Human tear fluid PDGF-BB, TNF-alpha and TGF-beta1 vs corneal haze and regeneration of corneal epithelium and subbasal nerve plexus after PRK.

The aim was to determine the association of tear fluid cytokine levels and post-PRK corneal haze evaluated by in vivo confocal microscopy. In addition, the possible association between subbasal neural regeneration and haze formation, or epithelial regeneration were investigated. Twenty eyes of 20 patients (16 women and four men, age 30.7 +/- 7.5 years, range 21-48 years) underwent a myopic PRK. The spherical equivalent (SE) of the intended correction was -4.7 +/- 1.5 D (range -2.75 to -9.00 D). ELISA-methods were used to assess tear fluid concentrations of TGF-beta1, PDGF-BB and TNF-alpha pre-operatively, and post-operatively on day 2 and at 3 months. Tear fluid flow in the collection capillary was recorded, and rates of cytokine release (= tear fluid flow-corrected concentrations) were calculated. In vivo confocal microscopy was performed at 3 months to evaluate the corneal morphology and to determine numerical haze estimate. There was wide interindividual variation between pre-operative and post-operative concentrations and rates of release of TGF-beta1, PDGF-BB and TNF-alpha. Subepithelial haze was observed in all corneas and the mean haze estimate was 506 +/- 401 U (100-1410 U). However, no association was found between tear fluid cytokine levels and post-PRK haze. Regenerating subbasal nerve plexus was found in 18 out of 20 corneas; in two corneas it was absent or could not be visualized due to subepithelial haze. The density of the subbasal nerve fiber bundles had a positive correlation with the epithelial thickness (Pearson correlation, r = 0.56, P = 0.011), but not with the haze estimate or the thickness of the haze area. At 3 months post-PRK, haze could be observed in all patients. The results suggest that tear fluid cytokine analysis, as measured, may not be suitable for screening the potential candidates for haze formation. We did not find any correlation between haze and regeneration of subbasal nerve plexus, but we demonstrated that the regeneration of subbasal nerve plexus might have significant influence on regulation of epithelial healing.

Corneal morphology and sensitivity in lattice dystrophy type II (familial amyloidosis, Finnish type).

PURPOSE: To describe the corneal abnormalities and to measure different modalities of corneal sensitivity in corneal lattice dystrophy type II (familial amyloidosis, Finnish type, also known as gelsolin-related amyloidosis and originally as Meretoja syndrome). METHODS: Twenty eyes of 20 patients were examined by in vivo confocal microscopy and noncontact gas esthesiometry. RESULTS: Pleomorphism of, and dense deposits between or posterior to, the basal epithelial cells were frequently observed, as well as a reduction of long nerve fiber bundles in the subbasal nerve plexus. The anterior stroma was altered in most cases, with fibrosis and abnormal extracellular matrix. In 15 corneas, thick anterior and midstromal filaments, corresponding to lattice lines, and in 11 corneas, thin undulated structures were observed. The average mechanical sensitivity threshold of 12 subjects was increased, and in the remaining 8 subjects there was no response, even to the highest intensity of stimuli used. Three patients did not respond to CO(2), 11 to heat, and 2 to cold, but those patients who responded had normal thresholds. Patients with more long nerve fiber bundles per confocal microscopic image had better mechanical and cold sensitivity than patients with fewer nerve fiber bundles. CONCLUSIONS: Lattice lines seem to be related to amyloid material and not to corneal nerves. However, the subbasal nerve density appears reduced, which results mainly in a decrease in mechanical and, to a lesser extent, thermal sensitivity. The location of stromal filaments and undulated structures changes with increasing age.

Photorefractive keratectomy in ophthalmic residents.

PURPOSE: To find out how ophthalmologists themselves experience the correction of myopia after photorefractive keratectomy. Visuomotor functions were of special interest. METHODS: Four ophthalmology residents and one medical engineer underwent photorefractive keratectomy for myopia. Objective measurements including refraction, corneal topography, perimetry, contrast sensitivity, pattern visual evoked potentials, in vivo confocal microscopy, and a car driving simulator test were performed preoperatively, postoperatively, and at 6 months. Subjective evaluation was reported. RESULTS: Performing ophthalmological examinations and microsurgery without spectacles was easier postoperatively and was appreciated by the four ophthalmology residents. Minimal haze formation, good accuracy, and normal performance in the car driving simulator were also observed. Visual fields, contrast sensitivity, and pattern visual evoked potentials did not show changes. Negative observations included postoperative pain for 2 to 4 days, dry eye symptoms, a period of anisometropia between operations, and hypersensitivity of the lids. CONCLUSIONS: The four ophthalmic residents were satisfied with the outcome of their refractive surgery. Low to moderate myopic correction did not affect the objective measurements of high and low contrast sensitivity, pattern visual evoked potentials, or simulated car driving in dark illumination.

Laser in situ keratomileusis flap margin: wound healing and complications imaged by in vivo confocal microscopy.

