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.

Postoperative inflammation, microbial complications, and wound healing following laser in situ keratomileusis.

Although the biology of corneal wound healing is only partly understood, healing after photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) differs in many respects, and the mechanisms appear to be differently controlled. There is less of an inflammatory and healing response after LASIK, but a longer period of sensory denervation. The cellular, molecular, and neural regulatory phenomena associated with postoperative inflammation and wound healing are likely to be involved in the adverse effects after LASIK, such as flap melt, epithelial ingrowth, and regression. Interface opacities in the early postoperative period include diffuse lamellar keratitis (DLK), microbial keratitis, epithelial cells, and interface opacities. Diffuse lamellar keratitis (sands of the Sahara syndrome) describes an apparently noninfectious diffuse interface inflammation after lamellar corneal surgery probably caused by an allergic or a toxic inflammatory reaction. Noninfectious keratitis must be distinguished from microbial keratitis to avoid aggressive management and treatment with antimicrobial drugs. Microbial keratitis is a serious complication after LASIK, but a good visual outcome can be achieved following prompt and appropriate treatment.

Effects of oleoresin capsicum pepper spray on human corneal morphology and sensitivity.

PURPOSE: To examine the potential harmful effects on corneal structure, innervation, and sensitivity of a spray containing the neurotoxin capsaicin (oleoresin capsicum, OC). METHODS: Ten police officers who volunteered for the study were exposed to OC. Clinical signs were assessed. Corneal sensitivity was measured using a Cochet-Bonnet or a noncontact esthesiometer that provides separate measurements of mechanical, chemical, and thermal sensitivity. Tear fluid nerve growth factor (NGF) was measured. Corneal cell layers and subbasal nerves were examined by in vivo confocal microscopy. The subjects were examined before application and 30 minutes, 1 day, 1 week, and 1 month after OC exposure. RESULTS: OC spray produced occasional areas of focal epithelial cell damage that healed within 1 day. Each eye showed conjunctival hyperemia and in two subjects, mild chemosis. All except one eye had unchanged best corrected visual acuity (BCVA). A transient decrease (day 1) of mechanical sensitivity was observed with the Cochet-Bonnet esthesiometer. With the gas esthesiometer, mechanical sensitivity remained below normal values for 7 days. Chemical sensitivity to CO2 was high for as much as 1 day and decreased below normal 1 week later, whereas sensitivity to cold was unaffected. Two subjects had measurable tear NGF that increased after exposure. Basal epithelial cell morphology suggested temporary corneal epithelial swelling, whereas keratocytes, endothelial cells, and subbasal nerves remained unchanged. CONCLUSIONS: Although OC causes immediate changes in mechanical and chemical sensitivity that may persist for a week, a single exposure to OC appears harmless to corneal tissues. The changes are possibly associated with damage of corneal nerve terminals of mainly unmyelinated polymodal nociceptor fibers.

Immunolocalization of transforming growth factor-beta1 and tenascin in human secondary cataract.

PURPOSE: This study was carried out to investigate the distribution of transforming growth factor-beta1 (TGF-beta1) and the extracellular matrix (ECM) glycoprotein tenascin in secondary cataract and anterior subcapsular cataract. METHODS: Twenty-four pseudophakic human eyes with secondary cataract, obtained at autopsy 1 d to 10 yr (mean 2.4 yr) after cataract surgery, were studied. Additionally, a specimen from an anterior subcapsular cataract was included. Immunohistochemistry was used to localize TGF-beta1 and tenascin in secondary cataract and in anterior subcapsular cataract. RESULTS: Polyclonal antibody to TGF-beta1 immunolabelled spindle-shaped cells in the plaques of secondary cataract in all eyes. Instead, the cells present in Soemmering's ring cataract were not labelled. The ECM in the plaques of secondary cataract was immunoreactive for tenascin in all eyes. In anterior subcapsular cataract spindle-shaped cells and ECM showed similar immunoreactivity. CONCLUSION: Spindle-shaped cells that are immunolabelled with TGF-beta1 and ECM showing tenascin-like-immunoreactivity are present in secondary cataract and in anterior subcapsular cataract, thus implicating a possible role in secondary cataract.

