Effect of upper eyelid blepharoplasty on corneal biomechanical, topographic and tomographic parameters 4 weeks after surgery.

PURPOSE: To investigate the effect of "skin-only" upper eyelid blepharoplasty on corneal biomechanics and central as well as peripheral topographic/tomographic parameters before and 4 weeks after surgery. METHODS: In a prospective study, the corneal hysteresis (CH) and corneal resistance factor (CRF) were evaluated before and after blepharoplasty. Corneal topographic (maximum simulated keratometry value, inferior-superior value, index of surface variance, index of vertical asymmetry, index of height asymmetry, index of height decentration) and tomographic parameters (corneal thickness, corneal astigmatism and mean 5-mm- and 7-mm-zone keratometry value) were measured by the Pentacam HR. Statistical analysis was performed using a linear mixed model considering correlated data of both eyes. RESULTS: This study included 42 eyes of 35 patients (mean age: 64.5 years, range 52-82 years). Four weeks after surgery CH and CRF increased (9.4 ± 2.3 to 10.2 ± 2.2 mmHg and 9.7 ± 2.1 to 10.5 ± 2.2 mmHg) but did not reach statistical significance (P = 0.100 and P = 0.072). A significant increase in central maximum simulated keratometry value (Kmax) from 45.0 ± 2.3 to 45.4 ± 2.2 diopters (D) was observed (P = 0.004). Inferior-superior value (I-S) and index of surface variance (ISV) showed significant changes from 0.32 ± 0.98 to 0.10 ± 0.98 D (P = 0.02) and from 19.98 ± 9.84 to 22.93 ± 11.23 (P = 0.009), respectively. These alterations did not affect the subjective spherical equivalent (-0.09 ± 4.71 to -0.04 ± 4.51 D; P = 0.437) and the best-corrected distance visual acuity of patients (0.11 ± 0.14 to 0.15 ± 0.15 logMAR; P = 0.142). Age, gender and corneal thickness were not correlated with pre and postoperative differences of CH, CRF, corneal compensated IOP, Kmax, corneal astigmatism or I-S. CONCLUSION: The trend of increasing CH and CRF values might indicate a rise of corneal damping capacity. Despite statistically significant differences of Kmax, I-S and ISV, all other tomographical and topographical parameters did not change 4 weeks after surgery. The corneal steepening with a mean change of 0.4 diopters and the decrease of I-S with a mean of 0.22 diopters do not seem to have a clinically relevant effect for blepharoplasty patients in daily practice.

Comparison of waveform-derived corneal stiffness and stress-strain extensometry-derived corneal stiffness using different cross-linking irradiances: an experimental study with air-puff applanation of ex vivo porcine eyes.

PURPOSE: To assess corneal stiffening of standard (S-CXL) and accelerated (A-CXL) cross-linking protocols by dynamic corneal response parameters and corneal bending stiffness (Kc[mean/linear]) derived from Corvis (CVS) Scheimpflug-based tonometry. These investigations were validated by corneal tensile stiffness (K[ts]), derived from stress-strain extensometry in ex vivo porcine eyes. METHODS: Seventy-two fresh-enucleated and de-epithelized porcine eyes were soaked in 0.1% riboflavin solution including 10% dextran for 10 min. The eyes were separated into four groups: controls (n = 18), S-CXL (intensity in mW/cm2*time in min; 3*30) (n = 18), A-CXL (9*10) (n = 18), and A-CXL (18*5) (n = 18), respectively. CXL was performed using CCL Vario. CVS measurements were performed on all eyes. Subsequently, corneal strips were extracted by a double-bladed scalpel and used for stress-strain measurements. K[ts] was calculated from a force-displacement curve. Mean corneal stiffness (Kc[mean]) and constant corneal stiffness (Kc[linear]) were calculated from raw CVS data. RESULTS: In CVS, biomechanical effects of cross-linking were shown to have a significantly decreased deflection amplitude as well as integrated radius, an increased IOP, and SP A1 (P < 0.05). Kc[mean]/Kc[linear] were significantly increased after CXL (P < 0.05). In the range from 2 to 6% strain, K[ts] was significantly higher in S-CXL (3*30) compared to A-CXL (9*10), A-CXL (18*5), and controls (P < 0.05). At 8% to 10% strain, all protocols induced a higher stiffness than controls (P < 0.05). CONCLUSION: Several CVS parameters and Kc[mean] as well as Kc[linear] verify corneal stiffening effect after CXL on porcine eyes. S-CXL seems to have a higher tendency of stiffening than A-CXL protocols have, which was demonstrated by Scheimpflug-based tonometry and stress-strain extensometry.

