Collagen cross-linkage: a comprehensive review and directions for future research.

Individuals with keratoconus form a significant proportion of patients for a practitioner specialising in corneal diseases. Yet it is a disease where the pathogenesis is poorly understood, and until recently, there has been no treatment apart from transplantation that could be offered that was curative or even capable of slowing the progression of the disease. Collagen cross-linking treatment using riboflavin and UV light has been developed to address this need, and the initial results are promising. The purpose of this review is to critically evaluate this treatment in light of the scientific basis for cross-linking, to highlight the strengths and limitations of the evidence in terms of efficacy and long-term safety, and finally to identify areas for future research in this area with a significant potential to change the way we treat our keratoconus patients. In addition, we hope that our unbiased review for the first time would bring together, in a concise fashion, scientific information for a practitioner contemplating on offering this treatment and to help inform their patients of its potential risks and benefits.

Non-resolving Descemet folds 2 years following deep anterior lamellar keratoplasty: the impact on visual outcome.

We describe 2 case reports of keratoconus patients showing Descemet's folds persisting more than 2 years after deep anterior lamellar keratoplasty (DLK). One patient had worse best corrected visual acuity postoperatively whereas the other did not improve beyond 6/24. We attribute this to the Descemet's folds. We believe the latter to be a complication following DLK consequent to the degree of localised corneal tissue expansion that may occur in advanced keratoconus in a pathological setting. We also discuss possible mechanisms to try to prevent such a complication.

Penetrating keratoplasty in adults with congenital glaucomas.

PURPOSE: To report the problems and outcomes of penetrating keratoplasty (PKP) in patients with corneal oedema due to congenital glaucoma. METHOD: Case note review of nine such consecutive patients treated with PKP. A Kaplan - Meir survival curve was plotted for the series. RESULTS: Nine patients, ranging in age from 27 to 71 years at the time of their surgery were followed up for a mean of 28 months. All had undergone previous surgery for glaucoma. Preoperatively all patients had controlled intraocular pressures, and four were on ocular hypotensives. Two patients developed corneal graft failure at 15 and 41 months postoperative. Raised pressure was the cause in both. Final visual acuity improved in five patients, was the same in three, and deteriorated in only one. Complications included raised intraocular pressure, cataract, retinal detachment, and endophthalmitis secondary to an infected Molteno implant. CONCLUSION: Despite multiple impediments to good postoperative vision including previous surgery and pre-existing glaucomatous damage, a successful outcome was achieved by careful patient selection, and an awareness of, and meticulous attention to postoperative complications.

Re-epithelialization in cornea organ culture after chemical burns and excimer laser treatment.

OBJECTIVE: To describe the epithelial healing rates observed in freshly cultured rabbit corneas chemically burned with high-concentration hydrochloric acid (HCl) and sodium hydroxide (NaOH) and subsequently treated with phototherapeutic keratectomy (PTK). METHODS: We obtained 126 fresh corneoscleral rims from cadaveric New Zealand white rabbits. Each cornea was exposed to 4-mm cellulose sponges soaked in a solution of topical 0.9% isotonic sodium chloride solution, 2M HCl, or 0.5M NaOH. A transepithelial PTK (6-mm zone; 100-microm ablation depth) was then performed using the excimer laser (150-mJ/cm(2) energy pulse; 20 nanosecond duration; and 10-Hz frequency). Corneas were placed in tissue culture, and 1 cornea from each group was taken out of culture each day after treatment. Re-epithelialization was monitored by means of fluorescein staining, slitlamp photography, and histopathological analysis. RESULTS: Corneas treated with HCl and NaOH exhibited immediate epithelial defects that slowly healed over time. In PTK-treated corneas, the re-epithelialization rate was accelerated compared with that of controls (P =.003 for the HCl group, and P<.001 for the NaOH group). The new epithelial layers were smoother in PTK-treated corneas, as confirmed by results of histopathological analysis. CONCLUSION: Corneal damage caused by HCl and NaOH may be modulated in vitro by PTK in this rabbit model. CLINICAL RELEVANCE: After corneal chemical damage, 193-nm excimer laser PTK accelerates epithelial wound healing.

