well as standard clinical tests for signs and symptoms of DED were performed. Results: Mean TFT and LLT at baseline were comparable between the 2 treatment groups. After a single drop instillation, perfluorohexyloctane eye drops temporarily increased TFT immediately. After multiple dosing, perfluorohexyloctane eye drops gradually increased TFT over time with a maximum effect at the end of the study (least square mean difference: 6.42%; P = 0.0142 at week 4). LLT values measured before drop instillation showed a more prominent increase in LLT for perfluorohexyloctane eye drops (13.36% ± 26.33% vs. 3.21% ± 28.65%). All other parameters got better in both treatment groups with no statistical difference between groups. Conclusions: These results demonstrate that perfluorohexyloctane eye drops increase TFT as well as LLT over time. These tear film reestablishing attributes are in line with the mode of action of perfluorohexyloctane eye drops to avoid evaporation through stabilization of the lipid layer.

Characterization of dry eye disease in a mouse model by optical coherence tomography and fluorescein staining.

A custom-built ultrahigh-resolution optical coherence tomography (UHR-OCT) system and fluorescein staining were employed for investigation of a scopolamine induced dry eye mouse model. Acquired data was used to evaluate common and complementary findings of the two modalities. Central corneal thickness as measured by UHR-OCT increased significantly over the study period of 24 hours, from 89.0 ± 3.57 µm to 92.2 ± 4.07 µm. Both techniques were able to show corneal lesions with a large range of severity. Localized fluorescein staining was detected in 5% and diffuse staining in 45% of cases where no epithelial damage was visible with OCT. However, OCT revealed stromal defects in 6% and endothelial defects in 18% of the cases, which could not be visualized via fluorescein staining. Thus, while fluorescein staining widely detected defects of the corneal surface in a mouse model of dry eye disease, OCT non-invasively revealed additional information about defect depth and involvement of particular layers.

Light-induced changes of the subretinal space of the temporal retina observed via optical coherence tomography.

Photoreceptor function is impaired in many retinal diseases like age-related macular degeneration. Currently, assessment of the photoreceptor function for the early diagnosis and monitoring of these diseases is either subjective, as in visual field testing, requires contact with the eye, like in electroretinography, or relies on research prototypes with acquisition speeds unattained by conventional imaging systems. We developed an objective, noncontact method to monitor photoreceptor function using a standard optical coherence tomography system. This method can be used with various white light sources for stimulation. The technique was applied in five volunteers and detected a decrease of volume of the subretinal space associated with light adaptation processes of the retina.

Relationship Between the Parameters of Corneal and Fundus Pulse Signals Acquired With a Combined Ultrasound and Laser Interferometry Technique.

PURPOSE: To estimate the relationship between the characteristics of the corneal pulse (CP) signal and those of the fundus pulse (FP) signal measured with a combined noncontact ultrasonic and laser interferometry technique in healthy subjects. METHODS: Twenty-two healthy subjects participated in experiments that included measurements of intraocular pressure, ocular pulse amplitude, ocular biometry, blood pressure, and heart rate. Additionally, simultaneous recordings of CP and FP signals were acquired with a noncontact ultrasonic device combined with laser interferometry. Subsequently, ocular perfusion pressure (OPP) and the time and spectral parameters of CP and FP signals were computed. A system model was proposed to relate the FP signal to the CP signal. RESULTS: The system model revealed that the eye globe transfers information between signals of the posterior and anterior eye, relatively amplifying higher spectral harmonics. The amplitude of the second CP harmonic is predicted by FPRMS and OPP (R 2 = 0.468, P = 0.002). Partial correlation analysis showed that the CP signal parameters are statistically significantly correlated with those of the FP signal and OPP, after correcting for age and sex. CONCLUSIONS: The eye globe can be viewed as a high pass filter, in which the CP characteristic changes in relation to the fundus pulsation. The FP signal and OPP have an impact on the variations of the CP signal morphology. TRANSLATIONAL RELEVANCE: Investigation of differences between the characteristics of the anterior and posterior tissue movements is a promising method for evaluating the role of circulatory and biomechanical components in the pathophysiology of ocular diseases.