Confocal microscopy evaluation of stromal fluorescence intensity after standard and accelerated iontophoresis-assisted corneal cross-linking.

The aim of this study is to determine modifications in stromal fluorescence intensity after different corneal cross-linking (CXL) procedures and to correlate stromal fluorescence to corneal biomechanical resistance. For confocal microscopy study, 15 human cadaver corneas were examined. Three served as control (group 1), three were just soaked with iontophoresis procedure (group 2), three were treated with standard epi-off technique (group 3), and six underwent iontophoresis imbibition. Three of later six were irradiated for 30 min with 3 mW/cm2 UVA (group 4) and three for 9 min at 10 mW/cm2 UVA (group 5). Confocal microscopy was performed to quantify the fluorescence intensity in the cornea at different stromal depths. For biomechanical study, 30 human cadaver corneas were randomly divided into five groups and treated as previously described. Static stress-strain measurements of the corneas were performed. Iontophoresis imbibition followed by 10mW/cm2 irradiation proved to increase stromal fluorescence into the corneal stroma and significant differences were revealed between group 3 and 5 both at 100 (p = 0.0171) and 250 µm (p = 0.0024), respectively. Biomechanical analysis showed an improvement of corneal resistance in group 5. Iontophoresis imbibition followed by accelerated irradiation increased the stromal fluorescence and is related to an improvement of biomechanical resistance. This approach may represent a new strategy to achieve greater concentrations of riboflavin without removing corneal epithelium and improve clinical results while reducing the side effects of CXL.

Analyzing biomechanical parameters of the cornea with glaucoma severity in open-angle glaucoma.

PURPOSE: The purpose was to investigate a possible association of corneal hysteresis (CH) and corneal resistance factor (CRF) with open-angle glaucoma and the severity of disease. METHODS: In this prospective cross-sectional study we recruited 86 open-angle glaucoma patients, 16 patients with ocular hypertension (OHT,) and 44 age-matched controls. Each participant had a complete glaucoma workup including measurements with the Ocular Response Analyzer and computerized perimetry with the Humphrey 30-2 SITA Standard program. Visual field damage was based on mean deviation (MD) and considered as early glaucomatous with a MD > - 6 dB, moderate glaucomatous between -6 and -12 dB and advanced glaucomatous < -12 dB. The association between ORA parameters, glaucoma, and disease severity was evaluated using univariate and multivariate linear regression analyses. RESULTS: There was a statistically significant correlation between the biomechanical parameters and intraocular pressure, central corneal thickness, axial length, and age. On average, glaucoma patients had the lowest adjusted CH (8.96 ± 1.43 mmHg) and CRF (9.07 ± 1.93 mmHg) values in comparison to OHT patients (CH: 10.2 ± 1.5 mmHg; CRF: 10.6 ± 2.1 mmHg) and controls (CH: 9.7 ± 1.4 mmHg; CRF: 10.2 ± 1.9 mmHg). This difference was statistically significant (CH: p = 0.003; CRF: p = 0.008). There was also a statistically significant difference in adjusted CH (p = 0.001) and CRF (p = 0.004) values between the controls and the visual field groups, with the lowest values being in the most advanced group. CONCLUSIONS: Before interpreting corneal biomechanical parameters, it seems important to adjust the measured data for their underlying influencing factors. Glaucoma patients with lower adjusted CH and CRF probably have more advanced disease and should, therefore, be treated more aggressively and monitored more carefully and frequently.

First proposed efficacy study of high versus standard irradiance and fractionated riboflavin/ultraviolet a cross-linking with equivalent energy exposure.