Precision and accuracy of an artificial anterior chamber system in obtaining corneal lenticules for lamellar keratoplasty.

PURPOSE: To determine the precision and accuracy of an artificial anterior chamber and a manual microkeratome in obtaining corneal lenticules for lamellar keratoplasty. SETTING: Department of Ophthalmology, Cornea, External Diseases and Refractive Surgery Service, University of California Irvine, Irvine, California, USA. METHODS: A lamellar keratectomy was performed in 47 human corneoscleral rims. Three lenticule thicknesses (180, 300, and 360 microm heads) and 3 diameters (7.0, 8.0, and 9.0 mm) were attempted. Diameters and thicknesses were measured by planimetry and pachymetry, respectively. RESULTS: Peripheral lenticule thickness was more likely to be within +/-50 microm of the intended depth in thinner cuts (180 microm, 9/15 corneas, 60%; 300 microm, 6/16 corneas, 40%; 360 microm, 3/12 corneas, 33.3%) (P = .045). Eighty percent (32/40 corneas) were within +/-0.5 mm of the expected diameter. Accuracy was best in the 8.0 mm group, with 47.1% (8/17 corneas) within +/-0.2 mm of the expected diameter. A thickness/diameter correlation was not observed (r(s) < or = 0.28). CONCLUSIONS: The precision and accuracy of this system varied according to the attempted thickness and diameter.

Simple organ cornea culture model for re-epithelialization after in vitro excimer laser ablation.

BACKGROUND AND OBJECTIVE: Most of the in vitro work to characterize the effects of clinical laser surgery on corneal tissues has concentrated on the effects on stromal keratocytes and endothelium with little attention being paid to corneal epithelium. Our purpose is to describe the epithelial healing rates observed in freshly cultured rabbit corneas treated with phototherapeutic keratectomy (PTK). STUDY DESIGN/MATERIALS AND METHODS: Corneas were placed in a simple organ culture system, with media change every 2 days. A clinical excimer laser was used to perform a 6 mm diameter, 100 microm depth transepithelial PTK on 24 cultured rabbit corneas, 1 day after culture initiation. For each post-treatment day, one experimental and one control cornea were removed from culture and stained with fluorescein, photographed, and fixed for histology. Epithelial defect area was measured with digital imaging software and analyzed statistically to assess the re-epithelialization rate. RESULTS: Control corneas, maintained in culture for 1-4 days, had no epithelial defects. Those corneas treated with PTK exhibited an immediate epithelial defect that slowly healed over 3 days. This was confirmed on histopathological analysis. A significant linear trend in re-epithelialization across the time points studied was found (F = 80.48, P = 0.0029). The slope of the linear regression model showed an estimate rate of re-epithelialization of -6.70 over the 3 days. CONCLUSION: We have described the development of a simple, whole organ, rabbit cornea culture model for re-epithelialization after PTK. Our rates of epithelial healing resemble those found in the literature in live rabbit models. Therefore, this model may possibly be used to monitor epithelial wound healing in different corneal diseases or injuries.

Excimer laser photorefractive keratectomy.

The excimer laser has proved to be a precise and versatile tool for treating refractive errors. Excimer laser photorefractive keratectomy for myopia is now an established safe and effective alternative to contact lenses and glasses, and should be part of the ophthalmologist's standard armamentarium for treating this condition. This article provides a brief overview of the basic principles underlying excimer laser technology, as well as the surgical technique and postoperative management of myopic correction.

Bacteriologic and clinical efficacy of ofloxacin 0.3% versus ciprofloxacin 0.3% ophthalmic solutions in the treatment of patients with culture-positive bacterial keratitis.