Automatic assessment of tear film and tear meniscus parameters in healthy subjects using ultrahigh-resolution optical coherence tomography.

Many different parameters exist for the investigation of tear film dynamics. We present a new tear meniscus segmentation algorithm which automatically extracts tear meniscus area (TMA), height (TMH), depth (TMD) and radius (TMR) from UHR-OCT measurements and apply it to a data set including repeated measurements from ten healthy subjects. Mean values and standard deviations are 0.0174 ± 0.007 mm2, 0.272 ± 0.069 mm, 0.191 ± 0.049 mm and 0.309 ± 0.123 mm for TMA, TMH, TMD and TMR, respectively. A significant correlation was found between all respective tear meniscus parameter pairs (all p < 0.001, all Pearson's r ≥ 0.657). Challenges, limitations and potential improvements related to the data acquisition and the algorithm itself are discussed. The automatic segmentation of tear meniscus measurements acquired with UHR-OCT might help in a clinical setting to further understand the tear film and related medical conditions like dry eye disease.

Ultrahigh-resolution anterior segment optical coherence tomography for analysis of corneal microarchitecture during wound healing.

PURPOSE: To employ ultrahigh-resolution (UHR) optical coherence tomography (OCT) for investigation of the early wound healing process in corneal epithelium. METHODS: A custom-built UHR-OCT system assessed epithelial healing in human keratoconic cornea after epi-off crosslinking (CXL) procedure and a wound healing model in rabbits with iatrogenic corneal injury. 3D OCT data sets enhanced obtaining epithelial thickness maps and evaluation of reepithelization stage. Accompanying changes in deeper corneal microarchitecture were analysed. RESULTS: The mean central corneal thickness in 40 eyes with keratoconus at baseline was 482.7 ± 38.2 µm, while mean central epithelial thickness (CET) was 43.8 ± 6.4 µm. At the final visit 20 ± 5 days post-CXL procedure, CET was 35.0 ± 5.8 µm, significantly thinner after reepithelization (p < 0.001). Surgical success was assessed at the final visit through the demarcation line (DL), identified at 43.7 ± 13.5% stromal depth. In rabbits, the mean CET in 20 eyes at baseline was 35.9 ± 2.6 µm. In rabbits that revealed complete wound closure (10/20 eyes) at the last study day at 72 hr, CET was significantly thinner compared to baseline (30.4 ± 2.8 µm versus 35.4 ± 2.9 µm, p = 0.005). An intra-stromal landmark indicating early keratocyte apoptosis was measured at 30.0 ± 5.1% stromal depth. Epithelial thickness maps showed the time-course of corneal healing. CONCLUSION: Ultrahigh-resolution (UHR)-OCT provided precise assessment of epithelial wound and its healing by 3D-mapping. In addition, microarchitectural changes in the cornea in early phases of epithelial healing were revealed.

Effect of Single Instillation of Two Hyaluronic Acid-Based Topical Lubricants on Tear Film Thickness in Patients with Dry Eye Syndrome.