PURPOSE: To document the first presented report in December 2008 of high irradiance riboflavin/ultraviolet A (UVA) corneal cross-linking in comparison with that of standard irradiance and of fractionated exposure to increase the time for oxygen diffusion into the cornea. METHODS: After in vitro studies of oxygen depletion and cross-linking density using type 1 human collagen gels, 36 ex vivo porcine globes were deepithelialized and exposed to 0.1% riboflavin drops in carboxymethylcellulose solution every 5 min for 3 initial doses and then throughout irradiation afterward. Six eyes each were irradiated with 370-nm UVA light at 2, 3, 9, and 15 mW/cm continuously and 15 mW/cm fractionated (with alternate cycles of 30 s "ON" and 30 s "OFF" exposure) using an equivalent radiant exposure of 5.4 mJ/cm. The final six eyes received no UVA exposure as a control. The exposed corneas were then dissected and subjected to extensiometry. Analysis of variance with Bonferroni post hoc test was performed between groups. RESULTS: The stress required to induce a 10% strain for the control eyes (no UVA) was 100.6±20.9×10 N/m in comparison with the stress of 3 mW/cm (standard irradiation) at 146.7±17.6×10 N/m (P=0.009). The stress at the other equidose irradiances of 2, 9, 15 continuously, and 15 mW/cm fractionated were 140±21.9, 162.8±70, 154.1±70, and 163.0±64×10 N/m, respectively. When comparing the irradiances of 15 mW/cm continuously and fractionated to the standard irradiation, the stress was not statistically different (P=0.799 and 0.643), respectively. CONCLUSION: High irradiance riboflavin/UVA cross-linking with equivalent energy exposure demonstrates comparable efficacy in stiffening corneal collagen with standard irradiance, but with considerably less exposure time. Over the past 6 years, since this report was first presented, the use of high irradiance cross-linking has been gaining popularity.

Association Between Corneal Changes and Retinal Oximetry in Diabetes Mellitus.

Purpose: Diabetes mellitus (DM) causes different corneal changes that are associated with the severity of diabetic retinopathy. To identify the pathophysiological reasons for this, corneal tomography and optical densitometry (COD) were combined with retinal oximetry. Methods: Patients with DM and healthy subjects were included in this pilot study. Spatially resolved corneal thickness and COD were assessed using the Pentacam HR (Oculus). The pachymetry difference (PACDiff) was calculated as an indicator of an increase in the peripheral corneal thickness. Oxygen saturation (SO2) of the retinal vessels was measured using the Retinal Vessel Analyzer (Imedos Systems UG). Subsequently, the associations between corneal and retinal parameters were analyzed. Results: Data from 30 patients with DM were compared with those from 30 age-matched healthy subjects. In DM, arterial (P = 0.048) and venous (P < 0.001) SO2 levels were increased, and arteriovenous SO2 difference was decreased (P < 0.001). In patients, PACDiff was higher than that in healthy subjects (P < 0.05), indicating a stronger increase in peripheral corneal thickness. The COD was reduced in DM (P = 0.004). The PACDiff of concentric rings with a diameter of 4 mm (r = -0.404; P = 0.033) to 8 mm (r = -0.522; P = 0.004) was inversely correlated with the arteriovenous SO2 difference. Furthermore, PACDiff 4 mm was negatively associated with arterial SO2 (r = -0.389; P = 0.041), and the COD of the peripheral corneal areas correlated positive with arterial SO2 (COD total 10-12 mm: r = 0.408; P = 0.025). Conclusion: These associations might indicate a common pathogenesis of corneal and retinal changes in DM, which could be caused by reduced oxygen supply, mitochondrial dysfunction, oxidative stress, and cytokine effects.

Retinal changes are particularly important for ophthalmologists in the management of diabetes mellitus. These are primarily consequences of diabetic vascular changes that can lead to a lack of oxygen. However, there is also evidence of significant changes in the cornea of patients with diabetes. In the present study, the associations between changes in corneal thickness profile, optical density of the cornea, and oxygen saturation of retinal vessels in diabetes mellitus were demonstrated for the first time. Therefore, this study could contribute to clarifying the possible causes of corneal changes in patients with diabetes.

Crosslinking with UV-A and riboflavin in progressive keratoconus: From laboratory to clinical practice - Developments over 25 years.

