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Biomedical subjects

R R Krueger

Publications and source records attributed to R R Krueger.

At least 37 records · Page 2Linked to original sources

Role of matrix metalloproteinases in failure to re-epithelialize after corneal injury.

Delayed re-epithelialization of the cornea after injury usually precedes stromal ulceration. Previous findings using a rat thermal injury model suggested that re-epithelialization is impeded by products of resident corneal cells, which destroy adhesive structures at the basement membrane zone. In this study, we provide additional evidence for this concept. Failure to re-epithelialize was found to correlate with an increase in the amounts of gelatinolytic matrix metalloproteinases present in the rat cornea. One of these gelatinases, gelatinase B, is synthesized by the resident corneal cells, and inhibitions of its synthesis correlated with inhibition of basement membrane dissolution. The matrix metalloproteinases collagenase and stromelysin are also synthesized by resident corneal cells in thermally injured corneas of rabbits, but the timing of bulk enzyme synthesis correlated more closely with deposition of repair tissue in the stroma than with failure to re-epithelialize. Nevertheless, in human corneas with repair defects, gelatinase B and collagenase are synthesized by cells in the basal layer of the epithelium directly adjacent to the basement membrane, suggesting that both could participate in dissolution of this structure. Importantly, treatment of thermally injured corneas with a synthetic inhibitor of matrix metalloproteinases significantly improved basement membrane integrity. These data support the concept that over-expression of matrix metalloproteinases by resident corneal cells impedes re-epithelialization after some types of corneal injury.

Animals↗

Clinical analysis of excimer laser photorefractive keratectomy using a multiple zone technique for severe myopia.

PURPOSE: In an investigational procedure, excimer laser photorefractive keratectomy for severe myopia was performed at three clinical trial centers to determine the effectiveness of the multiple zone technique. METHODS: A VisX Model Twenty/Twenty excimer laser (VisX, Santa Clara, California) was used to perform photorefractive keratectomy on 14 severely myopic eyes (-10.37 to -24.5 diopters) of 12 patients by using a multiple zone technique. Postoperative follow-up ranged from six months to two years; retreatments were performed on four patients, with a follow-up of at least nine months. RESULTS: At six months postoperatively, before retreatment, three of the 14 eyes were within 2 diopters and seven of the 14 eyes were within 4 diopters of attempted correction. Regression of effect to more severe myopia was worse in five eyes treated with nitrogen gas blowing. Retreatments also demonstrated considerable myopic regression. Three patients had loss of two or more lines of best-corrected visual acuity, and these patients also had moderate or severe levels of haze. CONCLUSION: Excimer laser photorefractive keratectomy for severe myopia using a multiple zone technique is associated with considerable regression, haze, and loss of best-corrected visual actuity, especially when performed in association with nitrogen gas blowing.

Adult↗

New stroboscopic light source for intraoperative retinal fluorescein angiography.

We tested a new stroboscopic light source for intraoperative fluorescein angiography. A pulsed xenon light source and narrow-band interference filters are coupled to a 20-gauge endoilluminator and a 35-mm camera mounted on a surgical microscope. The system was tested with good success in a patient undergoing penetrating keratoplasty. An 18- to 22-microJ flash of 2 milliseconds' duration is achieved, which is well below the safety threshold level. This new stroboscopic light source provides a flash of sufficient light intensity to perform high-resolution, high-contrast fluorescein angiography during operations for the removal of opacified media.

Aged↗

Corneal surface morphology following excimer laser ablation with humidified gases.

OBJECTIVE: To compare the effects of blowing dry (nitrogen or helium) and humidified gases over the corneal surface during photorefractive keratectomy. METHODS: Excimer laser myopic ablations were performed on porcine eyes (10 per group) using humidified and dry nitrogen and helium gas under ambient conditions. Surface smoothness was quantified with light and electron microscopy. RESULTS: Corneas that were ablated using humidified gas were smooth and equivalent to those ablated under ambient conditions. Dry nitrogen and helium blowing resulted in increased surface irregularity evident on light and electron microscopy (P < .001). The pseudomembranes in the humidified gas and ambient air groups had fewer surface discontinuities than did those in the nonhumdified gas groups and appeared to have a thinner electron-dense surface layer. CONCLUSIONS: The blowing of humidified gas during excimer laser corneal ablation produces a smoother surface than does the blowing of dry gas and is comparable to that produced under ambient (no blowing) conditions. Maintaining corneal moisture is important in photorefractive keratectomy. If blowing gas is necessary to remove debris from the surface, the gas should be humidified.