PURPOSE: To compare the efficacy and safety of ofloxacin 0.3% ophthalmic solution with ciprofloxacin 0.3% ophthalmic solution in patients with culture-positive bacterial keratitis. METHODS: Patients with a microbiologic diagnosis of bacterial keratitis were included in this double-masked, parallel-group study and were randomized to treatment with either ofloxacin 0.3% or ciprofloxacin 0.3% ophthalmic solution. One drop of the study medication was instilled during the daytime according to the following schedule: every half-hour on study day 1, every hour on days 2 through 4, and every 2 hours on days 5 through 21. Healing, the primary outcome measure, was defined as complete reepithelialization, accompanied by nonprogression of stromal infiltrate for 2 days. Secondary outcome measures included signs and symptoms of infection. Patients were monitored throughout the study period for any adverse events. RESULTS: A total of 217 patients completed the study: 112 were treated with ofloxacin and 105 were treated with ciprofloxacin. Streptococcus pneumoniae was the most commonly encountered pathogen in all patients. Complete corneal reepithelialization occurred in 85% of those treated with ofloxacin and in 77% of those treated with ciprofloxacin (p = 0.32). The average time to corneal ulcer healing was 13.7 days in those treated with ofloxacin and 14.4 days in those treated with ciprofloxacin. Both treatments were well tolerated with no patient discontinuing the study because of side effects. CONCLUSION: Ofloxacin 0.3% and ciprofloxacin 0.3% ophthalmic solutions are effective and safe in the treatment of patients with culture-positive bacterial keratitis.

Microkeratome-based limbal harvester for limbal stem cell transplantation: preliminary studies.

PURPOSE: To develop a mechanical device to perform limbal transplantation. This procedure is a valuable surgical technique for management of limbal stem cell deficiency. However, the freehand dissection to obtain donor tissue is laborious and time-consuming. METHODS: A manual microkeratome (LSK One; Moria/Microtech, Doylestown, Pennsylvania) using a redesigned head (200 microm thickness, 16-mm blade) was used to create partial thickness corneoscleral caps from human donor globes. RESULTS: Corneoscleral caps obtained from human donor globes included a ring of approximately 1 mm of perilimbal sclera in contiguity to the cornea. Scanning electron microscopy showed a smooth cut surface with some chatter at the margins. CONCLUSION: This instrument ensures an effective and straightforward method to dissect the area where stem cells have been indirectly localized.

Real time intraocular pressure and lamellar corneal flap thickness in keratomileusis.

BACKGROUND: To measure real time intraocular pressure (IOP) during keratomileusis and to determine variability of corneal flap thickness by using different suction ring pressure settings. METHODS: Eight human cadaver eyes, two groups of four each, were used. The suction ring of the UniversalKeratome was applied to each eye, the same as for a standard automated lamellar keratoplasty (ALK) or laser in situ keratomileusis (LASIK) procedure, to create a lamellar corneal flap. The pressure of the suction ring on each eye was raised to one of two different levels, four eyes to 488 and the remaining four to 600 mm Hg. IOP was continuously recorded by manometer, from application of the suction ring through the end of the passage of the microkeratome. Central corneal thickness was measured, both before and after each procedure. RESULTS: IOP increased by >90 mm Hg after application of the suction ring at a pressure of either 488 mm Hg or 600 mm Hg. Corneal flaps performed under a suction ring pressure of 488 mm Hg measured 159.98+/-5 microm, whereas flap thickness for the 600-mm Hg group varied 1.4-266.2 microm. Two flaps exhibited irregular configurations. CONCLUSIONS: Real time IOP can be measured during keratomileusis. The results demonstrated a significant increase in IOP during the procedure. Suction ring pressure setting is an important variable in determining consistent corneal flap thickness during the keratomileusis procedure.

In vitro effects of antiviral agents on human keratocytes.