Changes in the biomechanical and biochemical properties of the human cornea play an important role in the pathogenesis of ectatic diseases. A number of conditions in primarily acquired (keratoconus or pellucid marginal degeneration) or secondarily induced (iatrogenic keratectasia after refractive laser surgeries) ectatic disorders lead to decreased biomechanical stability. Corneal collagen cross-linking (CXL) represents a technique to slow or even halt the progression of ectatic pathologies. In this procedure, riboflavin is applied in combination with ultraviolet A radiation. This interaction induces the production of reactive oxygen species, which leads to the formation of additional covalent bonds between collagen molecules and subsequent biomechanical corneal strengthening. This procedure is so far the only method that partially interferes etiopathogenetically in the treatment of ectatic diseases that slows or stops the process of corneal destabilization, otherwise leading to the need for corneal transplantation. Besides, CXL process increases markedly resistance of collagenous matrix against digesting enzymes supporting its use in the treatment of corneal ulcers. Since the discovery of this therapeutic procedure and the first laboratory experiments, which confirmed the validity of this method, and the first clinical studies that proved the effectiveness and safety of the technique, it has been spread and adopted worldwide, even with further modifications. Making use of the Bunsen-Roscoe photochemical law it was possible to shorten the duration of this procedure in accelerated CXL and thus improve the clinical workflow and patient compliance while maintaining the efficacy and safety of the procedure. The indication spectrum of CXL can be further expanded by combining it with other vision-enhancing procedures such as individualized topographically-guided excimer ablation. Complementing both techniques will allow a patient with a biomechanically stable cornea to regularize it and improve visual acuity without the need for tissue transplantation, leading to a long-term improvement in quality of life.

The effect of acetazolamide on different ocular vascular beds.

PURPOSE: To assess the effect of acetazolamide (AZ) on different ocular vascular beds. METHODS: In a prospective study, 32 healthy volunteers (16 male, 16 female) with a mean age of 23.9 ± 3.3 years (20-39 years) were included. Before and after intravenous administration of 1,000 mg AZ (single dose), ocular microcirculation parameters were measured every 20 min for 2 h. Retinal vessel diameters (VD) were measured by the retina vessel analyzer, blood flow (BF) in the neuroretinal rim by the laser doppler flowmeter according to Riva, and the parapapillary retinal BF by the scanning laser Doppler flowmeter. Additionally, the Langham ocular blood flow system was used to determine the ocular pulse amplitude (OPA) and the pulsatile ocular blood flow (pOBF). The measurements were correlated with systemic blood pressure (BP), ocular perfusion pressure (OPP), capillary base excess parameters and serum AZ levels. RESULTS: Arterial and venous VD were significantly increased by about 4-5% each. Papillary BF increased significantly about 40%. Parapapillary retinal flow dropped significantly about 19% (120 min). OPA and pOBF showed no statistically significant changes. BP showed no significant changes, and OPP was significantly increased. There were no correlations with pH or systemic perfusion parameters. CONCLUSIONS: AZ leads to a dilatation of retinal VD, to an increase of BF in the optic nerve head, and to a decrease of parapapillary retinal BF. The different BF changes in different vascular beds might be due to different regulatory mechanisms, steal effects, or different distributions of the carbonic anhydrase.

Corneal crosslinking with riboflavin and ultraviolet A. I. Principles.

Changes in the biomechanical properties of the human cornea play an important role in the pathogenesis of corneal ectatic diseases. Biomechanical investigation shows significant differences between human ectatic corneas and normal corneas, including decreased stiffness and reduction of collagen crosslinks in the ectatic cornea. Induction of crosslinks is a well-established procedure in polymer chemistry to increase the elastic modulus of materials. Crosslinking (CXL) in connective tissue can occur during aging and as a side effect of diabetes mellitus. CXL has been used medically to increase stability and reduce the biodegradation of collagen-based biomaterials for bioprostheses. CXL of the cornea using riboflavin and UVA light with a wavelength of 370 nm and a dosage of 5.4 J/cm² is a new approach that increases the mechanical and biochemical stability of stromal tissue. This technique combines the principles of CXL (chemical and nonenzymatic) and the biochemical mechanisms of photo-oxidative CXL with riboflavin as a photosensitizer. In this review, the enrichment of riboflavin in the stroma by standard (epi-off) and transepithelial (epi-on) CXL is discussed. The theoretical and experimental measurements of the absorption of UV light explain the stronger CXL effect in the anterior stroma and its importance for the prevention of damage to the endothelial cells. UV devices are described. Changes of the physical properties after CXL, as well as the cellular changes, are discussed. From these basic investigations, treatment parameters for effective and safe CXL are identified.

