Cultivated human conjunctival epithelial transplantation for total limbal stem cell deficiency.

PURPOSE: To determine the feasibility of cultivated conjunctiva as a viable epithelial sheet for transplantation and corneal resurfacing in eyes with limbal stem cell deficiency (LSCD). METHODS: Human corneal epithelial (HCE) and human conjunctival epithelial (HCjE) cells were cultivated on human amniotic membrane (AM) to confluence and then air lifted to allow further stratification and differentiation. Denuded AM and cultivated HCE and cultivated HCjE cells were then transplanted into 18 eyes of rabbits with induced LSCD. The cultivated and engrafted epithelia were examined by transmission electron microscopy (TEM) and immunohistochemistry. Two weeks after transplantation, the eyes were examined by slit lamp biomicroscopy and scored on epithelial integrity, corneal haze, and corneal neovascularization. RESULTS: Both cultivated and engrafted HCjE sheets demonstrated confluent epithelial sheets with five to six layers of well-stratified epithelium. TEM examination of engrafted HCjE revealed numerous microvilli, desmosomes, and hemidesmosomes, identical with in vivo corneal epithelium. Immunohistochemical analysis of both HCjE and HCE cells showed the presence of CK3, CK4, and CK12, with absence of Muc5AC. Clinical outcomes for eyes receiving HCjE transplants and HCE transplants were comparable, with most having transparent, smooth corneas, free of epithelial defects. CONCLUSIONS: The study showed that microscopically, HCjE cells have features similar to HCE cells, with clinically equivalent outcomes. The ex vivo cultivation of conjunctiva to form transplantable epithelial sheets for corneal replacement is a promising new treatment modality in patients with LSCD.

Gundersen flaps in the management of ocular surface disease in an Asian population.

PURPOSE: To evaluate the indications, complications, and long-term outcomes with the use of a Gundersen flap for the treatment of various ocular surface conditions in an Asian population. DESIGN: Retrospective case series. STUDY SUBJECTS: Twenty-five eyes of 25 subjects who had undergone conjunctival flap surgery performed from 1999 to 2004 at the Singapore National Eye Centre. METHODS: Review of case records for indications and outcomes of patients undergoing Gundersen flap surgery. MAIN OUTCOME MEASURES: Surgical success was defined as attainment of a stable ocular surface with resolution of symptoms and no flap retraction or dehiscence resulting in reexposure of the corneal surface. Complications to be noted included infection, progression of the inflammatory or infectious disease process beneath the flap, flap loss from epithelial ingrowth, and epithelial cyst formation. RESULTS: Surgical success was achieved in all eyes. Intraoperative and postoperative complications were encountered in 6 patients (24%). One case had a buttonhole, 3 cases had partial flap retractions, and 2 had epithelial inclusion cysts. CONCLUSIONS: The Gundersen flap is still an important procedure and should be considered as a means of stabilizing globe integrity in the management of cases of severe ocular surface disease, particularly when visual potential is poor.

Distinct gene subsets in pterygia formation and recurrence: dissecting complex biological phenomenon using genome wide expression data.

BACKGROUND: Pterygium is a common ocular surface disease characterized by fibrovascular invasion of the cornea and is sight-threatening due to astigmatism, tear film disturbance, or occlusion of the visual axis. However, the mechanisms for formation and post-surgical recurrence of pterygium are not understood, and a valid animal model does not exist. Here, we investigated the possible mechanisms of pterygium pathogenesis and recurrence. METHODS: First we performed a genome wide expression analysis (human Affymetrix Genechip, >22000 genes) with principal component analysis and clustering techniques, and validated expression of key molecules with PCR. The controls for this study were the un-involved conjunctival tissue of the same eye obtained during the surgical resection of the lesions. Interesting molecules were further investigated with immunohistochemistry, Western blots, and comparison with tear proteins from pterygium patients. RESULTS: Principal component analysis in pterygium indicated a signature of matrix-related structural proteins, including fibronectin-1 (both splice-forms), collagen-1A2, keratin-12 and small proline rich protein-1. Immunofluorescence showed strong expression of keratin-6A in all layers, especially the superficial layers, of pterygium epithelium, but absent in the control, with up-regulation and nuclear accumulation of the cell adhesion molecule CD24 in the pterygium epithelium. Western blot shows increased protein expression of beta-microseminoprotein, a protein up-regulated in human cutaneous squamous cell carcinoma. Gene products of 22 up-regulated genes in pterygium have also been found by us in human tears using nano-electrospray-liquid chromatography/mass spectrometry after pterygium surgery. Recurrent disease was associated with up-regulation of sialophorin, a negative regulator of cell adhesion, and never in mitosis a-5, known to be involved in cell motility. CONCLUSION: Aberrant wound healing is therefore a key process in this disease, and strategies in wound remodeling may be appropriate in halting pterygium or its recurrence. For patients demonstrating a profile of 'recurrence', it may be necessary to manage as a poorer prognostic case and perhaps, more adjunctive treatment after resection of the primary lesion.