PURPOSE: To study the effects of antiviral agents on human keratocytes in vitro. METHODS: Cultured human keratocytes were incubated with either ganciclovir, idoxuridine, trifluridine, or cidofovir at concentrations from 0.0001 to 10 mg/mL. Phase-contrast microscopy and XTT (sodium [2,3-bis [2-methoxy-4-nitro-5-sulphophenyl]-2h-tetrazolium-5-carboxanilide, inner salt) colorimetric assay were performed after 24, 48, and 72 hours of incubation. RESULTS: When adjustments were made for time of incubation and concentration, trifluridine reduced cell viability significantly more than ganciclovir, idoxuridine, and cidofovir (p<0.001, three-way analysis of variance). There was significant time-and dose-dependent reduction of cell viability (p<0.001) with trifluridine and cidofovir. After a 72-hour incubation with ganciclovir or idoxuridine, cell viability was reduced as compared with 24- and 48-hour incubation (p<0.001); only the effects of the highest concentration tested (1.0 mg/mL) were significantly different from those of the lower concentrations (p<0.002). At a concentration of 1.0 mg/mL, trifluridine and cidofovir produced moderate to severe signs of cytotoxicity, whereas ganciclovir and idoxuridine displayed much less severe morphologic signs. CONCLUSIONS: Our results indicate that antiviral agents may have both time- and concentration-related toxic effects on stromal keratocytes. These findings may impact the selection of the most appropriate antiviral drug when it is needed to treat infections involving the corneal stroma.

The effectiveness of a topical antibiotic irrigating solution in a model of staphylococcal keratitis after lamellar keratectomy.

PURPOSE: To create a model of Staphylococcus aureus keratitis after lamellar keratectomy; to assess the toxicity of an antibiotic irrigating solution on the corneal stromal bed; and to test the chemotherapeutic effectiveness of a topical antibiotic, both alone and with an antibiotic-containing irrigating solution in preventing S. aureus keratitis after lamellar keratectomy. METHODS: The right eye of each of 38 rabbits were used in this study. In 18 eyes, a lamellar flap was created with a microkeratome, and an inoculum of S. aureus (either 1,000, 5,000, or 50,000 CFUs) was instilled under each flap; the eyes were examined for signs of infection and inflammation at 24 and 48 hours. In another five eyes, a lamellar flap was created in the same manner and the stromal bed was irrigated with 0.3% ofloxacin; the eyes were assessed for ocular inflammatory changes and evidence of crystalline deposits. Finally, in each of 15 additional eyes, 1,000 CFUs of S. aureus were instilled under a lamellar flap to create experimental infectious keratitis. The keratitis was treated according to three regimens: irrigation of the stromal bed with sterile balanced salt solution; irrigation of the stromal bed with 0.3% ofloxacin, followed by application of topical ofloxacin four times a day; application of topical ofloxacin only, four times a day. Eyes were examined for infection and ocular inflammatory changes at 24 and 48 hours. RESULTS: Staphylococcus aureus keratitis can consistently be produced under the stromal flap by inoculation of relatively few organisms. Irrigation of the stromal bed with commercial-strength topical ofloxacin does not appear to be toxic to the stromal bed, with no evidence of crystalline precipitates of the antibiotic. In our model of infectious keratitis after lamellar keratectomy, both topical ofloxacin alone and the combination of topical ofloxacin and irrigation of the stromal bed with 0.3% ofloxacin were effective at preventing S. aureus keratitis. However, the combined treatment of antibiotic irrigation plus topical antibiotic was more effective at preventing inflammation than topical ofloxacin alone. CONCLUSIONS: In this model of S. aureus keratitis after lamellar keratectomy, irrigation of the stromal bed with antibiotic plus topical antibiotic appears to be both safe and effective for preventing infection.

Surgical outcomes of photorefractive keratectomy and laser in situ keratomileusis by inexperienced surgeons.