Corneal crosslinking with riboflavin and ultraviolet A. Part II. Clinical indications and results.

Changes in the biomechanical properties of the human cornea play an important role in the pathogenesis of corneal ectatic diseases. A variety of conditions in primary acquired (keratoconus and pellucid marginal degeneration) or secondary induced (iatrogenic keratectasia after excimer refractive laser surgery) corneal ectatic disorders lead to reduced biomechanical resistance. Corneal collagen crosslinking (CXL) has emerged as a promising technique to slow or even to stop the progression of these corneal ectatic pathologies. In this procedure, riboflavin (vitamin B2) is administered in conjunction with ultraviolet A light (UVA, 365 nm). This interaction causes the formation of reactive oxygen species, leading to the formation of additional covalent bonds between collagen molecules, with consequent biomechanical stiffening of the cornea. Although this method is not yet accepted as an evidence-based medicine modality for the treatment of corneal primary or secondary ectasias, the results of prospective, randomized studies of CXL used in the treatment of these pathologic entities show significant changes in the properties of corneal tissue. This procedure is currently the only etiopathogenetic approach in ectatic eyes that can delay or stop the process of cornea destabilization, reducing the necessity for keratoplasty. Despite promising results, CXL is associated with issues that include long-term safety and duration of the stabilizing effect. Combination of CXL with vision-improving procedures, such as topography-guided custom ablation and implantation of intracorneal ring segments of phakic intraocular lenses, may expand the indications for this procedure.

Corneal Crosslinking With Riboflavin and UVA Light in Progressive Keratoconus: Fifteen-Year Results.

PURPOSE: To analyze the 15-year results of corneal crosslinking (CXL) in progressive keratoconus. DESIGN: Retrospective follow-up analysis of interventional study patients. METHODS: This study included keratoconic eyes with progressive disease treated from 2001 to 2006 at the Department of Ophthalmology, Carl Gustav Carus University Hospital, TU Dresden, Germany. CXL was performed by applying riboflavin and ultraviolet A (UVA) light according to a standard protocol. The best-corrected distance visual acuity (BCVA), slitlamp examination, and corneal topography as well as corneal thickness values were recorded preoperatively and 15 years after the treatment. RESULTS: A total of 42 eyes received a complete follow-up of 15 years. The mean age of the patients at baseline was 26.9 (95% CI: 25.0-28.8) years. The maximum keratometry was 61.6 (95% CI: 58.2 - 64.9) diopters (D) preoperatively and 55.1 (95% CI: 51.6-58.4) D postoperatively; the decrease was statistically significant (P < .001). The mean keratometry value changed from 50.3 (95% CI: 48.3-52.4) D to 47.5 (95% CI: 45.3-49.4) D (P < 0.001). Furthermore, the thinnest corneal thickness decreased statistically significantly by 40 (95% CI: 24-56) µm (P < .001). The BCVA improved statistically significantly from 0.4 to 0.2 logMAR after the treatment. Retreatment was needed in 14% of cases. Mild scarring of the superficial stromal corneal layers was observed in 36% of the eyes, and in 67% of them visual acuity was stable or even improved. CONCLUSIONS: The CXL procedure proved to be an effective method in the treatment of keratoconic eyes in the progressive stage of the disease, and achieved long-term stabilization without the occurrence of serious complications or side effects.

Evaluation of Biomechanical Changes After Accelerated Cross-Linking in Progressive Keratoconus: A Prospective Follow-Up Study.