Elevation of human alpha-defensins and S100 calcium-binding proteins A8 and A9 in tear fluid of patients with pterygium.

PURPOSE: The pathogenesis of pterygia is still not well understood. Recent studies suggest that it may be associated with inflammation and progressive proliferation triggered by ultraviolet radiation. In this study the authors determined that the inflammatory nature of pterygium is reflected in the protein components of tears. METHODS: Consent for this study was obtained from 12 patients (average age, 57 years; eight men, four women) with unilateral pterygium. Tears were collected from diseased eyes and contralateral healthy control eyes with the use of fire-polished 10-microL calibrated glass pipettes before pterygium surgery. Tear protein profiles obtained from diseased and control eyes (seven patient samples) were compared using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) technology. Tears from another five patients with pterygium were used for subsequent protein identification experiments with nano-LC coupled with nano-electrospray tandem mass spectrometry (nano-ESI-MS/MS). RESULTS: SELDI mass spectra showed that one peptide cluster with a molecular weight of 3.4 kDa and two proteins with molecular weights of 10.8 kDa and 12.7 kDa were elevated in tears of eyes with pterygium. Proteins of interest were identified by nano-LC-ESI-MS/MS as human alpha-defensins (human neutrophil peptide [HNP]-1, HNP-2, HNP-3) and S100 calcium-binding proteins A8 and A9. Mean concentrations (n = 7) of alpha-defensins were 1.33 +/- 0.47 microg/mL (HNP-1, P < 0.015) and 0.61 +/- 0.23 microg/mL (HNP-2, P < 0.012) for pterygium eyes and 0.17 +/- 0.12 microg/mL (HNP-1) and 0.02 +/- 0.02 microg/mL (HNP-2) for fellow control eyes. Compared with tears from eyes without pterygium or other abnormalities, the level of S100 A8 increased 1.4- to 13.4-fold (average fold change, 4.5) and S100 A9 increased 1.5- to 4.0-fold (average fold change, 2.3) in 4 of 7 patients. CONCLUSIONS: The upregulated expression of human alpha-defensins and S100 A8 and A9 in tear fluids of patients with pterygium indicates that they may be part of the response of the ocular surface to the formation of this fibrovascular tissue or the accompanying inflammation. They may also serve as a useful indicator for predicting recurrent pterygium.

Current understanding of the treatment and outcome of acute primary angle-closure glaucoma: an Asian perspective.

INTRODUCTION: Primary angle-closure glaucoma (PACG) is a major cause of blindness among Asians. A better understanding of the disease will improve the treatment and outcome of this condition. METHODS: A literature review of all recent publications on PACG was carried out. Articles were retrieved using a key word search of MEDLINE, PubMed and Science Citation Index databases. RESULTS: Following laser peripheral iritodomy for acute angle-closure, Asians were found to have a higher tendency to develop a subsequent rise in intraocular pressure compared to Caucasians. Furthermore, the extent and severity of visual field damage was more severe in Asians than Caucasians, particularly in eyes that presented insidiously with chronic PACG. Prophylactic laser iridotomy in the contralateral eye was found to be highly effective in preventing acute angle-closure attacks. CONCLUSION: PACG is more difficult to manage and is associated with more severe long-term visual morbidity in Asians than Caucasians. Regular follow-up of patients with PACG is important for the early detection of progression of the disease and visual field deterioration.