PURPOSE: To investigate the results of photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) procedures performed by novice ophthalmologists and compare the results with those of experienced refractive surgeons. SETTING: Doheny Eye Institute, University of Southern California, Los Angeles, and Department of Ophthalmology, University of California, Irvine, California, USA. METHODS: In this retrospective case series, data were examined from the first PRK procedures by 33 consecutive ophthalmologists and the first LASIK procedures by 19 consecutive ophthalmologists. Preoperative and postoperative uncorrected visual acuity (UCVA) and best spectacle-corrected visual acuity (BSCVA), as well as intraoperative and early postoperative complications, were recorded. RESULTS: In the PRK group, 33.3% of eyes achieved a UCVA of 20/20 and 87.8%, 20/40 or better; 54.5% were within +/-0.5 diopter (D) of emmetropia and 87.8%, within +/-1.0 D. Two eyes with a preoperative spherical equivalent of greater than -11.0 D lost 2 lines of BSCVA. If eyes with low myopia (

Effect of age on the refractive outcome of myopic photorefractive keratectomy.

PURPOSE: To evaluate the effect of age on the outcome of myopic photorefractive keratectomy (PRK). SETTING: University-based refractive surgery practice. METHODS: A retrospective analysis of 197 eyes that had PRK for myopia was performed. Four groups of patients were compared: Group 1, patients < 30 years (n=35); Group 2, patients 31 to 40 years (n=56); Group 3, patients 41 to 50 year s (n=47); Group 4, patients > 50 years (n=59). The percentage of eyes with an uncorrected visual acuity of 20/20 or better and 20/40 or better and the percentage of eyes with spherical correction within +/-0.5 and +/-1.0 diopter (D) of the attempted correction were derived for each age group. The percentage of patients overcorrected and undercorrected by 1.0 D or more in each age group was also calculated. The difference between the percentage of patients who achieved a visual acuity of 20/20 or better in Group 4 and in the younger groups was statistically significant at 3 months (P =.02) but not at 6 months (P =.70) and 12 months (P =.55). The difference between the percentage of patients who achieved an acuity of 20/40 or better was not significant at any time. The difference between the percentage of patients who were within +/-0.5 D of intended correction in Group 4 and in the younger groups was statistically significant at 3 months (P =.001), 6 months (P =.006), and 12 months (P =.008); the difference between the percentage of patients who were within +/-1.0 D of intended correction was not significant at any time. The difference between the percentage of patients overcorrected and undercorrected by 1.0 D or more in Group 4 and in the younger groups was not significant at any time. In the 3 younger groups, age did not significantly affect visual outcome or predictability. However, there was a significant difference between Group 4 and the 3 younger groups in predictability of the refractive outcome at 3, 6, and 12 months. Age may play a role in the outcome of refractive surgery.

Phototherapeutic keratectomy for superficial corneal fibrosis after radial keratotomy.

We report on 4 patients who developed superficial corneal fibrosis after radial keratotomy. All 4 were treated successfully with manual debridement of the epithelium and manual excision of the fibrous scar, followed by phototherapeutic keratectomy.

Corneal sensation after laser in situ keratomileusis.

PURPOSE: To report the time course for the return of corneal sensation following laser in situ keratomileusis (LASIK). SETTING: University-based retractive surgery practice. METHODS: Twenty-eight eyes of 18 patients having LASIK were evaluated. Preoperative and postoperative corneal sensation at the nasal flap hinge, at the central cornea, and within the temporal flap edge were measured before and after LASIK for a 3 week period using the Cochet-Bonnet esthesiometer (Luneau). RESULTS: Corneal sensation initially decreased in all 3 positions of the flap measured after LASIK; the greatest decrease was in the central cornea. Near preoperative corneal sensation returned by 3 weeks. The degree of sensation loss did not appear to correlate with the ablation depth. CONCLUSION: Corneal sensation is significantly decreased for approximately 2 to 3 weeks after LASIK, centrally greater than nasally at the flap hinge or temporally within the flap edge, but it generally returns to near the preoperative level by 3 weeks postoperatively.