Corneal endothelial cell damage after experimental diode laser thermal keratoplasty.

PURPOSE: To evaluate the safety of diode laser thermal keratoplasty (LTK) with respect to corneal endothelial cell damage. METHODS: In an in vitro animal model system, porcine eyes were irradiated with a continuously emitting laser diode at wavelengths (lambda) of 1.85 or 1.87 microm, corresponding to an absorption coefficient (micro(a)) of 1.1 or 2.0 mm(-1). Different irradiation and application parameters were tested serially. To determine the temperature threshold for endothelial damage, corneal buttons were analyzed separately in a waterbath experiment. The endothelial damage was assessed after trypan blue and alizarin red supravital staining under light microscopy. RESULTS: The thresholds for the 50% probability of thermal damage (ED50) were determined at corneal temperatures of 65 degrees C for a 10-second water-bath immersion, and 59 degrees C for 60 seconds. Coagulations that reached the deeper stromal layers revealed severe endothelial cellular alterations and areas of exposed Descemet's membrane. The thermally induced changes were dependent on laser power and the absorption coefficient (wavelength). Mean diameter of total endothelial cell damage was 245 +/- 154 microm (range, 0 to 594 microm) for an absorption coefficient of 1.1 mm(-1). The maximal lateral extent of endothelial cell damage induced by the laser exposure was 594 microm in diameter. Increasing the absorption coefficient decreased the penetration depth of the laser irradiation, creating a greater temperature rise within the corneal stroma and significantly less endothelial damage (P < .01), when the same laser power was applied. The calculated total area of damage for the paracentral human corneal endothelium ranged from 1.8% to 13.6%. CONCLUSION: Data obtained in this in vitro study were transferred to an endothelial cell damage nomogram, demonstrating that appropriate parameter improvements can minimize the adverse effects to the corneal endothelium. However, model adjustment to the human cornea indicates the potential for endothelial cell damage after diode laser thermal keratoplasty, and should be considered when performing this elective procedure.

Influence of temperature and time on thermally induced forces in corneal collagen and the effect on laser thermokeratoplasty.

PURPOSE: To investigate thermomechanical aspects of corneal collagen denaturation as a function of temperature and time and the effect of the induced forces on refractive changes with laser thermokeratoplasty (LTK). SETTING: Medical Laser Center Lübeck, Lübeck, Germany. METHODS: In a material-test setup, porcine corneal strips were denatured in paraffin oil at various constant temperatures for 10 and 500 seconds, and the temporal course of the contractive forces was studied under isometric conditions. Typical LTK lesions were performed in porcine eyes in vitro with a continuous-wave infrared laser diode at a wavelength of 1.87 microm for 10 and 60 seconds. The laser power was chosen to achieve comparable denatured volumes at both irradiation times. The refractive changes were measured and analyzed by histologic evaluations and temperature calculations. RESULTS: The time course of the induced forces was characterized by a maximal force, which increased almost linearly with temperature, and a residual lower force. After 500 seconds of heating, the highest force was achieved with a temperature of 75 degrees C. With a limited heating period of only 10 seconds, the forces steadily increased with temperature over the entire observation period. Laser thermokeratoplasty produced less refractive change after 10 seconds of irradiation than after 60 seconds, although the laser power was 25% higher in the short heating period. Polarization light microscopy of LTK lesions revealed different stages of thermal damage. CONCLUSION: The course of the contractive forces during and after heating is a complicated function of the spatial time/temperature profile. Laser thermokeratoplasty lesions produced with 2 irradiation times showed different stages of denaturation and induced refractive change.

[Thermal damage to the corneal endothelium in diode laser thermokeratoplasty]. / Thermische Schädigung des Hornhautendothels bei der Dioden-Laserthermokeratoplastik.

Laser thermokeratoplasty (LTK) can be applied for correction of hyperopia and astigmatism by means of concentrically placed coagulations. Because of the temperature rise during coagulation, damage of the endothelial cells directly below the irradiated areas is possible. In this study, we examined the extent of the thermally denatured zones for different laser parameters and the threshold of thermal endothelial damage as a function of of temperature and duration of elevated temperatures. The threshold for thermal damage of endothelial cells was determined in isotonic NaCl solution for temperature exposures of 10 s and 1 min in a water bath. To determine the damage zones, corneas were irradiated under standardized conditions with a continuously emitting infrared (cw-IR) laser diode at various wave-lengths and different power values and were stained after preparation with trypan blue and alizarine red. The extensions of the damage zones were compared with calculated isotherms. Fifty percent cell damage was found at temperatures of 65 degrees C for heating times of 10 s and at 59 degrees C for 1 min. With thicker corneas, less laser power and higher absorption coefficients, the damage zone was reduced. The damage range determined corresponded to the calculated isotherms of 60 degrees C and 70 degrees C. Regarding clinical LTK, a loss of endothelial cells can be predicted and minimized or totally avoided by choosing the appropriate irradiation parameters.