The Boston keratoprosthesis in Stevens-Johnson syndrome.

PURPOSE: To evaluate the use of the Boston keratoprosthesis (KPro) in patients with Stevens-Johnson syndrome (SJS). DESIGN: Retrospective, noncomparative, interventional case series. METHODS: Sixteen eyes of 15 patients with SJS underwent KPro surgery at the Massachusetts Eye and Ear Infirmary from January 2000 through December 2005. The preoperative, operative, and postoperative findings were recorded. All patients underwent either the type I or type II Boston KPro surgery by one surgeon (C.H.D.). Retention of the prosthesis, best-corrected visual acuity, the need for surgical revision, and postoperative complications were recorded. The outcomes were compared with those of an earlier group of patients from the 1990s. RESULTS: The mean age of patients was 50+/-18 years (range, 23 to 74 years), and the mean duration of their disease was 10+/-6.6 years. The mean follow-up period was 3.6+/-1.5 years (range, 10.2 months to 5.6 years). Ten eyes underwent type II KPro surgery, whereas six eyes underwent type I KPro surgery. Twelve eyes (75%) achieved a visual acuity of 20/200 or better after surgery, with eight eyes (50%) achieving excellent vision of 20/40 or better. Visual acuity was maintained at 20/200 or better over a mean period of 2.5+/-2.0 years. Preexisting glaucoma was found to be a significant risk factor for visual loss. There were no cases of KPro extrusion or endophthalmitis. CONCLUSIONS: KPro in SJS has improved, largely because of the introduction of vancomycin prophylaxis and better glaucoma treatment. It seems to be superior to standard penetrating keratoplasty, with or without allografted stem cell transplantation, as judged from the literature. However, the outcome of the KPro in SJS is still substantially less favorable than in nonautoimmune diseases.

Penetrating keratoplasty in Asian eyes: the Singapore Corneal Transplant Study.

PURPOSE: The Singapore Corneal Transplant Study (SCTS) is a 16-year prospective study of 2100 consecutive corneal transplants performed between January 1991 and November 2006 in patients from Southeast Asia at a single tertiary center. The indications, complications, long-term survival rates, and risk factors for graft failure of penetrating keratoplasty (PK) performed in Asian eyes are reported. DESIGN: Prospective cohort study. PARTICIPANTS: Of the 2100 corneal transplants, 1130 consecutive PKs were performed from January 1991 to December 2003. One graft per patient was selected, leaving 901 grafts for analysis. METHODS: Data were obtained from the Singapore Eye Bank's SCTS database. Cases were classified into optical, therapeutic, and tectonic indications and 9 corneal disease groups. Twenty-four demographic, preoperative, intraoperative, and donor risk factors were subjected to Kaplan-Meier survival analysis, univariate analysis, and multivariate analysis by Cox proportional hazards regression modeling. MAIN OUTCOME MEASURE: Graft failure, defined as the irreversible loss of optical clarity. RESULTS: Study patients were Asian, comprising Chinese (72.7%), Indian (11.54%), and Malay (11.1%) ethnicities (mean age, 56.65 years). The mean follow-up period was 36.8+/-35.5 months. Indications for surgery were optical (87.0%), therapeutic (8.1%), and tectonic (4.88%). Main diagnoses were pseudophakic/aphakic bullous keratopathy (23.4%), postinfectious scarring (12.9%), regrafts (12.4%), keratoconus (9.7%), and posttraumatic scarring (7.3%). Kaplan-Meier survival rates for optical grafts were 86.6%, 72.0%, 63.7%, and 52.0% at 1, 3, 5, and 10 years, respectively; survival rates for therapeutic grafts were 78.4%, 58.3%, and 37.3% at 1, 3, and 5 years, and those for tectonic grafts were 68.3% and 41.7% at 1 and 3 years. Endothelial rejection and late endothelial decompensation accounted for 50.51% of failures. Multivariate analysis revealed 9 predictors for graft failure: recipient gender, age, graft size, graft endothelial status, primary corneal disease, glaucoma, inflammation, perforation, and corneal vascularization. CONCLUSIONS: The long-term outcome for optical indications in Asian eyes follows a trend in endothelium-related attrition similar to that seen in the West. Tectonic and therapeutic keratoplasty for corneal infections and perforation, however, constitute a significant proportion of corneal transplantation performed in Asia and carry a graver prognosis in terms of graft survival.