Animals↗

Arachidonic acid metabolites after excimer laser corneal surgery.

OBJECTIVE: We investigated early mediators of inflammation following excimer laser ablation in a rabbit cornea model. The ability of topical anti-inflammatory agents to influence these responses was also examined. METHODS: Adult New Zealand white rabbits were subjected to photorefractive keratectomy with a 193-nm argon fluoride excimer laser. Prostaglandin E2 and leukotriene B4 levels were measured using an enzyme immunoassay, and leukocyte infiltration was determined histologically. RESULTS: Prostaglandin E2 production was rapid and sustained, but we were unable to detect the presence of leukotriene B4. Relative to control, postoperative topical diclofenac sodium treatment caused a significant decrease in prostaglandin E2 levels and a significant increase in corneal leukocytes at 10 hours. Fluorometholone treatment did not significantly alter prostaglandin E2 levels but markedly depressed leukocyte ingress. CONCLUSIONS: Diclofenac reduces prostaglandin E2 levels but not leukocyte infiltration in the cornea following photorefractive keratectomy and thus may be useful clinically to reduce postsurgical pain.

Animals↗

Photography of shock waves during excimer laser ablation of the cornea. Effect of helium gas on propagation velocity.

Shadow photography of shock waves excited by means of a xenon chloride excimer laser was performed to determine the shock wave propagation velocity in air, nitrogen and helium. Energy densities between 500 and 2,000 mJ/cm2 were used to ablate a rotating rubber cylindrical target and porcine corneas. In ablating the rubber cylinder, a shock wave velocity of 3.3 km/s was generated in air and nitrogen at 40 ns; this decreased to 1.4 km/s at 320 ns. When helium was blown on the target, the velocity increased by a factor of approximately two, to 5.9 km/s at 40 ns and 2.7 km/s at 320 ns. We suggest that blowing helium on the surface of the cornea during excimer laser ablation may speed the dissipation of high-energy acoustic waves and gaseous particles, and thus reduce the exposure and transfer of heat energy to the surrounding tissue.

Animals↗

Photokeratitis from 193 nm argon-fluoride laser radiation.

The threshold for photokeratitis at 193 nm was obtained for the rabbit cornea using an ArF excimer laser. Because ablation occurs at a level below that for photokeratitis, it was necessary to expose the cornea to a lengthy series of low-energy exposures. It is concluded that the 193 nm photons have such a shallow penetration depth, being limited to the outermost epithelial cells, that classical photokeratitis occurs from the fluorescence emitted at the corneal epithelial absorption site. An intact tear film may help to protect the cornea from low-level, scattered 193 nm laser radiation.

Animals↗

Long-term healing of the central cornea after photorefractive keratectomy using an excimer laser.

Using an excimer laser at 193 nm, 3-mm diameter discs were ablated from the optical zone of monkey corneas at a variety of depths up to 130 micron. Tissue was examined by light and electron microscopy 4 and 5 weeks and 6 and 8 months postoperatively. With the exception of the deepest ablation, all corneas were clear immediately postoperatively. At 1 month, ablations of 40 micron remained clear, whereas all others showed some degree of haze. By 3 months, discs at 60 micron depth were clear, and there was less haze in others. Haze was progressively lost over 6 months, but in the deepest discs it could still be discerned on slit-lamp examination. Microscopic observation showed that reepithelialization had occurred within 24 to 48 hours. Over subsequent months, a normal morphology was maintained in this layer with the exception that basal cells were slightly more elevated particularly at the disc margins where the epithelium contained more cell layers. Stromal reorganization was accompanied by an initial phase of vacuolation and invasion by keratocytes, but by 6 months postoperatively almost all vacuolation had disappeared and keratocyte numbers had almost returned to normal. By 8 months, the morphology was near normal with the exception that Bowman's membrane was absent and there was still a degree of disorder in the immediate subepithelial stromal fibers. The basement membrane of epithelial cells was reestablished but slightly more undulant than in nonirradiated areas. This undulation was retained in the 8-month postoperative specimens and at this time a normal thickness of 60 nm was displayed.

Animals↗

High-speed photography of excimer laser ablation of the cornea.

