Cell viability and shock wave amplitudes in the endothelium of porcine cornea exposed to ultrashort laser pulses.

PURPOSE: Some forms of keratoplasty assisted by ultrashort-pulse lasers require performing laser cuts close to the endothelium, which requires the knowledge of "safe" values concerning incision depth and pulse energy preserving endothelial cell viability. Our study aims to determine the thresholds for cell death in porcine corneas exposed to ultrashort laser pulses, in terms of laser pulse energy and nearness of the impacts to the endothelium. METHODS: Using a laboratory laser set-up, lamellar cuts were induced while varying pulse energies and distances from the endothelium. A fluorescent staining protocol was used to determine the percentage of surviving endothelial cells. Numerical simulations of the Euler equations for compressible fluids provided pressure level and axial and radial pressure gradient estimates at the endothelium. RESULTS: Ninety percent of the endothelial cells survived when using 16.5 µJ pulses no closer than 200 µm to the endothelium, or pulses not exceeding 2 µJ at a distance of 50 µm. The comparison of the observed percentage of surviving cells with the estimates of the shock wave amplitudes and gradients generated by the laser pulses yielded cell death thresholds at amplitudes in the megapascal range, or gradients of the order of 108 Pa/m. CONCLUSIONS: Our results provide limits in terms of pulse energy and distance of the incision from the endothelium within which endothelial cell viability is preserved. Current forms of corneal laser surgery are compatible with these limits. However, these limits will need to be considered for the development of future laser routines working in close proximity to the endothelium.

Wavelength optimization in femtosecond laser corneal surgery.

PURPOSE: To evaluate the influence of wavelength on penetration depth and quality of femtosecond laser corneal incisions in view of optimizing procedures in corneal surgery assisted by ultrashort pulse lasers. METHODS: We performed penetrating and lamellar incisions on eye bank corneas using several ultrashort pulse laser sources. Several wavelengths within the near-infrared and shortwave-infrared wavelength range were used and the pulse energy was varied. The corneas were subsequently analyzed using light microscopy as well as transmission and scanning electron microscopy. RESULTS: We found higher penetration depths and improved incision quality when using wavelengths close to λ = 1650 nm rather than the wavelength of λ = 1030 nm typical in current clinical systems. Optical micrographs show an improvement of the penetration depth by a factor of 2 to 3 while maintaining a good incision quality when using the longer wavelength. These results were confirmed with micrographs obtained with transmission and scanning electron microscopy. CONCLUSIONS: A wavelength change from the standard 1030 nm to 1650 nm in corneal surgery assisted by ultrashort pulse laser considerably reduces light scattering within the tissue. This results in a better preservation of the laser beam quality in the volume of the tissue, particularly when working at depths required for deep lamellar or penetrating keratoplasty. Using this wavelength yields improved penetration depths into the tissue; it permits use of lower energies for any given depth and thus reduces unwanted side effects as thermal effects.

Effect of incident light wavelength and corneal edema on light scattering and penetration: laboratory study of human corneas.

PURPOSE: The outcome of ultrashort pulse laser surgery of the cornea is strongly influenced by the light scattering properties of the tissue, for which little data are available. The purpose of the present study is to provide quantitative values for light scattering and its relation to the degree of edema. METHODS: An experimental optical measuring setup based on confocal geometry was used to measure the unscattered and scattered fractions of light transmitted by eye bank corneas presenting various degrees of edema. From these measurements, the effective light penetration depth in the cornea was calculated as a function of wavelength. RESULTS: Corneal transparency depends on the pathological state of the cornea and on wavelength. It may be predicted as a function of corneal thickness, ie, the degree of edema. In healthy and edematous cornea, the percentage of scattered light decreases with increasing wavelength. The total penetration depths at the wavelengths of ~1050 nm (which is used in typical clinical systems) and 1650 nm (which is recommended for future devices) are comparable; however, the former is limited by scattering, which degrades the laser beam quality, whereas the latter is only limited by optical absorption, which may be compensated for. CONCLUSIONS: The use of longer wavelengths should help improve the surgical outcome in ultrashort pulse laser surgery of the cornea when working on pathological tissue. A wavelength of approximately 1650 nm appears to be a good compromise, as it allows for reduced light scattering while keeping optical absorption reasonably low.