[Diode laser thermokeratoplasty. Initial clinical experiences]. / Dioden-Laserthermokeratoplastik. Erste klinische Erfahrungen.

PURPOSE: Pulsed holmium lasers are currently used to correct hyperopia by means of laser thermokeratoplasty (LTK). Series of microsecond laser pulses are applied with a high repetition rate to induce shrinkage of corneal collagen fibers. The pulsed energy application results in intrastromal temperature peaks of up to 200 degrees C. A continuously emitting laser diode can--as we demonstrated recently in an invivo study on minipigs--be used for LTK and may be of advantage because the temperature rise is more steady. The aim of this study was to examine the safety, amount, and stability of hyperopic correction of diode LTK on blind human eyes. METHODS: We used a laserdiode that was set to continuously emit light at lambda = 1.854 microns/mu a = 1.04 mm-1 (group I, n = 4) or 1.87 microns/mu a = 1.92 mm-1 (group II, n = 4). Radiation energy was 100 to 150 mW for 10 s per coagulation. Eight coagulations on a single ring (group I) and 16 coagulations on a double ring (group II) diameter were applied in the cornea concentric to the entrance pupil by means of a vacuum-fixed application mask (group I = conjunctival fixation; group II = corneal fixation) and a handpiece with a focusing optic. Preoperatively as well as 1 week, 1, 2, 3, 6 12 and 18 months postoperative ophthalmologic controls were performed and the corneal refractive power was measured. RESULTS: In group I initial refractive changes of up to +4.9 D were achieved (1 week postoperative). However, due to the great penetration depth of the laser irradiation, large endothelial defects resulted beneath the stromal coagulations. In group II an initial refractive change of up to +6.8 D was achieved and as a result of the reduced penetration depth, the endothelial cell damage was much reduced. Partial regression of the refractive effect occurred in all subjects, which continued in higher refractive changes during the 2nd postoperative year. The refractive effect at 12 months was +0.6 to +1.5 D in group I and +0.9 to +5.7 D in group II. At 12 months the induced astigmatism was 0.5 to 2.2 D in group I and 0.3 to 1.6 D in group II. No serious adverse effects were noticed. CONCLUSION: A continously emitting laser diode working at a wavelength of 1.87 microns can be used to correct hyperopia by means of LTK safely and effectively. Regression occurs predominantly in the first 6 postoperative months. Further studies must be conducted to determine the importance of patient inherent parameters such as age in establishing a nomogram.

Continuous-wave diode laser thermokeratoplasty: first clinical experience in blind human eyes.

PURPOSE: To evaluate the safety and stability of laser thermokeratoplasty (LTK) with a continuous-wave diode laser in blind human eyes and to optimize parameters for a study in sighted eyes. SETTING: Department of Ophthalmology, Medical University Lübeck, Germany. METHODS: A continuous-wave diode laser was set to emit radiation with a wavelength of 1.854 microns (Group 1, n = 4) or 1.870 microns (Group 2, n = 4) and 100 to 150 mW power for 10 seconds. A focusing handpiece was coupled with an application mask and fixed by partial vacuum to the conjunctiva or cornea. The radiation was focused into the corneal stroma between 400 and 600 microns in Group 1 and set to 1000 microns in Group 2. Eight (Group 1, single ring) or 16 (Group 2, double ring) coagulations were applied. RESULTS: The refractive change increased with higher laser power and smaller ring diameters. Two rings of coagulations provided higher and more stable refractive changes of up to 5.66 diopters (D) than a single ring. The refractive effect stabilized between 3 and 6 months postoperatively. At 1 year, mean refractive change was +0.99 D +/- 0.39 (SD) in Group 1 and +2.32 +/- 2.24 D in Group 2. Extensive endothelial damage occurred in Group 1 but was minimal in Group 2. CONCLUSIONS: Diode LTK was used to treat hyperopia safely and effectively. Regression occurred mainly in the first 3 postoperative months. With a wavelength of 1.870 microns, corneal endothelial damage was limited.

Diode laser thermokeratoplasty: application strategy and dosimetry.

