In vitro quantification of the stiffening effect of corneal cross-linking in the human cornea using radial shearing speckle pattern interferometry.

PURPOSE: To quantify the magnitude of the stiffening effect of corneal cross-linking (CXL) by studying intact human corneas exposed to physiological pressure transients. METHODS: Nine organ-cultured human corneas mounted in artificial anterior chambers were studied. A radial shearing speckle pattern interferometer was used to measure changes in corneal strain following an increase in artificial anterior chamber pressure from 15.0 to 15.5 mmHg before and after treatment. Measurements were taken from all corneas with their epithelium intact before all underwent epithelial debridement. Three specimens were used as controls and did not receive any further treatment; three underwent riboflavin/ultraviolet A (UVA) CXL using 30 minutes of 370-nm irradiation at 3 mW/cm2 following epithelial removal and saturation with 0.1% riboflavin; and three were fixed with the cross-linking agent 2.5% glutaraldehyde. Strain measurements were repeated after these treatments. Young's moduli of individual corneas were calculated by mathematical analysis. RESULTS: Mean donor age was 81.7 years. Before treatment, the mean Young's moduli of the control, riboflavin/UVA CXL, and glutaraldehyde-fixed corneas did not differ significantly: 0.46±0.03, 0.48±0.03, and 0.47±0.03 MPa, respectively. Following treatment these values changed to 0.46±0.2, 2.06±0.22, and 3.48±0.41 MPa, respectively. In proportional terms, this was equivalent to an increase in corneal Young's modulus by a factor of 4.3 (P<.05) following riboflavin/UVA CXL and 7.3 (P<.05) after glutaraldehyde fixation. CONCLUSIONS: Riboflavin-UVA CXL increases the stiffness of organ-cultured corneas by a factor of more than four. This finding quantifies the efficacy of CXL in a physiologic configuration.

Effects of variation in depth and side cut angulations in LASIK and thin-flap LASIK using a femtosecond laser: a biomechanical study.

PURPOSE: To study the corneal biomechanical effects of varying LASIK flap depth and side cut angulations and evaluate the relative contribution of the lamellar and side cuts using a femtosecond laser and radial shearing speckle pattern interferometry (RSSPI). METHODS: Forty-two organ-cultured human corneas were divided into a control group and three investigative groups, each undergoing different incision types at both 90- and 160-µm depth using a femtosecond laser. In the first group, typical LASIK flaps were created; in the second group, only the bed was cut (delamination); and in the third group, side cuts alone were affected. Corneal strain was measured using RSSPI before and after treatment following an increase in hydrostatic pressure from 15.0 to 15.5 mmHg and again after 1 week of incubation in culture medium. RESULTS: The flap group demonstrated a weakening of strength related to the depth of cut, with strain increasing by 9% and 32% at 90 and 160 µm, respectively. Similar changes, 9% and 33%, were observed following execution of side cuts to the same depths. By contrast, strain increase following delamination showed no relationship with depth, increasing by 5% in both instances. When the side cut angle was made more acute, strain decreased with a 2% strain increase being measured after a 90-µm, 150° side cut was created. No significant changes occurred during the period of organ culture. CONCLUSIONS: Vertical side cuts through corneal lamellae rather than horizontal delamination incisions contribute to the loss of structural integrity during LASIK flap creation. Angulating side cuts such that the stromal diameter of the flap exceeds its epithelial diameter can decrease this effect.

An interferometric ex vivo study of corneal biomechanics under physiologically representative loading, highlighting the role of the limbus in pressure compensation.

BACKGROUND: The mechanical properties of the cornea are complex and regionally variable. This paper uses an original method to investigate the biomechanics of the cornea in response to hydrostatic loading over the typical physiological range of intra-ocular pressure (IOP) fluctuations thereby increasing understanding of clinically relevant corneal biomechanical properties and their contributions to the refractive properties of the cornea. METHODS: Displacement speckle pattern interferometry (DSPI) was used to measure the total surface displacement of 40 porcine and 6 human corneal-scleral specimens in response to pressure variations up to 1 mmHg from a baseline of 16.5 mmHg. All specimens were mounted in a modified artificial anterior chamber (AAC) and loaded hydrostatically. Areas of high strain in response to loading were identified by comparing the displacements across different regions. RESULTS: The nature of the response of the corneal surface to loading demonstrated high regional topographic variation. Mechanical properties were shown to be asymmetrical, and deformation of the limbal and pre-limbal regions dominated these responses respectively with over 90% (N-T) and 60% (S-I) of the total maximum displacement occurring in these regions indicating high-strain. In contrast, the curvature of the central cornea remained relatively unchanged merely translating in position. CONCLUSIONS: The limbal and pre-limbal regions of the cornea appear to be fundamental to the absorption of small pressure fluctuations facilitating the curvature of the central cornea to remain relatively unchanged. The differential mechanical properties of this region could have important implications for the application of corneal surgery and corneal crosslinking, warranting further investigation.

Age-related differences in the elasticity of the human cornea.

PURPOSE: The goal of this study was to determine age-related variation in the elasticity of the human cornea using nondestructive means. METHODS: Organ cultured human corneoscleral buttons were studied. Changes in strain were measured with a radial shearing speckle pattern interferometer after an increase in intraocular pressure from 15.0 to 15.5 mm Hg. Changes in central corneal displacement were calculated by integration, and a bulk corneal Young's modulus was derived by mathematical analysis. RESULTS: Fifty corneas, including 17 pairs, were studied. Donors were aged between 24 and 102 years (mean, 73.1); 29 (58%) specimens were from male donors and 21 from female donors. Young's modulus of the cornea increased with age, with the line of best fit indicating an approximate doubling from 0.27 MPa at age 20 years (95% confidence interval, 0.22-0.31) to 0.52 (0.50-0.54) MPa at age 100 years (R² = 0.70). CONCLUSIONS: The stiffness of the human cornea increases by a factor of approximately two between the ages of 20 and 100 years. This variation is relevant to the algorithms used to predict the response to incisional and ablative refractive surgery and will also affect the formulas used to calculate intraocular pressure by applanation.

Nonperturbing measurements of spatially distributed underwater acoustic fields using a scanning laser Doppler vibrometer.

Localized changes in the density of water induced by the presence of an acoustic field cause perturbations in the localized refractive index. This relationship has given rise to a number of nonperturbing optical metrology techniques for recording measurement parameters from underwater acoustic fields. A method that has been recently developed involves the use of a Laser Doppler Vibrometer (LDV) targeted at a fixed, nonvibrating, plate through an underwater acoustic field. Measurements of the rate of change of optical pathlength along a line section enable the identification of the temporal and frequency characteristics of the acoustic wave front. This approach has been extended through the use of a scanning LDV, which facilitates the measurement of a range of spatially distributed parameters. A mathematical model is presented that relates the distribution of pressure amplitude and phase in a planar wave front with the rate of change of optical pathlength measured by the LDV along a specifically orientated laser line section. Measurements of a 1 MHz acoustic tone burst generated by a focused transducer are described and the results presented. Graphical depictions of the acoustic power and phase distribution recorded by the LDV are shown, together with images representing time history during the acoustic wave propagation.