Argon laser iridotomy-induced bullous keratopathy a growing problem in Japan.

AIMS: To describe the long-term risk of bullous keratopathy following argon laser iridotomy (ALI) in Japan and to compare it with other centres in the world. METHODS: We retrospectively reviewed the case records of all patients with ALI-induced bullous keratopathy that underwent penetrating keratoplasty at Kyoto Prefectural University of Medicine (KPUM) from January 2001 to December 2004. The results were compared with the other representative centres in Singapore and the UK. RESULTS: Thirty-nine eyes of 33 patients were included in the study. The mean age of patients was 73.3+/-6.9 years (range, 58 to 87 years). Patients developed bullous keratopathy at a mean duration of 6.9+/-4.9 years (range, 0.2 to 16 years) after the laser iridotomy procedure. The majority of eyes that developed bullous keratopathy (59.0%) occurred following prophylactic ALI. KPUM had the highest percentage of ALI-induced bullous keratopathy cases that underwent penetrating keratoplasties, as compared with other centres in Singapore and the UK (20.0%, 1.8% and 0%, respectively). CONCLUSION: Bullous keratopathy may arise many years following ALI, and is a growing problem in Asian countries. This condition is a major cause of ocular morbidity in Japan, which has seen a worrying increase in the number of cases in recent years.

Current concepts and techniques in pterygium treatment.

PURPOSE OF REVIEW: Pterygium is a common ocular disorder in many parts of the world. At present, there is a wide variety of surgical methods but very few clinical guidelines on the optimal treatment of primary or recurrent pterygium. The purpose of this review is to summarize the more recent and relevant studies on pterygium treatment. RECENT FINDINGS: The primary aim is to excise the pterygium and prevent its recurrence. As bare sclera excision is associated with a high recurrence rate, pterygium excision is often combined with conjunctival autograft, mitomycin C, beta-irradiation or other adjunctive therapies to reduce recurrence rates. There is currently, however, no consensus regarding the ideal treatment for the disease. Comparability between studies is also hampered by the various definitions of pterygium recurrence. SUMMARY: This article reviews the current concepts and techniques used for the treatment of pterygium. Conjunctival autografting and mitomycin C application are the most commonly used methods for preventing recurrences. The use of mitomycin C and beta-irradiation should be used judiciously because of the potential long-term risk of sight-threatening complications. Additional clinical trials should be performed to evaluate the relative efficacies and long-term safety of the various treatment modalities.

New grading system for the evaluation of chronic ocular manifestations in patients with Stevens-Johnson syndrome.

PURPOSE: To evaluate and grade the extent and severity of chronic ocular manifestations in Stevens-Johnson syndrome (SJS). DESIGN: Prospective multicenter case series. PARTICIPANTS: We enrolled 73 patients (138 eyes) with SJS seen between April 2003 and March 2005 at 3 tertiary referral centers. METHODS: Patients with a confirmed history of SJS and chronic ocular complications that persisted for at least 1 year from the onset of SJS were included. Their detailed medical history and ophthalmic examination results were recorded on an itemized data collection form. Complications were categorized as corneal, conjunctival, and eyelid complications, and 13 components were evaluated and graded on a scale from 0 to 3 according to their severity. MAIN OUTCOME MEASURES: These were broadly classified as corneal (superficial punctate keratopathy, epithelial defect, loss of the palisades of Vogt, conjunctivalization, neovascularization, opacification, keratinization), conjunctival (hyperemia, symblepharon formation), and eyelid (trichiasis, mucocutaneous junction involvement, meibomian gland involvement, punctal damage) complications. RESULTS: The most severely affected complication components were loss of the palisades of Vogt (114 eyes; 82.6%) and meibomian gland involvement (102 eyes; 73.9%). Visual acuity in 74 of the 138 eyes (53.6%) was worse than 20/200. The severity of corneal, conjunctival, and eyelid complications was significantly correlated with visual loss. All 13 complications were correlated significantly with logarithm of the minimum angle of resolution (logMAR) visual acuity; the correlation coefficient (R) ranged from 0.359 to 0.810 (P<0.0001); for corneal epithelial defects, R was 0.169 (P = 0.0473). Eyes with a higher total score for the 3 complication categories had poorer vision (R = 0.806; P<0.0001). Multivariate regression analysis showed that corneal neovascularization, opacification, keratinization, and cataracts significantly affected logMAR visual acuity (P<0.0001, P<0.0001, P = 0.0142, P = 0.0375, respectively). CONCLUSIONS: The authors describe a new method for grading the extent and severity of ocular involvement in patients with SJS and demonstrate that the severity of ocular involvement is correlated significantly with the final visual outcome. This new grading system provides a more objective method for evaluating SJS patients and may be adapted for use in other cicatricial ocular surface diseases.