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.

Corneal stromal changes induced by myopic LASIK.

UNLABELLED: PURPOSE. Despite the rapidly growing popularity of laser in situ keratomileusis (LASIK) in correction of myopia, the tissue responses have not been thoroughly investigated. The aim was to characterize morphologic changes induced by myopic LASIK in human corneal stroma. METHODS: Sixty-two myopic eyes were examined once at 3 days to 2 years after LASIK using in vivo confocal microscopy for measurement of flap thickness, keratocyte response zones, and objective grading of haze. RESULTS: Confocal microscopy revealed corneal flap interface particles in 100% of eyes and microfolds at the Bowman's layer in 96.8%. The flaps were thinner (112 +/- 25 microm) than intended (160 microm). The keratocyte activation in the stromal bed was greatest on the third postoperative day. Patients with increased interface reflectivity due to abnormal extracellular matrix or activated keratocytes at > or = 1 month (n = 9) had significantly thinner flaps than patients with normal interface reflectivity (n = 18; 114 +/- 12 versus 132 +/- 22 microm, P = 0.027). After 6 months the mean density of the most anterior layer of flap keratocytes was decreased. CONCLUSIONS: Keratocyte activation induced by LASIK was of short duration compared with that reported after photorefractive keratectomy. The flaps were thinner than expected, and microfolds and interface particles were common complications. The new findings such as increased interface reflectivity associated with thin flaps and the apparent loss of keratocytes in the most anterior flap 6 months to 2 years after surgery may have important clinical relevance.

Soluble Fas and Fas ligand in human tear fluid after photorefractive keratectomy.

BACKGROUND/AIMS: The Fas-Fas ligand system is thought to be involved in stromal cell apoptosis after corneal wounding. The aim was to measure changes in human tear fluid levels of soluble Fas (sFas) and Fas ligand (sFasL) following myopic photorefractive keratectomy (PRK). METHODS: Tear samples of 59 patients were collected preoperatively, and 1 or 2 days after PRK. Tear fluid sFas or sFasL concentrations were determined using sandwich ELISAs. Subsequently, tear flow corrected concentrations (releases) were calculated to compensate for the postoperative tear hypersecretion. RESULTS: The preoperative tear fluid flows (TFF) were 6.4 (1.7) microl/min (mean (SEM)) in sFas group (n = 18), and 7.5 (1.5) microl/min in sFasL group (n = 39). Postoperatively TFFs increased to 37.9 (10.9) microl/min (p = 0.003) and 58.3 (7.0) microl/min (p = 0.000), respectively. The mean preoperative sFas concentration (24.4 (11.6) U/ml) decreased to 9.7 (4.1) U/ml (p = 0.001) postoperatively, and the mean sFasL concentration (299.1 (28.8) ng/l) to 118.7 (15.9) ng/l (p = 0.000). However, the release of both substances increased significantly: sFas from 87.3 (29.4) mU/min to 229.4 (82.9) mU/min (p = 0.002) and sFasL from 1620.6 (226.4) fg/min to 4777.1 (596.1) fg/min (p = 0.000). CONCLUSIONS: Both sFas and sFasL are normal constituents of human tears. Despite a decrease in concentrations related to reflex tears, the release of sFas and sFasL increases significantly after excimer laser photorefractive keratectomy, which suggests that they are involved in corneal healing after PRK in humans.

Tumour necrosis factor alpha (TNFalpha) in tears of atopic patients after conjunctival allergen challenge.