PURPOSE: This study investigated the effect of single administration of 2 different hyaluronic acid-based topical lubricants on tear film thickness (TFT) in patients with moderate-to-severe dry eye disease (DED). METHODS: Sixty patients were randomized to receive eye drops containing unpreserved sodium hyaluronate, triglycerides, and phospholipids (HTP), unpreserved sodium hyaluronate (HA), or unpreserved sodium chloride (NaCl). TFT and lipid layer thickness (LLT) were assessed before and at defined time points after single instillation with a custom-built ultrahigh-resolution optical coherence tomography system and a white light interferometer. Standard tests for DED such as a visual analogue scale (VAS), corneal fluorescein staining (CFS), Schirmer I test, and tear break-up time (BUT) were performed. RESULTS: The time course in TFT was significantly different between the 3 administered agents (P < 0.001). Single administration of HTP significantly increased TFT over a period of 40 min, an increase in TFT over 20 min was observed after instillation of HA, whereas NaCl had no effect. No significant difference in the change in BUT, CFS, or LLT was observed between the 3 groups (P = 0.57, 0.97, and 0.86, respectively). A significant improvement in subjective symptoms (VAS) was found after single instillation of HTP (P = 0.03 vs. baseline) and HA, but not after administration of NaCl (P = 0.03, P < 0.01, and P = 0.08 vs. baseline, respectively, P = 0.57 between groups). CONCLUSIONS: Single administration of HTP and HA significantly increased TFT showing a longer residence time and improved subjective symptoms, whereas instillation of NaCl had no effect. Long-term studies with multiple instillations will be necessary to further investigate the effect of the tested products on the clinical outcome. ClinicalTrials.gov ID: NCT03161080.

Anterior segment optical coherence tomography.

Optical coherence tomography (OCT) provides non-contact, rapid in vivo imaging of ocular structures, and has become a key part of evaluating the anterior segment of the eye. Over the years, improvements to technology have increased the speed of capture and resolution of images, leading to the increasing impact of anterior segment OCT imaging on clinical practice. In this review, we summarize the historical development of anterior segment OCT, and provide an update on the research and clinical applications of imaging the ocular surface, cornea, anterior chamber structures, aqueous outflow system, and most recently anterior segment vessels. We also describe advancements in anterior segment OCT technology that have improved understanding with greater detail, such as tear film in dry eye disease evaluation, intra-operative real-time imaging for anterior segment surgery, and aqueous outflow with angle assessment for glaucoma. Improvements to image processing and software have also improved the ease and utility of interpreting anterior segment OCT images in everyday clinical practice. Future developments include refinement of assessing vascular networks for the anterior segment, in vivo ultra-high resolution anterior segment optical coherence tomography with histology-like detail, en-face image with 3-dimensional reconstruction as well as functional extensions of the technique.

Ultrahigh-resolution OCT imaging of the human cornea.

We present imaging of corneal pathologies using optical coherence tomography (OCT) with high resolution. To this end, an ultrahigh-resolution spectral domain OCT (UHR-OCT) system based on a broad bandwidth Ti:sapphire laser is employed. With a central wavelength of 800 nm, the imaging device allows to acquire OCT data at the central, paracentral and peripheral cornea as well as the limbal region with 1.2 µm x 20 µm (axial x lateral) resolution at a rate of 140 000 A-scans/s. Structures of the anterior segment of the eye, not accessible with commercial OCT systems, are visualized. These include corneal nerves, limbal palisades of Vogt as well as several corneal pathologies. Cases such as keratoconus and Fuchs's endothelial dystrophy as well as infectious changes caused by diseases like Acanthamoeba keratitis and scarring after herpetic keratitis are presented. We also demonstrate the applicability of our system to visualize epithelial erosion and intracorneal foreign body after corneal trauma as well as chemical burns. Finally, results after Descemet's membrane endothelial keratoplasty (DMEK) are imaged. These clinical cases show the potential of UHR-OCT to help in clinical decision-making and follow-up. Our results and experience indicate that UHR-OCT of the cornea is a promising technique for the use in clinical practice, but can also help to gain novel insight in the physiology and pathophysiology of the human cornea.

Effect of Diffuse Luminance Flicker Light Stimulation on Total Retinal Blood Flow Assessed With Dual-Beam Bidirectional Doppler OCT.