LASIK in the presbyopic age group: safety, efficacy, and predictability in 40- to 69-year-old patients.

OBJECTIVE: To report the safety, efficacy, and predictability of LASIK in a presbyopic population and to examine possible differences between age groups. DESIGN: Retrospective, descriptive, comparative consecutive case series. PARTICIPANTS: Seven hundred ten eyes of 424 patients between 40 and 69 years of age who underwent LASIK by the same surgeon between January 1999 and September 2005. Patients had manifest refractive spherical errors ranging from -10.5 to +6 diopters (D) and cylinder of up to 2.50 D. METHODS: LASIK was performed with IntraLase femtosecond laser or Hansatome microkeratomes and VISX Star (S4 or S2; VISX Inc., Santa Clara, CA) or Baush & Lomb Technolas 217z (Zyoptix or PlanoScan; Bausch & Lomb, Claremont, CA) excimer lasers. There were 511 myopic eyes (spherical equivalent [SE], -5.03+/-2.02 D) and 199 hyperopic eyes (SE, +2.21+/-1.21 D). Patients were divided into 3 groups: group 1 (40 to 49 years old; n = 359 eyes), group 2 (50 to 59 years old; n = 293 eyes), and group 3 (60 to 69 years old; n = 58 eyes). MAIN OUTCOME MEASURES: Uncorrected visual acuity (UCVA) and best spectacle-corrected visual acuity (BSCVA), retreatment rates, safety, efficacy, and predictability. RESULTS: The mean age of patients in groups 1, 2, and 3 was 45, 53, and 63 years, respectively. With increasing age, there was a trend toward higher retreatment rates and more myopic end points. Safety, efficacy, and predictability were comparable between all groups. At the final follow-up (mean+/-standard deviation, 10.4+/-7.4 months), 80% to 100% of eyes had 20/30 or better UCVA and 81% to 90% were within +/-1.00 D. No eyes lost more than 2 lines of BSCVA, and only 9 eyes (1.3%) lost 2 lines. Subgroup analysis of eyes with a follow-up of 12 months or more was performed. The results were compared with those with shorter follow-up. Both subgroups had comparable outcomes; the duration of follow-up did not affect the visual outcomes. CONCLUSIONS: Despite a trend toward worse final BSCVA and higher retreatment rates in older patients, a greater risk of visual loss after LASIK was not observed. LASIK for myopia and hyperopia has reasonable safety, efficacy, and predictability profiles in the 40- to 69-year-old presbyopic population.

A comparison between cultivated and conventional limbal stem cell transplantation for Stevens-Johnson syndrome.