PURPOSE: To investigate suitable application parameters for efficient hyperopic correction by laser thermokeratoplasty (LTK) using mid-infrared laser diodes. SETTING: Medical Laser Center Lübeck, Lübeck, Germany. METHOD: A tunable continuous-wave laser diode in the spectral range between 1.845 and 1.871 microns was used. Transmitted by waveguides, the laser energy was used to induce coagulations on freshly enucleated porcine eyes to increase corneal curvature. The coagulations were equidistantly applied by a fiber-cornea contact and a noncontact focusing device that were adjusted on a ring concentric to the corneal apex. Different laser parameters and application geometries were evaluated. Refractive changes were measured by computer-assisted corneal topography before and after treatment. Polarization light microscopy and temperature calculations were used to analyze the coagulations. RESULTS: Because of the tunability of the laser diode, the influence of the corneal absorption coefficient (between 0.9 and 1.6 mm-1) on the refractive change could be measured. A laser power between 125 and 200 mW was adequate to achieve refractive changes up to 10.0 diopters. In the preferable focusing device, the refractive change increased almost logarithmically with the irradiation time up to 15 seconds. The number of coagulations on a fixed application ring showed no significant influence on refractive change; however, it showed an almost linear decrease with increasing ring diameter from 5.0 to 10.0 mm. Histological analysis revealed 3 stages of thermal damage. CONCLUSION: Diode LTK provided defined and uniform coagulations when using a well-adapted focusing device, resulting in sufficient refractive change. The results indicate that diode LTK is superior to pulsed holmium LTK.

[Imaging of laser thermokeratoplasty lesions by optical low coherence tomography and polarization microscopy after Sirius Red staining]. / Darstellung von LTK-Läsionen durch optische Kurzkohärenztomographie (OCT) und Polarisationsmikroskopie nach Sirius-Rot-Färbung.

BACKGROUND: Information on the extent and degree of the thermal effect produced is of great importance for control of the laser dosage in laser thermokeratoplasty (LTK) and for postoperative follow-up. We investigated on acute LTK effects which information images obtained by optical low coherence tomography (OCT) offer compared to those obtained by polarization microscopy. METHODS: Porcine eyes were irradiated through a 400 microns quartz fiber using light from a laser diode emitting up to 300 mW at a wavelength of 1.86 microns. Thermal lesions of varying strength were scanned using an experimental OCT device with about 25 microns lateral and 20 microns axial resolution. Histologic evaluation of the scanned areas was done by polarization microscopy after Sirlus-Red staining, and similar lesions were also analyzed by TEM. RESULTS: Both methods differentiated three damage zones a transition zone, a zone of moderate coagulation, and a central zone of strong coagulation. In the transition zone, increased birefringence was seen in polarization microscopy, which correlated with increased light scattering seen in the OCT images. In the moderately coagulated zone, a decrease in birefringence was associated with an even stronger increase of the OCT signal. In the central zone, a loss of the fibrillar tissue structure was observed, which led to a complete loss of birefringence and a strong reduction of the OCT signal. CONCLUSIONS: Although OCT does not provide the detailed information on thermal changes of tissue seen by the histologic method, it offers information on the extent and degree of tissue changes without preparation artifacts and provides a non-invasive method of immediate and follow-up control of LTK lesions. A quantitative analysis of changes in corneal thickness and curvature is much simpler than by a slit lamp. Time-resolved measurements of corneal light scattering may be used for on-line control of the laser-light dosage during LTK.

[Optical coherence tomography of the cornea and the anterior eye segment]. / Optische Kohärenztomographie der Kornea und des vorderen Augenabschnitts.

TARGET: The method of optical coherence tomography (OCT) was investigated regarding its suitability and limits for measuring the cornea and the anterior segment of the eye. Furthermore, the stromal expansion of thermally induced lesions in the cornea directly after irradiation was determined within the scope of the laser thermokeratoplasty (LTK). MATERIAL AND METHODS: With the experimental scanning OCT system, x-z sections of the anterior eye segment were made with an optical resolution of about 20 microns axially and 25 microns laterally. Freshly enucleated, tonicized porcine eyes were used as model eyes. Thermal lesions were applied with a continuously emitting laser diode (lambda = 1.86 microns) and various radiation parameters. Before and after coagulation, the cornea was viewed from limbus to limbus in a central OCT scan and the individual coagulation source was measured. RESULTS: Global and local changes of the thickness of the cornea as well as the distance between cornea and lens were measured with high precision. Thermal lesions in their expansion can be dearly presented and matching well with the histologically stained sections, but are not as exactly defined at the edges due to the limited optical resolution, as known from histological preparations. CONCLUSION: With the OCT method quantitative measuring of the anterior eye segment can be performed in vitro and with reduced resolutions also in vivo. Due to the qualitatively good correspondence regarding the dimensions of thermal damage of the cornea with histologically obtained morphometric results, this method can be used for supervision of coagulation directly after LTK as well as for examination of the individual healing process.

[Comparative studies with various corneal topography measurement devices]. / Vergleichende Untersuchungen verschiedener kornealer Topographiemessgeräte.