Purpose: We assess the increase in total retinal blood flow (TRBF) induced by flicker stimulation of the human retina in vivo and investigate the flicker induced hyperemia by means of a vascular flow model of the retinal circulation to study neurovascular coupling (NC). Methods: In six healthy subjects, TRBF was measured before and during stimulation with diffuse luminance flicker. Blood flow velocities in retinal vessels were measured via dual-beam bidirectional Doppler Fourier-domain optical coherence tomography (FD-OCT), retinal vessel diameters were assessed based on FD-OCT phase data. This allowed for the calculation of TRBF before and during visual stimulation. Additionally, a mathematical flow model for the retinal vasculature was adapted to study the implications of diameter variations on retinal perfusion. Measured and simulated perfusion was compared to draw conclusions on the diameter variations in different layers of the vascular tree. Results: The measured mean baseline flow was 36.4 ± 6.5 µl/min while the mean flow during flicker stimulation was 53.4% ± 8.3 µl/min. The individual increase in TRBF during flicker stimulation ranged between 34% and 66%. The average increase in TRBF over all measured subjects was 47.6% ± 12.6%. Conclusions: Dual-beam bidirectional Doppler FD-OCT allowed quantifying NC in the human retina in vivo and may be a promising method for monitoring alterations in NC caused by various pathologies. The comparison of the measured data with the results obtained in the simulated vasculature indicates that the vasodilation induced by NC is more pronounced in smaller vessels.

Effect of different lubricant eye gels on tear film thickness as measured with ultrahigh-resolution optical coherence tomography.

PURPOSE: To compare the effect of a single drop of different lubricant eye gels on tear film thickness (TFT) as measured with ultrahigh-resolution optical coherence tomography (UHR-OCT) in patients with mild-to-moderate dry eye disease (DED). METHODS: The study followed a randomized, single-masked, observer-blinded parallel group design. Patients received a single dose of either unpreserved trehalose 3% + hyaluronic acid 0.15% (TH), hyaluronic acid 0.2% (HA) or polyethylene glycol 0.4% + propylene glycol 0.3% (PP) eye drops. In total, 60 patients were included and TFT was measured using a custom-built UHR-OCT system. RESULTS: The mean TFT at baseline was 3.5 ± 0.7 µm. There was a significant difference in the time-course of TFT between the three groups (p = 0.001 between groups). Ten minutes after instillation, TFT increased by 155.8 ± 86.6%, 65.7 ± 71.5% and 33.4 ± 19.6% in the PP, TH and HA group, respectively (p < 0.001 between groups). Thirty minutes after instillation, the effect of all three different agents on TFT was comparable. Sixty and 120 min after administration, a significant increase in TFT was only seen for the TH group, but not for the other products (60 min: p < 0.021 between groups; 120 min: p < 0.037 between groups). CONCLUSION: In conclusion, we observed differences in TFT after administration of the lubricant gels. Ten minutes after instillation, a pronounced increase in TFT was observed in all groups. As compared to the other products, the combination of trehalose 3% + hyaluronic acid 0.15% offers a significantly longer increase in TFT indicating for a longer residence time.

Effect of a Matrix Therapy Agent on Corneal Epithelial Healing After Standard Collagen Cross-linking in Patients With Keratoconus: A Randomized Clinical Trial.