PURPOSE: To compare the resolution of inflammation and long-term results of cultivated and conventional limbal stem cell transplantation (LSCT) in a patient with Stevens-Johnson syndrome (SJS). DESIGN: Interventional case report. METHODS: A 32-year-old man with SJS and bilateral total limbal stem cell deficiency underwent cultivated LSCT in the right eye, followed by conventional LSCT in the left eye three weeks later. The postoperative medication included dexamethasone 0.1% and ofloxacin 0.3% eyedrops and a tapering dose of systemic corticosteroid, cyclosporine, and cyclophosphamide. Tear samples were collected and analyzed for interleukin (IL) 8 levels. RESULTS: Complete corneal epithelialization was achieved 48 hours after cultivated LSCT, compared with three weeks after conventional LSCT. Ocular inflammation and IL-8 levels decreased more rapidly in the eye with cultivated LSCT. Four years after surgery, more severe corneal scarring and opacification were noted in the conventional LSCT eye. CONCLUSIONS: Cultivated LSCT resulted in a better clinical result and vision, with less stromal scarring compared with conventional LSCT.

Modified automated lamellar therapeutic keratoplasty for keratoconus: a new technique.

PURPOSE: To describe a modified automated lamellar therapeutic keratoplasty (ALTK) technique for the treatment of keratoconus. METHODS: The ALTK procedure was modified to include the use of a Hanna trephine system to control donor and recipient diameters and was performed in 2 stages. The recipient corneal lamellar bed was first prepared by creating a laser in situ keratomileusis (LASIK) flap at a preset depth that was left to heal over. The second stage involved the use of a Hanna trephination system to perform central trephination within the lamellar flap and transplantation of a donor lamellar graft that was fashioned to precisely match the recipient corneal bed, again using the Hanna donor punch to create a central trephination within the lamellar button obtained with the ALTK unit. RESULTS: The modified ALTK procedure effectively treated a patient with moderate keratoconus with a best-corrected visual acuity of 20/80, despite rigid gas-permeable contact lenses. Postoperatively, there was a significant reduction in the irregular astigmatism, and visual acuity improved to 20/25. There were no complications, and the ALTK interface remained clear throughout the 20-month follow-up period. CONCLUSIONS: ALTK combined with Hanna trephination enables precise control of both depth and diameter of lamellar dissection during surgery, removes the need for any form of manual dissection, and may therefore improve optical and visual outcomes in lamellar keratoplasty.

Autologous serum-derived cultivated oral epithelial transplants for severe ocular surface disease.

OBJECTIVE: To evaluate the use of autologous serum (AS)-derived cultivated oral epithelial transplants for the treatment of severe ocular surface disease. METHODS: We used AS from 10 patients with severe ocular surface disease and total limbal stem cell deficiency to develop autologous cultivated oral epithelial equivalents. These were compared with epithelial equivalents derived from conventional fetal bovine serum-supplemented medium. Surgery involved removal of the corneal pannus and surrounding diseased tissue and transplantation of the AS-derived epithelial equivalents. The oral equivalents were analyzed by review of histologic and immunohistochemical findings. RESULTS: Oral epithelial sheets cultivated in AS- and fetal bovine serum-supplemented media were similar in morphology, and both formed basement membrane assembly proteins important for maintaining graft integrity. Complete corneal epithelialization was achieved within 2 to 5 days postoperatively. The ocular surface remained stable without major complications in all eyes during a mean +/- SD follow-up of 12.6 +/- 3.9 months. The visual acuity improved by more than 2 lines in 9 of 10 eyes, with transplanted oral epithelium surviving up to 19 months. CONCLUSION: The successful use of an AS-derived oral epithelial equivalent to treat severe ocular surface disease represents an important advance in the pursuit of completely autologous xenobiotic-free bioengineered ocular equivalents for clinical transplantation.

Orbital cysts lined with both stratified squamous and columnar epithelia: a late complication of silicone implants.

Two patients presented with orbital cysts 5 and 7 years after orbital blowout fracture repair with silicone plate implants. The orbital cysts caused significant exophthalmos and restriction in ocular motility. Surgical excision revealed thick-walled cysts that were displacing the globe and encapsulating the silicone implant. On histopathologic examination, the cysts were lined with both stratified squamous and ciliated columnar (respiratory) epithelia. We propose that squamous and respiratory epithelial cells may have been deposited during surgery from the conjunctival and sinus epithelia, respectively. This case series illustrates that although an uncommon complication, epithelium-lined inclusion cysts may develop several years after orbital fracture repair with a silicone implant. A transconjunctival surgical approach is a possible risk factor.