PURPOSE: The aim of this study was to analyze the biomechanical effect of accelerated corneal cross-linking (9*10) in progressive keratoconus (KC) in comparison to untreated fellow eyes using Scheimpflug-based tonometry (Corvis ST, CVS). METHODS: Forty-three eyes of 43 patients with KC showed progressive KC and were treated using accelerated corneal cross-linking. Twenty-five untreated fellow eyes were used as the control group. All eyes were examined biomechanically (CVS) and tomographically (Pentacam) at baseline, after 1-month, 6-month, and 12-month follow-up. Statistical analysis was performed using a linear mixed model. A logistic regression was performed to attribute the effects of changes in each parameter to treatment status (treated or untreated). RESULTS: Maximum keratometry values decreased statistically significantly at 12 months by -1.1 D (95 confidence interval: -2.0 to -0.1, P = 0.025) compared with baseline. Thinnest corneal thickness decreased significantly after 1 month ( P < 0.001) and recovered to baseline after 12 months ( P = 0.752). In the corneal cross-linking (CXL) group, biomechanical changes were observed by an increased bIOP, a shorter A2 time, and a lower integrated radius after 1 month (all P < 0.05). No biomechanical and tomographical changes were observed in the control group (all P > 0.05). Logistic regression pointed out that treated eyes can be separated from untreated eyes by differences in bIOP, corneal thickness, A1 velocity, integrated radius, and Kc mean at 1, 6, and 12 months. CONCLUSIONS: The alterations in biomechanical parameters indicated a corneal stiffening effect after CXL treatment, which was mostly detectable 1 month after treatment, although corneal thickness was reduced. The logistic regression model showed an adequate separation between CXL-treated and untreated eyes.

Estimating pulsatile ocular blood volume from intraocular pressure, ocular pulse amplitude, and axial length.

The purpose of this study was to develop a method of estimating pulsatile ocular blood volume (POBV) from measurements taken during an ophthalmic exam, including axial length and using a tonometer capable of measuring intraocular pressure (IOP) and ocular pulse amplitude (OPA). Unpublished OPA data from a previous invasive study was used in the derivation, along with central corneal thickness (CCT) and axial length (AL), as well as IOP from the PASCAL dynamic contour tonometer (DCT) and intracameral (ICM) measurements of IOP for 60 cataract patients. Intracameral mean pressure was set to 15, 20, and 35 mmHg (randomized sequence) in the supine position, using a fluid-filled manometer. IOP and OPA measurements were acquired at each manometric setpoint (DCT and ICM simultaneously). In the current study, ocular rigidity (OR) was estimated using a published significant relationship of OR to the natural log of AL in which OR was invasively measured through fluid injection. Friedenwald's original pressure volume relationship was then used to derive the estimated POBV, delivered to the choroid with each heartbeat as a function of OR, systolic IOP (IOPsys), diastolic IOP (IOPdia), and OPA, according to the derived equation POBV = log (IOPsys/IOPdia) / OR. Linear regression analyses were performed comparing OPA to OR and calculated POBV at each of the three manometric setpoints. POBV was also compared to OPA/IOPdia with all data points combined. Significance threshold was p < 0.05. OR estimated from AL showed a significant positive relationship to OPA for both DCT (p < 0.011) and ICM (p < 0.006) at all three manometric pressure setpoints, with a greater slope for lower IOP. Calculated POBV also showed a significant positive relationship to OPA (p < 0.001) at all three setpoints with greater slope at lower IOP, and a significant negative relationship with IOPdia. In the combined analysis, POBV showed a significant positive relationship to OPA/ IOPdia (p < 0.001) in both ICM and DCT measurements with R2 = 0.9685, and R2 = 0.9589, respectively. POBV provides a straight-forward, clinically applicable method to estimate ocular blood supply noninvasively. Higher IOP in combination with lower OPA results in the lowest values of POBV. The simplified ratio, OPA/ IOPdia, may also provide a useful clinical tool for evaluating changes in ocular blood supply in diseases with a vascular component, such as diabetic retinopathy and normal tension glaucoma. Future studies are warranted.

Biomechanical and morphological differences between the sclera canal ring and a peripheral sclera ring in the porcine eye.

AIM: To investigate a possible association between the biomechanical load and unload behaviour and the elastin content of the sclera canal ring (SCR) and a superiorly localized sclera ring (SPS) in the porcine eye. METHODS: Two sclera rings were trephined from each of 40 porcine eyes, one containing the SCR and the other an SPS. The load and the unload curves were measured in the extension range of 0-2.0 mm by a biomaterial tester. Hysteresis was determined from the area enclosed by the loading and unloading curve. Histochemical staining with resorcin-fuchsin and morphometric analysis of paraffin-embedded sections of both rings were performed to detect the area occupied by elastin fibres. RESULTS: At 1 mm extension, the mean load of the SCR was 0.89 ± 0.22 N and that of the SPS 1.13 ± 0.19 N, which was not significantly different between both rings (p > 0.05). Mean hysteresis in the SCR was 1.55 ± 0.30 N × mm and 1.90 ± 0.18 N × mm in the SPS, which was significantly different between both rings (p = 0.01). Mean sclera thickness was 986 µm in the SCR (range: 900-1,060 µm) and 971 µm in the SPS (range: 800-1,200 µm) without a statistically significant difference between both sclera rings (p = 0.78). The area occupied by elastin fibres was 15.5 ± 3.4% in the SCR and 4.5 ± 1.5% in the SPS, which was significantly different between both rings (p = 0.0001). CONCLUSION: Hysteresis in the SCR was significantly lower than in the SPS, indicating a higher elasticity of the SCR in the porcine eye. This effect could be explained by a higher content of elastin in the surrounding ring of the peripapillary optic nerve head providing reversible contraction in cases of intra-ocular pressure variations.