We have used laser-based high-speed photography to investigate excimer laser ablation of the cornea. Photographs of the ablation plume were obtained 500 ns to 150 microseconds after incidence of a 193- or 248-nm excimer laser pulse on the surface of the cornea. Ejection of material from the cornea begins on a time scale of nanoseconds and continues for 5 to 15 microseconds following the excimer pulse. At 193 nm the ablation plume resembles a burst of smoke, and individual particles are too small to be optically resolved with our apparatus. At 248 nm the plume resembles a spray of larger, discrete droplets. Material is ejected from the cornea at supersonic velocity but decelerates rapidly; the velocity for the first 500 ns following the excimer pulse averages 400 m/s at 193 nm. Plume size and velocity increase with increasing fluence.

Animals↗

A comparative study of corneal incisions induced by diamond and steel knives and two ultraviolet radiations from an excimer laser.

This paper reviews the potential role of excimer lasers in corneal surgery. The morphology of incisions induced by two wavelengths of excimer laser radiation, 193 nm and 248 nm, are compared with the morphology of incisions produced by diamond and steel knives. Analysis suggests that ablation induced by excimer laser results from highly localised photochemical reactions and that 193 nm is the optimal wavelength for surgery. The only significant complication of laser surgery is loss of endothelial cells when incisions are within 40 micron of Descemet's membrane.

Animals↗

Excimer laser therapy for experimental Candida keratitis.

In vivo treatment of experimental Candida albicans keratitis with the excimer laser achieved sterilization on culture and histopathologically in all corneas in which clinically visible infiltration was removed with the laser at the 193-nm wavelength. Treatment at the 248-nm wavelength was not successful in eradicating infection compared with untreated controls. The 193-nm wavelength is highly effective, probably because infected tissue is totally removed by ablative photodecomposition. At the 248-nm wavelength, thermal effects become more dominant and, therefore, fungal elements are incompletely ablated. Light microscopy two days after treatment at the 193-nm wavelength showed healing of ulcerated areas, fine basophilic stippling at the epithelial-stromal interface, and undamaged underlying stroma.

Animals↗

Quantitation of corneal ablation by ultraviolet laser light.

A surgical role has been suggested for the excimer laser. This followed the discovery of precision etching of biological materials by the laser's ultraviolet emissions. In studying the amount of tissue removed as the laser energy density (irradiance) was varied, we found significant differences for the 193-, 249-, and 308-nm laser emission lines. The most efficient cutting irradiance for each pulse appeared to be about 200 millijoules/sq cm for 193 nm, 1,000 millijoules/sq cm for 249 nm, and 1,500 millijoules/sq cm for 308 nm. The flat slope of the etch depth curve for 193 nm suggests it will be a stable frequency when used clinically.

Animals↗

Argon laser coagulation of blood for the anastomosis of small vessels.

A new technique is introduced in which an argon laser coagulates blood to form an adherent sleeve for the anastomosis of small vessels. The argon laser solidifies the blood into a dark tensile substance which supports the site of repair until vascular continuity is achieved. Laser pulses of 0.75 W and 0.5 seconds exposure are used, and damage to the underlying vessel wall is minimal. A histological analysis is performed and the results are examined qualitatively. The technique shows advantages of speed, reduced vascular trauma, and external protection at the site of vascular repair.

Animals↗

Interaction of ultraviolet laser light with the cornea.

The effect of pulsed ultraviolet (UV) laser light on the cornea depends on wavelength (photon energy), irradiance (photon flux), and pulse firing rate. At the available excimer laser wavelengths of 193, 249, 308, and 351 nanometers, the authors have varied the irradiance per pulse (10 to 2000 mj/cm2) as well a pulse frequency (1, 10, 25 Hz) and determined the thresholds for coagulation and ablation of the corneal stroma. The latter ablative action creates a groove resembling an incision and was present at all wavelengths studied. The threshold for ablation increased for longer wavelengths and lower pulse frequencies, except for 193-nm exposure, which was characterized by a constant threshold independent of laser pulse rate. The grooves at 193 nm were both biomicroscopically and histologically smooth and no coagulation effects were noted. Some degree of coagulation of adjacent tissues was noted at 249, 308, and 351 nm.

Animals↗

Ultrastructure of picosecond laser intrastromal photodisruption.