Transplantation of autologous serum-derived cultivated corneal epithelial equivalents for the treatment of severe ocular surface disease.

PURPOSE: To evaluate the use of autologous serum (AS)-derived cultivated corneal epithelial transplantation for the treatment of severe ocular surface disease. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Nine eyes from 9 patients with total limbal stem cell deficiency were studied. These consisted of 2 eyes with Stevens-Johnson syndrome, 1 with chemical injury, 1 with ocular cicatricial pemphigoid, 1 with Salzmann corneal dystrophy, 1 with aniridia, 1 with graft-versus-host disease, and 2 with idiopathic ocular surface disease. METHODS: Autologous serum obtained from patients was used for cultivating corneal epithelial cells on an amniotic membrane substrate. These AS-derived corneal epithelial equivalents were compared with those derived from fetal bovine serum (FBS)-supplemented medium. At the time of surgery, complete removal of the corneal pannus and conjunctiva up to 3 mm from the limbus was performed. Allogeneic (7 cases) and autologous (2 cases) AS-derived cultivated corneal epithelial equivalents were transplanted onto the ocular surface. Postoperative follow-up included serial slit-lamp examinations with fluorescein staining, as well as photographic documentation. MAIN OUTCOME MEASURES: Ocular surface reconstruction with corneal epithelialization, graft integrity, visual acuity, and postoperative complications. RESULTS: The corneal epithelial sheets cultivated in AS- and FBS-supplemented media were morphologically similar, and demonstrated the normal expression of tissue-specific keratins and junctional specialization assembly proteins. After transplantation, complete corneal epithelialization was achieved within 2 to 5 days. All eyes demonstrated an improvement in visual acuity by > or =2 lines. During the follow-up period of 14.6+/-4.36 months, the corneal surface of all patients remained stable and transparent, without significant complications. CONCLUSIONS: Transplantation of AS-derived cultivated corneal epithelial equivalents was shown to be a feasible method of treating patients with severe ocular surface disease. The use of AS is of clinical importance in the development of autologous xenobiotic-free bioengineered ocular surface equivalents for clinical transplantation.

Establishment of a cultivated human conjunctival epithelium as an alternative tissue source for autologous corneal epithelial transplantation.

PURPOSE: The corneal epithelium is essential for maintaining corneal transparency, and efforts have been made to develop improved techniques for corneal epithelial transplantation in patients with total limbal failure. We evaluated the suitability of transplanted cultivated human conjunctival epithelium (HCjE) as a corneal epithelium replacement in rabbits with total corneal and limbal deficiency. METHODS: HCjE cells, cultivated on human amniotic membrane (AM) to confluence and exposed to an air-liquid interface (air-lifted), were transplanted onto denuded rabbit corneas and monitored for 2 weeks. The cultivated HCjE sheet and the engrafted epithelium were analyzed by immunohistochemistry and transmission electron microscopy (TEM). RESULTS: The transplanted HCjE remained transparent, smooth, and without epithelial defects during the follow-up period. Both the cultivated HCjE cells and the engrafted epithelium manifested five to six layers of stratified squamous epithelium similar in morphology to normal corneal epithelium. The basal cells expressed the putative stem cell markers (ABCG2 and P63) and hemidesmosome and desmosome component proteins. The cytokeratins (CK4, CK13, CK3, and CK12) and MUC4 were found in the engrafted epithelium. However, MUC5AC was not expressed. The results indicate that HCjE cultivated on AM has the potential to be used as an alternative corneal epithelium. CONCLUSIONS: The transplantation of cultivated HCjE sheets is a promising technique for the treatment of eyes with limbal failure.

Inferior corneal decompensation following laser peripheral iridotomy in the superior iris.