BACKGROUND: Proinflammatory cytokines, such as TNFalpha are involved in the pathogenesis of allergic rhinitis and asthma. OBJECTIVE: Our aim was to study the tear fluid TNFalpha levels in atopic patients before and after topical ocular allergen challenge. METHODS: Thirteen patients were first challenged with topical allergen in the left eye and then dilution buffer in the right eye. Tear fluid samples were collected before the challenges and after 5, 30 and 60 min. Clinical symptoms were scored. Tear fluid TNFalpha concentrations were measured using a double-antibody radioimmunoassay. TNFalpha release rates were calculated using the tear fluid flow in the collection capillary as an estimate for the tear secretion rate. RESULTS: The mean baseline TNFalpha concentrations (ng/L) were 1310 (allergen-challenged eye) and 967 (control eye), and release rates (pg/min) 1.81 (allergen-challenged eye) and 1.39 (control eye), respectively. In the allergen-challenged eye, TNFalpha concentrations and release rates were 1479 and 4.30 (5 min), 1367 and 3.20 (30 min) and 1426 and 3.80 (60 min). In the control eye, TNFalpha concentrations and release rates were 746 and 0.67 (5 min), 1001 and 0.92 (30 min) and 1504 and 1.05 (60 min). The release rates of the allergen-challenged eye were significantly increased at all time points after the challenge when compared with the baseline or the control eye. CONCLUSION: Increased TNFalpha release rates suggest that this cytokine is an early mediator of allergy after conjunctival challenge.

Prospective evaluation of external ocular microbial growth and aqueous humor contamination during cataract surgery.

PURPOSE: To analyze the route of aqueous humor contamination leading to the development of postoperative endophthalmitis. SETTING: Department of Ophthalmology, University of Helsinki, Finland. METHODS: Forty-nine eyes of 49 patients (31 having phacoemulsification and 18 extracapsular cataract extraction [ECCE]) participated in the study. Four bacterial cultures were taken: preoperative conjunctival swab, lid margin culture, intraoperative lacrimal lake sample, and immediate postoperative anterior chamber fluid sample. RESULTS: Preoperative lid margin cultures were positive in 59.2% of eyes, conjunctival cultures in 69.4%, and lacrimal lake cultures in 24.9%. Four aqueous humor samples (8.2%) showed bacterial growth in the anterior chamber aspirate: 3 in the phacoemulsification and 1 in the ECCE group. The bacteria isolated in this study, Staphylococcus epidermidis and Propionibacterium acnes (2 positive isolates each) were sensitive to the preoperative topical antibiotics used. No aqueous humor sample or any from other locations showed gram-negative microbe growth. The most frequently recovered microbes in all samples collected from the 3 other sources were S epidermidis and other coagulase-negative staphylococcus species, followed by P acnes and other propionibacterium species. Staphylococcus aureus, and diptheroids. CONCLUSION: The ocular surface significantly contributed to the transmission of microbes into the eye during cataract surgery. These microbes could not be eradicated by topical preoperative antibiotics. However, no patient developed postoperative endophthalmitis. Natural defense mechanisms appear to fend off a minor inoculum with these microbes of relatively low pathogenicity.

Corneal wound healing after laser in situ keratomileusis in rabbits.

BACKGROUND: The aim of this study was to characterize the cell biology of wound healing in rabbit corneas subjected to laser in situ keratomileusis (LASIK). METHODS: Rabbit corneas underwent LASIK with various multizone photoablations or only a lamellar keratotomy followed by repositioning of the flap. We looked for indications for an active wound healing process. Immunohistochemistry for the extradomain A cellular fibronectin (EDA-cFn) or tenascin (Tn) and routine histology were examined. RESULTS: Four days after LASIK or lamellar keratotomy followed by repositioning of the flap, epithelial plugs and prominent keratocytes as well as Tn and EDA-cFn immunoreactions-indicative of a wound-healing process-appeared in the wound margins. Epithelial plugs were less conspicous, and prominent, presumably activated, keratocytes were no longer identified at the wound margin at 2.5 and 5 months after wounding. However, EDA-cFn and Tn immunoreactivities could still be observed. Only the stromal cells located in the periphery of the flap and in relatively close contact with the epithelium were surrounded by scar tissue expressing immunoreactivity for EDA-cFn or Tn. The central corneal stroma was devoid of scar tissue. CONCLUSION: Results indicate that the wound healing reaction after LASIK takes place only at the periphery of the microkeratome wound, leaving the central optical zone clear.

Results of photorefractive keratectomy for low, moderate, and high myopia.