IMPORTANCE: Corneal abrasions are frequent after standard (epithelium-off [epi-off]) corneal collagen cross-linking (CXL) in patients with progressive keratoconus. A new matrix therapy agent (ReGeneraTing Agent [RGTA]) has been developed to promote corneal wound healing. OBJECTIVE: To assess the effect of the new type of matrix therapy agent on corneal wound healing after epi-off CXL in patients with keratoconus. DESIGN, SETTING, AND PARTICIPANTS: This double-masked randomized clinical trial enrolled 40 patients with keratoconus undergoing epi-off CXL from July 18, 2014, to October 21, 2015, when the last follow-up was completed. The analysis of the intention-to-treat population was performed at the Department of Clinical Pharmacology in cooperation with the Center for Medical Physics and Biomedical Engineering and the Department of Ophthalmology and Optometry of the Medical University of Vienna. INTERVENTIONS: Patients were randomized to receive the matrix therapy agent or hyaluronic acid-containing eyedrops, 0.1%, every other day starting immediately after surgery. The size of the corneal defect was measured using ultrahigh-resolution optical coherence tomography (OCT) and slitlamp photography (SLP) with fluorescein staining. MAIN OUTCOMES AND MEASURES: Corneal wound healing rate, defined as the size of the defect over time. RESULTS: Among the 40 patients undergoing epi-off CXL (31 men; 9 women; mean [SD] age, 31 [10] years), wound healing was significantly faster in the matrix therapy agent group compared with the hyaluronic acid group (4.4 vs 6.1 days; mean difference, 1.7 days; 95% CI, 0.25-3.15 days; P = .008). The defect size was smaller in the matrix therapy agent group than in the hyaluronic acid group as measured with OCT (12.4 vs 23.9 mm2; mean difference, 11.6 mm2; 95% CI, 0.8-23.5 mm2; P = .045) and SLP (11.9 vs 23.5 mm2; mean difference, 11. 6 mm2; 95% CI, 1.3-22.9 mm2; P = .03). A correlation between the defect size measured with OCT and SLP was found (r = 0.89; P < .001). No ocular or serious adverse events occurred. CONCLUSIONS AND RELEVANCE: Use of a new matrix therapy agent appears to improve corneal wound healing after CXL in patients with keratoconus. Monitoring of corneal wound healing using ultrahigh-resolution OCT might be an attractive alternative to SLP because OCT provides an objective and 3-dimensional evaluation of the corneal defect. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT02119039.

In vivo tear film thickness measurement and tear film dynamics visualization using spectral domain optical coherence tomography.

Dry eye syndrome is a highly prevalent disease of the ocular surface characterized by an instability of the tear film. Traditional methods used for the evaluation of tear film stability are invasive or show limited repeatability. Here we propose a new non-invasive fully automated approach to measure tear film thickness based on spectral domain optical coherence tomography and on an efficient delay estimator. Silicon wafer phantom were used to validate the thickness measurement. The technique was applied in vivo in healthy subjects. Series of tear film thickness maps were generated, allowing for the visualization of tear film dynamics. Our results show that the in vivo central tear film thickness measurements are precise and repeatable with a coefficient of variation of about 0.65% and that repeatable tear film dynamics can be observed. The presented approach could be used in clinical setting to study patients with dry eye disease and monitor their treatments.

Effect of hyaluronic acid on tear film thickness as assessed with ultra-high resolution optical coherence tomography.

PURPOSE: The aim of this study was to assess the effect of a single drop of hyaluronic acid on tear film thickness (TFT) in healthy subjects. METHODS: Sixteen healthy subjects (eight male/eight female) aged between 20 and 36 years were included in this randomized, double-masked placebo-controlled study. One eye received a single dose of hyaluronic acid (Olixia pure(®) ; Croma Pharma, Korneuburg, Austria) eye drops, and the fellow eye received physiologic saline solution as placebo control. The study eye was chosen randomly. TFT as measured with a custom-built Fourier-domain optical coherence tomography (FD-OCT) system was the main outcome variable and measured before and every 10 min until 1 hr after topical administration. RESULTS: Baseline TFT was 4.8 ± 0.5 µm in the study eye and 5.0 ± 0.4 µm in the control eyes. Hyaluronic acid significantly increased TFT (p = 0.008 versus placebo) with a maximum effect 10 min after instillation (13.9 ± 11.9%). Post hoc analysis revealed that an increase in TFT was seen until 30 min after administration compared to placebo. Data in the placebo group show high reproducibility with an intraclass correlation coefficient of 0.93 and a coefficient of variation of 5.4 ± 3.3%. CONCLUSION: The data of this study indicate that hyaluronic acid increases TFT for as long as 30 min in healthy subjects. In addition, our data provide evidence that our custom-built OCT system is capable of measuring residence time of lubricants on the ocular surface.