PURPOSE: To describe a previously unreported clinical entity of inferior corneal decompensation occurring as a long-term complication of superior laser peripheral iridotomy. DESIGN: Retrospective, non-comparative observational case series. METHODS: Chart review of 14 subjects (16 eyes) with inferior corneal decompensation. RESULTS: All patients were Chinese, most were females (71.4%), and the mean age was 69.5 +/- 8.1 year. The mean interval from iridotomy to the onset of inferior corneal decompensation was 66.1 +/- 34.0 months. The most common indication for laser iridotomy was for prophylaxis, either in the fellow eye of acute angle closure (43.8%) or for occludable angles (12.5%). CONCLUSIONS: Corneal decompensation in the inferior part of the cornea at a site remote from the iridotomy is a rare but important complication of laser iridotomy. Because many of the laser iridotomies were performed prophylactically in eyes without glaucoma, patients may need to be warned of this risk.

Characterisation of human tear proteins using high-resolution mass spectrometry.

INTRODUCTION: The proteins found in tears play an important role in maintaining the ocular surface and changes in tear protein components may reflect changes in the health of the ocular surface. Proteomics provides a comprehensive approach for cataloguing all the proteins of the tear proteome, which will help to elucidate disease pathogenesis, make clinical diagnoses and evaluate the influence of medications on the structure, composition and secretion of tear proteins. In this study, an alternative proteomic strategy was investigated to explore the human tear proteome. MATERIALS AND METHODS: Tear samples were obtained from patients who had pterygium and were collected on the first day and third day after pterygium surgery. Tears pooled from 6 patients were used in the analysis. Reverse-phase high-pressure liquid chromatograph (RPHPLC) was used as the first step to separate intact proteins into 21 peaks. Each fraction was then tryptic-digested and analysed by nanoLC-nano-ESI-MS/MS to characterise the protein components in each fraction. RESULTS: In total, 60 tear proteins were identified with high confidence, including well-known abundant tear proteins, and tear-specific proteins such as lacritin and proline-rich proteins. Among them, proline-rich protein 5 was found for the first time in tear fluid. A large number of plasma proteins were also observed in tear fluid. CONCLUSIONS: The results showed that the proteomic strategy used in this study was successfully applied to analyse tear proteome.

Long-term outcomes in fellow eyes after acute primary angle closure in the contralateral eye.

PURPOSE: To determine the long-term outcome of the contralateral eye in Asian persons with a unilateral attack of acute primary angle closure (APAC). DESIGN: Cross-sectional observational case series. PARTICIPANTS: Seventy-nine individuals who were examined from 4 to 10 years after a unilateral episode of APAC at 2 Singapore hospitals. METHODS: All subjects underwent a complete eye examination. The optic discs were graded clinically and photographically for the presence of glaucomatous optic neuropathy, and automated visual field tests were assessed for damage. All visual fields and optic nerve photographs (when available) underwent a second evaluation by a masked glaucoma specialist, who assessed whether the changes were compatible with glaucoma. MAIN OUTCOME MEASURES: Glaucomatous optic neuropathy, intraocular pressure (IOP), and visual impairment (defined as best-corrected visual acuity of worse than 6/12). RESULTS: Ninety of 138 eligible patients (65.2%) with APAC were examined, 79 with unilateral attacks. Subjects were predominantly Chinese (n = 68; 86%). There were 54 females (68%), and mean age was 68.5+/-8.9 years (standard deviation) at the time of APAC, with a mean duration of 6.3+/-1.5 years from the time of the APAC episode to the study examination. The contralateral eyes of 7 patients (8.9%) had definite or probable glaucoma, 2 of whom were felt to have glaucoma in that eye at the time of the attack. Three eyes had markedly cupped optic discs (cup-to-disc ratio > or = 0.9). Thirteen eyes (16.9%) had best-corrected vision worse than 6/12, due to cataract in almost half the cases. Only 1 eye had vision < 6/60, the cause of which was corneal decompensation. One other patient had only a central island remaining with vision of 6/12. Mean IOP of the study participants was 15.7+/-4.7 mmHg, with 6 subjects (7.6%) having undergone trabeculectomy. CONCLUSIONS: Definite or probable glaucoma was present at the time of diagnosis in 2 (2.5%) fellow eyes and developed in an additional 5 (6.5%) with a mean follow-up of 6 years. More than 80% of this cohort retained good vision in the contralateral eye, in contrast to the eye that underwent APAC. Unoperated cataract accounted for most of the visual impairment in this group.