PURPOSE: To compare the predictability, safety, and stability of photorefractive keratectomy (PRK) in patient groups with low, moderate, and high myopia. The astigmatic component of these corrections was also evaluated. METHODS: In 110 patients, 110 consecutive eyes were operated with a VISX 20/20 excimer laser using standard settings and a 6 mm ablation zone for PRK. The eyes were divided into three groups. In low myopia group (N = 52) the intended correction ranged from -1.50 to -6.00 D (mean -4.28 +/- 1.29 D), in the moderate myopia group (N = 34) from -6.10 to -8.00 D (mean -7.05 +/- 0.70 D), and in the high myopia group (N = 24) from -8.10 to -11.50 D (mean -9.40 +/- 1.13 D). Twenty-seven eyes had 0.75 D of preoperative myopic astigmatism. All patients were followed for 12 months and 36% (N = 40) were available for the 24-month follow-up examination. RESULTS: At 12 months the spherical equivalent manifest refraction was within 0.50 D of intended refraction in 58% (N = 30) of eyes in the low myopia group, 50% (N = 17) in the moderate myopia group, and 29% (N = 7) in the high myopia group. Eighty-seven percent (N = 45) in low, 79% (N = 27) in moderate, and 67% (N = 16) in the high myopia group were within 1.00 D of intended refraction. At 12 months, 88% (N = 46) of the eyes with low myopia, 68% (N = 23) with moderate myopia, and 68% (N = 16) with high myopia achieved uncorrected visual acuity of 20/40 or better, while 6% (N = 16) of the eyes lost and 4% (N = 4) gained two or more lines of Snellen visual acuity. Uncorrected visual acuity of 20/20 or better at 12 months was achieved by 58% (N = 30) of low myopes, 26% (N = 9) of moderate myopes, and 33% (N = 8) of high myopes; 20/25 or better by 71% (N = 37) of low myopes, 39% (N = 13) of moderate myopes, and 42% (N = 10) of high myopes. Overcorrection (> 0.25 D) was detected at 12 months in 31% (N = 16) with low, 38% (N = 13) with moderate, and 67% (N = 16) with high baseline myopia, with statistically significant differences between the low and high myopia groups. CONCLUSION: In all groups, the safety of PRK was satisfactory, without major complications; in the high myopia group where corrections exceeded 8.0 D (mean -9.40 +/- 1.13 D) the procedure was only slightly more unpredictable than in low and moderate myopia at 1 year.

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.

Recovery of corneal nerve morphology following laser in situ keratomileusis.

Morphological changes in the corneal nerves after laser in situ keratomileusis (LASIK) were investigated and the changes were compared with those observed after creation of the keratectomy flap without subsequent photoablation. After creating the hinged flap, a multizone excimer laser photoablation (myopic correction from 6.00 to 6.66 D; diameter 6 mm) was performed on 27 rabbit corneas. Seven of these 27 rabbits received an automated keratectomy without laser photoablation on the fellow eye. A histochemical acetylcholinesterase reaction was used to demonstrate the changes in the morphology of the corneal nerves 3 days, 2.5 and 5 months after the operations. In all specimens the deepest stromal nerve bundles showed normal morphology. Cut nerve trunks were found at the wound margins and at the level of the flap interphase in the stromal bed. At 3 days, both epithelial and basal epithelial/subepithelial nerves were found at the hinge of the flap but the rest of the flap showed a major loss of epithelial, basal epithelial/subepithelial and superficial stromal nerves. A few new regenerating thin nerve fibers were found to emerge from the cut stromal nerve trunks. They appeared to pass the wound margin into the flap area below the epithelium. At 2.5 and 5 months an increasing number of regenerating nerve leashes were observed to emerge from the cut stromal nerve trunks. They appeared to send anastomosing fibers among the neighboring stromal nerves. By this time the epithelial, basal epithelial/subepithelial and anterior stromal innervation had gained an almost normal nerve density and architecture. In the corneas with the flap only, the epithelial innervation was slightly better spared in the center of the flap, and the stromal changes were somewhat less prominent compared with the LASIK corneas.