PURPOSE: To investigate the ultrastructure of the corneal stroma after picosecond intrastromal photodisruption with a neodymium-doped yttrium-lithium-fluoride (Nd:YLF) laser. METHODS: We performed picosecond intrastromal photodisruption on six human eye-bank eyes using a lamellar technique. Thirty picosecond pulses at 1000 Hz and 20 to 25 mJ per pulse were placed in an expanding spiral pattern, the pulses separated by 15 microns. Three layers were placed in the anterior stroma, separated from each other by 15 microns. In addition, intrastromal radial and arcuate incisions were generated in two living rabbit eyes in a plane perpendicular to the corneal surface. After the procedure, the corneas were processed for scanning and transmission electron microscopy. RESULTS: Scanning electron microscopy of the eye-bank eyes demonstrated multiple, coalescing intrastromal cavities forming a layer oriented parallel to the corneal surface. These cavities had smooth inner walls. Transmission electron microscopy demonstrated tissue loss surrounding some cavities, with the terminated ends of collagen fibrils clearly evident. Other cavities were formed by separation of lamellae, with little evidence of tissue loss. A pseudomembrane was present along the margin of some cavities. Although there was occasional underlying tissue disruption along the border of a cavity, there was no evidence of thermal damage or tissue necrosis. The perpendicular photodisruptions demonstrated intrastromal cleavage of corneal collagen similar to diamond-knife incisions, with the exception of intact overlying Bowman's and epithelial layers. CONCLUSION: Intrastromal photodisruption with a Nd:YLF picosecond laser induced no thermal necrosis or coagulative change in the region of tissue interaction. Lamellar intrastromal photodisruption demonstrated both tissue loss and lamellar separation when performed with the current treatment parameters, possibly limiting ablation efficiency and predictability.

Animals↗

Picosecond laser in situ keratomileusis with a 1053-nm Nd:YLF laser.

BACKGROUND: Excimer laser in situ keratomileusis requires a microkeratome to generate an anterior corneal flap, plus an excimer laser to ablate the underlying stromal tissue. In this paper we introduce the concepts of laser flap formation and in situ keratomileusis using a picosecond laser. METHODS: A neodymium-doped yttrium-lithium-fluoride (Nd:YLF) laser with a plano-plano quartz applanation lens was used to generate various patterns of intrastromal photodisruption in human donor eyes to fashion anterior corneal flaps and generate intrastromal lenticules. RESULTS: Smooth intrastromal dissections, 6 mm in diameter, were generated 160 microns below the corneal surface when the laser delivered pulses at 1 kHz with energies of either 40 microJ/pulse or 60 microJ/pulse, placed 20 microns apart in an expanding spiral. This enabled us to fashion anterior corneal flaps. The ease of the surgery and quality of the dissection corresponded well, and it was evident that both deteriorated noticeably when the laser pulses were separated by 25 microns or 30 microns, regardless of pulse energy. Using 40 microJ laser pulses placed 20 microns apart we also created a 5-mm diameter, 320 microns thick (130 microns-450 microns deep) stromal lenticule below a corneal flap that was easily extracted when the flap was raised. CONCLUSIONS: Anterior corneal flaps were easily fashioned using a Nd:YLF laser. Picosecond laser in situ keratomileusis with a Nd:YLF laser could offer a favorable alternative to combined microkeratome/excimer laser in situ keratomileusis.

Cornea↗

Retrephination keratoplasty for high astigmatism after penetrating keratoplasty.

PURPOSE: We report preliminary results of a new procedure for correcting high astigmatism after penetrating keratoplasty. METHODS: The procedure entails full-thickness trephination along the original donor-recipient junction with careful suturing in a combined interrupted and running fashion. Four eyes of four patients with severe astigmatism and myopia after penetrating keratoplasty underwent the procedure. RESULTS: High preoperative cylinder ranging from 4.50 to 16.00 D (mean 9.00 D) was reduced to 0.50 to 3.50 D (mean 1.90 D) at the last examination (between 3 to 6 months). Spherical equivalent myopia ranging from -2.00 to -10.25 D (mean -4.90 D) was essentially unchanged at plano to -9.00 D (-4.70 D) at the last examination. Overall, there was a mean refractive cylinder reduction of 7.10 D (79%). CONCLUSION: Retrephination after penetrating keratoplasty appears to be an acceptable alternative for correcting high astigmatism, and had only a small effect on the level of myopia.

Astigmatism↗