Diabetes mellitus affects biomechanical properties of the optic nerve head in the rat.

BACKGROUND: To investigate the effect of diabetes on the biomechanical behavior of the optic nerve head (ONH) and the peripapillary sclera (ppSc) in streptozocine-induced diabetic rats. METHODS: Diabetes mellitus was induced in 20 Wistar rats using streptozocine. Twenty-five nondiabetic rats served as controls. Eyes were enucleated after 12 weeks and 2 strips of one eye were prepared containing ONH or ppSc. The stress-strain relation was measured in the stress range of 0.05-10 MPa using a biomaterial tester. RESULTS: At 5% strain the stress of the ONH in diabetic rats was 897±295 kPa and in the control group it was 671±246 kPa; there was a significant difference between both groups (p=0.011). The stress of the diabetic ppSc (574±185 kPa) increased compared to that of the nondiabetic ppSc (477±171 kPa), but this did not reach statistical significance (p=0.174). The calculated tangent modulus at 5% strain was 11.79 MPa in the diabetic ONH and 8.77 MPa in the nondiabetic ONH; there was a significant difference between both groups (p=0.006). The calculated tangent modulus at 5% strain was 7.17 MPa in the diabetic ppSc and 6.12 MPa in the nondiabetic ppSc, without a statistically significant difference (p=0.09). CONCLUSION: In contrast to the ppSc, the ONH of diabetic rats showed a significant increase in stiffness compared to nondiabetic rats, which might be explained by nonenzymatic collagen cross-linking mediated by advanced glycation end products due to high blood glucose levels in diabetes. Further studies are needed to investigate if these biomechanical changes represent a detrimental risk factor for intraocular pressure regulation in diabetic glaucoma patients.

Statistical Evaluation of Correlated Measurement Data in Longitudinal Setting Based on Bilateral Corneal Cross-Linking.

PURPOSE: In ophthalmology, data from both eyes of a person are frequently included in the statistical evaluation. This violates the requirement of data independence for classical statistical tests (e.g. t-Test or analysis of variance (ANOVA)) because it is correlated data. Linear mixed models (LMM) were used as a possibility to include the data of both eyes in the statistical evaluation. METHODS: The LMM is available for a variety of statistical software such as SPSS or R. The application was applied to a retrospective longitudinal analysis of an accelerated corneal cross-linking (ACXL (9*10)) treatment in progressive keratoconus (KC) with a follow-up period of 36 months. Forty eyes of 20 patients were included, whereas sequential bilateral CXL treatment was performed within 12 months. LMM and ANOVA for repeated measurements were used for statistical evaluation of topographical and tomographical data measured by Pentacam (Oculus, Wetzlar, Germany). RESULTS: Both eyes were classified into a worse and better eye concerning corneal topography. Visual acuity, keratometric values and minimal corneal thickness were statistically significant between them at baseline (p < 0.05). A significant correlation between worse and better eye was shown (p < 0.05). Therefore, analyzing the data at each follow-up visit using ANOVA partially led to an overestimation of the statistical effect that could be avoided by using LMM. After 36 months, ACXL has significantly improved BCVA and flattened the cornea. CONCLUSION: The evaluation of data of both eyes without considering their correlation using classical statistical tests leads to an overestimation of the statistical effect, which can be avoided by using the LMM.

The Impact of Repetitive and Prolonged Eye Rubbing on Corneal Biomechanics.