PURPOSE: It is difficult to measure the topography of the cornea with high resolution and visualize it on a map displaying refraction. This is demonstrated by the ongoing improvement and further development of different methods and by the fact that users of these techniques are not always satisfied. MATERIALS AND METHODS: Five different ring projectors were compared. Spherical and aspherical ball standards were used to measure the standard deviation of the refraction of the system. A patient group of eight people with healthy eyes was used to compare the measurement accuracy and operational errors. Some patients were also measured after cataract surgery. The lateral and axial range of the devices was determined. RESULTS: The measurement accuracy for ball standards for the five devices was below 1/8 D. For the in vivo case in the control group it was below 1/4 D for four devices. Furthermore, the reproducibility of the results, the accuracy of angle determination and the influence of mechanical and optical design will be discussed. The study demonstrates applications and limits of this measurement method.

Photofragmentation of lens nuclei using the Er: YAG laser: preliminary report of an in vitro study.

The energy of the erbium: YAG laser (2.940-nm wave-length) can be used for minimally traumatic photoablation due to its high absorption at the tissue water and its consequently low penetration depth. Laser sclerostomy ab externo, an application of this principle, has undergone advanced clinical investigation. Another potential application is photofragmentation of the lens for cataract extraction. A laboratory model Er: YAG laser (flashlamp-pumped. 200-microseconds pulse length, 5-Hz repetition frequency) was coupled to a short low-OH quartz fiber (400 microns in diameter). The laser energy was applied by direct contact of the fiber tip to human lenses with very dense cataract. The lenses rested in a small cuvette filled with an aqueous-humor-analogous fluid. The fragmentablation rate was evaluated in relation to the number of pulses and the pulse energy. A laser-triggered flash-photography unit was engaged to visualize the ablation dynamics. We found tissue-ablation rates to range from 4 to 19 micrograms/pulse, depending on the nucleus density and ulse energy. The maximal size of the removed fragments was always below 500 microns. During ablation, rapidly increasing and collapsing cavitation bubbles were photographed at the distal tip of the application fiber. The impact radius of these cavitation effects markedly exceeded the pure penetration depth of laser radiation at a 2.9-microns wavelength. A clinical application of the method should be possible as judged by the results obtained for tissue-ablation rate and fragment size. Cavitation-bubble dynamics seems to be responsible for the high fragmentation efficiency. Special application probes have to be developed to optimize ablation and to prevent inadvertent destruction of the posterior lens capsule by cavitation effects.

[Use of the confocal laser scanning method for determining corneal topography and corneal tissue effects in refractive corneal surgery]. / Verwendung des konfokalen Laserscanverfahrens zur Bestimmung der Hornhauttopographie und der kornealen Gewebseffekte bei der refraktiven Hornhautchirurgie.

Refraction of the cornea head been generally measured with ophthalmometers or computer disk keratometers. We therefore used a confocal laser scanning system for measurement of the corneal topography. Enucleated tonicized pig eyes were measured before and after laser thermokeratoplasty (LTK). The topographical data were used to determine refraction and refractive change; the data were stored digitally. The single images and their differences were displayed on a PC. Unlike conventional ophthalmometry, confocal laser scanning can demonstrate the topographical shape, showing the overall topography of the cornea and local corneal effects, e.g., coagulation, mechanical lesions or high-energy laser effects. Topographical laser scanning has proven to be a generally useful method of determining refraction and surface alterations in corneal refractive surgery.

Morphological and biomorphometrical observations on laser thermal keratoplasty. Histological and biomorphometrical examination of the relationship between refractive change and the volume following Cr:Tm:Ho:YAG laser treatment.

Laser thermal keratoplasty (LTK) is currently under clinical trial for the correction of hyperopia and hyperopic astigmatism by means of collagen coagulation in the peripheral cornea. The purpose of our study was to optimize the ratio between the volume of damaged corneal stroma and the refractive effect so as to minimize potential side effects such as endothelial damage or induction of glare phenomena. We therefore performed histological and morphometrical examinations of enucleated pig eyes to determine the relationship between the coagulated stromal volume and the refractive change after LTK using a pulsed Cr:Tm:Ho:YAG laser (wavelength 2.12 microns) on enucleated pig eyes. The refractive change was documented with a Littman ophthalmometer. Morphometrical analysis was performed using polarized light microscopy of sirius red-stained specimens. This special stain separated the thermally changed stroma into a dark nonbirefringent center and a birefringent peripheral zone. The volume of both zones was positively correlated with the refractive change induced. The volume was in turn influenced by the choice of laser parameters. From the ratio of the volume to the refractive change it was found that pulse energies above 30 mJ let to an enlargement of the coagulation volume without increasing the refractive change effectively. The use of high pulse energies did not improve the effect of LTK but only increased the risk of unwanted side effects. However, an increase in the laser repetition rate at a constant pulse number per spot led to refractive changes with minimal coagulation volume. The highest relative refractive change was achieved with a dark central zone and a birefringent zone, each having a volume of about 50-80 x 10(-3) mm3.