PURPOSE: To examine the healing response of laser in situ keratomileusis flap margin in vivo. METHODS: Forty-three eyes of 43 patients who had undergone myopic (n = 39) or hyperopic (n = 4) laser in situ keratomileusis were examined once after surgery. The flap margin was imaged by in vivo confocal microscopy at various depths, and the wound healing response, flap alignment, and complications were evaluated. Ten eyes were examined on day 3 postoperatively, 13 eyes at 1 to 2 weeks, 10 eyes at 1 to 2 months, five eyes at 3 months, and five eyes at 6 months or later. RESULTS: At 3 days after laser in situ keratomileusis, the surface epithelium and basal epithelium appeared normal. Keratocyte activation was strongest at 1 to 2 weeks and 1 to 2 months, and an increased amount of haze was observed correspondingly. Intrastromal epithelial cells forming a plug could occasionally be perceived in the wound gape. Wound constriction was completed in most cases by 3 to 6 months or later. Good alignment was observed in 12 of 43 flaps (27.9%) and moderate and poor alignment in 17 of 43 flaps (39.5%) and 13 of 43 flaps (30.2%), respectively. Poor alignment was not associated with lamellar epithelial ingrowth. Epithelial ingrowth was associated with dense haze at the interface. Diffuse lamellar keratitis was imaged in two corneas after hyperopic laser in situ keratomileusis. CONCLUSIONS: The laser in situ keratomileusis incision wound at the flap margin appears to heal after the sequence observed in incisional wounds in nonhuman primates. Complications, such as lamellar epithelial in growth and diffuse lamellar keratitis, were often observed, particularly after hyperopic laser in situ keratomileusis.

Confocal microscopy of a patient with irregular astigmatism after LASIK reoperations and relaxation incisions.

PURPOSE: Laser-assisted in situ keratomileusis (LASIK) is widely used for correcting refractive errors. If the predicted refractive result is not achieved after the first operation, a re-operation can be performed by ablating more stromal tissue after reopening the flap. The goal of this study was to analyze, by using in vivo confocal microscopy, the morphologic changes associated with repeated LASIKs. METHODS: Clinical examination, computed corneal topography, and real-time in vivo confocal microscopy were performed on both eyes of a 50-year-old patient with induced irregular astigmatism leading to decreased best-corrected vision in the left eye after LASIK. The left cornea had been operated on 5 times (LASIK with two reoperations followed by two relaxing incisions), and the right cornea twice (LASIK with one reoperation). RESULTS: Microfolds at the level of the Bowman's layer and highly reflective particles at the flap interface were observed in both corneas. The subbasal nerve plexus was severed in the left eye. In addition, we identified epithelial material in the flap margin and inside one of the two relaxing incisions placed inferotemporally. CONCLUSION: Repeated LASIKs may stretch the flap and result in microfolding at the Bowman's layer. This and deposition of particles in the flap interface may increase with the number of reoperations, challenging the healing response. Microfolding and occurrence of foreign material in the interface may add to the irregular astigmatism and poor visual outcome after LASIK. Clinical in vivo confocal microscopy offers new possibilities for the assessment of ultrastructural changes after corneal refractive surgery.

Tenascin and cytokines in tear fluid after photorefractive keratectomy.

BACKGROUND: Basic knowledge of the substances involved in wound healing after photorefractive keratectomy (PRK) is essential for development of pharmacological intervention. We present preoperative and postoperative analysis of tear fluid extracellular matrix proteins and cytokines after PRK. METHODS: Tear fluid samples from 70 patients (72 eyes) who had PRK (38 women and 32 men, mean age 31.5 yr) were studied. Samples from 18 patients (18 eyes) were analyzed in two different studies. RESULTS: Mean preoperative tear fluid flow in the collection capillary (volume divided by tear collection time) varied from 4.5 to 22.5 microliters/min in five series of patients. It increased significantly during the first two postoperative days (range of means, 55.5 to 88.8 microliters/min, p < 0.01), and decreased to the preoperative level by day 7 (range of means, 9.7 to 18.2 microliters/min). The tenascin and cytokine release rates increased significantly during the first two days after PRK and returned to the preoperative level by day 7. Mean +/- standard error for tenascin: day 0 (5.2 +/- 1.9 ng/min); day 2 (22.7 +/- 6.1 ng/min; p = 0.02). Mean +/- standard error for HGF: day 0 (3.2 +/- 0.7 pg/min); day 1 (22.8 +/- 4.2 pg/min; p = 0.0003). Mean +/- standard error for TGF-beta 1: day 0 (63.3 +/- 19.6 pg/min); days 1-2 (826.2 +/- 253.7 pg/min; p = 0.001). Mean +/- standard error for VEGF: day 0 (166.0 +/- 29.6 pg/min); days 1-2 (824.4 +/- 165.1 pg/min; p = 0.0007). Mean +/- standard error for PDGF-BB: day 0 (0.42 +/- 0.19 pg/min); day 2 (27.6 +/- 5.8 pg/min; p = 0.0000). Mean +/- standard error for TNF-alpha: day 0 (9.5 +/- 2.6 pg/min); day 2 (28.6 +/- 5.9 pg/min; p = 0.003). Excluding PDGF-BB, all substances studied were present in normal human tear fluid. PDGF-BB was present in only 17% of the preoperative samples. CONCLUSION: Corneal wounding induces an increased release of several growth modulating cytokines which may be involved in healing processes.