PURPOSE: To evaluate the effect of simulated repetitive eye rubbing on the corneal biomechanics of porcine eyes using an ex vivo model system. METHODS: The average rubbing force that patients with keratoconus apply to their eyelids was previously determined. Fresh porcine eyes with eyelids were either exposed to 10,500 rub cycles from a custom-built eye rubbing machine that rubbed with a similar force to knuckle human eye rubbing (n = 33) or no rubbing at all (control; n = 37). A total of 10,500 rubs are equivalent to 1 year of rubbing six times daily, five movements per rub. The corneal biomechanical properties of these eyes were then tested by measuring the elastic modulus of 5-mm strips. RESULTS: The elastic modulus at the range of 1% and 5% of strain was 1.219 ± 0.284 and 1.218 ± 0.304 N/mm2 in the eye rubbing group and the no-rub control group, respectively. Corneal stiffness was similar in both groups (P = .984). CONCLUSIONS: The threshold to induce biomechanical changes (purely by eye rubbing) must be higher than 10,500 rubbing movements, suggesting that occasional eye rubbing may not affect corneal biomechanics in normal eyes, and likely only triggers keratoconus progression in predisposed corneas. Further in vivo studies assessing the impact eye rubbing has on inflammatory activity and the biomechanical properties of weakened corneas is warranted. [J Refract Surg. 2022;38(9):610-616.].

Laboratory measurement of the absorption coefficient of riboflavin for ultraviolet light (365 nm).

PURPOSE: Corneal cross-linking (CXL) is an increasingly used treatment technique for stabilizing the cornea in keratoconus. Cross-linking (polymerization) between collagen fibrils is induced by riboflavin (vitamin B2) and ultraviolet light (365 nm). Although reported to reach a constant value at higher riboflavin concentrations, the Lambert-Beer law predicts a linear increase in the absorption coefficient. This work was carried out to determine absorption behavior at different riboflavin concentrations and to further investigate the purported plateau absorption coefficient value of riboflavin and to identify possible bleaching effects. METHODS: The Lambert-Beer law was used to calculate the absorption coefficient at various riboflavin concentrations. The following investigated concentrations of riboflavin solutions were prepared using a mixture of 0.5% riboflavin and 20% Dextran T500 dissolved in 0.9% sodium chloride solution: 0%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.08%, 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%, and were investigated with and without aperture plate implementation. An additional test series measured the transmitted power at selected riboflavin concentrations over time. RESULTS: In diluted solutions, a linear correlation exists between the absorption coefficient and riboflavin concentration. The absorption coefficient reaches a plateau, but this occurs at a higher riboflavin concentration (0.1%) than previously reported (just above 0.04%). Transmitted light power increases over time, indicating a bleaching effect of riboflavin. CONCLUSIONS: The riboflavin concentration can be effectively varied as a treatment parameter in a considerably broader range than previously thought.

Detection of biomechanical changes after corneal cross-linking using Ocular Response Analyzer software.

PURPOSE: To investigate biomechanical changes after corneal cross-linking (CXL) with riboflavin/ultraviolet-A (UVA) in keratoconus using the recently developed Ocular Response Analyzer (ORA, Reichert Technologies) software. METHODS: Through use of the new ORA software (version 2.04), 37 new parameters derived from the best measurement signal with the highest wavescore of 4 measurements from 50 eyes of 46 patients with keratoconus were obtained before and 1 year after CXL. The parameters of 96 eyes from 96 age-matched, healthy individuals with a spherical equivalent refraction <3.00 diopters served as controls. RESULTS: Corneal hysteresis (CH) and corneal resistance factor (CRF) before CXL were 7.38±1.42 mmHg and 6.16±1.42 mmHg, respectively, compared to 7.37±1.26 mmHg (P=.971) and 6.16±1.50 mmHg after CXL (P=.997), respectively. Based on these 37 new parameters, the area under peak 2 (p2area) showed a statistically significant increase from 1262.3±623.1 before CXL to 1704.3±732.3 1 year after CXL (35%; P=.001). The related value for the p2area of the healthy control group was 3374.9±1099.9. A significant negative correlation was observed between the p2area and the difference in CH-CRF values (r=-0.29, P=.001). CONCLUSIONS: The area under peak 2 appears to be a more sensitive parameter to detect biomechanical changes after CXL than CH or CRF alone. After CXL, keratoconic corneas display altered biomechanical properties, which remain different to those observed in healthy corneas.